Relationship Between Serum Vitamin D Levels and Acute Pneumonia in Children Aged 1-59 Months in Nigeria.

INTRODUCTION: Acute pneumonia is a leading infectious cause of death among children under 5 years globally and in Nigeria. Despite various existing strategies and interventions, pneumonia mortality remains unacceptably high. Novel interventions like improving vitamin D status may be needed as optimal vitamin D status may facilitate the ability of immune cells to fight against infections like pneumonia. We investigated the relationship between serum vitamin D [25(OH)D] levels and acute pneumonia in children younger than 5 years in Nigeria. SUBJECTS AND METHODS: This cross-sectional study involved 135 children with pneumonia and 135 apparently healthy controls. Acute pneumonia was diagnosed using the revised World Health Organization criteria (2012) and chest radiological signs. Serum 25(OH)D concentrations were determined using a vitamin D ELISA kit. The mean serum 25(OH)D levels in both groups were compared and also determined odds ratio (OR) of pneumonia. RESULTS: The mean serum 25(OH)D level of children with pneumonia (52.14 ± 21.87 nmol/l) was significantly lower than that of controls (60.91 ± 32.65 nmol/l), p = 0.010. The proportion of children with low serum vitamin D levels (≤75.0 nmol/l) was significantly higher in the pneumonia group (n = 123, 91.1%) than the control group (n = 97, 71.9%), p < 0.001. After adjusting for confounders, serum 25(OH)D levels of greater than 75 nmol/l was associated with decreased odds of acute pneumonia (adjusted OR = 0.33, p = 0.007). CONCLUSION: A low vitamin D level was associated with increased risk of acute pneumonia. Lay summary. INTRODUCTION: Chest infection (pneumonia) is a leading cause of death in children younger than 5 years of age globally and also in Nigeria. Pneumonia death is still very high despite all the existing efforts at reducing it. New methods may still be needed to drastically reduce this problem. One of these new methods may include improving the vitamin D status of an individual because optimal vitamin D levels may help the body to fight against infections like pneumonia. We investigated the relationship between blood levels of vitamin D and pneumonia in children younger than 5 years. SUBJECTS AND METHODS: Vitamin D levels of 135 children with pneumonia were measured and compared with vitamin D levels of another 135 healthy children without pneumonia. We diagnosed pneumonia by using both revised World Health Organization criteria (2012) and chest X-rays signs. Blood levels of vitamin D were measured using a vitamin D ELISA kit. RESULTS: The average blood vitamin D level of children with pneumonia (52.14 ± 21.87 nmol/l) was low compared with that of children without pneumonia (60.91 ± 32.65 nmol/l), p = 0.010. The number of children with low blood vitamin D levels (≤75.0 nmol/l) was more in the pneumonia group (n = 123, 91.1%) than in children without pneumonia (n = 97, 71.9%), p < 0.001. After adjusting for other potential risk factors, blood level of vitamin D >75 nmol/l was associated with lower risk of having pneumonia, (adjusted OR = 0.33, p = 0.007). CONCLUSION: A low vitamin D level was associated with increased risk of acute pneumonia.

Relationship between serum level of ionized calcium, magnesium, phosphate, vitamin D and parathyroid hormone with stages of CKD

Background: Chronic kidney disease is defined as either damage or a decreased Glomerular Filtration Rate of less than 60ml/min/1.73m2 for 3 or more months. There is destruction of renal mass with irreversible sclerosis and loss of nephron leading to a progressive decline in GFR.Secondary hyperparathyroidism hyperphosphataemia, hypocalcaemia and vitamin-D deficiency are common complications of CKD. Objective: To determine relationship between serum level of ionised calcium, magnesium, phosphate, vitamin-D and parathyroid hormone with stages of CKD. Method: This study was conducted at ABUTH Zaria. 125 consecutive adult patients in various stages of CKD who presented were enrolled and 125 apparently healthy matched for sex and age controls were also recruited. Results: 9% of patients were in stage-1, 16% in stage-2, 22% in stage-3, 12% in stage-4 and 41% in stage-5. Serum ionised calcium, vitamin-D and eCrCl showed a progressive decline as the stage of CKD advances, while serum phosphate, creatinine and iPTH showed a progressive increase as the stage of CKD advances. Changes in serum magnesium showed a slight change with advancing stages of CKD. The difference in mean serum levels of calcium, phosphorus, vitamin-D, parathyroid hormone, creatinine and eCrCl with different stages of CKD were statistically significant. eCrCl correlated negatively with phosphate and iPTH while serum creatinine correlated negatively with calcium and positively with phosphate and iPTH. Conclusion: Majority of CKD patients were in late stage. Correlation of analytes with stages was more in late stages and biochemical derangements occurred in late, rather than early stages of CKD

Anti-hyperglycaemic Effects of Rutin on Blood Glucose, Oxidative Stress Biomarkers and Lipid Peroxidation in Alloxan-induced Hyperglycaemic Wistar Rats.

The present study investigated the anti-hyperglycaemic effect of rutin on blood glucose, oxidative stressbiomarkers and lipid peroxidation in alloxan induced hyperglycaemic wistar rats. Diabetes was induced in rats by anintraperitoneal (i.p) injection of alloxan monohydrate 150 mg/kg body weight. Twenty five wistar rats were divided asfollows; Group1 served as diabetic control received distilled water 2 mg/kg, Group served as positive control received 2mg/kg glibenclamide, 3, 4 and 5 received rutin at 50, 100 and 200 mg/kg body weight for 28 days respectively. At the end of the treatment, rats were sacrificed and the blood and serum were used for the analysis of blood glucose and oxidativestress biomarkers respectively. The determinations of blood glucose levels were carried out at intervals of 7, 14, 21 and 28days respectively Serum oxidative stress biomarkers lipid peroxidation, were done on the 28 days. Administrations of rutinat the three different doses 50,100 and 200 mg/kg to diabetic rats significantly (p<0.05) decreased the blood glucose levelsas compared to diabetic control. The dose of 200 mg/kg exhibited a maximum glucose lowering effect with blood glucoseof 102.8± 0.06 as compared to diabetic control 346.2±0.16. Furthermore, in relation to the oxidative stress biomarkers therewas a significant (p<0.05) increased in the levels of gluthathione peroxidase, superoxide dismutase and catalase as comparedto control. However, there was also a significant decreased in the malondialdehyde levels as compared to control. It may beconcluded that oral administration of Rutin for 28 days decreases blood glucose levels and prevented oxidative stress andantioxidant status in hyperglycaemic rats.

Cardiac Troponin T and creatine kinase MB fraction levels among patients with acute ischemic stroke in Nigeria.

BACKGROUND: Stroke has been a global burden, with increasing morbidity and mortality. Serum cardiac troponin t (cTnT) and creatine kinase (CK-MB) fraction are reported to be elevated in patients admitted with acute ischaemic stroke and high level of these biomarkers indicated more severe stroke and neurologic deficit in some of the patients. OBJECTIVE: To evaluate the serum levels cardiac troponin t (cTnT) and creatine kinase MB fraction (CK-MB) in patients with acute ischaemic stroke and relate the analytes to severity of stroke. METHOD: Patients with clinical diagnosis of ischaemic stroke diagnosed, confirmed by brain Computerized Tomography scan and equal number of apparently healthy age and sex-matched were recruited. Serum cardiac troponin t (cTnT) and creatine kinase MB fraction (CK-MB) were analysed using ELISA method and Stroke severity was determined using National Institute of Health Stroke Score (NIHSS). RESULTS: Mean serum cardiac troponin t (cTnT) and creatine kinase MB fraction (CK-MB) in stroke patients were found to be higher than age sex matched control (p<0.05). NIHS Score of 12.2 ± 5.43 and 9.78 ± 3.97 were observed in Patients with elevated and normal cTnT respectively (p=0.009) while NIHS Score were similar in patients with elevated and normal CK-MB (p = 0.772). CONCLUSION: The mean values of serum cTnT and CK-MB were higher in acute ischaemic stroke patients compared to controls. Serum cardiac Troponin t level may be a significant biomarker of the severity of stroke.

Assessment of biochemical liver function tests in relation to age among steady state sickle cell anemia patients.

BACKGROUND AND OBJECTIVE: Multiorgan failure including liver dysfunction is a common finding in sickle cell anemia (SCA) patients, the cause of which is multifactorial with advancing age said to be a major determinant. There is a paucity of data on liver function among SCA patients in relation to age in northern Nigerian hospitals, including Ahmadu Bello University Teaching Hospital (ABUTH), Zaria. This study was to assess the biochemical liver function tests (LFTs) as they relate to age among SCA patients in steady state, with a view to improving the overall monitoring of these patients. SUBJECTS AND METHODS: This study was carried out in ABUTH, Zaria, Northern Nigeria. LFTs were carried out in 100 SCA and 100 apparently healthy participants (controls). The SCA group was made up of fifty adults and fifty children diagnosed of SCA, whereas the control group was made up of fifty adults and fifty children who were apparently healthy and had hemoglobin AA. Paired two-tailed Student's t-test for matched samples and Pearson's linear correlation statistical methods were employed for the data analysis using Microsoft Office Excel 2007. A P ≤ 0.05 was considered as statistically significant. RESULTS: The serum concentrations of total bilirubin (TB), alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP), and AST/ALT ratio were significantly higher in SCA patients compared to the controls (P = 0.001, P = 0.001, P = 0.05, P = 0.05 and P = 0.001, respectively). Serum total protein (TP) and ALB were significantly lower (P = 0.01 and P < 0.05, respectively) in SCA patients compared with the controls. The levels of TB, ALT, AST, ALP, and AST/ALT were significantly lower in SCA adults compared to SCA children, whereas TP and ALB were higher in SCA adults compared to the SCA children. There were significant negative correlations between age and each of TB, ALT, AST, ALP, and AST/ALT, and significant positive correlations between age and each of TP and ALB in SCA patients. CONCLUSION: There are mild LFTs derangements in SCA patients even in steady state with the extent of the abnormalities decreasing with advancing age of the patients.

Tuberculosis of the Breast: An Initial Presentation of the Metabolic Syndrome with Type 2 Diabetes Mellitus in a Young Nigerian Woman.

Breast tuberculosis is an uncommon presentation of extra pulmonary tuberculosis. A 40-year-old obese woman presented with a right breast abscess which had failed to heal after surgical drainage. There was no family history of breast disease. Biopsy and histology of the lesion showed chronic granulomatous inflammation with positive stains for acid fast bacilli compatible with tuberculosis. Further evaluation confirmed metabolic syndrome with type 2 diabetes mellitus. She was placed on antituberculosis chemotherapy and appropriate therapy for diabetes mellitus with complete resolution of the lesion. We report this case because of its rarity and to highlight the association between tuberculosis an infectious disease and overnutrition in diabetes mellitus, a noncommunicable disease.

Psychosocial factors and obesity in 17 high-, middle- and low-income countries: the Prospective Urban Rural Epidemiologic study.

BACKGROUND/OBJECTIVES: Psychosocial stress has been proposed to contribute to obesity, particularly abdominal, or central obesity, through chronic activation of the neuroendocrine systems. However, these putative relationships are complex and dependent on country and cultural context. We investigated the association between psychosocial factors and general and abdominal obesity in the Prospective Urban Rural Epidemiologic study. SUBJECTS/METHODS: This observational, cross-sectional study enrolled 151 966 individuals aged 35-70 years from 628 urban and rural communities in 17 high-, middle- and low-income countries. Data were collected for 125 290 individuals regarding education, anthropometrics, hypertension/diabetes, tobacco/alcohol use, diet and psychosocial factors (self-perceived stress and depression). RESULTS: After standardization for age, sex, country income and urban/rural location, the proportion with obesity (body mass index ≥30 kg m(-)(2)) increased from 15.7% in 40 831 individuals with no stress to 20.5% in 7720 individuals with permanent stress, with corresponding proportions for ethnicity- and sex-specific central obesity of 48.6% and 53.5%, respectively (P<0.0001 for both). Associations between stress and hypertension/diabetes tended to be inverse. Estimating the total effect of permanent stress with age, sex, physical activity, education and region as confounders, no relationship between stress and obesity persisted (adjusted prevalence ratio (PR) for obesity 1.04 (95% confidence interval: 0.99-1.10)). There was no relationship between ethnicity- and sex-specific central obesity (adjusted PR 1.00 (0.97-1.02)). Stratification by region yielded inconsistent associations. Depression was weakly but independently linked to obesity (PR 1.08 (1.04-1.12)), and very marginally to abdominal obesity (PR 1.01 (1.00-1.03)). CONCLUSIONS: Although individuals with permanent stress tended to be slightly more obese, there was no overall independent effect and no evidence that abdominal obesity or its consequences (hypertension, diabetes) increased with higher levels of stress or depression. This study does not support a causal link between psychosocial factors and abdominal obesity.

Psychosocial factors and obesity in 17 high-, middle- andlow-income countries: the Prospective Urban RuralEpidemiologic study

Psychosocial stress has been proposed to contribute to obesity, particularly abdominal, or centralobesity, through chronic activation of the neuroendocrine systems. However, these putative relationships are complex anddependent on country and cultural context. We investigated the association between psychosocial factors and general andabdominal obesity in the Prospective Urban Rural Epidemiologic study.SUBJECTS/METHODS: This observational, cross-sectional study enrolled 151 966 individuals aged 35–70 years from 628 urban andrural communities in 17 high-, middle- and low-income countries. Data were collected for 125 290 individuals regarding education,anthropometrics, hypertension/diabetes, tobacco/alcohol use, diet and psychosocial factors (self-perceived stress and depression).RESULTS: After standardization for age, sex, country income and urban/rural location, the proportion with obesity (body massindex ⩾ 30 kgm−2) increased from 15.7% in 40 831 individuals with no stress to 20.5% in 7720 individuals with permanent stress,with corresponding proportions for ethnicity- and sex-specific central obesity of 48.6% and 53.5%, respectively (Po0.0001 forboth). Associations between stress and hypertension/diabetes tended to be inverse. Estimating the total effect of permanent stresswith age, sex, physical activity, education and region as confounders, no relationship between stress and obesity persisted(adjusted prevalence ratio (PR) for obesity 1.04 (95% confidence interval: 0.99–1.10)). There was no relationship between ethnicityandsex-specific central obesity (adjusted PR 1.00 (0.97–1.02)). Stratification by region yielded inconsistent associations. Depressionwas weakly but independently linked to obesity (PR 1.08 (1.04–1.12)), and very marginally to abdominal obesity (PR 1.01(1.00–1.03)).

Fecal indicators and bacterial pathogens in bottled water from Dhaka, Bangladesh.

Forty-six bottled water samples representing 16 brands from Dhaka, Bangladesh were tested for the numbers of total coliforms, fecal indicator bacteria (i.e., thermotolerant Escherichia coli and Enterococcus spp.) and potential bacterial pathogens (i.e., Aeromonas hydrophila, Pseudomonas aeruginosa, Salmonella spp., and Shigella spp.). Among the 16 brands tested, 14 (86%), ten (63%) and seven (44%) were positive for total coliforms, E. coil and Enterococcus spp., respectively. Additionally, a further nine (56%), eight (50%), six (37%), and four (25%) brands were PCR positive for A. hydrophila lip, P. aeruginosa ETA, Salmonella spp. invA, and Shigella spp. ipaH genes, respectively. The numbers of bacterial pathogens in bottled water samples ranged from 28 ± 12 to 600 ± 45 (A. hydrophila lip gene), 180 ± 40 to 900 ± 200 (Salmonella spp. invA gene), 180 ± 40 to 1,300 ± 400 (P. aeruginosa ETA gene) genomic units per L of water. Shigella spp. ipaH gene was not quantifiable. Discrepancies were observed in terms of the occurrence of fecal indicators and bacterial pathogens. No correlations were observed between fecal indicators numbers and presence/absence of A. hydrophila lip (p = 0.245), Salmonella spp. invA (p = 0.433), Shigella spp. ipaH gene (p = 0.078), and P. aeruginosa ETA (p = 0.059) genes. Our results suggest that microbiological quality of bottled waters sold in Dhaka, Bangladesh is highly variable. To protect public health, stringent quality control is recommended for the bottled water industry in Bangladesh.

Sensitivity of assays for the detection of HPA-1a antibodies: results of an international workshop demonstrating the impact of cation chelation from integrin αIIbß3 on three widely used assays.

BACKGROUND AND OBJECTIVES: HPA-1a antibodies account for 70-80% of cases of fetal-neonatal alloimmune thrombocytopenia (FNAIT) in Caucasians. However, numerous workshops have demonstrated variability in their detection. We recently showed that exposure of αIIbß3 to ethylene diamine tetraacetic acid (EDTA) affected binding of many anti-αIIbß3 monoclonal, and HPA-1a allo-, antibodies; this adversely affected sensitivity of the monoclonal antibody-specific immobilization of platelet antigens (MAIPA) assay and indirect platelet immunofluorescence test (PIFT). This study presents results from an international workshop studying the impact of cation chelation on HPA-1a antibody detection in routine diagnostic laboratories. MATERIALS AND METHODS: Serum and EDTA-anticoagulated plasma samples containing anti-HPA-1a were distributed to 39 laboratories. Participants were asked to detect and identify any HPA antibodies present. RESULTS: 2/39 (5.1%) participants were able to detect and identify anti-HPA-1a in the serum, but not in the plasma sample. EDTA plasma reduced MAIPA assay sensitivity by ≥ 20% in 17/24 (70.8%) laboratories and by ≥ 50% in 9/24 (37.5%) when using HPA-1a1a platelets (mean: 27.7%, range 0-85.1%); when using HPA-1a1b platelets 3/4 (75%), participants reported ≥ 50% loss of sensitivity (mean 65.6%, range 0-96.6%). A small but significant increase in optical densities was observed in antigen capture ELISA assays when using plasma (mean difference: 0.081, P < 0.01). Insufficient PIFT data were returned to draw firm conclusions. CONCLUSION: Use of EDTA plasma significantly affects the sensitivity of the MAIPA assay and can affect detection of even potent, FNAIT-causing examples of anti-HPA-1a. These data highlight the importance of use of αIIbß3 in an appropriate conformation for the sensitive detection of anti-HPA-1a.

Fecal indicators and bacterial pathogens in bottled water from Dhaka, Bangladesh

Forty-six bottled water samples representing 16 brands from Dhaka, Bangladesh were tested for the numbers of total coliforms, fecal indicator bacteria (i.e., thermotolerant Escherichia coli and Enterococcus spp.) and potential bacterial pathogens (i.e., Aeromonas hydrophil, Pseudomonas aeruginos, Salmonella spp., and Shigella spp.). Among the 16 brands tested, 14 (86%), ten (63%) and seven (44%) were positive for total coliforms, E. coil and Enterococcus spp., respectively. Additionally, a further nine (56%), eight (50%), six (37%), and four (25%) brands were PCR positive for A. hydrophila lip, P. aeruginosa ETA, Salmonella spp. invA, and Shigella spp. ipaH genes, respectively. The numbers of bacterial pathogens in bottled water samples ranged from 28 ± 12 to 600 ± 45 (A. hydrophila lip gene), 180 ± 40 to 900 ± 200 (Salmonella spp. invA gene), 180 ± 40 to 1,300 ± 400 (P. aeruginosa ETA gene) genomic units per L of water. Shigella spp. ipaH gene was not quantifiable. Discrepancies were observed in terms of the occurrence of fecal indicators and bacterial pathogens. No correlations were observed between fecal indicators numbers and presence/absence of A. hydrophila lip (p = 0.245), Salmonella spp. invA (p = 0.433), Shigella spp. ipaH gene (p = 0.078), and P. aeruginosa ETA (p = 0.059) genes. Our results suggest that microbiological quality of bottled waters sold in Dhaka, Bangladesh is highly variable. To protect public health, stringent quality control is recommended for the bottled water industry in Bangladesh.

Autosplenectomy of sickle cell disease in zaria, Nigeria: an ultrasonographic assessment.

OBJECTIVES: During infancy and early childhood, the spleen commonly enlarges in patients with sickle cell anemia (SCA), and it thereafter undergoes progressive atrophy due to repeated episodes of vaso-occlusion and infarction, leading to autosplenectomy in adult life. However, this may not always be the case as some studies have reported splenomegaly persisting into adult life. This study aims to determine and review the prevalence of autosplenectomy by abdominal ultrasonography in sickle cell anemic patients in Zaria, Nigeria. METHODS: An ex-post-facto cross study of 74 subjects was carried out between May to July in 2010. Hematological parameters were determined by an analyzer while B mode Ultrasonography was used to determine the craniocaudal length of the spleen, if visualized. RESULTS: The mean age of the sickle cell subjects was 23.2 ±5.3 years, while that of the controls was 22.7±12.4 years. Of the 74 sickle cell subjects, 55.4% were females; while of the 20 controls, 50% were females. Forty one subjects (55.4%) had autosplenectomy and a significant difference existed in the mean splenic size compared with the control (p<0.0001). Only 3 (4.05%) subjects had splenomegaly, while 23 (31%) had a shrunken spleen. CONCLUSION: Anatomical autosplenectomy is not an uncommon finding in SCA patients. This may be related to inadequate clinical care due to the lack of good health education, ignorance, poverty, and poor standard of care, as well as the lack of newer therapeutic agents.

Autologous SCT with a dose-reduced BU and CY regimen in older patients with non-Hodgkin's lymphoma.

Autologous SCT is a potentially curative procedure for patients with relapsed lymphoma (NHL). We analyzed the outcomes of 34 patients > or =60 years old, including eight patients > or =70 years old, who received BU and CY and SCT for NHL. Patients received BU 0.8 mg/kg i.v. (n=25) or 1 mg/kg p.o. (n=9) q 6 h x 14 doses and CY 60 mg/kg i.v. q day x 2 days. The median age was 66 (range, 60-78) years. Twenty-two patients had large cell, 10 follicular and two-mantle cell lymphoma. Fifteen patients were in a second or greater CR and 19 patients were in a PR. The median days to ANC >500/microl and platelet count >50,000/microl were 10 and 13 days respectively. The 100-day transplant-related mortality was 0%. Toxicities included interstitial lung disease (n=2), seizures in a patient with CNS lymphoma (n=1), mild veno-occlusive disease (n=2), and transient atrial fibrillation (n=4). With a median follow-up of 40 months, the 2-year overall survival and PFS were 67 and 54% respectively. BU/CY is a well-tolerated conditioning regimen for older patients with NHL. Age alone should not be used as an exclusion criterion for autologous SCT.

Utility of osteopontin as a biomarker in recurrent epithelial ovarian cancer.

OBJECTIVE: Osteopontin (OPN) is overexpressed in tumors and serum of ovarian cancer patients and may serve as a biomarker. To evaluate the utility of serum osteopontin in monitoring disease status, we evaluated 234 serum samples from post-oophorectomy patients with ovarian cancer and 38 samples from healthy controls. METHODS: Serum samples were collected from 203 women with recurrent ovarian cancer and 31 newly diagnosed women participating in an experimental chemotherapeutic clinical trial. Controls included 11 young healthy women and 27 peri- or postmenopausal women without ovarian cancer. Samples were assayed for osteopontin using an enzyme-linked immunosorbent assay (ELISA) kit. Statistical analyses for group comparisons of biomarker distribution used the nonparametric Wilcoxon's rank sum test for two-group comparisons and the Kruskal-Wallis test for three-group comparisons. RESULTS: Osteopontin values ranged from 25 to 1463 ng/ml for patients and 25 to 617 ng/ml for controls. Mean patient levels were lower than mean control levels (74 ng/ml vs. 147 ng/ml, respectively, P = 0.0006). Serum osteopontin levels correlated with recurrent disease versus remission (68 ng/ml vs. 34 ng/ml, P = 0.0034), presence of ascites versus absence (71 ng/ml vs. 53 ng/ml, P = 0.0002), and bulky disease vs. nonbulky disease (75 ng/ml vs. 38 ng/ml, P = 0.0005). CA-125 values yielded the same trends with greater statistical difference. CONCLUSIONS: These results demonstrate that serum osteopontin concentrations in post-oophorectomy patients with recurrent ovarian cancer are not greater than in healthy controls. Nevertheless, within this heterogeneous patient population, the values do correlate with bulk of disease. The potential utility of this assay in monitoring women with CA-125 negative disease is worthy of exploration.

Paclitaxel resistance: molecular mechanisms and pharmacologic manipulation.

It has been approximately ten years since the Food and Drug Administration (FDA) approved paclitaxel for the treatment of platinum resistant epithelial ovarian carcinoma. Since the approval, the drug has found therapeutic applications in a variety of schedules and in a wide variety of epithelial malignancies. Its novel mechanism of action provided the hope that it would demonstrate anti-neoplastic activity in multidrug resistant tumor cells. Unfortunately, as with other chemotherapeutic drugs, resistance is commonly seen. Laboratory investigation has defined a wide variety of resistance mechanisms including overexpression of multidrug resistance (MDR-1) gene, molecular changes in the target molecule (betatubulin), changes in apoptotic regulatory and mitosis checkpoint proteins, and more recently changes in lipid composition and potentially the overexpression of interleukin 6 (IL-6). This review describes the in vitro molecular data that define and support the various mechanisms of resistance and critically evaluates the evidence for the participation of these mechanisms in clinically relevant paclitaxel resistance. This review also explores pharmacologic attempts to modulate paclitaxel resistance, principally through inhibition of the MDR-1 drug efflux pump. Future avenues for drug resistance research and its pharmacologic manipulation in the clinic are discussed.

Pharmacokinetics of fluvoxamine in relation to CYP2C19 phenotype and genotype.

OBJECTIVE: To evaluate the pharmacokinetics of fluvoxamine (FLV) in poor metabolizers (PMs) versus extensive metabolizers (EMs) of cytochrome P450 (CYP)2C19. METHODS: This was a prospective, open-label study conducted at the Clinical Research Unit School of Pharmacy. Fifty-seven healthy, nonsmoking volunteers aged 21-40 years participated. Subjects abstained from caffeinated products 12 hours prior to and during each testing period. To assess CYP2C19 activity, blood samples were collected from each subject prior to and two hours after a single dose of omeprazole 20 mg. Once PMs were identified, a sample population of EMs were selected for comparison between the two groups regarding FLV disposition. A single 100 mg FLV dose was given to EMs and PMs; blood samples for FLV analysis were obtained prior to drug administration and 0.5, 1, 2, 3, 4 6, 8, 12 and 24 hours later. A blood sample one day prior to FLV administration was also obtained for CYP2C 19 and CYP2D6. RESULTS: Four PMs were identified with the omeprazole phenotype probe and had a mean +/- SD hydroxylation index of 1.335 +/- 0.271. Nine EMs were selected based upon a hydroxylation index between 0.100 and 0.400 (mean 0.193 +/- 0.079). FLV pharmacokinetic parameters (AUC, elimination half-life, Cmax and Tmax) did not significantly differ between the two groups. Genotype analysis for CYP2C19 revealed a mutant allele for the *2 which confirmed phenotype detection of PM status. Genotype analysis for CYP2D6*3 and *4 alleles showed that all PMs of CYP2C19 were EMs of CONCLUSIONS: FLV disposition and dosing is unlikely to be affected by CYP2C19 polymorphism.

Overexpression of human phosphoglycerate kinase 1 (PGK1) induces a multidrug resistance phenotype.

BACKGROUND: Multidrug resistance is a significant barrier to the development of successful cancer treatment. To identify genetic alterations that are directly involved in paclitaxel resistance, a functional cloning strategy was developed. MATERIALS AND METHODS: Using mRNA from paclitaxel resistant human ovarian cancer cell line SW626TR, a cDNA library was established in a pCMV-Script vector that permits expression of cDNA inserts in mammalian cells. Transfection of the pCMV-Script/SW626TR cDNA library into the paclitaxel-sensitive human osteogenic sarcoma cell line, U-20S, resulted in several paclitaxel-resistant clones. RESULTS: DNA sequencing of clone C16 demonstrates complete homology to human phosphoglycerate kinase 1 (PGK1). Retransfection of the PGK1 insert into U-20S confers a multidrug resistant phenotype, characterized by a 30-fold increase in paclitaxel resistance, and cross-resistance to vincristine; adriamycin and mitoxantrone, but not methotrexate or cisplatin. Enzymatic analysis of the PGK1 transfectants demonstrates an increase in PGK1 activity as compared to the parental cell line, U-20S. Northern and Western analysis of PGK1 transfectants reveals no change in MDR-1 expression compared with the parental cell line. In addition, co-culture of PGK1 transfectants with verapamil only partially reverses the multidrug resistant phenotype. Rhodamine 123 studies are also consistent with an MDR-1 independent mechanism of increased drug efflux. CONCLUSION: Together this data suggests that PGK1 can induce a multidrug resistant phenotype through an MDR-1 independent mechanism.

Cytogenetic studies of spontaneous miscarriages: a seven year study to compare significance of primary vs. secondary culture methods for assessment of fetal karyotype yield and maternal cell contamination.

Primary in-situ culture (PIC) and secondary trypsinized culture (STC) are the two currently used methods for culturing chorionic villi in order to cytogenetically evaluate products of conception (POC) from spontaneous miscarriages. We compare these culture techniques in our laboratory over a period of seven years to evaluate fetal karyotype yield and maternal cell contamination. Data from a total of 2077 cases from 1992-1999 was entered into a data entry program created in Epi Info version 6. Analysis, using the chi square test of significance, was performed in the same program. Our data demonstrated a statistically significant excess of normal female karyotype detected by the STC method and a statistically significant excess of abnormal karyotypes detected by the PIC method. We attribute these findings to the greater risk of maternal cell contamination with the STC method. We conclude that the PIC method is more accurate in detecting the fetal karyotype and the STC method has a higher risk of maternal cell contamination. We suggest that the PIC method should be adopted as the method of choice when evaluating POC by culturing chorionic villi.

Losing God.

Shortly before his death in 1995, Kenneth B. Schwartz, a cancer patient at Massachusetts General Hospital (MGH), founded the Kenneth B. Schwartz Center. The Schwartz Center is a non-profit organization dedicated to supporting and advancing compassionate health care delivery, which provides hope to the patient, support to caregivers, and sustenance to the healing process. The center sponsors the Schwartz Center Rounds, a monthly multidisciplinary forum where caregivers reflect on important psychosocial issues faced by patients, their families, and their caregivers, and gain insight and support from fellow staff members. Nebulous language, distrust, and dogma confound spiritual aspects of cancer care. However, existential well being is an important determinant of quality of life: finding meaning and purpose make suffering more tolerable. The case presented is of a patient who experienced "losing God" as a Hodgkin's disease survivor with metastatic prostate cancer and severe coronary artery disease. His caregivers were able to provide the sense of community in which he could re-establish his faith. Health care providers do not have to be religious in order to help patients to deal with a spiritual crisis. The clinical skills of compassion need to be deployed to diagnose and respond to spiritual suffering. Acknowledging and addressing anger or guilt, common sources of suffering, are essential to adjustment. Simply being there for the patient and being open to their hurt can help resolve their spiritual crisis, a responsibility that is shared by the whole health care team.