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Biomedical subjects

N Ahmad

Publications and source records attributed to N Ahmad.

At least 37 records · Page 2Linked to original sources

Sephadex and sephadex ion-exchange filtration improves the quality and freezability of low-grade buffalo semen ejaculates.

The effect of sephadex and sephadex ion-exchange filtration on the improvement in quality and freezability of low-grade buffalo semen ejaculates was assessed. Two types of filtration columns were used: one containing only sephadex G-10 (FS) and the other sephadex G-10 along with ion-exchangers (diethyl amino ethane-52 (DEAE-52) cellulose and carboxy methyl-52 (CM-52) cellulose; FS+IE). Unfiltered samples served as controls. Semen ejaculates extended in Tris-citric acid (1:4) (n=16; initial motility 40-50%) were filtered at the rate of 1.5 ml/min under negative pressure at room temperature (28-30 degrees C). The mean recovery rate (%) of motile spermatozoa in the FS (85.9+/-1.51) and FS+IE (77.10+/-2.28) filtrates did not differ significantly. Percentages of sperm motility, normal acrosomes, and intact plasma membranes were highest (P<0.05) in FS+IE, intermediate (P<0.05) in FS and lowest (P<0.05) in controls at the three stages of cryopreservation (postfiltration final dilution, after equilibration, and after freezing). Mean sperm abnormalities were lowest (P<0.05) in the filtrates of FS+IE, moderate (P<0.05) in FS and highest in controls at all stages of freezing. Compared to dilution and equilibration, freezing greatly reduced (P<0.05) the overall percent motility, normal acrosomes and intact plasma membranes. The spermatozoa eluted through FS+IE columns proved more resistant (P<0.05) in bearing dilution, equilibration, freezing and thawing stresses than the spermatozoa from FS and control samples. It is concluded that filtration systems containing an FS+IE column can effectively enhance the quality and freezability of extended, low quality buffalo semen.

Acrosome↗

Caesium concentration in the Pakistani diet.

Daily dietary intakes of radioactive and non-radioactive caesium for the Pakistani population were measured. Food samples were collected on market basket method and were analysed using Instrumental Neutron Activation Analysis technique. The radioactive caesium ((137)Cs) in these samples was below the detectable limit, i.e. 1 mBq g(-1). The geometric mean of the mass fraction of stable caesium was 9.56 x 1.53 ng g(-1). The estimated daily dietary intake of caesium was 5.65 x 1.53 microg d(-1) or 0.088 microg kg(-1) of body weight of the reference Pakistani man weighing 64 kg. These values are 38.2% smaller than the recommended ICRP values for a 70 kg standard man. However, caesium concentration in the Pakistani diet is comparable with other reported values in the literature.

Cesium Radioisotopes↗

Nutritional and radiological impact of dietary potassium on the Pakistani population.

The nutritional status of dietary potassium and the radiological impact of its isotope (40)K were estimated for the Pakistani population. Potassium concentration was determined in dietary samples by atomic absorption spectrometry. An average potassium concentration was 4.54+/-0.89 mg/g, which leads to potassium dietary intake of 2.69+/-0.54 g/day and the radiological hazard due to (40)K activity was 79.94 Bq/day. The annual and lifetime effective doses calculated for the reference Pakistani man weighing 64 kg due to measured activity of (40)K were found to be 1.65 x 10(-4) Sv y(-1) and 8.27 x 10(-3) Sv, respectively. Measured data showed that the potassium concentration in the Pakistani diet is adequate and safe according to international standards.

Adult↗

An observational study investigating the health provision for children with epilepsy within secondary schools in the East Midlands, UK.

BACKGROUND: Epilepsy is a common medical condition affecting many children in mainstream education. The practice of local secondary schools and the health service with regard to pupils with epilepsy was examined. The extent to which these children had activities at schools restricted and the schools' perception of the health service with regard to epilepsy was explored. METHOD: A structured interview was conducted with selected staff members from mainstream secondary schools in East Midlands region, UK. RESULTS: Fourteen out of 17 schools participated. Fifty-seven children were identified from a total pupil population of 14 534. Seven schools used the DOH/DfEE (The Department of Health/Department for Education and Employment) 'Supporting Pupils with Medical Needs' publication as a resource. No schools created individual health care plans for children with epilepsy. Six out of seven schools reported appropriate provision for children during swimming activities. The commonest restriction was related to computer use. Nine schools stated a need for more training for staff members with regard to seizure management. CONCLUSIONS: Schools rely heavily on obtaining medical information from non-medical sources. Individual health care plans for children with epilepsy are not commonly undertaken. Few children had any restriction or special provision placed upon them because of their epilepsy except in the area of computer use. Further action is needed to consider how the system for providing for such children can be improved so that each child's individual health needs can be satisfactorily met.

Adolescent↗

Thorium concentration in Pakistani diet.

To evaluate radiation doses due to ingestion of thorium activity by members of the public, baseline analytical data on thorium concentration in a typical Pakistani diet was generated. Food samples were collected from major districts/ cities of the country using the market basket method, and typical daily diets were prepared. Thorium concentration was determined by instrumental neutron activation analysis. It was found to vary from 1.6 ng g(-1) to 12 ng g(-1). The median value was 4.6 ng g(-1). This leads to median daily thorium intake of 2.7 microg d(-1). The geometric mean was 2.6 microg d(-1), which was 13% less than the ICRP value. Based on these measurements the average intake of 232Th activity was 10 mBq d(-1), and the average annual and committed effective doses for adults were estimated to be 8.8 x 10(-7) Sv y(-1) and 4.4 x 10(-5) Sv, respectively.

Diet↗

Do private doctors follow national guidelines for managing pulmonary tuberculosis in Pakistan?

As private medical practitioners play a major role of in providing care to pulmonary tuberculosis (TB) patients, a survey was made of knowledge and practice in 2 cities in Pakistan. Only 1 of the 245 physicians was aware that cough > 3 weeks alone is the main symptom suggesting pulmonary TB. The majority diagnosed (80%) and treated (83%) cases themselves without referral. Less than 1% relied on sputum microscopy alone for diagnosis. None of the practitioners were following National TB Control guidelines for prescribing drugs and none ensured compliance with anti-TB treatment under supervision of a doctor/health worker. Only 3% kept records of pulmonary TB patients. None of the physicians assessed the effectiveness of treatment with sputum microscopy alone; the majority (76%) used only clinical assessment.

Adult↗

Intercomparison of INAA and ICP-MS results for thorium determination in Pakistani diet.

A pilot study on ingestion and organ content of trace elements of importance in radiological protection was to be carried out in Pakistan. Baseline analytical data on daily dietary intake of thorium was to be measured using Instrumental Neutron Activation Analysis (INAA) technique. To determine the accuracy and reliability of our technique, some samples were measured in Pakistan using INAA and in Japan using Inductively Coupled Plasma-Mass Spectrometry. For intercomparison of results overall mean Z-scores were calculated. The results showed validity of our technique. Mean value of 232Th concentration in Pakistani diet samples using INAA technique is 0.0062 +/- 0.0028 microg/g and with ICP-MS technique is 0.0069 +/- 0.0032 microg/g.

Diet↗

Biological characterization of HIV type 1 envelope V3 regions from mothers and infants associated with perinatal transmission.

Our previous study has shown that the human immunodeficiency virus type 1 (HIV-1) envelope V3 region minor genotypes of infected mothers were transmitted to their infants and predominated initially as a homogeneous virus population in the infants (Ahmad N, Baroudy BM, Baker RC, et al.: J Virol 1995;69:1001-1012). Here we have characterized the biological properties, including cellular tropism, replication efficiency, cytopathic effects, and coreceptor utilization, of these V3 region isolates from mothers and infants. Nineteen V3 region sequences from three mother-infant pairs, including the minor variants of mothers and the major variants of infants as characterized in our previous study, were reciprocally inserted into an HIV-1 infectious molecular clone, pNL4-3, and chimeric viruses were generated by DNA transfections into HeLa cells. Equal amounts of chimeric viruses were then used to infect T lymphocyte cell lines (A3.01 and MT-2), primary blood lymphocytes (PBLs), primary monocyte-derived macrophages (MDMs), and coreceptor cell lines. We found that the V3 region chimeras failed to replicate in T lymphocyte cell lines but replicated in MDMs and PBLs, albeit at reduced levels compared with R5 laboratory HIV-1 strains. In addition, the V3 region chimeras were able to infect the HOS-CD4(+)CCR5(+) cell line, suggesting CCR5 coreceptor utilization. Moreover, the V3 region chimeras were unable to induce syncytia in MT-2 cells, indicative of non-syncytium-inducing (NSI) phenotypes. In conclusion, the HIV-1 minor genotypes of infected mothers with macrophage-tropic and NSI or R5 phenotypes are transmitted to their infants and are initially maintained with the same properties.

Amino Acid Sequence↗

Genetic characterization of HIV type 1 gag p17 matrix genes in isolates from infected mothers lacking perinatal transmission.

The gag p17 matrix sequences of human immunodeficiency virus type 1 (HIV-1) were analyzed from three nontransmitting mothers (mothers who failed to transmit HIV-1 to their infants in the absence of antiretroviral therapy), including multiple deliveries in the case of mother 3. There was a low degree of heterogeneity of gag p17 matrix sequences in nontransmitting mothers compared with our previously analyzed mother-infant pairs' sequences. Whereas most of the functional domains essential for p17 matrix function were generally conserved, the polymerization site was less conserved. Several amino acid motifs, including KIEEEQN (positions 103-109) at the major antibody-binding site, were variable and the C-terminal 6-mer QVSQNY, a lysine or glutamine at position 15, an alanine at position 54, a lysine at position 76, a valine at position 104, and an aspartic acid at positions 102 and 121 were conserved in nontransmitting mothers' sequences compared with transmitting mothers' sequences. Phylogenetic analyses of 82 p17 matrix sequences revealed distinct clusters for each nontransmitting mother. Some of these motifs in gag p17 matrix sequences that are present in nontransmitting mothers and absent in transmitting mothers could be used as new targets for the development of preventive strategies for perinatal transmission.

Adult↗

Involvement of the retinoblastoma (pRb)-E2F/DP pathway during antiproliferative effects of resveratrol in human epidermoid carcinoma (A431) cells.

Resveratrol (trans-3,4',5-trihydroxystilbene), a polyphenolic phytoalexin found in grapes, nuts, many other fruits, and red wine, is a potent antioxidant with anti-inflammatory and cancer-preventive properties. The mechanism(s) by which resveratrol imparts cancer chemopreventive effects has not been clearly defined. Earlier, we have shown that resveratrol treatment results in an induction of the cyclin kinase inhibitor WAF1/CIP1/p21 which, by inhibiting cyclin (E, D1, and D2) and cyclin-dependent kinases (cdk2, cdk4, and cdk6), results in a G0/G1-phase arrest followed by apoptosis of A431 human epidermoid carcinoma cells (Ahmad et al., Clin. Cancer Res. 7, 1466-1473, 2001). Retinoblastoma (pRb) and the E2F family of transcription factors are important proteins, which regulate the progression of the cell cycle at and near the G1-->S phase transition. Here we provide evidence for the involvement of the pRb-E2F/DP pathway as an important contributor of resveratrol-mediated cell cycle arrest and apoptosis. Immunoblot analysis demonstrated that resveratrol treatment of A431 cells results in a dose- as well as time-dependent decrease in the hyperphosphorylated form of pRb with a relative increase in hypophosphorylated pRb. This response was accompanied by downregulation of protein expression of all five E2F ( 1 - 5 ) family members of transcription factors studied and their heterodimeric partners DP1 and DP2. This suggests that resveratrol causes a downregulation of hyperphosphorylated pRb protein with a relative increase in hypophosphorylated pRb that, in turn, compromises with the availability of free E2F. We suggest that this series of events results in a stoppage of the cell cycle progression at the G1-->S phase transition thereby leading to a G0/G1 arrest and subsequent apoptotic cell death. To our knowledge, this is the first study showing the involvement of the pRb-E2F/DP pathway as a mechanism of the cancer-chemopreventive effects of resveratrol.

Antineoplastic Agents, Phytogenic↗

Legume lectin family, the 'natural mutants of the quaternary state', provide insights into the relationship between protein stability and oligomerization.

Legume lectins family of proteins, despite having the same 'jelly roll' tertiary structural fold at monomeric level, exhibit considerable variation in their quaternary structure arising out of small changes in their sequence. Nevertheless, their folding behavior and stability correlates very well with their patterns of assembly into dimers and tetramers. A conservation of their fold during evolution, its wide distribution in many protein families together with the availability of structural information on them make them interesting as proteins to explore the effect of inter- versus intra-subunit interactions in the stability of multimeric proteins. Additionally, as 'natural mutants' of quaternary association, proteins of legume lectin family provide interesting paradigms for studies addressing the effect of subunit oligomerization on the stability, folding and function as well as the evolution of multimeric structures.

Amino Acid Sequence↗

Inhibition of prostate carcinogenesis in TRAMP mice by oral infusion of green tea polyphenols.

Development of effective chemopreventive agents against prostate cancer (CaP) for humans requires conclusive evidence of their efficacy in animal models that closely emulates human disease. The autochthonous transgenic adenocarcinoma of the mouse prostate (TRAMP) model, which spontaneously develops metastatic CaP, is one such model that mimics progressive forms of human disease. Employing male TRAMP mice, we show that oral infusion of a polyphenolic fraction isolated from green tea (GTP) at a human achievable dose (equivalent to six cups of green tea per day) significantly inhibits CaP development and increases survival in these mice. In two separate experiments, the cumulative incidence of palpable tumors at 32 weeks of age in 20 untreated mice was 100% (20 of 20). In these mice, 95% (19 of 20), 65% (13 of 20), 40% (8 of 20), and 25% (5 of 20) of the animals exhibited distant site metastases to lymph nodes, lungs, liver, and bone, respectively. However, 0.1% GTP (wt/vol) provided as the sole source of drinking fluid to TRAMP mice from 8 to 32 weeks of age resulted in (i) significant delay in primary tumor incidence and tumor burden as assessed sequentially by MRI, (ii) significant decrease in prostate (64%) and genitourinary (GU) (72%) weight, (iii) significant inhibition in serum insulin-like growth factor-I and restoration of insulin-like growth factor binding protein-3 levels, and (iv) marked reduction in the protein expression of proliferating cell nuclear antigen (PCNA) in the prostate compared with water-fed TRAMP mice. The striking observation of this study was that GTP infusion resulted in almost complete inhibition of distant site metastases. Furthermore, GTP consumption caused significant apoptosis of CaP cells, which possibly resulted in reduced dissemination of cancer cells, thereby causing inhibition of prostate cancer development, progression, and metastasis of CaP to distant organ sites.

Administration, Oral↗

Molecular characterization of HIV type 1 vpu genes from mothers and infants after perinatal transmission.

We compared 162 vpu sequences of human immunodeficiency virus type 1 (HIV-1) in peripheral blood mononuclear cell DNA of 6 infected mother-infant pairs after perinatal transmission, and found a 90.12% frequency of intact vpu open reading frames. The heterogeneity of vpu genes between epidemiologically linked mother-infant pairs was lower compared with epidemiologically unlinked individuals. However, the variability of vpu genes was higher than that seen for other HIV-1 genes, including vif, vpr, tat, and gag p17 from the same mother-infant pairs. Moreover, the infants' sequences displayed patterns similar to those seen in their mothers. The functional domains essential for Vpu activity, including efficient release of virus particles from infected cells and CD4 degradation, were conserved in most of the sequences. In a phylogenetic analysis, the 162 sequences from 6 mother-infant pairs formed distinct clusters for each mother-infant pair sequences and grouped with subtype B sequences. These data support the importance of vpu in HIV-1 replication of mother-infant isolates that are involved in perinatal transmission.

Adult↗

A policy analysis of major decisions relating to Creutzfeldt-Jakob disease and the blood supply.

Creutzfeldt-Jakob disease (CJD) is the first major challenge that the blood system has faced since the completion of the Krever inquiry in 1997. We report the results of a detailed policy analysis comparing 2 CJD-related decisions: a 1995 recall of blood from a donor with classic CJD and the 1999 decision to defer donations from individuals with a 6-month travel history to the UK between 1980 and 1996 due to concerns related to variant CJD. Overall, we observed that decision-making improved significantly from 1995 to 1999. In 1998/99 the potential threat of variant CJD was identified at an early stage, and a systematic risk assessment process was initiated. Decision-making was consultative and involved consumers. However, the perception existed that further improvement could take place in the areas of transparency of process and interaction of organizations. We observed that the presence of a second operator had an important impact on decision-making in 1998/99.

Blood Donors↗

Low conservation of functional domains of HIV type 1 vif and vpr genes in infected mothers correlates with lack of vertical transmission.

Human immunodeficiency virus type 1 (HIV-1) vif and vpr sequences were analyzed from four nontransmitting mothers (infected mothers who failed to transmit HIV-1 to their infants mainly in the absence of anti-retroviral therapy), including a mother with multiple deliveries, and compared with the vif and vpr sequences of five and six previously analyzed transmitting mothers, respectively. In contrast to a high functional conservation of vif and vpr genes in transmitting mother isolates, we found that there was a low degree of conservation of functional domains of these genes in nontransmitting (NT) mother isolates. For vif sequences, NT-2 contained stop codons and no initiation codons, whereas NT-1 sequences carried a substitution of a highly conserved tyrosine to histidine at position 30. In addition, NT-3 and NT-4 sequences contained additional substitutions, including asparagine at position 22, lysine at position 77 and histidine at position 110, that were absent in transmitting mother and consensus subtype B sequences. Similarly, the vpr sequences of NT-2 contained stop codons and no initiation codons, NT-4 contained a substitution of serine in place of alanine at position 30, some NT-1 sequences substituted arginine in place of glycine at position 75, and NT-3 sequences presented a deletion in the C terminus that was absent in transmitting mother and consensus subtype B sequences and is essential for Vpr function. Furthermore, vif and vpr sequences of nontransmitting mothers were less heterogeneous compared with transmitting mother sequences. In conclusion, a low degree of conservation of functional domains and heterogeneity of HIV-1 vif and vpr genes in these infected mothers correlates with lack of vertical transmission.

Adult↗

Liposome mediated antigen delivery leads to induction of CD8+ T lymphocyte and antibody responses against the V3 loop region of HIV gp120.

There is general consensus that the use of whole viruses for the development of a vaccine against human immunodeficiency virus (HIV) would be unsafe. While currently available nonreplicating vaccines, composed of synthetic peptides or purified subunit antigens, can help in tricking the humoral immune responses, they fail to incite the other major arm of the immune defense system, i.e., cell mediated immunity (CMI). To overcome the difficulty in generating CMI, we have entrapped an immunodominant HIV envelope glycoprotein peptide in liposomes made up of fusogenic lipids isolated from Escherichia coli. We have established the role of fusogenic liposomes in stimulation of HIV-specific CD8+ cytotoxic T lymphocytes. Interestingly, the same liposomes elicit strong HIV-specific antibody production as well. Moreover, untoward manifestations such as skin damage or antibody production against lipid components were also not observed. Thus, E. coli lipid liposomes (escheriosomes) could prove to be a potent candidate vaccine, capable of eliciting both humoral and cell mediated immune responses against HIV infection.

AIDS Vaccines↗

Characterization of HIV type 1 tat sequences associated with perinatal transmission.

Human immunodeficiency virus type 1 (HIV-1) tat exon I sequences were analyzed from six mother-infant pairs after perinatal transmission. The tat open reading frame was maintained in 140 of the 154 clones analyzed, with a 90.9% frequency of intact tat open reading frames. In addition, a low degree of heterogeneity was observed in tat sequences within mothers, within infants, and between epidemiologically linked mother-infant pairs. However, the distances of tat sequences between epidemiologically unlinked individuals were greater than in epidemiologically linked mother-infant pairs. The infant sequences showed amino acid sequence patterns similar to those present in their respective mothers. The functional domains required for Tat function, including amino-terminal, cysteine-rich, core and basic regions, which constitute domains for activation and RNA binding, were highly conserved in most of the sequences. Phylogenetic analysis of 154 mother-infant tat sequences showed that they formed distinct clusters for each mother-infant pair and grouped with subtype B sequence. These findings suggest that an intact and functional tat gene is conserved in HIV-1 mother-infant isolates that are involved in perinatal transmission.

Adult↗