Development of <Emphasis Type="Italic">T</Emphasis><Subscript><Emphasis Type="Italic">m</Emphasis></Subscript>-shift genotyping method for detection of cat-derived <Emphasis Type="Italic">Giardia lamblia</Emphasis>

To develop T _m -shift genotyping method for detection of cat-derived Giardia lamblia, two sets of primers with two GC-rich tails of unequal length attached to their 5′-end were designed according to two SNPs (BG434 and BG170) of β-giardin (bg) gene, and specific PCR products were identified by inspection of a melting curve on real-time PCR thermocycler. A series of experiments on the stability, sensitivity, and accuracy of T _m -shift method was tested, and clinical samples were also detected. The results showed that two sets of primers based on SNP could distinguish accurately between assemblages A and F. Coefficient of variation of T _m values of assemblage A and F was 0.14 and 0.07% in BG434 and 0.10 and 0.11% in BG170, respectively. The lowest detection concentration was 4.52 × 10^?5 and 4.88 × 10^?5 ng/μL samples of assemblage A and F standard plasmids. The T _m -shift genotyping results of ten DNA samples from the cat-derived G. lamblia were consistent with their known genotypes. The detection rate of clinical samples by T _m -shift was higher than that by microscopy, and their genotyping results were in complete accordance with sequencing results. It is concluded that the T _m -shift genotyping method is rapid, specific, and sensitive and may provide a new technological mean for molecular detection and epidemiological investigation of the cat-derived G. lamblia.     

Late onset immune pancytopenia following bone marrow transplantation

A 17-year-old boy developed autoimmune pancytopenia in the absence of chronic graft-versus-host disease 170 d after allogeneic bone marrow transplantation (BMT) from his HLA identical brother. The anaemia and thrombocytopenia responded to conventional immunosuppressive treatment, but the neutropenia was refractory to this and to splenectomy and subsequent removal of splenic remnant. Following total lymphoid irradiation the neutrophil count rose to low normal levels but thrombocytopenia and anaemia secondary to marrow hypoplasia required transfusion support. Bone marrow function was finally normalized by an additional transfusion of donor marrow without prior immunosuppressive therapy. We conclude that late onset immune pancytopenia post BMT caused by antibodies of probable donor origin may be life threatening in the absence of chronic graft-versus-host disease.     

Use of corticosteroids to suppress drug toxicity in complicated tuberculosis

Four cases of tuberculosis complicated by allergic or toxic reactions to antibiotic treatment are presented. In each, it was considered essential to suppress the reactions in order to give effective chemotherapy. This was achieved by using prednisolone generally in a dose of 40 mg/day or less during the continuation phase of therapy. Reactions to essential treatment are an important indication for corticosteroid treatment in tuberculosis.     

Deciphering the immuno-pathological role of FLT,and evaluation of a novel dual inhibitor of topoisomerases and mutant-FLT3 for treating leukemia

Acute myeloid leukemia (AML) is a type of leukemia with an aggressive phenotype, that commonly occurs in adults and with disappointing treatment outcomes. Genetic alterations were implicated in the etiology of cancers and form the basis for defining patient prognoses and guiding targeted therapies. In the present study, we leveraged bulk and single-cell RNA sequencing datasets from AML patients to determine the clinical significance of Fms-related receptor tyrosine kinase 3 (FLT3) alterations on the T-cell phenotype and immune response of AML patients. Subsequently, we evaluated the therapeutic potential of Lwk-n019, a novel small-molecule derivative of thiochromeno[2,3-c]quinolin-12-one. Our results suggested that FLT3 plays an important role in the progression, aggressive phenotype, and worse immune response of patients. An FLT3 mutation was associated with dysfunctional T-cell phenotypes, and high risk and shorter survival of AML patients. Our findings further suggested that the aggressiveness of AML and the prognostic role of FLT3 are associated with the co-occurrence of NPM1 and DNMT3A mutations. Lwk-n019 demonstrated dose-dependent anticancer activities against various leukemia cancer cell lines. Lwk-n019 demonstrated highly selective kinase inhibitory activities against the wild-type FLT3 (D835V) and mutant FLT3 (internal tandem duplication (ITD), D835V) with >95% and 99% inhibitory levels, respectively. Moreover, the compound demonstrated the best binding constant (Kd value) of 0.77 µM against FLT3 (ITD, 835V). In addition, Lwk-n019 significantly inhibited the activities of both the topoisomerase I (TOPI) and TOPII enzymes, with higher TOPI inhibitory activity than camptothecin, a clinical inhibitor. While the jejunum, duodenum, cecum, and colon were prime sites of absorption, Lwk-n019 achieved maximum concentration (Cmax), Vd, blood/plasma ratio, time to maximum concentration (Tmax), area under the receiver operating concentration curve (AUC)_(0-24), and AUC_(0-∞) values of 0.665 µg/mL, 5.21 Vc, L/kg, 1.5 h, 6634.7, and 6909.2, respectively. In conclusion, Lwk-n019 demonstrated anticancer activities via multi-target inhibition of TOPs and kinases with high inhibition preference for mutant ITD-FLT3. The present pioneer study provides a basis for advanced optimization of drug potency, selectivity, specificity, and other properties desired of anticancer drug leads. Studies are ongoing to determine the full therapeutic properties of Lwk-n019 and the detailed mechanisms of FLT3 in TOP inhibition.     

Regulation of male sex hormones strongly heightens copulation behavior indices in rat's model administered Cocos nucifera husks extract

BackgroundCocos nucifera (Coconut, Arecaceae family) is consumed as dietary fruit but little is known of it as sex enhancer.ObjectiveTo investigate male sex hormones and copulation behavior indices in male Wistar rats administered husks extract.Materials and MethodsRats were grouped into control A, and treatment B to E (n=4); and administered husks extract at varying doses: 125mg/kg, 250mg/kg, 375mg/kg and 500mg/kg for 48days by oral intubation. Copulation behavior was assessed by introducing female rats to males (1:1) in a rectangular Plexiglas chamber and monitored by veterinary Anatomists. Blood samples for male sex hormones were collected and assayed by ELISA method.ResultsCopulation behavior (500mg/kg treatment): mounting (9.66±0.79 to 29.08±2.16), ejaculation (7.66±0.38 to 16.93±0.76), intromission (22.01±1.67 to 38.11±3.35) frequencies, and ejaculation latency (7.92±0.43 to 12.28±0.41) increased remarkably (Group E). Correspondingly, mounting (133.31±1.18 to 93.39±0.43) and, intromission (88.13±3.12 to 74.55±1.19) latencies; post-ejaculation (3.16±0.14 to 2.18±0.34), inter-intromission (19.48±0.26 to 14.32±1.65) intervals reduced significantly (P≤0.05). Testosterone levels (Group E) increased: 3.82±0.6ng/dL to 5.14±0.3ng/dL while, LH and FSH values reduced: 2.92±0.6IU/L to 2.13±0.3IU/L and 2.28±0.7IU/L to 1.58±0.3IU/L significantly (p ≤ 0.05) while changes were dose-related.ConclusionC. nucifera husks extract improves sexuality indices by regulating male sex hormones in male Wistar rats.     

The severity, outcome and challenges of breast cancer in Nigeria

Breast cancer is a very common disease in Nigeria. It is often associated with a poor prognosis for a variety of reasons. This study was designed to investigate the challenges, severity, outcome and factors influencing the outcome of the management of breast cancer in a Nigerian Teaching Hospital with a view to finding ways to improve the current dismal outlook for patients with the disease. The clinical records of patients seen with breast cancer over an 8-year period (1996-2003) in the two units of the Teaching hospital were reviewed. The two units serve the urban, semi-urban and rural communities of some parts of southwestern Nigeria. Two hundred and twelve patients with breast cancer were seen over the 8-year period of the study. The mean age was 48 years (23-85 years). There were 211 female and one male. One hundred and three patients (48.7%) had either postprimary or tertiary education. A proportion of 66.7% were premenopausal, 79.2% had pregnancy early in life and were multi-parous. These also gave a history of prolonged breast-feeding of their children. The tumour was self-detected in 195 (92%). The mean duration of symptoms was 11.2 months (9 days-7 years). Pain in 100 patients (47%) was the most common symptom and the cancer was in the left breast in 113 (53.3%). Localized cancer was in the upper outer quadrant in 85 (40%), whereas the whole breast was involved in 55 patients (26%). Loco-regional features of advanced cancer were seen in 157 patients (74%). The tumour was fungating in 83 (39%) and there was clinical evidence of systemic metastasis in 28 patients (13%). One hundred and seventy-four (80.6%) patients had advanced disease (stages 3 and 4). Definitive surgery was possible in 185 patients (87.3%), neoadjuvant chemotherapy was required in 65 (30.6%), postoperative adjuvant chemotherapy in 178 (84%; drug combinations were CMF-cyclophosphamide, methotrexate, 5-fluorouracil, CMFP-CMF plus prednisone, and CAF-cyclophosphamide, adriamycin, 5-fluorouracil), tamoxifen was administered in all the patients. Only 70 (33.2%) patients were known to have received radiotherapy among those referred to the Radiotherapy unit, with associated fair treatment compliance after surgery. Outpatient clinic attendance was also very poor, only 27 (12.7%) were still being seen in the clinic, 83 patients (39%) were known to be dead and 102 patients were lost to follow-up. The mean follow-up period was 8.4 months (1 week-6 years). In conclusion, breast cancer is very common in our area of practice in Nigeria; the majority of our patients were young and premenopausal women presenting in the advanced stages of cancer. Treatment compliance was very poor. The majority of the patients were dead or lost to follow-up within a year of diagnosis.     

An ethnobotanical survey of Bauchi State herbal plants and their antimicrobial activity

A survey of medicinal plants used locally in the treatment of various diseases was carried out in Bauchi State-Nigeria. A total of 84 medicinal plants were listed. Preliminary antimicrobial activity of the aqueous extracts of the plants was investigated. The results indicated that out of 84 plants, 75 exhibited antimicrobial activity against one or more of the test organisms at a concentration of 200 mg/ml. The extracts were found to show potentially interesting activity against Proteus mirabilis, Pseudomonas aeruginosa, Staphylococcus aureus and Escherichia coli.     

Site factors may be more important than participant factors in explaining HIV test acceptance in the prevention of mother-to-child HIV transmission programme in Kenya, 2005

Objective  To determine the role of participant factors on the acceptance of a Prevention-of-Mother-to-Child (PMTCT) HIV test programme in a situation with an opt-out testing strategy.
Methods  We analysed antenatal clinic (ANC) HIV sentinel surveillance data. All 43 sites in the 2005 round of Kenya's ANC surveillance offered opt-out PMTCT services and recorded if women were offered PMTCT HIV testing and whether they accepted or refused. Logistic regression was used to determine the role of participant-level factors on PMTCT acceptance.
Results  During the period of sentinel surveillance, 13 026 women attended ANC and testing was offered to 12 030 women. Of those offered testing, 9690 (80.5%) accepted, with a large variation in the percent of acceptors by site. Age, residence and educational status were significant determinants of PMTCT acceptance. However, after adjusting for site none of the participant-level factors were significant determinants of PMTCT acceptance.
Conclusions  Participant level factors were not significant determinants of PMTCT HIV test acceptance after adjusting for sites. PMTCT programmes should collect and evaluate the role of site-level (provider and testing service) factors on PMTCT acceptance. Improvement of site-level factors could improve PMTCT uptake.