Quality and Quantity of Nucleic Acids Extracted from Formalin-Fixed Paraffin-Embedded Lymphoma Biopsies from Nigerian Archived Biopsy.

BACKGROUND: Integrity of nucleic acids derived from archived formalin-fixed paraffin-embedded (FFPE) cancer specimens affects diagnosis, prognosis, and therapy. Several factors affect the quality and quantity of extracted nucleic acids and one of such factors is storage period. AIM: We investigated the impact of storage duration on the quality and quantity of nucleic acids extracted from archived FFPE lymphoma biopsies in Nigeria. MATERIALS AND METHODS: A total of 53 FFPE biopsies diagnosed as lymphoma stored over several years (2008-2019) were analyzed. They were 22 chronic lymphocytic leukemia (CLL) cases, 17 Hodgkin lymphoma (HL) cases, and 14 diffuse large B-cell lymphoma, not otherwise specified (DLBCL, NOS). DNA was extracted from all the lymphoma samples which were analyzed for integrity and amplifiability using the four pairs of control genes polymerase chain reaction (PCR) primers of BIOMED-2 protocol, whereas RNA extraction was from 6 CLL cases used for qPCR analysis of RNU43. RESULTS: For CLL, the mean DNA yield was 193.6 ng/µl (range: 3.0-533.0 ng/µl), whereas the mean A260/A280 ratio was 1.7 (1.2-1.9). For DLBCL, NOS, and HL, 255.5 ng/µl (range: 32.9-605.4 ng/µl), 1.8 (1.5-2.0) and 242.7 ng/µl (range: 1.3-886.0 ng/µl), and 1.7 (0.9-1.8), respectively. The extracted DNA gave amplifiable products of at least 200bp, whereas the RNA analysis showed CT values of <38 in all the samples. The mean RNA yield was 462.2 ng/µl (range: 74.7-1082.1), whereas the mean A260/A280 was 1.7 (1.5-1.8). CONCLUSION: Quantity and quality of nucleic acids from FFPE tissues stored for different time periods showed no significant difference in yield and quality.

Phytosterols demonstrate selective inhibition of COX-2: In-vivo and in-silico studies of Nicotiana tabacum.

The inhibition of cyclooxygenase-1 (COX-1) enzyme by Nonsteroidal anti-inflammatory drugs (NSAIDs) exposes the gastrointestinal mucosa to peptic injuries. Selective inhibition of COX-2 generates surpassing anti-inflammatory drug candidates with reduced side effects over current NSAIDs. Phytosterols consumption is reported to decrease the risk of cardiovascular problems. Reports on the selective inhibition of COX-2 by phytosterols are scarce. The present study assesses the anti-inflammatory potentials of phytosterols from Nicotiana tabacum (of the family Solanaceae) through selective inhibition of COX-1 and/or COX-2. Virtual High Throughput Screening (vHTS) and Molecular Docking of phytochemicals from Nicotiana tabacum against the catalytic pockets of COX-1 and COX-2 were used to identify the lead bioactive(s) components of the plant. The hit phytosterols were isolated, histopathological examination of the stomach, in-vivo COX-1/COX-2 mRNAs expression patterns in the liver through reverse transcription-polymerase chain reactions, and enzymes activities of Nicotiana tabacum phytosterol isolates (NTPI) in HCl/ethanol-induced inflammation in Wistar rats were all investigated. Formation of hydrogen bonds favour selective inhibition of COX-2 while hydrophobic interactions favour selective inhibition of COX-1. NTPI demonstrates inhibition of COX-2 by down-regulate the expression of COX-2 mRNA and were ineffective against the expression COX-1 mRNA. NTPI demonstrates hepatoprotective abilities by improving the antioxidant defense system of the liver. Histopathological analyses show NTPI at 50 mg/kg bodyweight regenerates the parietal cells and maintain the gastrointestinal architecture. Drug likeness prediction and ADME toxicity screening show that phytosterols possess good oral bioavailability with no side effects. Phytosterols are selective inhibitors of COX-2, they are hepatoprotective, regenerate parietal cells, and non-toxic.

Evaluation of the modulatory effects of Piper guineense leaves and seeds on egg albumin-induced inflammation in experimental rat models.

ETHNOPHARMACOLOGICAL RELEVANCE: Inflammation processes are implicated in many degenerative diseases. Piper guineense, a West African spice belonging to the Piperaceae family has been reported to contain anti-inflammatory agents. AIM OF STUDY: This study determined the modulatory effects of methanolic extracts of Piper guineense leaves and seeds on egg albumin-induced inflammation in rats. STUDY DESIGN AND METHODS: Inflammation in the hind paw was induced by injecting 0.1ml egg albumin subcutaneously. Treatments including diclofenac were given orally. Rectal temperature and paw size were monitored hourly for the first 3 h' post-induction of inflammation and then at the 6th and 24th hour. Serum levels of CRP, MDA, LDH and GGT activities were determined at these hours. RESULTS: Results showed that egg albumin-induced inflammation caused a significant (p < 0.05) increase in paw size and rectal temperature. It further showed that treatment with the leaves and seed extracts reversed the effect of inflammation on serum levels of CRP and MDA, and on LDH and GGT activities similar to diclofenac in rats. CONCLUSION: Extracts of the Piper guineense seed and leaves have potentials of being used as an anti-inflammatory agent but further studies need to be done to determine their toxicity and effects on immunological markers of inflammation.

Efficacy of silver nanoparticles against multidrug resistant clinical Staphylococcus aureus isolates from Nigeria.

Multidrug resistant (MDR) S. aureus infections continue to account for excess mortality in hospital and community settings and constitute a rising global health problem. However, data on the efficacy and mechanism of actions of alternative solutions like silver nanoparticles in developing countries are lacking. This study investigated anti-staphylococcal activity of silver nanoparticles (AgNP) against local strains from Nigeria. A total 119 clinical isolates of S. aureus from five Nigerian laboratories categorized as MRSA (n = 52) and MSSA (n = 67) by PCR were studied. The MIC of AgNP produced by chemical reduction method and characterized by surface plasmon resonance absorbance and size equivalence by scanning electron microscopy was determined by microbroth dilution method. Its effect on protease activity and plasmids were also investigated. Baseline characteristics of the isolates revealed MDR phenotype of the isolates, carriage of diverse plasmids (15-32 kb) among the MDR MSSA, and mean extracellular protease activity of 24.8-55.7 U/mL. The chemically synthesized AgNP had a peak absorbance at 400 nm with a size equivalence of 4.58 nm. The MICs of AgNP against the isolates were 4.7 µg/mL and 4.9 µg/mL, respectively, for MRSA and MSSA (P > 0.05). The bactericidal effect of AgNP at 2.5-5 µg/mL on the MSSA and MRSA isolates was observed at 2.7-5.5 h post exposure in vitro. Further analysis revealed plasmid eviction in the MDR MSSA isolates exposed to 5 µg/mL AgNP and dose-dependent reduction in extracellular protease activity by 84.6-93.1%. Hemolysis of human erythrocytes by AgNP was not observed at the MIC range. Conclusion: This study revealed safety and efficacy of AgNP against clinical MDR S. aureus isolates from Nigeria, using plasmid eviction and protease inhibition as mechanisms of action.

Cerium oxide nanoparticle elicits oxidative stress, endocrine imbalance and lowers sperm characteristics in testes of balb/c mice.

The toxicity of metallic nanoparticles is a growing concern due to its application in industries and homes. We investigated the toxicity of cerium oxide nanoparticles (CeO2 NPs) on reproductive system in male balb/c mice. Twenty mice were divided into four groups of five animals each and treated thus: normal saline (control), 100, 200 and 300 µg/kg CeO2 NPs (i.p.,) thrice in a week for five consecutive weeks. Results showed that CeO2 NPs significantly reduced the levels of haemoglobin, PCV and RBC count relative to controls. In addition, luteinising and follicle-stimulating hormones (FSH and LH) and prolactin were significantly reduced in the mice. Specifically, CeO2 NPs at 100 µg/kg decreased testosterone by 23%, while CeO2 NPs at 200 µg/kg decreased FSH, LH and prolactin by 25%, 26% and 13%, respectively. Testicular malondialdehyde was increased by 103%, 106% and 135% in mice treated with 100, 200 and 300 µg/kg CeO2 NPs, respectively. CeO2 NPs caused a significant reduction in activities of antioxidant enzymes and levels of reduced glutathione and total nitric oxide. Moreso, CeO2 NPs decreased sperm motility and count and increased total sperm abnormality in mice. Histology revealed congestion and degeneration of seminiferous tubules. Overall, CeO2 NPs induces testicular dysfunction via disruption of antioxidant/oxidant balance and endocrine suppression.

Doxorubicin induced neuro- and cardiotoxicities in experimental rats: Protection against oxidative damage by Theobroma cacao Stem bark.

80 rats, randomly selected, were divided into 3 treatment groups: pre-, co- and post-treatment; consisting of 6 sub-groups each (5 rats per sub-group): baseline, normal saline (2 mL), α-lipoic acid (20 mg/kg body weight), 200 mg/kg, 400 mg/kg or 800 mg/kg body weight Theobroma cacao stem bark aqueous extract (TCAE). All rats except for baseline group were intoxicated with 20 mg/kg body weight doxorubicin (DOX) intraperitoneally. The animals in pre- or post-treatment group received a single dose of DOX (20 mg/kg body weight) intraperitoneally 24 h before or after 7 days' oral administration with TCAE respectively while those in co-treatment group were co-administered 2.86 mg/kg body weight of DOX with either normal saline, α- lipoic acid or TCAE orally for 7 days. Animals were sacrificed (pre- and post- treatment groups were sacrificed on the ninth day while the co-treatment group sacrificed on the 8th day). Brain and heart tissue samples were harvested for enzyme markers of toxicity, oxidative stress and histopathological examinations. DOX intoxication caused significant decrease in activities of LDH and ACP, and increase in γGT and ALP activities in brain tissues while causing a significant increase in LDH, ACP, γGT activities and decrease in ALP activity in the cardiac tissues. DOX intoxication caused a significant increase in concentrations of H2O2 generated, MDA and PC, XO, MPx and NOX activities with concomitant decrease in CAT, SOD, GPx and GST activities, and in concentrations of GSH, AsA and α-Toc in brain and cardiac tissues. Pre-, co- and post-treatment with TCAE at either 200 mg/kg, 400 mg/kg or 800 mg/kg body weight significantly reversed the oxidative damage to the organs induced by DOX-intoxication. The result affirmed that T. cacao stem bark aqueous extract protected against DOX induced oxidative damage in brain and cardiac tissues of experimental rats.

Diagnostic Value of Procalcitonin in Neonatal Sepsis

Introduction: Neonatal sepsis is a major cause of mortality in developing countries. Accurate and quick diagnosis are difficult because clinical presentation are non-specific; bacterial cultures are time-consuming and other laboratory tests lack sensitivity and specificity. Serum procalcitonin (PCT) has been proposed as an early marker of infections in neonates. Objectives: This study investigated the value of PCT in the diagnosis of Neonatal Sepsis.Methods: Neonates undergoing sepsis evaluation at the Special Baby Care Unit; Federal Medical Centre; Abeokuta; Nigeria between January and April 2013 were included. Blood samples were obtained for white cell count; blood cultures; serum CRP and PCT analysis. Neonates were categorised into Proven Sepsis; Suspected Sepsis and Clinical Sepsis groups on the basis of laboratory findings and risk factors. A control group with no clinical and biological data of infection was also included. Predictive values and area under the receiver operating characteristic curve (AUC) of PCT were evaluated.Result: Of the 85 neonates; 19 (22.4%) had positive blood culture. PCT level was significantly higher in neonates in all sepsis groups in comparison with those in the control group (P 0.05). At a cut-off of 0.5 ng/ml; the negative predictive value (NPV) of PCT was 80% and the positive predictive value (PPV) 39%. There were no significant statistical difference between the AUC values of PCT in Early onset and Late onset sepsis; as well between AUC in Preterm and term cases. A higher percentage of neonates who died (96%) had elevated PCT levels compared to those who survived (46%).Conclusion: These findings support the usefulness of the PCT in diagnosis of Neonatal sepsis

Red Palm Oil Attenuates Lead Acetate Induced Testicular Damage in Adult Male Sprague-Dawley Rats.

To study the protective effect of Red Palm Oil (RPO) on testicular damage induced by administration of lead acetate on male Sprague-Dawley rats, 28 rats divided into four groups of 7 animals each were used. They were administered orally with RPO (1 mL and 2 mL) and lead acetate (i.p.) 6 mg/kg body weight/day, respectively. Treatment was conducted for 8 weeks, and 24 hrs after the last treatment the rats were sacrificed using cervical dislocation. Sperms collected from epididymis were used for seminal fluid analyses; while the testes sample was used for ROS and oxidative enzyme activities assessment. Statistical analysis was carried out using GraphPad Prism 5.02 statistical analysis package. Administration of lead acetate increased generation of reactive oxygen species (ROS) significantly (p < 0.05) as evidenced by the elevated value of H2O2 and LPO and decreased GSH level. Also there was reduced epididymal sperm count, poor grade of sperm motility, and lower percentage of normal sperm morphology significantly. Coadministration with RPO, however, has a protective effect against lead toxicity by decreasing H2O2 production, increased GSH level, and increased sperm qualities especially. This shows that RPO has a potential to attenuate the toxic effect of lead on testicular cells preventing possible resultant male infertility.

Serum Testosterone, 17ß-Estradiol and PSA Levels in Subjects with Prostate Disorders.

Prostate carcinoma is the most frequently diagnosed malignancy and the second leading cause of death as a result of cancer in men in the US and other parts of the world. There are conflicting reports on the serum levels of testosterone and 17ß-estradiol (E2) in benign prostatic hyperplasia (BPH) and prostate cancer. This study was designed to evaluate the serum concentrations of these hormones in patients with these disorders. Serum levels of prostate specific antigen (PSA), total testosterone and estradiol were determined in 228 subjects comprising of 116 subjects with BPH, 62 subjects with prostate cancer (CaP) and 50 age-matched apparently healthy controls, using ELISA methods. PSA levels were significantly elevated (p < 0.05) in BPH subjects than controls, while there was no significant difference (p > 0.05) in testosterone and estradiol levels of these subjects. PSA and estradiol levels were significantly higher (p < 0.05) in CaP subjects than in controls, while there was no observed significant difference (p > 0.05) in testosterone levels. CaP subjects had significantly raised PSA, testosterone, and estradiol levels than BPH subjects. The mean molar ratio of testosterone: E2 was lowest among CaP patients (134:1) and highest among controls (166:1). Significant positive correlation between PSA and 17ß-estradiol was observed in prostate disorders (BPH and CaP patients: r = 0.347; p = 0.000). Significant negative correlations between testosterone and PSA were also observed among BPH patients (r = -0.221, p = 0.049) and control subjects (r = -0.490, p = 0.000). No significant correlation existed between testosterone and PSA in CaP patients (r = 0.051, p = 0.693). Correlations between age and estradiol in both BPH and CaP were not significant (p > 0.05). This study has shown that, there was a significant increase in serum estradiol in CaP subjects, while the testosterone levels in both BPH and CaP subjects were not different from those of controls.

Coexistence of meconium ileus with duodenal atresia and trisomy 21 in a newborn: a case report.

The coexistence of duodenal atresia (DA) may mask the antenatal ultrasound findings of meconium ileus (MI) and delay its postnatal diagnosis. We report a rare case of MI in a newborn infant diagnosed antenatally to have trisomy 21 and DA. The diagnosis of MI was only established intraoperatively after the patient showed persistent signs of intestinal obstruction following the surgical repair of the DA.

Variation in CAG and GGN repeat lengths and CAG/GGN haplotype in androgen receptor gene polymorphism and prostate carcinoma in Nigerians.

Prostate cancer has become the most common cancer in Nigerian men. The growth of the prostate gland depends on circulating androgens and intracellular steroid signalling pathways. The effects of androgens are mediated through the androgen receptor (AR), a nuclear transcription factor encoded by the AR gene. The common polymorphisms, CAG and GGN repeats, in exon 1 of this gene have been implicated as possible risk factors. Thus far, existing supporting data are scanty and none are from sub-Saharan African populations. Therefore, this study investigates the possible association between AR polymorphism repeat length (CAG and GGN) and prostate cancer in Nigerians. A total of 261 subjects (70 with prostate cancer, 68 with benign prostate hyperplasia [BPH], 123 age-matched apparently normal subjects as controls) were studied. CAG and GGN repeats length were determined by fragment length analysis using GeneScan. The CAG repeat length in prostate cancer and in BPH compared to the controls was significantly different (P < 0.05) with reduce length of CAG repeats showing a significant odds ratio (OR) in both cases. However, this was not observed in GGN repeat length, which showed no significant difference between cases and controls (P > 0.05). CAG and GGN haplotype variation showed no significant difference between cases and controls (P > 0.05), except that the haplotypes CAG > or =21 and GGN < or =21 were more common in the control group. The results of this study, the first from sub-Saharan Africa, supports the hypothesis that reduced CAG repeat length is a risk factor for prostate cancer, and also suggests an association with BPH.

Effects of contraceptives on serum trace elements, calcium and phosphorus levels / Efectos de los contraceptivos sobre los elementos traza en suero, y los niveles de calcio y fósforo

BACKGROUND: Women on different contraceptive methods have been linked with the development of various diseases and possible changes in serum trace elements and vitamins of women on contraceptives have been postulated. Therefore, the relationship between contraceptive use and trace elements needs to be investigated. Methods:This is a cross-sectional randomized study. After informed consent was obtained, blood samples were collected from a total of 100 women of child-bearing age on different contraceptive methods: 50 on oral contraceptives, 25 on injectables and another 25 on intra-uterine device. Blood samples were also collected from another 50 age-matched non-contraceptive users to serve as control. Serum was analysed using atomic absorption spectrophotometer for zinc, copper, manganese, iron, selenium, cadmium, lead and magnesium while colorimetric method was used for phosphorus and calcium. Body mass index (BMI) was calculated as weight in kilogram/height in meter squared. Results obtained from laboratory analysis and anthropometric measurements were analysed using computer SPSS package. Results:The mean serum zinc, selenium, phosphorus and magnesium levels obtained from subjects on contraceptives were significantly lower (p < 0.01, p < 0.05, p < 0.05 and p < 0.05 respectively) than those of the control group. However, the mean serum copper, iron, calcium and cadmium levels were significantly higher (p < 0.05) in participants on contraceptive when compared with the control group. Manganese and lead levels were similar in participants and control groups. Correlation analysis shows significant association between some trace elements and the duration ofcontraception and body mass index ofthe participants. CONCLUSION: The study showed and confirmed reduced levels of trace elements in women on contraceptives. The reduction is proportional to the duration ofcontraceptive use.

ANTECEDENTES: Mujeres que usan diferentes métodos anticonceptivos han sido vinculadas al desarrollo de varias enfermedades, y se han presupuesto cambios en los elementos traza en suero y las vitaminas de mujeres en torno a los contraceptivos. Por lo tanto, se hace necesario investigar la relación entre el uso de contraceptivos y los elementos traza. MÉTODOS: Éste es un estudio transversal randomizado. Después de obtener el consentimiento infor mado, se recogieron muestras de sangre de un total de 100 mujeres de edad de maternidad en relación con diferentes métodos anticonceptivos: 50 con contraceptivos orales, 25 con inyectables, y otras 25 con dispositivos intrauterinos. También se tomaron muestras de sangre de otras 50 mujeres pareadas por edad y no usuarias de anticonceptivos, para servir de control. Se analizó el suero usando el espectrofotómetro de absorción atómico para zinc, cobre, manganeso, hierro, selenio, cadmio, plomo, y magnesio, mientras que el método colorimétrico se usó para el fósforo y el calcio. El índice de masa corporal (IMC) fue calculado como el peso en altura/kilogramo en metro cuadrado. Los resultados obtenidos del análisis de laboratorio así como las dimensiones antropométricas, se analizaron usando el paquete computacional SPSS. RESULTADOS: Los niveles promedio en suero de zinc, selenio, fósforo y los niveles de magnesio obtenidos de sujetos bajo el uso de contraceptivos fueron significativamente más bajos (p < 0.01, p < 0.05, p < 0.05 y p < 0.05 respectivamente) que aquéllos del grupo de control. Sin embargo, los niveles promedio en suero de cobre, hierro, calcio y cadmio fueron significativamente más altos (p < 0.05) en los participantes bajo el uso de contraceptivos, en comparación con el grupo de control. Los niveles de manganeso y plomo fueron similares en los participantes y los grupos de control. El análisis de la correlación muestra una asociación significativa entre algunos elementos traza, y la duración de la contracepción y el índice de masa corporal de los participantes. CONCLUSIÓN: El estudio mostró y confirmó niveles reducidos de elementos traza en las mujeres bajo contraceptivos. La reducción es proporcional a la duración del uso de contraceptivos.

Effects of contraceptives on serum trace elements, calcium and phosphorus levels.

BACKGROUND: Women on different contraceptive methods have been linked with the development of various diseases and possible changes in serum trace elements and vitamins of women on contraceptives have been postulated. Therefore, the relationship between contraceptive use and trace elements needs to be investigated. METHODS: This is a cross-sectional randomized study. After informed consent was obtained, blood samples were collected from a total of 100 women of child-bearing age on different contraceptive methods: 50 on oral contraceptives, 25 on injectables and another 25 on intra-uterine device. Blood samples were also collected from another 50 age-matched non-contraceptive users to serve as control. Serum was analysed using atomic absorption spectrophotometer for zinc, copper manganese, iron, selenium, cadmium, lead and magnesium while colorimetric method was used for phosphorus and calcium. Body mass index (BMI) was calculated as weight in kilogram/height in meter squared. Results obtained from laboratory analysis and anthropometric measurements were analysed using computer SPSS package. RESULTS: The mean serum zinc, selenium, phosphorus and magnesium levels obtained from subjects on contraceptives were significantly lower (p < 0.01, p < 0.05, p < 0.05 and p < 0.05 respectively) than those of the control group. However, the mean serum copper iron, calcium and cadmium levels were significantly higher (p < 0.05) in participants on contraceptive when compared with the control group. Manganese and lead levels were similar in participants and control groups. Correlation analysis shows significant association between some trace elements and the duration of contraception and body mass index of the participants. CONCLUSION: The study showed and confirmed reduced levels of trace elements in women on contraceptives. The reduction is proportional to the duration of contraceptive use.

Antioxidant Levels of Acute Leukaemia Patients in Nigeria

The incorporation of nutritional screening and comprehensive assessments of oxidative stress is increasingly recognised as imperative in the development of standards for quality care in oncology. This study evaluated the levels of nitric oxide (NO); some essential trace metals (Zn; Cu; Fe; and Se); superoxide dismutase (SOD) activity and malondialdehyde (MDA) in twenty five (25) patients with acute leukaemia and 25 apparently healthy controls. The mean levels of plasma Zinc (Zn); Iron (Fe) and Selenium (Se) were not significantly elevated (p 0.05) in leukaemia patients compared with controls. Also; slightly lower level of plasma Cu was observed in leukaemia patients compared with the controls. However; nitric oxide was significantly increased (p 0.05) in leukaemia patients compared with controls. The implication of the present finding is that intervention to increase antioxidant status in patients with Acute Lymphoblastic Leukaemia (ALL) should be considered

Trace elements and vitamin E status in Nigerian patients with prostate cancer.

BACKGROUND: Prostate cancer is the most common type of cancer among men. OBJECTIVES: To investigate the trace elements (Se, Zn, Cu and Cd) and vitamin E status of some Nigerian prostate cancer (PCa) patients relative to their prostate-specific antigen (PSA) values. METHODS: Prostate cancer patients were assigned into groups 1, 2 and 3 with PSA of 5-10 ng/ml, 11-20 ng/ml and > 20 ng/ml, respectively. RESULTS: The results showed that the levels of whole blood superoxide dismutase (SOD) and serum Se and Zn were significantly lower (p< 0.05) in the PCa patients. Specifically, levels of SOD, Se and Zn decreased by 67%, 30% and 35%; 70%, 52% and 41%; 81%, 58% and 47%, in subjects with PSA of 5-10 ng/ml, 11-20 ng/ml and > 20 ng/ml, respectively. There were no significant differences (p> 0.05) in levels of Cu and Cd. Serum Cu/ Zn ratio were significantly higher in PCa patients. The Cu/ Zn ratios were 1: 1.2: 1.3 for subjects in groups 1, 2 and 3, respectively. Vitamin E levels in PCa patients were significantly lower and followed the order; normal > PSA (5-10) > PSA (11-20) > PSA (> 20). CONCLUSIONS: Deficiency of vitamin E, Zn and Se may be risk factors for development of PCa.

Androgen receptor gene CAG and GGN polymorphisms in infertile Nigerian men.

The human androgen receptor gene (AR) is an important regulator of male sexual development including spermatogenesis. Exon 1 of this gene encodes the N terminal domain, which controls transcriptional activity of the receptor and the two polymorphic repeats CAG and GGN. Many studies have reported association of the expanded CAG repeat length with male infertility, although this is still controversial. The GGN repeat, in contrast, has been less thoroughly studied. Thus far, only scanty studies have been reported from African populations and none from Nigeria. Therefore, we have investigated the possible association between AR polymorphism repeats length (CAG and GGN) and reduced spermatogenesis in infertile Nigerian men (no.=60) consisting of 20 non-obstructive azoospermic and 40 oligozoospermic subjects compared with controls with normozoospermia and proven evidence of fertility (no.=38). In addition, 48 volunteers with normal spermatogenesis were recruited from a German population. CAG and GGN repeats length were determined by fragment length analysis using GeneScan. The CAG and GGN repeats length of infertile compared to fertile populations were not significantly different (p>0.05). We found a unique AR GGN allele distribution with 20-23 GGN repeats predominant in the Nigerian study population. Our results show that CAG and GGN repeats polymorphisms are not a critical index of male infertility. While we do not find a relationship with CAG and GGN repeats haplotypes and male infertility, we report for the first time a unique and wider distribution of the GGN allele in the Nigerian population which is significantly different from the Caucasian population. The functional relevance of this variance to male fertility warrants in-depth elucidation.

The Use of Rap-Pcr in Studying Mycobacterium Tuberculosis Intracellular Gene During Macrophage Infection

Mycobacterium tuberculosis is the second leading cause of death from infectious agent. This study sought to detect M. tuberculosis genes; which were specifically expressed; or upregulated during intracellular infection of J774 murine macrophages; as such genes may be potential targets for novel drug action. J774 murine macrophage cell line was infected with M. tuberculosis (H37Rv strain) at 10:1 multiplicity of infection (MOI). RNA was differentially extracted from M. tuberculosis infecting J774 macrophage cell line. The control in this case was RNA from extracellular broth grown bacteria. Approximately 50 ng of RNA from intracellular derived bacteria and extracellular derived bacteria (control) were subjected to random arbitrarily primed PCR (RAP-PCR) using 50 primer combinations. Eleven differential RAP-PCR products were observed. All RAP-PCR products were cloned into pCRr2.1 and sequenced in order to determine the identity of the products. Four of the eleven products were derived from macrophage genes and another 4 products were derived from the M. tuberculosis rRNA genes (three 23S and one 16S rRNA). The 3 remaining RAP-PCR products were found to be mycobacterial genes other than ribosomal genes. The three products were genes encoding enzyme involving in a shikimate pathway; a putative carboxyphosphonoenolpyruvate phosphonomutase and a serine protease with homology to HtrA. Of the 3 mycobacterial genes other than ribosomal genes detected; none were specifically expressed during intracellular infection but bacilli

Mutation analysis of the X-chromosome linked, testis-specific TAF7L gene in spermatogenic failure.

The precise temporal and spatial expressions of specific transcription regulation factors (TRF) have long been considered essential for spermatogenesis. Recently, it has been speculated that mammals have evolved more specialised TRF genes. In the human, the TAF7L gene may be essential for maintenance of spermatogenesis. In this study, we investigated the possible role of the TAF7L gene located on the X chromosome in testicular function and spermatogenic failure. In a case-controlled retrospective study, we recruited 16 infertile males with consistent, nonobstructive azoospermia and with normal serum follicle-stimulating hormone (FSH) levels. Twenty age-matched men with normal spermatogenesis with the same ethnic background (Caucasian) were recruited as controls. Their genomic DNA was screened for sequence changes in the coding regions and part of the flanking introns of the TAF7L gene by direct sequencing. Amino acid sequence was compared with the NCBI standard sequence (BC043391). Semen analysis and hormone evaluation were performed. We observed six sequence variations in four patients, consisting of two point mutations, one each in exon 9 and 13 and one six-basepair deletion in exon 13 with concomitant changes in amino acid. One additional nucleotide exchange was observed in intron 8. Most of these changes were also found in eight controls with the exception of changes in exon 13. A meta-analysis including the present study and literature data suggests a possible association of the point mutation in exon 13 with infertility. There was no association or relationship with reproductive hormones. In conclusion, the sequence variants in the cDNA sequence observed are common polymorphisms. The changes in intron 8 appear novel. We report for the first time that most of the alterations are not associated with gonadal dysfunction, while the sequence variant in exon 13 may represent a risk factor for spermatogenic failure.

Asymptomatic bacteriuria of pregnancy in Ibadan, Nigeria: a re-assessment.

Asymptomatic bacteriuria in pregnancy is the major risk factor for developing symptomatic urinary tract infection during pregnancy. In the present study, 300 pregnant women are screened for significant asymptomatic bacteriuria in order to provide an insight into the prevalence in developing countries, reassessment of some predisposing factors and aetiological agents and their susceptibility tests. The mean age of the patients in the study is 26.8 years (SD: 5.8 years, range: 16-40 years). Using 10(3) organisms/mL as a significant level of bacteriuria, the prevalence was found to be 21.0%. One hundred and fifty-eight samples had no pus cells, with 25 showing significant bacteriuria, 116 samples contained 1-4 pus cells/high power field (hpf) with 25 showing significant bacteriuria, while 26 samples had > or = 5 pus cells/hpf with 13 showing significant bacteriuria. There was no particular trend associated with age and rate of infection. However, there was a decline in the rate of infection in the 26-30 age group, with a sharp increase as age increased. There was high incidence of bacteriuria during the third trimester of pregnancy (21.9%) compared with that in the first trimester (7.7%), while the level in the second trimester was 22.5%. Multiparity is associated with increased bacteriuria in pregnancy. Thirty-one (49.2%) isolates grew Gram-negative bacilli; 27 (42.9%) grew Gram-positive cocci and the remainder (7.9%) grew yeast-like cells. Staphylococcus aureus was the most frequent pathogen (41.3%), followed by Klebsiella species (33.3%) and Escherichia coli (11.1%). Bacterial isolates from this study were most sensitive to ceftazidime, followed by ceftriazone, and least susceptible to co-trimoxazole.

Possible antidiabetic and antihyperlipidaemic effect of fermented Parkia biglobosa (JACQ) extract in alloxan-induced diabetic rats.

1. The hypoglycaemic effect of fermented seeds of Parkia biglobosa (PB; African locust bean), a natural nutritional condiment that features frequently in some African diets as a spice, was investigated in the present study in alloxan-induced diabetic rats. Its effect was compared with that of glibenclamide (Daonil; Sanofi-Aventis, Paris, France), a reference antidiabetic drug. The effects of PB on lipid profiles were also examined. 2. In order to assess the hypoglycaemic and hypolipidaemic effects of aqueous and methanolic extracts of PB on experimental animals, fasting plasma glucose (FPG), total cholesterol, triglyceride, high-density lipoprotein (HDL) and low-density lipoprotein (LDL) were determined. In addition, the weight of each animal was determined to assess any possible weight gain or loss in the experimental animals (diabetic rats treated with Daonil (group C), the aqueous extract of PB (group D) or the methanolic extract of PB (group E)) compared with control groups (non-diabetic (group A) and non-treated diabetic (group B)). 3. A single dose of 120 mg/kg, i.v., alloxan administered to rats resulted in significant increases in the FPG (P < 0.001) of test animals compared with controls. However, dietary supplementation with PB (6 g/kg extract for 4 weeks administered orally using an intragastric tube) ameliorated the alloxan-induced diabetes in a manner comparable with that of the reference antidiabetic drug glibenclamide. Aqueous and methanolic extracts of PB (6% w/w) elicited 69.2% and 64.4% reductions, respectively, in FPG compared with 70.4% in 0.01 mg/150 g glibenclamide-treated rats. 4. Although animals treated with the aqueous extract of PB gained weight in manner similar to normal controls, animals given the methanolic extract and glibenclamide lost weight in manner similar to non-treated diabetic rats. In addition, high levels of HDL and low LDL were observed in animals treated with the aqueous extract of PB, a pattern similar to that seen in normal controls. Low levels of HDL and high levels of LDL were observed in animals treated with the methanolic extract of PB, a pattern similar to that seen in non-treated diabetic controls. 5. The results of the present study demonstrate that both aqueous and methanolic extracts of fermented seeds of PB exert a hypoglycaemic effect; hence, PB has an antidiabetic property. However, only the aqueous extract of PB ameliorated the loss of bodyweight usually associated with diabetes. Although the aqueous extract has a favourable lipid profile, which is probably an indication of its possible anti-arteriogenic property (hypertension and ischaemic heart diseases being common complications in diabetes mellitus), the methanolic extract shows possible contraindication to ischaemic heart diseases.