Introduction: Hybridoma technology plays significant role for the production of both monoclonal and polyclonal antibody. Here, only monoclonal antibody production will be discussed. In this case, a normal B-cell and a myeloma cell are used. B-cell has antibody production ability and myeloma cell has immortality and high proliferation rate. Produced antibodies are specific in action. So, Identical antibodies production process is known as hybridoma technology.
The process includes six stages.
1.Immunization :First, mice is immunized. Then antibody is produced against the immunization in the mice’s body. When the amount of antibody is optimum in the mice. It is sacrificed and spleenocytes are taken out from it. Spleenocyte contains antibody producing B-cell.
2. Fusion :Here, the spleenocyte is fused with myeloma cell.50% polyethylene glycol(PEG) is used to fuse the cell. Fused cell is referred as hybridoma cell.
3.Selection: Selection is done in hypoxanthineaminopterinthymidine(HAT) medium. Here, mainly threetypes of cell are found.
*unfused B-cell: which will die in the medium in short time as it has short life span.
*unfused myeloma cell: which will die in the medium because of synthesis stoppage as it is HGPRT- and Ig-.
*hybridoma cell: it will survive the medium because of B-cell activity.
So, hybridoma cell is selected by this way.
4.screening :It is done by ELISA method. The selected cells are transferred to 96 plastic well plates. Each well contains one cell. Specific antigens are adsorbed at the bottom of the plates. If the cells produce desired antibody, it will bind to that antigens. this antibody is then detected by immunoconjugate which contains two component. Onecomponent of the immunoconjugate is specific for epitopeand antibody is immobilized because of this component. Another component is enzyme which brings color to the well. After incubation enzyme activity is stopped and optical density is measured by ELISA reader.
5.Cloning :After screening, cloning of the antibody can be done in interleukin-6 media for further growth and production of the antibodies.
6.Characterization and storage: After characterization the antibodies can be stored in liquid nitrogen media.

Prostate cancer is considered to be next in rank to the first reason of cancer throughout the whole globe. According to a research 29,000 men was predicted to be dead due to prostate cancer in 2014. It was found that testosterone suppression with hormone therapy paved a milestone in the treatment of patients who were having severe metastatic disease. The initial attempt to reduce the amount of testosterone and modifying the AR axis was believed to be a controlling factor for this disease. (Sternberg, Petrylak, Madan & Parker, 2014)
Being the second purpose of death prostate cancer is common mainly in male. The scopes of treatment for such cancer are ADT (Androgen deprivation therapy) or castrating surgically or chemically. Though these treatment options are highly effective but after some times CRPC is seen in the patients. Because of the CRPC implementations of other therapies have been made and androgen axis can be blocked. So, this has made the androgens and AR signaling pathway a very significant in prostate cancer advancement. In addition to their effects on the prostate tumor cells the ADTs show impact on the immune system too. (Gamat & McNeel, 2017)
For the treatment of repeated prostate cancer the ADTs are the basic options. The ADTs have systemic immunomodulatory effect and cases thymopoiesis. In the modern time the combination of ADTs with immunotherapy has been showing the ability to improve the output of prostate cancer immunotherapy. Clinical trials of such combination therapies are already being investigated but there remain some doubts on 1) the best androgen-targeted-agents, 2) best series of therapies and 3) the applicability of these combination therapies. It is expected that within the next 5-10 years we might be able to clarify these doubts and ultimately it will lead us to design a safer and sustainable treatment therapy of prostate cancer. (Gamat & McNeel, 2017)

The core objectives of this particular study on “Anti-androgen therapies in prostate cancer” are:
To have an insight of prostate cancer.

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To understand how anti-androgen therapies are given.

To know the pharmacokinetics of anti-androgens.

To know about ongoing researches.

To know about few cases.

Prostate Cancer:
The androgen testosterone are the elementary agonists for the AR while the DHT( dihydrotestosterone) being the most useful one. These can attach on the AR-LBD and can cause a conformational change and as a result the foldsome can be moved to a new location and the AR can dislocate inside the nucleus. Then dimerization of the AR takes place and the AR interact with DNA androgen-responsive elements(AREs) and finally cell proliferation takes place. (cano et al., 2013). The additional transactivation of AR can be done through growth factors and cytokines. The sophisticated activity and biology of the AR is also related with AR signaling that is started from the MISS which are the binding sites and the plasma membrane being the location of the MISS. The MISS can be further explained in multiple cell lines, even the cells produced from prostate cancer and it is separable from the observable effects that are related to the NISS. According to Pelekanou et al. it is stated that “membrane androgen binding sites have been described in cell lines and prostate specimens, even in the absence of the full length receptor FL-AR.” (Pelekanou & Castanas, 2016)
Though there’s been a major improvement in the drug development of prostate cancer castration resistance disease in found in the majority of the cancer patients of prostate cancer. Firstly we need to understand what CPRC is. CPRC can be defined as a condition where the androgen signaling remains prominent by the cells and in addition to that they can evade cell death or apoptosis even under the ADT. In order to elucidate the disease condition such term CPRC is applied even if the testosterone level in the circulation persists in castration range (50 ng/dl, alternative to 280-1100 ng/dl which is considered as normal in males), on the other hand tumor growth remains at large due to the AR signaling. According to few researches the castration resistance can be done based few mechanisms such as, 1) AR and ligand dependency, 2) AR dependency and ligand independency, 3) AR and ligand independency. (Pelekanou ; Castanas, 2016)

The imprimatur of abiraterone and enzalutamide has been a break through event in the medication of advanced CRPC which in turn pushed the level of management of prostate cancer to a whole new level. And the advantages were found to be in the recoil of the disease, clinical profit, and better tolerance. Even though with this much huge advancement the ADT resistance is being a significant problem for the enzalutamide and abiramide. According to Narayanan et al,. a study from 2016 stated that, “nevertheless, these two drugs have provided strong evidence on the importance of AR targeting, even under castration resistance”. Abiraterone and enzalutamide both have been able to show elevated containment of this cancer and such facts obvious. These two drugs have been able to provide the mechanisms that refer to the AR axis, drug resistance improvements, tumor cell anointment even under the ADT. (Pelekanou ; Castanas, 2016) Among the other achievements that these two drugs have earned are:
Influencing glucocorticoids with AR.

Differentiating treatment induced-neuroendocrine.

Activating AR variants steadily.

Figure: AR signaling axis and mechanisms of AR targeted inhibition.
Here we can see the mechanism of AR signaling and how the prostate cancer is being developed and several AR target inhibitors. The CYP17A1 is the main thing that is causing the conversion of androgen precursors to DHEA and the AD is made from DHEA and this is done by HSD3?1. Testosterone is made from AD with the help of AKR1C3 and ultimately testosterone is converted into DHT by 5?-reductase. When the AR is activated by the DHT a structural modification is seen at the location where the dimerization of AR takes place and after that the translocation of AR occurs towards the nucleus.

And in the nucleus the AR binds with the DNA and then the DNA under goes transcription which results in biologic responses such elevated growth of the prostate, increasing the PSA and the survival rate of the cancer cells.

Anti-androgens are the name given to a diverse group of medicines that counteract the effects of the male sex hormones, testosterone and dihydrotestosterone. The basic principle is some anti-androgens work by lowering the body’s production of androgens while others block androgen receptors, limiting the body’s ability to make use of the androgens produced.

The term ADT is referred to the decrease or inhibition of the gonadal androgen manufacture or signaling. It has been 70 years since this has been a portion of the norm of endeavor for prostate cancer patient. (Huggins and Hodges, 2002). In the early stages medical castration done and after that GnRH agonists were used. Deduction of LH, FSH secretion and testicular testosterone production are the most known disadvantage of such agonists. In addition to that glucocorticoids are used in order to suppress androgen synthesis. Through the invention of AR antagonists which are also known as anti-androgens the activity of androgens can be inhibited. Bicalutamide, flutamide, nilutamide being the first gen antagonists while the enzalutamide is the second gen antagonist. It is seen that Ketoconazole can also block the Cytochrome P450 enzymes by which the testosterone is biosynthesized. Because of being most accurate and powerful blocker of androgen synthesis Abiraterone is being used even though it has been established newly. The synthesis of androgens can be contained in the testes, besides in the cancer cells of prostate gland by Abiraterone. (Pelekanou ; Castanas, 2016)
Among the anti-androgens Enzalutamide belongs to the second generation of drugs and acts of several steps in the AR signaling pathway. Its main action is to inhibit the androgen from attaching to AR and this inhibition in don in competitive manner. The major advantages of Enzalutamide over the 1st gen of anti-androgens are that the translocation of AR can also be blocked by it as well as DNA attachment and co-activator recruitment. The identification of Enzalutamide was made through the quest of finding nonsteroidal anti-androgens that can show activity even when the AR expression amount is very high. (Rodriguez-Vida, Chowdhury, Sternberg, Rudman ; Galazi, 2018)
Confocal microscopy that was done in live LNCaP cells by researchers found that when enzalutamide was used the ratio of nuclear versus cytoplasmic AR was decreased to about fivefold comparing to bicalutamide. They also demonstrated that impairment of nuclear translocation of AR was also done by Enzalutamide. Another study was also done, in this case a VCap cell lines having endogenous AR gene enhancement and CRPC were used. The results were quite good relative to bicalutamide. Apoptosis and growth were found to be subdue by enzalutamide which tells us that enzalutamide was able to show the blocked effect on AR even in CRPC containing situations. (Rodriguez-Vida, Chowdhury, Sternberg, Rudman ; Galazi, 2018)

Figure: Mechanism of action of enzalutamide in the androgen receptor signaling pathway. Obtained from:
Here we can see how enzalutamide is giving its anti-androgen effect. Firstly it is competitively inhibiting the testosterone to bind with the AR. The secondly it is inhibiting the nuclear translocation of the AR into the nucleus. Then again lastly enzalutamide is also inhibiting the transcription process of DNA which ultimately leading to the death of the cancer cells.

Introduction: Heavy metals are present in abundance amount in soil, water, food stuffs, trash, sewage, and soft drinks as well released by industries, commercials, residentials. Presence of these metals cause exposure to the humans, animals, plant and other biotic factors like air, water and environment and make them polluted. Detection of these metals can be done by various electrochemical methods and machineries. There are several analytical testing methods for detection of heavy metals. HM contaminants in solid waste often originate from multiple sources, including natural (weathering and erosion of parent rocks) and anthropogenic sources (mining, metallurgy, manufacturing of HM-containing product, vehicle emissions), (Weiss D J, Rehkämper M, Schoenberg R, et al. 2008, 42: 655–664). The few of them are AAS (Atomic Absorption Spectrometry), AES/AFS (Atomic emission fluorescence spectrometry), ICP-MS (Inductively coupled plasma mass spectrometry), ICP-OES (Inductively coupled plasma optical emission spectrometry), NAA (Neutron activation analysis), (XRF) X-Ray Fluorescence and AVS (Anodic striping voltammetry), MP-AES (Microwave Plasma Atomic Emission Spectroscopy). In this review of literature number of research papers are to be taken as reference to study amount of heavy metals in different fields and the detection methods for them.
Key Words: Heavy metals (HM), AAS, AES, ICP-MS, ICP-OES, NAA, MP-AES, X-Ray.
Review of Studies:
Boron, Phosphorus and Molybdenum content was determined in biosludges samples using MP-AES where microwave assisted acid digestion method was utilized to extract B, P, and Mo from biosludge. The limit of quantification and detection of B, P, and Mo in the extracts were determined; the linear regression coefficient was greater than 0.998 for all the three techniques, (Srinivasulu vudugandha, Nadavalasiva kumar March 2017).
Ijaz Ahmad, 2017 used Perkin Elmer Atomic absorption (AA) spectrophotometer for determination of heavy metals like Cd, Cr, Cu, Fe, Mn, Ni, & Zn in milk of camel, sheep, cattle, buffalo. They found high levels of Zn (5.150 ± 0.021 Mg/Kg), Mn (0.094 ± 0.530 Mg/ Kg) & Fe (1.580 ±0.530 Mg/Kg) in milk of camel while higher levels of noxious heavy metals such as Cu (0.223 ± 0.010 Mg/Kg) & Cd (0117 ± 0.086 Mg/Kg) were detected from goat milk. Moreover, it can be say that camel and buffalo have equal higher concentration of heavy metals.
Acid extraction was done under microwave irradiation in closed vessels using two method MP-AES & ICP-OES. The results using MP-AES was best alternatives in compare to ICP-OES for Cd, Cr, Cu, Ni, Pb and Zn. Some elements such as Cd, Cr, Pb and Zn were not influenced by applied acid treatment. But they extracted Cr, Ni in higher rate using aqua regia in compare to nitric acid, (V.Y zapryonova and E.N. Piskova (2016, dec19)).
The heavy metals determination in 25 tea samples was done in china by (Wen-si-zhong et. al, 2015, 18 July) using continuum sourced graphite furnace atomic absorption spectroscopy for quantitative analysis. They established method for sample preparation, digestion and quantified analysis of lead content in.
The study on fishes such as Chitalachitola Barbonymus Goninatus & Tachysuras Maculatus was carried out by (Rohasliney Hashim et. al 2014) using graphite furnace atomic absorption spectrometer. They detected heavy metals Cd in Chitala (0.076 Mg/Kg) while in Barbonymus Gonionatus & Tachysuras Muchulatus were already serious levels. That Ni level in all fish sample were under permissible limit. The concentrations of those heavy metal were higher in wet season, even they compared omnivorous fish & carnivorous fish with higher concentration of Pb in carnivorous fish. Moreover, they compared fish weight with concentration of Pb & Cd which were positively correlated overall, they conducted that Kelantan river fishes not exceed the EC &FAQ or WHO guidelines.
Study on water sample &fish from Tasikmutiura, puchongo for elemental analysis using ICP-OES.Only Al, Cu, Fe and Zn were detected whereas as, Cd, Pb were below detection limit. The level of heavy metals in fish sample was serious matter compare to water sample. That could be attributed to low Dissolved Oxygen (DO) levels in the lake, (Ismaniza&Idaliza Mat Salch (2012,)).
L. Dospatliev, K. Kostadinov, G. Mihaylova, N. Katrandzhiev in the year 2012 found Heavy metals Pb, Zn, Cd and Ni in the eggplant sample in the municipality of Plovdiv. Sample preparation was done by microwave system Mileston 1200 MEGA. ICP – OES was used for the determination.
Ma Lin1, Wu Chunhua, Zhao Yang in 2012 developed a method for the determination of extractable heavy metals in leather and textiles by MP-AES. Samples were extracted by artificial perspiration. The result showed that the limit of detection was 0.3?28.8?g/L, the average recoveries ranged from 88%?108%,and the relative standard deviations are lower than 1.5%.

Study of heavy metal content in 15 cosmetic products both imported and locally manufacture products were analyzed using AAS. The concentration of Pb, Cu was higher and if such products used for long time then may pose threat to human health, (Shumaila Zubair, Muhammad Adnan and Ijaz ahmad (2013sep11)).
(M Bettinellia,G.M Beoneb, S Speziaa and C Baffib in 1 December 2000) has done the measurement of the entire substance and the entire example by the blend of nitric acid and hydrochloric acid with the molar proportion of 1:3 which is otherwise called Aqua regia is a yellow-orange fuming fluid. They consolidated the microwave absorption system with the inductively coupled plasma nuclear outflow spectrometry (ICP-OES). Notwithstanding utilizing the customary reflux framework, they utilized the closed vessel framework for the assurance of Aqua regia blend. As the CRM they have utilized the CRM 141R ‘Calcareous Loam Soil’, CRM 142 ‘Light Sandy Soil’, CRM 143 ‘Sewage Sludge Amended Soil’, CRM 277 ‘Estuarine Sediment’ and CRM 320 ‘Waterway Sediment’. The test was completed in two of the research laboratories by utilizing the microwave plasma and the ICP-OES. To check the execution of the two of the laboratories, they did the confirmatory testing of the repeatability, reproducibility and recovery. The investigation was finished by the nested plan and it was confirmed that the changeability in the outcomes were from ICP-OES and not from the Microwave Plasma assurance.
Michalski Rajmund, Muntean Edward, Konczyk Joanna, used A Microwave Plasma – Atomic Emission Spectroscopy method for the determination of selected metals and metalloids (Cr, Mn, Fe, Ni, Cu, Zn, Cd, Al, Pb, Co) in herbal tea infusions the limits of detection were in the range of 0.01-0.12 mg L-1.
The concentration of heavy metal elements of environmental significance in the leachate from landfills of urban solid wastes was determined using Inductively-Coupled Plasma Atomic Emission Spectrometry (ICP-AES). Both highly toxic elements (As, Cd, Co, Cr, Pb and V) and elements present at major concentrations (Al, B, Cu, Fe, Mn, Ni and Zn) were quantified at different levels within and under the landfill (A.Alimonti, S.Caroli, L.Musmeci determined).

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The incidence of tuberculosis (TB) is increasing in Egypt, as well as other developing countries, especially among the immune compromised population 1. The global incidence of TB is estimated to be 10.4 million (90% adults; 65% males) new cases per year 2. The standard, antituberculosis therapy (ATT) shows efficacy for treatment of TB; it consists of isoniazid (INH), rifampin (RIF), pyrazinamide (PZA), and ethambutol (EMB). However, the rate of therapeutic failure among the Egyptian patients with pulmonary TB ranges between 23% and 29% 3, 4. Vitamin D is one of the potential factors that may be involved in TB 5.
Vitamin D deficiency is estimated to affect between 24% and 77% of the general population in Egypt 6, 7. Studies exploring the effect of vitamin D on the therapeutic outcome of ATT revealed conflicting results 8. While it was reported that supplementation of vitamin D accelerated both clinical and radiographic improvement in all patients with TB 9, such supplementation was not successful in improving the rate of sputum clearance from Mycobacterium tuberculosis (Mtb) 10. Vitamin D binds to vitamin D receptor (VDR), activates VDR signaling, and induces a series of antimicrobial responses including induction of autophagy, phagolysosomal fusion, release and activation of the antimicrobial peptide “cathelicidin”, and killing of intracellular Mtb 11, 12.
Our research hypothesis is that vitamin D deficiency is prevalent among the general population in our locality. Vitamin D deficiency may contribute to the therapeutic failure of ATT; consequently, augmenting the standard ATT with cholecalciferol (vitamin D3) supplementation may lead to an improved therapeutic response of the patients with pulmonary TB compared to the standard ATT. We explored the role of adding cholecalciferol to the standard ATT in improving the therapeutic outcome among the naïve patients with pulmonary TB.

Patients and methods:
Study design:
A randomized, controlled, clinical study was conducted.
Study location:
The study population was recruited from the patients attending the outpatient clinics and admitted to the inpatient sectors of the department of Chest Diseases and the department of Tropical Medicine & Gastroenterology (Fever Unit), Assiut University, Egypt.
Study duration:
The study population was recruited during the period from October 2014 to August 2017.
Inclusion criteria:
The study included 500 naïve, patients with pulmonary TB eligible for ATT who accepted to be enrolled, consecutively. The patients were randomly allocated, by a computer-generated stratified, random assignments list (block randomization), to two groups. Group-A included 250 patients who received only ATT, while group-B included another 250 patients who received ATT with cholecalciferol supplementation (cholecalciferol-augmented ATT).
Pulmonary TB was diagnosed based on positive sputum culture on Lowenstein-Jensen medium performed by an experienced microbiologist, with or without the radiological evidence on chest radiography 13.

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Exclusion criteria:
Pregnant or lactating patients, patients less than 18 years old, those with previous ATT, and those with extrapulmonary TB were excluded from being enrolled. In addition, we excluded patients with hypercalcemia, chronic liver disease and/or elevated serum levels of aminotransferases, chronic kidney disease and/or elevated serum level of creatinine, any alcohol intake, and those receiving hepatotoxic drugs other than ATT, as well as those receiving corticosteroids or antimetabolites for any other indication. Patients with human immunodeficiency virus (HIV) infection were not included in the study.
All the participating patients had pretreatment evaluation including clinical evaluation (medical history and physical examination with estimation of weight and height), imaging studies (chest radiography and abdominal ultrasonography), laboratory investigations (serum level of calcium, fasting serum level of glucose, liver chemistry panel, virology panel, kidney chemistry panel, and complete blood count). Body mass index (BMI) was calculated according to the following: body weight (kg) / height (m)2. Diabetes mellitus (DM) was defined as serum level of glucose of 7 mmol/L or more. Liver chemistry panel included estimation of serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase, bilirubin, and albumin. Virology panel included testing for serum antibody to hepatitis C virus (anti-HCV Ab), hepatitis B surface antigen (HBsAg), antibody to hepatitis B core (anti-HBc Ab), and antibody to human immunodeficiency virus (Anti-HIV Ab).
All the enrolled patients received the World Health Organization (WHO)-recommended, standard ATT therapy: INH (5 mg/kg/day; the maximum dose was 300 mg/day), RIF (10 mg/kg/day; the maximum dose was 600 mg/day), PZA (30 mg/kg/day; the maximum dose was 2000 mg/day), and (EMB) (20 mg/kg/day; the maximum dose was 1600 mg/day) for two months, followed by INH and RIF for extra four months 14. All patients received pyridoxine (vitamin B6, 50 mg/day) during the whole treatment period for prophylaxis against isoniazid-related peripheral neuropathy 15. Group-B patients, in addition to ATT, received cholecalciferol supplementation during the whole treatment period (600 IU/day, orally, immediately after lunch). The dose of cholecalciferol was determined according to the recommended dietary allowances for persons 19-70 years old 16. Higher doses of cholecalciferol were avoided to decrease the risk of hypercalcemia. Vitamin D3 was administered with the largest meal to improve its absorption 17. Cholecalciferol supplementation was produced by the same pharmaceutical company.
Follow up included clinical evaluation and laboratory investigations. Daily clinical evaluation during the period of hospital admission, followed by clinical evaluation and laboratory investigations (liver chemistry panel) at the outpatient clinic on weekly basis for the first two months of treatment period, and then monthly for rest of the period, were carried out. Sputum culture on Lowenstein-Jensen medium was performed twice, at the third month and at the fifth month of therapy, for all the patients. Early therapeutic response to ATT was defined as negative sputum culture at the third month during the treatment period, while therapeutic failure of ATT was de?ned as positive sputum culture at the fifth month 18.
For the patients who had elevated serum levels of ALT and/or AST (three times the upper limit of normal (ULN) or more with symptoms or five times the ULN with or without symptoms) during the first two months of ATT, RIF and PZA were discontinued immediately. Less hepatotoxic drug (streptomycin (SM)) was used in addition to INH and EMB until RIF and PZA were resumed after decline of ALT and/or AST serum levels to less than two times the ULN. The dose of SM was 15 mg/kg/day by intramuscular injection with a maximum daily dose of 1000 mg. For patients with the same criteria but after the first two months of ATT, RIF was replaced by EMB temporarily until it was resumed according to the previously mentioned rules 14.
Statistical analysis:
Data were analyzed using the Statistical Package for Social Sciences (IBM SPSS Statistics, version 22.0, release; IBM Corp, Armonk, New York, US) for Microsoft Windows® (64-bit version). Results were expressed as mean ± standard deviation or frequency (percentage) as appropriate. Student’s t-test or Mann-Whitney U test, and Yates’ corrected chi-squared test or Fischer’s exact test as appropriate were used to compare the variables between the study groups. A p value less than 0.05 was considered statistically significant.
The software G*Power version was used for a post hoc power analysis of the performed chi-square tests. An arbitrary effect size was chosen for the power analysis, which precisely was a Cohen’s w statistic of 0.3. This value conventionally corresponds to a medium sized effect. The power achieved was 0.84.
Ethical considerations:
The study was conducted after approval of the Clinical Research Ethical Committee of Assiut Faculty of Medicine, and was carried out according to the code of ethics of the World Medical Association (Declaration of Helsinki). All the participants signed a consent certi?cate after discussing in detail with the investigators the certi?cate subjects and the study aim. Participants were clearly informed that refusing to participate in the study will not affect having full bene?t of the available medical service. Data con?dentiality was respected.

After exclusion of three patients of group-A (due to development of severe ATT-related fulminant hepatopathy leading to death of one patient and acute liver failure of two more) and a single patient of group-B (died because of motor vehicle accident), the study included 496 patients (247 of group-A and 249 of group-B). Pretreatment demographic, clinical, radiological, and laboratory characteristics of the study population are shown in table-1. The two groups were matching. The mean age of the study population 32.5±10.8; the mean age of group-A patients was 31.9±10.4, while it was 33.2±11.1 for those of group-B. The female patients represented 55.6% of the study population; 53% of patients of group-A, and 58.2% of those of group-B. Among 5.8% of the study population, DM was detected (7% of patients of group-B and 5% of those of group-A. Fever was present among 66.1% of the study population (69.6% of group-A patients and 62.7% of those of group-B).
Table-2 shows the therapeutic outcome of ATT among the study population. The overall rate of therapeutic failure among the study population was 29.4%; the rate was significantly lower among patients of group-B compared to those of group-A (22.1% (95% CI 14.7-26.2) vs 38.1% (95% CI 31.5-46.1), p 0.036). The rate of early therapeutic response was significantly higher among patients of group-B compared to those of group-A (35.3% (95% CI 29.6-42.3) vs 19.4% (95% CI 15.1-24.6), p 0.041). Comparing the patients of group-B to those of group-A, the late therapeutic response was nearly equal among the patients of both groups. The therapeutic outcome of the study population is presented in figure-1. All the patients with therapeutic failure (positive sputum culture at the fifth month of ATT) had a positive sputum culture at the seventh month of ATT.
Regarding the adverse effects of ATT among our study population, the overall incidence rate was 19.3%; it was higher among patients of group-A compared to those of group-B (21.9% vs 16.9%). However, this difference was not statistically significant. The most frequent adverse effect among the study population was elevated liver chemistry (9.9%). Patients of group-A compared to those of group-B, had significantly higher rates of elevated liver chemistry (15% vs 4.8%, p 0.001), jaundice (2% vs 0.8%, p 0.008), and myalgia (4.1% vs 2.4%, p 0.047).

Cholecalciferol-augmented ATT needs to be considered as the standard of care therapy for the naïve patients with pulmonary TB. According to the results of our study, the naïve patients with pulmonary TB who received vitamin D3 combined with ATT achieved significantly lower rates of therapeutic failure compared to those who received only the standard ATT. Moreover, the patients on cholecalciferol-augmented ATT compared to those on the standard ATT, had significantly higher rates of early therapeutic response; the earlier the therapeutic response, the less frequent the possibility of spreading the infection. The advantage of the therapeutic outcome gained by adding vitamin D3 to ATT was not on the expense of the safety. On the contrary, the patients who received cholecalciferol-augmented ATT had lower incidence rates of adverse effects compared to those who received the standard ATT, although this was not statistically significant except for the hepatic and muscular side effects, which were significantly lower among the patients with cholecalciferol-augmented ATT compared to those with the standard ATT.
Compared to the reported rates of therapeutic failure for ATT among the Egyptian, naïve patients with pulmonary TB, which ranges between 23% and 29% 3, 4, the overall rate of therapeutic failure among our study population with naïve pulmonary TB was 29.4% which is equal to the highest rate reported. A lower therapeutic failure rate of ATT when combined with vitamin D3 compared to the standard ATT (19% vs 27%) was reported among the Egyptian patients with naïve pulmonary TB who received vitamin D3 to decrease the ATT-related hepatopathy. However, this difference was not statistically significant. This might be attributed to the larger size of our study population (496 vs 300 patients). In addition, our study revealed significantly higher rate of early therapeutic response among the patients who received cholecalciferol-augmented ATT compared to those who received the standard ATT; the early therapeutic response was not evaluated by the study of Hasanain, et al 4.
Sufficient vitamin D3 levels in serum protects against tuberculin skin test conversion 19. Vitamin D insufficiency is a risk factor for tuberculosis in men 20 and is associated with progression to active TB among healthy subjects 21. The SUCCINCT Study showed that high doses of vitamin D accelerated the improvement of patients with pulmonary TB 10. Also, this accelerated improvement was previously evident among the pediatric patients with pulmonary TB 22. The clearance rate of sputum among patients with moderately advanced pulmonary TB is associated with vitamin D supplementation 23. In addition, among the patients with HIV infection, vitamin D3 supplementation reduces the incidence of pulmonary TB 24.
A single, large, oral dose of ergocalciferol significantly reduces the growth of Mtb 25. Vitamin D metabolites contribute to Mtb elimination via enhancement of macrophage phagocytosis, activation of monocytes, cytokines modulation, limitation of intracellular growth of Mtb, and suppression of enzymes involved in pulmonary cavitation 25, 29. The immunomodulatory role of vitamin D in Mtb infection depends on the activation of cathelicidin-mediated mycobacterial destruction through binding with vitamin D receptors (VDR) 26. Vitamin D-mediated enhanced innate immunity of monocytes among patients with Mtb infection is related to the transcriptional activation of the antimicrobial peptide LL-37 11. Vitamin D supplementation enhances Mtb-induced interferon-gamma secretion in patients with vitamin D deficiency, leading to improved cell mediated immunity against Mtb 10. In addition, gene polymorphism of VDR contributes to TB susceptibility 27.
Disagreeing with our findings, vitamin D supplementation did not alter the therapeutic response among patients with pulmonary TB 28, possibly due to the high prevalence of obesity, alcohol use, and HIV infection among those patients. Also, among patients undergoing hemodialysis, vitamin D supplements was not associated with lower incidence of TB 29. This can be explained by the defective renal conversion of vitamin D to its active form 30. Two further studies reported lack of enhanced therapeutic outcome among patients with pulmonary TB who received high doses of vitamin D 31, 32. Nielsen, et al, reported the association of TB with both low and high levels of vitamin D 33. High levels of vitamin D may lead to downregulation of VDR expression resulting in their defective signaling 34.
The lower rates of hepatic adverse effects were previously reported among the Egyptian, naïve patients with pulmonary TB receiving vitamin D3 supplementation with ATT compared to those receiving the standard ATT 4. Deficiency of vitamin D can aggravate the damage to hepatocytes 35, 36.
The serum levels of vitamin D were not measured among our study population which is considered as a major limitation of the study. Also, antibiotic susceptibility testing for Mtb cultures was not available. However, this study included a large sample size and is considered the first of its type among the Egyptian, naïve, patients with pulmonary TB.
In conclusion, cholecalciferol-augmented ATT can be more efficacious in treating naïve patients with pulmonary TB compared to the standard ATT. In addition, adding vitamin D3 to ATT provides extra protection against the hepatic and muscular adverse effects of ATT. Considering vitamin D3 as a safe and inexpensive medication, it is recommended to be combined with the standard ATT. Further studies are encouraged to explore the different mechanisms by which vitamin D3 contributes to the improved therapeutic outcome of ATT.

This paper will give summary of what de Gruchy talks about in his book “A Theological Odyssey”, and critically engage some of the themes that are highlighted on the summary not all of the themes will be engage on this paper.
John de Gruchy in this book “A Theological Odyssey” the esteemed South African theologian takes us on a journey through his life as a theologian. This is a book that traces his own growth and understanding of the formative role that theology could play in the church but especially in society as a whole in spite of different and changing historical contexts. With the celebration of his 75th birthday in 2014 this book forms part of a celebration and recognition of his creative output through the years amidst the lean and better times of the country and church of his lifetime (de Gruchy, 2014, p. 1).
In his prologue John de Gruchy introduces himself as theologian who realizes that theology is about more than writing books, de Gruchy argues that (2014, p. 2), “Doing theology is about more than thinking great thoughts or writing books and learned articles, it is a way of being in the world, of engaging reality, an on-going quest, a form of prayer, a performance located in a particular time and space, and shared with fellow travellers”. In a creative and honest manner all these distinctions are found in the different chapters, in which he reworked the main themes of his own theological history. These themes are The Church Struggle; Doing Theology in Context; In Dialogue with Dietrich Bonhoeffer; Liberating Reformed Theology; Democracy, Reconciliation and Restoring Justice; Christianity, Art and Transformation; Confessions of a Christian Humanist; Led into Mystery. At the end of each chapter there is a bibliography, which can help the reader to pursue some of de Gruchy’s theological insights further.
What intrigues in reading these chapters is John de Gruchy’s understanding of theology as something alive and never stagnant. A theology that is constantly in dialogue with the context: “Studying theology is a necessary and important academic activity in which we engage as we explore and excavate tradition, doing theology is a faith practice, a committed engagement, a way of being, a passion, a contemporary and existential engagement with the gospel in the world of daily reality” (de Gruchy, 2014, p. 39). His theology then also flows from the interaction with his conversation partners, living and dead: From Calvin to Bonhoeffer, from modern science to his own late son Steve.
The Chapter on the “Church Struggle” is full of important historical background, which forms the context for “Doing Theology in Context”. The history is presented from his own experience and participation. It is from this context that Bonhoeffer and Mandela’s legacy is looked at in Chapter 3, and questions asked regarding the future of liberation? De Gruchy presents Bonhoeffer as his main conversation partner through the years in the formation of his own integral theology.
Especially important, for those from a Reformed background is the Chapter on the liberation dimension of Reformed Theology. Reformed theology has the potential for creating a just and compassionate society, for critical solidarity, if the proponents are honest and willing enough to retrieve and transform their own traditions. In Chapter 6 de Gruchy shares his own reflections on the importance of an aesthetic dimension in liturgy, the retrieving of symbols, the importance of worship and creativity: “Beauty as conveyed through the arts can become a way of encountering God” (de Gruchy, 2014, p. 121). The unleashing of artistic creativity helps us to reflect on the God of justice and peace. It reminds us that truth, goodness and beauty is integral to our living in this world in relationship with God.
In retrieving the reformed traditions de Gruchy opens up a whole new scope on understanding what it means to be reformed and how want can be Reformed and a humanist (chapter 7). His rethinking and tracing less well-known traditions of Christian humanism helps the reader to rethink the implication of justice and incarnation. To become truly human is a journey, which can erase divisions: “There is a profound sense of human solidarity and compassion that ignores the boundaries of religion and race, culture and country” (de Gruchy, 2014, p. 142).
In this “Odyssey” one does not only read about our history and the events of the past decades, one also reads about the play of beauty and tragedy in life. This book contains insights not just on the realities facing a democratic society but also the realness of sorrow and loss, which invites us into the mystery of God and life (Chapter 5, 8). In chapter 8 he invites us to places of imagination where our certainties, all of a sudden in the face of tragedy, loses its certainness: “The way of ‘unknowing’ begins when it dawns on us that God is beyond our knowing, and therefore that the answers to ultimate questions are also beyond our grasp” (de Gruchy, 2014, p. 153). One is not led into an understandable system but into mystery.

De Gruchy ends this “Odyssey” with reflections towards the future, looking at specific moments in the Western Cape welfare and rethinking global responsibility and resources for the future. He helps the reader to understand that gospel language, like “peace” is not a cliché but something that all are invited to strife and hope for.
De Gruchy’s education:
De Gruchy received an education to which only the white, privileged population of South Africa were entitled. He matriculated high school and began his undergraduate studies at Rhodes University from which he received his Bachelor of Arts, with Distinction and in 1960 he received his Bachelor of Divinity degree from Rhodes, having achieved First Class Honors. He was ordained to the ministry in the United Congregationalist Church in 1961, the same year he married Isobel Anita Dunstan. While serving a congregation in Durban in 1963 de Gruchy was awarded the World Council of Churches Fellowship which enabled him to spend one year at the Chicago Theological Seminary (de Gruchy, 2014, pp. 4-5). It was in Chicago Theological seminary that de Gruchy was able to attend lectures given by Paul Tillich. In his thesis de Gruchy used Tillich’s understanding of anxiety and the fear of change in Tillich’s “Courage to Be” (Tillich, 1980), to reflect on the South African Christian church but de Gruchy’s real interest was in Bonhoeffer and what he might have said regarding the church situation in South Africa. This growing interest in Bonhoeffer would eventually lead him to doctoral studies. The seeds of a rich theological career having been sown, de Gruchy left Chicago to return to South Africa with a Master of Theology degree having graduated Summa Cum Laude. Beginning in 1964, he worked for four years as a pastor in congregations in Durban and Johannesburg. In 1968 he began his doctoral studies at the University of South Africa where he set out to answer Bethge’s lectures at the University of Chicago in the early 60s on “The Challenge of Dietrich Bonhoeffer’s life and Theology” (de Gruchy, 2014, p. 5).

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Critically Engagement:
He resumed his duties as pastor in 1964 working in the mid-sized United Congregationalist church in Sea View, a town just outside Port Elizabeth in the Eastern Cape. It was during this time that de Gruchy befriended Christiaan Frederick Beyers Naudé, a minister in the DRC and the founding director of the Christian Institute (CI). With Naudé’s encouragement, de Gruchy joined the CI in 1965 having already contributed to Pro Veritate,13 the ecclesiastical newspaper started by Naudé in 1962. Pro Veritate provided a public voice for those wishing to express opinions and concerns regarding apartheid, especially those anti-apartheid activists worshipping and working in the mainline Christian churches. The paper was published from 1963 until 1977 when the CI was banned having been declared an ‘affected organization’ by the government. De Gruchy’s name appears for the first time on the masthead of Pro Veritate in October of 1966 where he is listed as a member of the editorial staff. His work on Pro Veritate was the beginning of a long publishing career. It is possible that, at this time, he began to understand the power of the published word to effect positive change in the Christian church. Before de Gruchy was known as a theologian, he had been an active participant in public protests and anti-apartheid demonstrations while a student at Rhodes University and subsequently as a pastor in the Congregationalist Church. He recounts two such events which illustrate his understanding of the strong connection between the Christian church and the anti-apartheid movement that began to grow in earnest during the 1960s.

One of these events was an anti-apartheid protest in the City Hall where he saw his own minister, Basil Brown, sitting on the stage. De Gruchy recalled this event forty-seven years later in his book Confessions of a Christian Humanist where he remembered “that this made a considerable impression on me. But it was at Rhodes University that political sensitivities were sharpened. I recall the first anti-apartheid protest march in which Isobel and I participated, along High Street in Grahamstown in 1959. Hewson, one of the most saintly people I have ever met (a true model of holiness) was among its leaders. Early the next year, on 21 March, the Sharpeville massacre sent shock waves around the country. That was a critical turning point for me as it was for many others, but few in the evangelical fundamentalist camp seemed to take much notice. Indeed, evangelical fundamentalists in South Africa refused to get involved in opposing apartheid, and many of them openly or tacitly supported it. Ideologically, they had identified themselves as right-wing supporters of the status quo” (de Gruchy, 2006, p. 74).
Protests and demonstrations provided a means of expressing the anger and frustration felt by those who lived under the deadening weight of apartheid’s legislation but they did not provide much opportunity for theological dialogue, at least not the kind de Gruchy envisioned. He was convinced that the clergy in the mainline churches were lacking in theological depth. He was also convinced that, if there were a proper mean of engaging in dialogue, the church struggle against apartheid could be more effective. Pro Veritate provided de Gruchy with a sense of the possibilities of theological engagement but he knew that this was not the newspaper’s primary purpose. He also knew that there was no adequate forum in South Africa where theologians could grapple with the pressing issues of the time. One such pressing issue was the conflict between certain Christian churches and the apartheid state which evolved into a situation where two formidable institutions became locked in a battle for truth and the imaginations of the people they claimed to serve. How did one talk about this with other theologians and church members? Pro Veritate was a newspaper focusing on news from the churches and about the churches in South Africa. It also provided a means by which news about critical issues regarding the church struggle against apartheid could be brought to the foreground and disseminated efficiently and inexpensively. It gave a public voice to the private longings and hopes of the lay and clergy who were associated with the CI but it was not the academic journal de Gruchy believed was necessary for the intellectual debate to flourish in South Africa. For the time being, he would find a theological and ecclesiastical path to walk with the South African Council of Churches.
The South African Council of Churches’ (SACC) subdued and unassuming entry onto the ecclesio-political playing field gave little hint as to its future role in the antiapartheid movement. As Bernard Spong wrote in his history of the SACC, it was a ‘quiet birth’. It came into being without a fanfare of trumpets or any special form of celebration. The event is simply recorded in the minutes of the seventeenth biennial meeting of the Christian Council of South Africa, held in the Observatory Congregational church in Cape Town on May 29, 1968. Spong commented on the event and includes the text of the minutes to bring attention to the contrast between the simplicity of the motion and the magnitude of the SACC’s influence in the church struggle. Spong wrote: “Name of the Council: It was agreed that the name of the Council should be changed to The South African Council of churches” (Spong; Mayson, 1993, p. 2). It was as important to the churches of South Africa as the later establishment of the World Council of Churches’ Program to Combat Racism was to the world church.

These were humble beginnings for an organization that enjoyed the leadership of people like Manas Buthelezi, Desmond Mpilo Tutu and Beyers Naudé and it would go on to be a unifying and powerful force against apartheid. The year 1967 and the birth of the SACC marks the beginning of new level of commitment on behalf of the participating churches in the struggle against apartheid. With its first publication, ‘A Message to the People of South Africa’, it declared itself to be a Christian voice speaking on behalf of the victims of apartheid, both black and white (de Gruchy, 2014, p. 22). The document received a mixed reaction from the participating churches in the SACC. Some of the churches felt that the Message went too far while others felt it did not go far enough. It is the sentence like the following that caught the attention of the government and Afrikaner churches as it seemed highly critical of the claim that God had somehow ordained apartheid and their way of life. De Gruchy notes that, “The central theme of the Message was the rejection of apartheid as a false gospel (de Gruchy, 2014, p. 22).
From gloomy beginnings, a champion of the people rose to meet the challenge of apartheid. In the same year de Gruchy moved with Isobel and their three young children to Johannesburg, leaving his congregational work to serve as the SACC’s first Director of Studies and Communications. It is worth remarking, for the sake of emphasis, on the growing importance of the SACC’s new-found role as church advocate in the struggle against apartheid. While it may be difficult to measure de Gruchy’s influence on the burgeoning Council, it may be useful to consider the fact that he participated in the SACC at the highest level during the important first years following its inception. He was present in the organization as the new constitution with a clear mission statement was crafted and entrenched. It was during this time that the SACC’s message was clearly articulated and it fell to de Gruchy to disseminate this message to its member congregations. As the SACC’s first Director of Studies and Communications, de Gruchy was responsible for publicizing “A Message to the People of South Africa” to the member churches. De Gruchy was not a member of the drafting group of A Message but he was a signatory and he was later asked to co-author a book about A Message which was published in 1968. With the publication of A Message, the SACC had declared the ‘South African way of life,’ or apartheid, to be a false Gospel. It may seem strange that the church would confront apartheid on theological grounds as the term apartheid does not hold any theological meaning. However, it may be useful to remember that apartheid was a social and political system based on an Afrikaner self-understanding, along with a perception of the black majority that was allegedly based on the Christian biblical texts and theological tenets that had become popular in some parts of Europe. The SACC’s inaugural publication revealed the theological direction it would follow for the next twenty-seven years of its involvement in the church struggle. For de Gruchy and the churches that gathered under the new banner, it was a new way of engaging the Nationalist regime and apartheid. De Gruchy had committed himself to the struggle against apartheid in his time as a theology student and pastor; now the SACC presented a larger forum within which to explore theological possibilities for making inroads into the debate and critique of the Republic’s21 racist policies. His experience in the church and in the SACC suggested the need for proper theological discourse around the issue of the Christian churches’ role in the struggle against apartheid.
De Gruchy entered the theological debate in the 1960s at the same time the participating churches in the SACC began to explore new ways of working together to combat state-sponsored apartheid. Like most South Africans, de Gruchy was influenced and his theology shaped, to some extent, by apartheid. His contributions to theological research, particularly in the areas of Bonhoeffer Studies and Reformed Theology, are recognized internationally but South Africa is where he served as pastor and educator. It is for this reason that Bonhoeffer’s work, especially which treated the theological foundations of the Confessing church in Germany, resonated so strongly with him. He found in Bonhoeffer a theologian of considerable skill and compassion who chose to work within the Christian church as the place where God and God’s people meet. For de Gruchy, Bonhoeffer was a kindred spirit in his struggle to discern God’s will for the church within the context of national crisis (de Gruchy, 2014, p. 110).
In a letter to his friend Bethge, Bonhoeffer asserts that, “the church stands, not at the point where human powers fail, at the boundaries, but in the center of the village.” (Bonhoeffer, 1997, p. 367) This quote expresses the importance of the theological context in Bonhoeffer’s, and subsequently, de Gruchy’s theological method. It is a reminder that theology is done where we nurture and are nurtured in community. Theology ought not to be relegated to the realm of mystery where humanity can no longer answer its own questions. Bonhoeffer is suggesting that the church is not the provider of answers to the questions for which we have no mortal reply; God is not a convenient for lifting us out of the world but the One who throws us back into the world where God’s self-revelation intersects with humanity living in the heat of the moment.
De Gruchy, Barth and Bonhoeffer:
The similarities between de Gruchy’s situation in South Africa and Bonhoeffer’s in Germany are not lost on the reader of de Gruchy’s work but they must not be overemphasized. When de Gruchy read Bethge’s lectures on Bonhoeffer, helped shape de Gruchy’s exploration of the church’s proper theological response to apartheid. As de Gruchy wrestled with this question, his insights and conclusions helped the SACC formulate and hold a biblically sound, theological stance against apartheid, thereby avoiding the pitfall of becoming just another consumable item in the marketplace of political ideas that were available in South Africa at the time. It helped the Christian church claim its place in South Africa and speak against apartheid from an alternate perspective. Bernard Spong says this about de Gruchy’s contributions in his book entitled Come Celebrate written on the occasion of the SACC’s twenty-fifth anniversary: “at the 1981 National Conference, Dr. John de Gruchy was to point to the need for the supporters of the liberation movement in the church to ensure that they relied on God’s word and sought God’s Spirit or face the danger of “becoming indistinguishable from any other political movement.” It was this kind of constant reminder that helped the Council maintain that necessary balance, the “wary path between”, in personal and social Gospel. A balance that none would claim to have been complete throughout all its work and witness, but a balance that has provided the vision for what the Council should be about and the blueprint in its planning” ( Spong; Mayson; 1993, pp. 82-83).
De Gruchy is also a recognized Barth scholar. In 2000 he was awarded the prestigious Karl Barth prize. In the JTSA, Beyers Naudé (2000, p. 1) wrote in a brief tribute to de Gruchy that “No other person in South Africa deserves the Karl Barth Prize more than Prof. John de Gruchy”. Naudé’s tribute to de Gruchy echoed the sentiment of many Barth scholars. Lyn Holness recorded the event in this way: “Marking the centenary of Karl Barth in 1986, the prize was created in order to ‘honour outstanding works on the Theological Declaration of Barmen and the tradition created by it’. The jury reached its decision in favor of John de Gruchy in recognition of his vision in the transmission of this tradition, as well as of the theological impetus of Dietrich Bonhoeffer and Karl Barth in the ecclesiastical and social contexts of South Africa. The citation continues: Through his Reformed theology John W. de Gruchy has contributed with prophetic impulses to the overcoming of apartheid mentality as well as to the democratization of South African society and the renewal of his church, thus playing an outstanding role for a culture of international and intercontinental theological exchanges” (Holness, 2003, pp. 41-42). The citation begins, “Through his Reformed theology” This suggested that the jury recognized de Gruchy’s unique contribution to the reclamation of the Reformed tradition in South Africa. Barth was the pastor who became a theologian while Bonhoeffer was the theologian who became a pastor. This move from the university to the congregation placed Bonhoeffer in the midst of the faithful community struggling to find answers to the political questions of the day, questions that seemed to all but defy reasonable enquiry. De Gruchy remains both theologian and pastor and has done so throughout his entire career. His theology is grounded in the experience of the church, and of God’s people seeking the face of God in the other. His theological method pivots on the question by Bonhoeffer: ‘Who is Jesus Christ, for us, today?’ (de Gruchy, 2014, p. 54). It seeks to be a living critical engagement of events in a dynamic context rather than a snapshot, a static moment in time, frozen and immobile, ready for dispassionate examination. He worked alongside others in formulating a proper theological response to what some perceived as a heretical use of the Christian religion to support an oppressive political ideology.
Apartheid and the Church:
The question of identity has been part of South African social consciousness from the first time European sailors set foot in the Cape. Apartheid was the most pervasive and systematic attempt at defining identity in the history of the country and it was a theme that underpinned the theology that was done in South Africa. Beginning in 1948, the largely Afrikaner government took great pains to construct a national identity that would situate the white minority population firmly in the roots of the nation. The question of identity was no less important for the English-speaking population of South Africa. The term English-speaking refers to a small segment of the population whose origins are British. In terms of the church, the English-speaking churches are those whose roots are particular to Britain. De Gruchy explained: “A final consideration regarding the title ‘English-speaking churches’ is its exclusive character. It should include the Baptists, but it generally does not, especially after the Baptist Union withdrew from the South African Council of churches. It could include some of the Pentecostal churches, but their distinct character and lack of involvement in ecumenical groups and social issues excluded them. In some respects, Catholics and Lutherans have been in the vanguard of Christian witness and action in South Africa. Yet, because they are not of British origin, we cannot properly refer to them as ‘English-speaking'” (de Gruchy, 2005, p. 86). The English-speaking South Africans were a minority within the minority white population. It was difficult to find belonging within either the black majority or the larger white minority that supported apartheid.
Opposition to apartheid was always present and it had come to be expressed in different ways by the various Christian churches in South Africa. Often, at least within the Reformed tradition, individual congregations within the denominations were free to express their concerns in whichever way they chose. There was no real unified stance among the churches but the SACC served as a focal point for ecclesiastical resistance and opposition to apartheid. De Gruchy noted that the churches of the SACC provide a living example or model of a community in which black and contradict the policy, intention, and spirit of apartheid. Blacks and whites worked together in open defiance of the state’s discriminatory race laws but the disunity of the Christian church continued to hinder opposition. There was a general consensus among the member churches of the SACC and among the World Alliance of Reformed Churches (WARC). This was made clear in Ottawa in 1982 when the WARC declared apartheid to be a heresy (de Gruchy, 2014, p. 27).
The WARC had taken a stand in opposition to the DRC’s position regarding apartheid. The English-speaking churches had, for the most part, spoken against racial separation but seemed to be slow to make any unequivocal or unified statements about where the churches ultimately stood. Theologians and clerics sought clarity while the Christian churches struggled for identity in the face of division and strife. The DRC, on the other hand, was more certain about its support of the minority leadership of the National Party and faced no such crisis of identity (de Gruchy, 2014, p. 24).
Division was not new to the Christian church. The Great Schism of 1054 divided the church into the Roman Catholic and the Eastern Orthodox churches. The Reformation of the sixteenth century further segmented the church. South Africa inherited a divided European church, and any move toward unity seemed to be further thwarted by apartheid. De Gruchy, in commenting on the history of division in the Christian church, lamented the fact that, “while it is painfully true that the divisions which separated Christians from one another in Europe were transplanted into South Africa, it is equally true, and more painful, that these confessional divisions have been exacerbated by separation along racial, cultural and ethnic lines. These issues, normally regarded as non-theological, must now be seen as equally confessional, because they have to do with the truth of the Gospel as much as those that, for example, traditionally separate Catholics from Calvinists. If the churches seriously begin to confess Jesus Christ as Lord in South Africa in terms that relate to the critical issues of our society, that is, the real issues which divide them, they will begin to discover their unity in a new way. There is a confessing movement in South Africa, and one which includes Christians from virtually all denominations who regard apartheid as a heresy and who strive for true justice and peace” (de Gruchy & Villa- Vicencio, 1983, p. 80). The ‘confessing movement’ to which de Gruchy referred was the status confessionis which was forming around the issue of church-sponsored apartheid. What were once non-theological issues were now being thrust upon the churches and were considered among those theological matters which concerned the Gospel message of equality and unity. De Gruchy’s concern was that the lack of a properly articulated theology, one that was able to challenge the theology of apartheid had created a situation in which the Christian church needed to confess its having rejected God’s rule of justice. De Gruchy’s assumption was that the Christian church, by seeking guidance from within a worshipping and contextually-relevant community, informed by the preaching of the biblical texts and celebrating the sacraments, would understand more fully God’s call to bear witness to God’s Reign of justice and grace. But the church had become divided over the issues of apartheid (de Gruchy, 2014, pp. 66-67).
De Gruchy was also concerned that the church, in its willingness to divide along political lines, had usurped the will of God for the church. Apartheid society meant an apartheid church. According to de Gruchy, nothing could be farther from the idea of the Reign of God. Apartheid had become part of the social landscape and the systematic separation of black, coloureds, asians, and whites was now official policy in the DRC. It appeared that the Nationalist government would need to maintain a hold on the political imagination of the country in the face of national and international criticism. Perhaps one of the ways to do this was nurture the nascent civil religion that would make it easier to accept minority rule as God’s plan for South Africa. A civil religion could also maintain the appearance of justice, allowing the English-speaking community to stave-off a critical examination of rather questionable social and economic structures that always favored whites over black.
Another pillar of the Reformation was the idea of the liberty of conscience. It implied that the ideal for any society was for a free church within a free society, unencumbered by the demands of the civil authority. There are two parts to Calvin’s conception of the liberty of conscience. First, as de Gruchy points-out, the role of the government is to cause the Christian church to respect the individual’s liberty of conscience. Second, the government must give way to the Sovereign Conscience. De Gruchy expressed this same idea by suggesting that, the sovereignty of God implies the liberty of conscience. Indeed, conscience, he maintained, can never be subject to man but always and ever to God Almighty. Humanity may enjoy liberty of conscience when it comes to matters of doctrine or teachings of the church that cannot be biblically substantiated but it is never free from God’s authority over humanity. At the same time, Calvin’s emphasis on the importance of the right of the individual conscience to assert itself under the Sovereignty of God cannot be overstated. De Gruchy took this notion one step further arguing, that ‘the struggle for liberty is not only declared permissible, but it is made a duty for each individual in his own sphere (de Gruchy, 2014, pp. 75-76).
De Gruchy raises the question, ‘Can a Christian within the Reformed tradition support the use of violence against an unjust government?’ This was a question that stayed with de Gruchy for many years. He supported the Calvinist proposition that condoned the Christian’s resistance to the tyrant. On obedience and authority, Calvin asserted that God restrains the fury of tyrants’ in two ways; “either by raising up from among their own subjects open avengers, who rid the people of their tyranny, or by employing for that purpose the rage of men whose thoughts and contrivances are totally different, thus overturning one tyranny by means of another. There have been many instances where theologians within the Reformed tradition supported the Christian’s right to resist an oppressive and unjust government
De Gruchy chose to stay in South Africa when many of those who were able to leave the country did so. He had quickly risen to prominence in the academic world and he would have had opportunities for employment outside South Africa. In a passage from his book Confessions of a Christian Humanist, he did suggest that leaving had been a possibility: “I, like many of my peers, never felt as our parents did that we belonged in some way to Britain, but were equally uncertain about belonging to apartheid South Africa. For that reason, and out of a fear for the future, many left to build their homes elsewhere. Those who could go but stayed, again like myself, and not least those who became conscientious objectors, did so because we had a sense of loyalty to South Africa in a much broader sense, and longed for the day when we could be proud of our country as other people were of theirs” (de Gruchy, 2006, p. 191). There were others in de Gruchy’s life who, at the time, realized that it would not be easy for an outspoken critic of apartheid to work with any real freedom in South Africa. There were enough academic opportunities outside South Africa for a theologian as capable as de Gruchy but he was, first and foremost, a South African. His call to serve the people of God in South Africa was a compelling impetus for his doing theology in the first place. The church community was where he found God active in the world and nowhere more so than in South Africa. Like Bonhoeffer, de Gruchy chose to contribute to his country from within while looking forward to the time when he could be proud to call himself South African.
De Gruchy described belonging as locating oneself within a larger, corporate narrative that knits together one’s own fragmented story with similar fragments of those with whom you share a nation.39 One such fragment is the story of black South Africans paying the cost of white privilege. De Gruchy lived in just such a privileged environment of the white European community. The question ‘What is my nation?’ was not easily answered. The question ‘Who is my nation?’ was more easily answered by white South Africans than by black, coloured, or asian South Africans. The expression ‘nonwhites,’ used to describe the black, coloured, and asian segments of the population was in use in South Africa until recently. When a group is described only in the negative it becomes difficult to assert a positive or historical place in the nation. It is as if the group has no history or land which they may call their own. De Gruchy was aware of the privilege of being able to trace his ancestry when so many South Africans could not. He recognized that it was a privilege to know where he came from and that there ” were many others who came to Cape Town as slaves in the eighteenth and nineteenth centuries, wrenched from the soil, families and communities in which they had been nurtured in East Africa and the Dutch East Indies” (de Gruchy, 2006, p. 10).

While de Gruchy may have lived the majority of his life as a privileged white person, he was keenly aware that his privilege came at a cost. Nelson Mandela, for example, was not so privileged. While de Gruchy was completing high school and preparing himself for university, Nelson Mandela, along with other African National Congress activists, was arrested under laws entrenched in the civil code that were relevant for black men and women, only (de Gruchy, 2014, p. 5). Again, a particular segment of the nation was identified by laws that applied to that segment alone. It is a way of segregating a population within a nation by describing who they are not, to the point of annihilation. De Gruchy was one of the few privileged white South Africans who both voiced his concerns and acted upon them. He had decided early on that the cost of white privilege was simply too high.
I believe de Gruchy’s theology is what I perceive to be his understanding of God’s reconciling work through Christ, as made manifest by the Christian community that worships, preaches and celebrates the sacraments together. I maintain that, for de Gruchy, Christology and ecclesiology are done together for they seem to be inseparable. The Christian church is the institution that exists for others. The church acts, not as refuge for the oppressed and marginalized in society, but as their advocate. De Gruchy developed and shaped his ecclesiology within this context. He rejects the idea that theology could provide a strategic process of compromise and he affirms instead the church’s duty to provide a critical and prophetic appraisal of the relationship between church and state, illuminated by biblical texts and guided by the principles and priorities of the Reign of God as made manifest in the church.
For anybody who would like to engage with the theology of one man’s lifetime this is a must read, especially for those who are convinced that theology is a constantly lively engagement with God in context. The words that act as a refrain right through all these different chapters and quests are justice, beauty, love, being truly human and restoration. All these are building blocks of his writings that still inspire and will keep on inspiring.

Sugarcane juice is a luscious natural drink, full of nutrients.It is produced by pressing the raw, decorticated sugarcane stems in a crushing process. Sugarcane juice is essentially a mix of water and saccharose and it holds all the nutrients from sugarcane. It is a natural energy drink with electrolytes, antioxidants and complex carbohydrates which has a very low effect on sugar levels of blood. There are numerous healthful advantages and medicative properties of this drink many of which are unknown to the people. It makes the abdomen, kidneys, digestive system, heart, eyes, and brain sturdy. The juice can be extremely useful in case of weak teeth by providing a form of exercise to the teeth and makes them sturdy. It additionally helps keep the teeth clean and increasing their life.
This sweet, appetizing beverage is enjoyed in many cultures of the globe. In Pakistan, sugar cane juice has become a popular ingredient at local juice vendors. This is definitely because of the sweetness it saturates in juice mixtures without the loss of processed sugars which might be harmful to the body.

History and Background:
The origin of the sugarcane juice consumption is linked to the exploration of the sugarcane. About 195 countries grow the crop to produce 1.5 billion tonnes. The world’s largest producer of sugar cane out and away is Brazil followed by India. Uses of sugar cane include the production of sugar, sugarcane juice, Falernum, molasses, rum, soda, and ethanol for fuel.

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Sugarcane is grown in Pakistan from the past times attributed to the mighty river Indus and its tributaries. The region, referred to as Indus valley civilization historically had the knowledge of sugarcane production and also the extraction of juice. Traditionally sugarcane juice and pealed sugarcane cut in small pieces for chewing is used round the year.
Sugarcane Production in Pakistan:
Sugarcane juice is greatly appreciated in Pakistan, particularly in Sindh and Punjab, where the sugarcane culture has always been dominant, not only due to its flavor, but also because of its great energy worth. Pakistan captures a vital position among the countries which produce sugarcane. It is on fifth position in sugarcane production. Area and Cane yield of some cane producing countries is given in the following table:

Sugarcane production trends:
Sugarcane is grown on an area of more or less million hectares in Pakistan. The Punjab shares 62%, Sindh 26% and N.W.F.P. shares 16% of the entire area. The national average cane yield is 47 t ha-1.Sindh with 53 t ha-1 is the chief province followed by N.W.F.P. (45 t ha-1) and Punjab (40 t ha-1).
Cane yield potential of some commercial sugarcane varieties in the country:

Marks & Spencer (M&S) had been leading the retailing business of commodities including clothing and food for 20 years. However, M&S has been losing business, thus changes in its operation was proposed. In this essay, the changes in operation, which is segmentation of the clothing market will be introduced. Moreover, the order winners and qualifiers of each sub-brand will be analysed. Lastly, the logistic performance objectives of the brands will be discussed.
1. Segmentation of clothing
In the past, M&S had been holding onto its single brand name, St Michael. But as times change, the company realized the need of changing its strategy and switch to dividing markets into three segments: The Perfect Collection, the Classic Collection (which the above two are of the same market segment), the Autograph Collection, and the Per Una Collection.
1.1 The Perfect and Classic Collections
The Perfect and Classic Collections serve consumers who looks for high quality and highly functional clothing at reasonable prices. The consumers targeted in this market range are those who have a busy lifestyle, therefore wanting to save time and money on shopping for clothes. Moreover, they desire clothes that functional in terms of fashion, which should be timeless as well. The clothes from the collection should also be functional in terms of maintenance e.g. machine washable, colours are less likely to fade, can be tumble-dried, etc. At the same time, the collection fits all body shapes and sizes, for both men and women. The clothes also target the market segment composed of a mature audience that concerns the functionality more than fashion, but also wants to be smart or elegant.
1.2 The Autograph range
The Autograph range targets people who are more fashion-oriented, also cares about quality but emphasises less on the price of acquiring clothes. The designs change seasonally and focus on spring and summer, catering to the needs of more fashion-savvy consumers than those who purchases the Perfect and Classic Collections. The collection is designed by some of the best designers, including Sonja Nuttall, Philip Treacy and Julien Macdonald. The collection includes both menswear and womenswear, with an addition of accessories. Furthermore, the collection is only offered at selected boutique stores and sells at high street prices. Therefore, the autograph range serves the market segment of those who values fashion and quality more than costs of clothing.
1.3 The Per Una range
The Per Una range serves a very specific market segment of fashion-conscious women aged between 25 and 35, sizes 8-18 (Studymoose, 2016). Since women belong in this particular market segment are not in favour of having wearing the same outfit as others, the clothes in this range are limited-edition items with a small number in stock. Moreover, the targeted women would also buy clothes often to catch up with the trends, therefore the designs in stores are changed monthly. As a result, the price and quality of this range is the highest among all the ranges and collections offered by M&S, which the price was 10% more expensive than M&S’s normal range.
2. Order winners and order qualifiers
The three ranges M;S divided their products into catered to the needs and wants of different consumers, providing a number of order winners and order qualifiers as an effort to improve business. According to Terry Hill (2017), order qualifiers are the ‘criteria that a company must meet to consider it as a possible supplier’. To be considered by consumers, the company needs to maintain one criterion to several criteria in order to stay in competition. Meanwhile, order winners are those criteria that win the order. In the above ranges, they all have several order winners and order qualifiers, and will be presented in the following paragraphs.
2.1 Perfect and Classic Collection
It is expected that the collections have sizes available for all body shapes and sizes, therefore this aspect is an order qualifier. In the case, it was presented that both respondents expected their sizes are available in the collections’ standard items. Another order qualifier would be having quality equaling the price charged i.e. reasonable pricing. Regarding order winners, the collection should require no specialist washing, meaning it should friendly towards machine washing and tumble-drying. Furthermore, the size and colour should not be altered after washing. Lastly, having a variety in colour choices would be an order winner as well.
2.2 Autograph range
An exclusive design would be the most important order qualifier. Since the Autograph range focuses on providing unique designs, it is reasonable for consumers to consider the Autograph range only when it provides a special design. Regarding order winners, quality is an order winner because some of the range’s buyers believe a higher price equals a higher quality but others believe it would not be crucial.
2.3 Per Una Range
For this high-quality range, one order qualifier is that the designs are limited-edition items that showcase the latest trends. Furthermore, the items are expected to be high-quality, creating the other order qualifier. Yet, the Per Una range should be at affordable prices, despite the items are of brilliant design. Therefore, although the market segment of Per Una is willing to pay 10% above the normal range for M;S, price is a more important attribute to this segment rather than those in the Autograph range. As a result, one order winner of the Per Una Range is prices being affordable. Furthermore, another order winner would be the design changes more often than the Autograph range, resulting in higher excludability and specialness for the consumers.
3. Logistics performance objectives
Regarding the nature of this essay is to discuss the properties of M;S’s change in operations, logistic performance objectives are important. In order to recognize the logistic performance objectives in the three ranges respectively, the following table will list several aspects for analysing.
Perfect and Classic ranges Autograph range Per Una range
Product range The range is limited due to the fact that they should be timeless basic items. The range is larger than the perfect and classic range, focus is on spring and summer items. The range focuses on high quality fashion that matches the latest trends.
Design changes Designs are rarely changed as they are long-lasting basic timeless items. Designs are changed seasonally. Designs are changed every three months.
Price The prices are reasonable. The prices are at high-street level. The prices are marked at 10% higher than the main range of M&S.
Quality High quality High quality High quality
Sales Volume SKU More than the other ranges. Less than the other ranges. Sales volume is limited.
Order Winners High quality. High quality. Affordable price.
Order Qualifiers Availability of sizes. Exclusive unique designs. Exclusive unique designs.
Operations priorities Mass sales to core customers. Sell cutting-edge items. Sell limited edition items to fashion-savvy consumers.
Based on the above information, much like the decision M&S made, the Perfect and Classic ranges should be available in all M&S stores since they are products for the middle class and are standardized; the Autograph range should be available in less stores than the Perfect and Classic ranges because the designs should be exclusive; in order to further increase the exclusivity of the Per Una range, the range should also be only selling in a number of selective stores. Due to the fact that the fashion industry requires a responsive supply chain, the above product distribution would be efficient regarding the nature of customers each range serves (Christopher, Lowson &Peck, 2004). Additionally, supply chain costs and interaction between consumers and suppliers would increase.
4. Conclusion
The changes in market segmentation and supply chain would imrpove the company’s product quality, appeal towards customers and availability of products for customers. This is because the number of suppliers decrease while increasing responsiveness towards customers’ needs. On the whole, products have a larger variety and the supply chain is more efficient, leading to better company performance.

Introduction :
Bacterial pathogens have been reported on fresh cucumbers and other vegetables used for commercial fermentation. The Food and Drug Administration currently has a 5-log reduction standard for?E. coli?O157:H7 and other vegetative pathogens in acidified pickle products. For fermented vegetables, which are acid foods, there is little data documenting the conditions needed to kill acid resistant pathogens. To address this knowledge gap, we obtained 10 different cucumber fermentation brines at different stages of fermentation from 5 domestic commercial plants. Cucumber brines were used to represent vegetable fermentations because cabbage and other vegetables may have inhibitory compounds that may affect survival. The 5-log reduction times for?E. coli?O157:H7 strains in the commercial brines were found to be positively correlated with brine pH, and ranged from 3 to 24 d for pH values of 3.2 to 4.6, respectively. In a laboratory cucumber juice medium that had been previously fermented with?Lactobacillus plantarum?or?Leuconostoc mesenteroides?(pH 3.9), a 5-log reduction was achieved within 1 to 16 d depending on pH, acid concentration, and temperature. During competitive growth at 30 °C in the presence of?L. plantarum?or?L. mesenteroides?in cucumber juice,?E. coli?O157:H7 cell numbers were reduced to below the level of detection within 2 to 3 d. These data may be used to aid manufacturers of fermented vegetable products determine safe production practices based on fermentation pH and temperature. PRACTICAL APPLICATION:? Disease causing strains of the bacterium?E. coli?may be present on fresh vegetables. Our investigation determined the time needed to kill?E. coli?in cucumber fermentation brines and how?E. coli?strains are killed in competition with naturally present lactic acid bacteria. Our results showed how brine pH and other brine conditions affected the killing of?E. coli?strains. These data can be used by producers of fermented vegetable products to help assure consumer safety.

E. coli bacteria: what are they, where did they come from, and why are some so dangerous?
Escherichia coli (E. coli) are members of a large group of bacterial germs that inhabit the intestinal tract of humans and other warm-blooded animals (mammals, birds). Newborns have a sterile alimentary tract, which within two days becomes colonized with E. coli.
More than 700 serotypes of E. coli have been identified. The “O” and “H” antigens on their bodies and flagella distinguish the different E. coli serotypes, respectively. The E. coli serotypes that are responsible for the numerous reports of outbreaks traced to the consumption of contaminated foods and beverages are those that produce Shiga toxin (Stx), so called because the toxin is virtually identical to that produced by another bacteria known as Shigella dysenteria type 1 (that also causes bloody diarrhea and hemolytic uremic syndrome HUS in emerging countries like Bangladesh) (Griffin & Tauxe, 1991, p. 60, 73). The best-known and most notorious Stx-producing E. coli is E. coli O157:H7. It is important to remember that most kinds of E. coli bacteria do not cause disease in humans, indeed, some are beneficial, and some cause infections other than gastrointestinal infections, such urinary tract infections. This section deals specifically with Stx-producing E. coli, including specifically E. coli O157:H7.
Shiga toxin is one of the most potent toxins known to man, so much so that the Centers for Disease Control and Prevention (CDC) lists it as a potential bioterrorist agent (CDC, n.d.). It seems likely that DNA from Shiga toxin-producing Shigella bacteria was transferred by a bacteriophage (a virus that infects bacteria) to otherwise harmless E. coli bacteria, thereby providing them with the genetic material to produce Shiga toxin.
Although E. coli O157:H7 is responsible for the majority of human illnesses attributed to E. coli, there are additional Stx-producing E. coli (e.g., E. coli O121:H19) that can also cause hemorrhagic colitis and post-diarrheal hemolytic uremic syndrome (D+HUS). HUS is a syndrome that is defined by the trilogy of hemolytic anemia (destruction of red blood cells), thrombocytopenia (low platelet count), and acute kidney failure.
Stx-producing E. coli organisms have several characteristics that make them so dangerous. They are hardy organisms that can survive several weeks on surfaces such as counter tops, and up to a year in some materials like compost. They have a very low infectious dose meaning that only a relatively small number of bacteria (fewer than 50) are needed “to set-up housekeeping” in a victim’s intestinal tract and cause infection.
The Centers for Disease Control and Prevention (CDC) estimates that every year at least 2000 Americans are hospitalized, and about 60 die as a direct result of E. coli infection and its complications. A recent study estimated the annual cost of E. coli O157:H7 illnesses to be $405 million (in 2003 dollars), which included $370 million for premature deaths, $30 million for medical care, and $5 million for lost productivity (Frenzen, Drake, and Angulo, 2005).
E.coli in Cucumbers
A major news story over the weekend concerned death and serious illness in Germany by an infection with the bacteria E.coli ) O157, allegedly from imported cucumbers.
Escherichia coli (E.coli) are common bacteria that live in the intestines of warm-blooded animals. Some strains of E.coli do not cause any ill-effects but some, like O157, can cause illness. This can range from mild diarrhoea to death. The infective dose is very low meaning that only a few bacteria can cause infection.
Infection can occur from consuming food or water that has been contaminated by faeces from infected animals or through contact with them e.g. on farms and premises with animals open to the public.
According to the Food Standards Agency there is no evidence that cucumbers from the alleged sources have been distributed to the U.K.
However, we wish to reiterate the importance of thorough hand washing before eating and after contact with animals. It is also good practice to wash fruit and vegetables before eating.
Germany’s federal agriculture minister conceded Tuesday that “Spanish cucumbers are not the cause of the E. coli outbreak,” according to the Guardian. Germany is now scrambling to find out the real source of the outbreak.
Spain said it was considering suing German officials for blaming its cucumbers for the E. coli outbreak, and for the damage it has likely already done to its food export and farm business, Reuters reported. Spanish farmers have said they are losing around $286m per week in lost sales.
The number dead has now risen to 16 people, and the number sickened 1,500 in Germany, Sweden and other countries. 365 new cases were also reported on Wednesday. The Robert Koch Institute, Germany’s disease control center, said one quarter of the latest cases involved a serious complication called hemolyti-uremic syndrome, which affects the blood and kidneys, according to al-Jazeera.
E. coli cucumber scare: Cases likely to increase

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The Andalucian agriculture minister was keen to show her confidence in the region’s cucumbers
A deadly E. coli outbreak in Europe is expected to worsen in coming days, a senior German scientist has said.
Fourteen people have died in Germany and one woman has now died in Sweden after a trip to Germany.
“We hope the number of cases will go down but we fear it will worsen,” said Oliver Grieve, of the University Medical Centre Schleswig-Holstein, where many victims are being treated.
It is thought cucumbers from Spain caused the outbreak.
But Spanish officials have refused to accept the blame, saying it is still unclear exactly when and where the vegetables were contaminated.
The president of Spain’s fruit and vegetable export federation has urged the government to deal with the outbreak, saying it was costing Spanish exporters $200m (£120m) a week.
Asked which countries had stopped buying Spanish produce, Jorge Brotons reportedly told a news conference: “Almost all Europe. There is a domino effect on all vegetables and fruits.”
Travel linkThe World Health Organization (WHO) has described the outbreak as “very large and very severe” and has urged countries to work together to find the source of contamination.
The Robert Koch Institute (RKI), Germany’s national disease institute, has confirmed 329 cases in the country – though some reports have mentioned as many as 1,200 cases.

How the cucumber crisis affects Europe

Country Action
Germany Consumers told not to eat cucumbers, lettuces and raw tomatoes. 373 cases of E.coli confirmed; 14 deaths
Spain Top European cucumber producer – threatens to seek compensation from the European Union for lost vegetables sales
Russia Ban on all imports of cucumbers, tomatoes and fresh salad from Spain and Germany pending further notice

This mainly involves adequate hygiene to prevent contamination of food and person to person spread. It particularly applies to farms, abattoirs and those working in healthcare, nurseries and food provision.

Correct food preparation is also important:
• Washing of raw vegetables.
• Adequate cooking of meat especially beefburgers.
• Pasteurisation of dairy products.
Advice to the public from the HPA is:10
• Safe food preparation:
o Fully cook minced meat products like beefburgers or meat loaf so that they are coloured all the way through, and no blood runs from them.
o Keep cooked and uncooked meats separately; store uncooked meat on the bottom shelf of the fridge to avoid dripping raw meat juices on to other food.
o Never put cooked food back on a plate which has had fresh uncooked meat on it.
o Thoroughly wash all salads and vegetables that are to be eaten raw.
o Avoid eating and drinking unpasteurised milk and dairy products.
o Boil any drinking water if you are unsure of its source.
• Do not swim in water that may be contaminated.
• Hygiene:
o Thoroughly wash hands after using the toilet, handling raw meat, before meals and after contact with animals.
o Ensure children wash their hands with warm water and soap after contact with animals, particularly while on farm visits.
o If someone has E. coli infection, wash all dirty clothes, bedding and towels in the washing machine on the hottest cycle possible. Clean toilet seats, toilet bowls, flush handles, taps and wash hand basins after use, with detergent and hot water, followed by a household disinfectant.
o If you have E. coli infection you should not prepare food for others.
• Some sources suggest that ‘probiotics’ (e.g. certain strains of lactobacilli) may help prevent gastro-intestinal infections, because they colonize the gastrointestinal tract and theoretically prevent pathogenic organisms from infecting the gut.12




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