Health care providers acceptance of default prescribing of TB preventive treatment for people living with HIV in Malawi: a qualitative study.

BACKGROUND: Tuberculosis (TB) preventive treatment (TPT) substantially reduces the risk of developing active TB for people living with HIV (PLHIV). We utilized a novel implementation strategy based on choice architecture (CAT) which makes TPT prescribing the default option. Through CAT, health care workers (HCWs) need to "opt-out" when choosing not to prescribe TPT to PLHIV. We assessed the prospective, concurrent, and retrospective acceptability of TPT prescribing among HCWs in Malawi who worked in clinics participating in a cluster randomized trial of the CAT intervention. METHODS: 28 in-depth semi-structured interviews were conducted with HCWs from control (standard prescribing approach) and intervention (CAT approach) clinics. The CAT approach was facilitated in intervention clinics using a default prescribing module built into the point-of-care HIV Electronic Medical Record (EMR) system. An interview guide for the qualitative CAT assessment was developed based on the theoretical framework of acceptability and on the normalization process theory. Thematic analysis was used to code the data, using NVivo 12 software. RESULTS: We identified eight themes belonging to the three chronological constructs of acceptability. HCWs expressed no tension for changing the standard approach to TPT prescribing (prospective acceptability); however, those exposed to CAT described several advantages, including that it served as a reminder to prescribe TPT and routinized TPT prescribing (concurrent acceptability). Some felt that CAT may reduce HCW´s autonomy and might lead to inappropriate TPT prescribing (retrospective acceptability). CONCLUSIONS: The default prescribing module for TPT has now been incorporated into the point-of-care EMR system nationally in Malawi. This seems to fit the acceptability of the HCWs. Moving forward, it is important to train HCWs on how the EMR can be leveraged to determine who is eligible for TPT and who is not, while acknowledging the autonomy of HCWs.

Glutathione-S-transferase T1 and M1 gene polymorphisms in Greek patients with multiple sclerosis: a pilot study.

Oxidative stress has been implicated in the pathogenesis of multiple sclerosis (MS). Glutathione-S-transferases (GSTs) are detoxification enzymes, evolved to protect cells against reactive oxygen metabolites. Both GSTT1 and GSTM1 genes exhibit a homozygous deletion polymorphism (null genotype) leading to abolished enzyme activity. We studied the impact of the GSTT1 and GSTM1 polymorphisms on MS susceptibility in a case-control study of 47 Greek patients and 165 controls. Correlations between genotype, gender and disability status were also investigated. The incidence of both GSTT1 and GSTM1 genotypes did not differ significantly between controls and patients. A significantly increased frequency of GSTM1 null genotype was found amongst female patients (65.5%) as compared with males (33.3%, P =0.04). The results suggest that GSTT1 and GSTM1 have no major pathogenetic role on the MS occurrence, nor any strong modifying effect on the disability status. The higher incidence of GSTM1 null genotype observed in female patients, suggests a possible role of the GSTM1 detoxification pathway in a gender-dependent manner.

Genomic gain at 6p21: a new cryptic molecular rearrangement in secondary myelodysplastic syndrome and acute myeloid leukemia.

Fluorescence in situ hybridization and comparative genomic hybridization characterized 6p rearrangements in eight primary and in 10 secondary myeloid disorders (including one patient with Fanconi anemia) and found different molecular lesions in each group. In primary disorders, 6p abnormalities, isolated in six patients, were highly heterogeneous with different breakpoints along the 6p arm. Reciprocal translocations were found in seven. In the 10 patients with secondary acute myeloid leukemia/myelodysplastic syndrome (AML/MDS), the short arm of chromosome 6 was involved in unbalanced translocations in 7. The other three patients showed full or partial trisomy of the 6p arm, that is, i(6)(p10) (one patient) and dup(6)(p) (two patients). In 5/7 patients with unbalanced translocations, DNA sequences were overrepresented at band 6p21 as either cryptic duplications (three patients) or cryptic low-copy gains (two patients). In the eight patients with cytogenetic or cryptic 6p gains, we identified a common overrepresented region extending for 5-6 megabases from the TNF gene to the ETV-7 gene. 6p abnormalities were isolated karyotype changes in four patients. Consequently, in secondary AML/MDS, we hypothesize that 6p gains are major pathogenetic events arising from acquired and/or congenital genomic instability.

Clonal chromosome rearrangements in hairy cell leukemia: personal experience and review of literature.

Cytogenetic studies in hairy cell leukemia (HCL) are rare. In the present report, cytogenetic investigations were performed on marrow cells obtained from 21 HCL male patients with a mean age of 57 years and active disease. Karyotypic analysis was successful in 18 of the 21 patients, either at diagnosis or in relapse after treatment with IFNa. Clonal chromosome abnormalities were detected in eight of 18 cases. The chromosome most frequently involved in the rearranged karyotypes was chromosome 14. Results are discussed with respect to 79 abnormal HCL cases obtained from an extensive review of the literature from 1978 to 2000.

Trisomy 6 and double minute chromosomes in a case of chronic myelomonocytic leukemia.

A 65-year-old woman with chronic myelomonocytic leukemia was shown to have trisomy 6 and multiple double minute chromosomes. The patient had no history of prior exposure to any mutagenic or carcinogenic agents. To our knowledge, this is the first report for presence of only these two aberrations. The expression of several oncoproteins and onco-related proteins was detected immunohistochemically in bone marrow cells. Among them, only the bcl-2 oncoprotein was positively stained in 100% of myeloblasts. Although the c-myc oncogene is frequently reported to be overexpressed in myeloid disorders with double minutes and associated with chemotherapy resistance and disease aggressiveness, in our case, the c-myc oncoprotein was not positively expressed. The patient received chemotherapy and complete hematological remission was successfully achieved.

Human chromosome 19 confers the CD2/E rosette receptor phenotype in interspecific cell hybrids.

Interspecific cell hybrids between Chinese Hamster Ovary (CHO) and phytohaemagglutinin (PHA) stimulated human T lymphocytes were purified by preparative rosetting with sheep red blood cells (SRBC). The hybrid cell clone used in the present study consisted of cells containing a complete set of the 20 CHO chromosomes and one extra human chromosome, No 19. Hybrid cells constitutively expressed high levels of human CD2 surface receptor and formed multilayer rosettes with SRBC and human erythrocytes. In addition to CD2 they produced low levels of a small number of human extracellular proteins. These findings suggest that the factor(s) responsible for CD2 expression are produced by the hybrid and that genes responsible for CD2 expression are located on chromosome 19. However, the present work cannot exclude that material of chromosome 1, where the CD2 gene has been assigned previously, is integrated somewhere in the hybrid karyotype. Further work is needed to clarify this point.

Stimulatory effect of low dose X-irradiation on the expression of the human T lymphocyte CD2 surface antigen.

We investigated the early effects of low doses of ionizing radiation on the CD2 gene expression in normal human T lymphocytes in order to clarify if low-dose ionizing radiation has an enhancing and/or stimulatory effect on the immune response. The results indicate that even low doses of X-irradiation strongly enhance the appearance of CD2 antigen, both in PHA-stimulated and in resting T lymphocytes as demonstrated by a rosette assay or by immunofluorescence. Moreover, an accumulation of CD2 mRNA is observed in X-irradiated cells compared with non-irradiated, a fact that is attributed mainly to transcriptional activation of the CD2 gene and not to stabilization of preformed mRNA.

Effects of low-doses of X-rays on the expression of human cell surface CD2 antigen in CD2+ CHO cells.

Thioguanine resistant CHO cells (HPRT-) were stably cotransfected with pSV2-gpt and pi H3-CD2 vectors using the calcium phosphate coprecipitation technique. The effects of single low doses of ionizing radiation were studied in a CD2+ CHO clone. The CD2+ phenotype responsible for binding sheep erythrocytes and rosette formation, was not affected by X-rays doses in the range 2-6 cGy. However, after 10 cGy of X-irradiation, 50% of the cells lost the CD2+ phenotype. These results suggest that this CD2+ clone might be a very sensitive indicator of very low X-ray doses. The implications of the phenotypic changes, observed after very low doses of irradiation, are discussed.

Biological dosimetry of absorbed radiation by C-banding of interphase chromosomes in peripheral blood lymphocytes.

Induction of premature chromosome condensation enables direct observation of radiation-induced cytogenetic damage in non-stimulated, interphase, human peripheral blood lymphocytes. This phenomenon can be explored in radiation protection for biological dosimetry in instances of accidental exposure to ionizing radiation. Quantification of an exposure by means of this approach has been limited so far mainly to the analysis of chromosome fragments. This limitation is due to the fact that conventional Giemsa staining of prematurely condensed chromosomes (PCCs) does not allow visualization of the centromeric regions and, as a result, the identification of dicentrics, centric rings and acentric fragments. In the present report a C-banding procedure, refined to avoid swelling and chromosome distortion of freshly prepared PCCs spreads, is used to identify such aberrations in non-stimulated human lymphocytes. The method allows immediate banding of the centromeric regions and enables scoring of aberrations within a time interval (3-4 h after blood sample withdrawal) that is only a fraction of that normally required when cells stimulated to proliferate are analysed at metaphase. The dose-response for dicentrics and centric rings measured in interphase lymphocytes was found to be similar to that obtained at metaphase. Measurement of dicentrics and centric rings in prematurely condensed chromosomes of human lymphocytes would provide valuable information on radiation dose estimates, especially in cases of extreme urgency.

On the formation of steroidal amidoesters of 4-[N,N-bis(2-chloroethyl)amino]benzoic acid and their cytotoxic activity.

The 4-[N,N-bis(2-chloroethyl)amino]benzoate of 17beta-acetamido-5alpha-androstan-3beta-ol, 17beta-acetamido-5-androsten-3beta-ol, 3beta-acetamido-5alpha-androstan-17beta-ol and 3alpha-acetamido-5beta-androstan-17beta-ol have been prepared and their antineoplastic effect evaluated against MIA Pa-Ca-2 pancreatic carcinoma, T47D breast carcinoma and A431 squamus cell carcinoma. Among the compounds tested, the compound 17beta-acetamido-3beta-hydroxy-5-androsten-4-[N, N-bis(2-chloroethyl)amino]benzoate appeared to possess a significant cytotoxic effect against A431 cells.

Constitutional pericentric inversion of chromosome 9 and hematopoietic recovery after allogeneic stem cell transplantation.

Recent reports suggest that hemopoietic stem cells with constitutional pericentric inversion of chromosome 9 [inv(9)] may be related to delayed engraftment or hemopoietic defect after stem cell transplantation (SCT). We conducted a retrospective study on five allogeneic SCT in which constitutional inv(9) was detected either in the donor or the recipient. The results showed that hematologic recovery was within the expected time range for all our patients. However, one patient exhibited decreasing blood counts between day +45 and +272 after transplantation, possibly due to protracted cytomegalovirus (CMV) infection and gansiclovir and imatinib treatment. Our findings suggest that constitutional inv(9) may not be associated with delayed hemopoietic recovery after SCT.

Formation of multilayer rosettes in phytohemagglutinin-stimulated lymphocytes: correlation with chromatin condensation conformation in premature chromosome condensation.

We found that the formation of multilayer rosettes by transformed human blood lymphocytes after phytohemagglutinin (PHA) stimulation is correlated with conformational changes of the chromatin as seen by premature chromosome condensation (PCC). The frequency distribution of grades of PCC and multilayer rosette formation suggests that changes in chromatin are a prerequisite for rosette formation. Rosette formation was most pronounced for 24-h and 48-h cultures. Chromatin decondensation and rosette formation showed identical patterns. The possibility that multilayer rosette formation is directly dependent on conformational changes of chromatin is discussed.

Effects of homo-aza-steroids on acute non-lymphocytic leukaemia cell proliferation in vitro.

Homo-aza-steroids (modified steroid molecules) in their esterified forms have been used extensively as carrier molecules of alkylating agents against several neoplastic malignancies in vivo and in vitro. We studied the effects of two homo-aza-steroid carrier molecules alone, namely 3 beta-hydroxy-13 alpha-amino-13,17-seco-5 alpha-androstan-17-oic-13, 17-lactam (compound 1) and 13 alpha-amino-13,17-seco-1,3,5-estratrien-17-oic- 13,17-lactam (compound 2), on human acute non-lymphocytic leukaemia cell proliferation in vitro. We used peripheral blood samples from 27 untreated ANLL patients (eight M1, four M2, two M3, six M4, three M5a, two M5b and two M6, according to FAB criteria). Proliferative activity was estimated by using thymidine uptake and the percentage of cells in metaphase in 24, 48 and 72 h of culture. Exposure of human leukaemic blasts with either of the two compounds resulted in enhanced cell proliferation in M1, M2, M4, M6 and M5a (only by compound 2) cases, whereas there was no significant effect in the M3 and M5b cases. Our results indicate that the two compounds tested exhibit stimulatory effect on cell proliferation, particularly in blast cells possessing a relatively smaller degree of differentiation (M1 and M6 cases exhibiting CD34 and CD7). Further research is needed to study the cell growth effect and the therapeutic potential of these steroid molecules in human blood malignancies in vitro and in vivo.