Evaluation of the wound healing activity of formulated ointments and water preparation from Sida rhombifolia leaf extract

Background: Several plants including Sida rhombifolia Linn. (Malvaceae) which are said to be used by traditional health practitioners in Tanzania for wound treatment have not yet been evaluated. The objective of this study was to investigate the ointment formulation of S. rhombifolia leaves for its potential wound healing activities.Methods: Wound healing activity of S. rhombifolia leaves was investigated in mice using 50%, 33% and 25% formulated 80% ethanolic leaves extract ointment and water preparations. Excisional and incisional wound-induction models were used with 6 albino mice in each group. The wound diameter (for contraction assessment), duration of re-epithelisation in days, percentage tensile strength as well as the degree of collagenisation and fibrosis were investigated.Results: S. rhombifolia leaves extract had significant mean percentage wound closure for all ointment formulations used and for the water preparation from day 7. A significant percentage tensile strength on day 10 for all formulations used was observed. The 50% ointment had a mean of 64.1±1.7 (p=1.2-09), 33% ointment had a mean of 64.0±3.2 (p=2.4-08) and the 25% ointment had a mean of 53.1±4.0 (p=1.3-06). A remarkable fibrosis and collagenisation for the 50% ointment and the water preparation was observed.Conclusion: The formulated ointments and the water preparations of S. rhombifolia leaves have a potential benefit in enhancing wound healing. A bioassay guided fractionation is recommended to allow identification of its active compound(s) with wound healing activity for drug development

KSHV/HHV-8 and HIV infection in Kaposi's sarcoma development

Kaposi's sarcoma (KS) is a highly and abnormally vascularized tumor-like lesion affecting the skin, lymphnodes and viscera, which develops from early inflammatory stages of patch/plaque to late, nodular tumors composed predominant of spindle cells (SC). These SC are infected with the Kaposi's sarcoma-associated herpesvirus or human herpesvirus-8 (KSHV/HHV-8). KS is promoted during HIV infection by various angiogenic and pro-inflammatory factors including HIV-Tat. The latency associated nuclear antigen type 1 (LANA-1) protein is well expressed in SC, highly immunogenic and considered important in the generation and maintenance of HHV-8 associated malignancies. Various studies favour an endothelial origin of the KS SC, expressing "mixed" lymphatic and vascular endothelial cell markers, possibly representing hybrid phenotypes of endothelial cells (EC). A significant number of SC during KS development are apparently not HHV8 infected, which heterogeneity in viral permissiveness may indicate that non-infected SC may continuously be recruited in to the lesion from progenitor cells and locally triggered to develop permissiveness to HHV8 infection. In the present study various aspects of KS pathogenesis are discussed, focusing on the histopathological as well as cytogenetic and molecular genetic changes in KS

A Review on the Status of Breast Cancer Care in Tanzania

This research article published by Multidisciplinary Cancer Investigation, 2020Tanzania continues to enjoy stability and growth in different sectors similar to other countries; however, some challenges persist in the health sector, especially in the area of cancer care. The current study aimed at reviewing social economic status (SES), as well as factors contributing to the increased burden of breast cancer (BC) in Tanzania. The current study reviewed different literature ranging from nationally/ internationally published statistics, academic publications, health information of non-governmental organizations, academic researchers, and other sources of health information to better understand the socioeconomic and BC care status in Tanzania. The current review showed that BC is still the second most prevalent malignant disease in Tanzania, and most of the patients referring to medical centers are in the advanced stage of the disease due to shortage and unaffordability of health care services. The majority of health-related interventions and investments target infectious diseases, including HIV-AIDS, tuberculosis, malaria, as well as maternal and child health-related conditions, compared to noncommunicable diseases (NCDs) such as BC. In spite of some setbacks and improvements in healthcare facilities (e g, novel techniques for early detection), the best way to address BC care is affordable fees for clinical and laboratory investigations, accessible treatment, palliative care, follow-up, rehabilitative care, and better management and allocation of resources

Tanzanian malignant lymphomas: WHO classification, presentation, ploidy, proliferation and HIV/EBV association

<p>Abstract</p> <p>Background</p> <p>In Tanzania, the International Working Formulation [WF] rather than the WHO Classification is still being used in diagnosing malignant lymphomas (ML) and the biological characterization including the HIV/EBV association is sketchy, thus restraining comparison, prognostication and application of established therapeutic protocols.</p> <p>Methods</p> <p>Archival, diagnostic ML biopsies (N = 336), available sera (N = 35) screened by ELISA for HIV antibodies and corresponding clinical/histological reports at Muhimbili National Hospital (MNH) in Tanzania between 1996 and 2006 were retrieved and evaluated. A fraction (N = 174) were analyzed by histopathology and immunohistochemistry (IHC). Selected biopsies were characterized by flow-cytometry (FC) for DNA ploidy (N = 60) and some by <it>in-situ </it>hybridization (ISH) for EBV-encoded RNA (EBER, N = 37).</p> <p>Results</p> <p>A third (38.8%, 109/281) of the ML patients with available clinical information had extranodal disease presentation. A total of 158 out of 174 biopsies selected for immunophenotyping were confirmed to be ML which were mostly (84. 8%, 134/158) non-Hodgkin lymphoma (NHL). Most (83.6%, 112/134) of NHL were B-cell lymphomas (BCL) (CD20+), of which 50.9%, (57/112) were diffuse large B-cell (DLBCL). Out of the 158 confirmed MLs, 22 (13.9%) were T-cell [CD3+] lymphomas (TCL) and 24 (15.2%) were Hodgkin lymphomas (HL) [CD30+]. Furthermore, out of the 60 FC analyzed ML cases, 27 (M:F ratio 2:1) were DLBCL, a slight majority (55.6%, 15/27) with activated B-cell like (ABC) and 45% (12/27) with germinal center B-cell like (GCB) immunophenotype. Overall, 40% (24/60) ML were aneuploid mostly (63.0%, 17/27) the DLBCL and TCL (54.5%, 6/11). DNA index (DI) of FC-analyzed ML ranged from 1.103-2.407 (median = 1.51) and most (75.0%) aneuploid cases showed high (>40%) cell proliferation by Ki-67 reactivity. The majority (51.4%, 19/37) of EBER ISH analyzed lymphoma biopsies were positive. Of the serologically tested MLs, 40.0% (14/35) were HIV positive, mostly with high (≥40.0%) Ki-67 reactivity.</p> <p>Conclusions</p> <p>According to the 2001 WHO Classification, most subtypes are represented in Tanzanian ML. Extranodal presentation was common among MNH lymphoma patients who also showed high aneuploidy, tumor proliferation (KI-67) and EBER positivity. DLBCL was frequent and phenotype heterogeneity appeared similar to observations in Western countries suggesting applicability of established intervention approaches. HIV was apparently associated with high ML cell proliferation but extended studies are needed to clarify this.</p

Malignant lymphomas (ML) and HIV infection in Tanzania

\ud HIV infection is reported to be associated with some malignant lymphomas (ML) so called AIDS-related lymphomas (ARL), with an aggressive behavior and poor prognosis. The ML frequency, pathogenicity, clinical patterns and possible association with AIDS in Tanzania, are not well documented impeding the development of preventive and therapeutic strategies. Sections of 176 archival formalin-fixed paraffin-embedded biopsies of ML patients at Muhimbili National Hospital (MNH)/Muhimbili University of Health and Allied Sciences (MUHAS), Tanzania from 1996-2001 were stained for hematoxylin and eosin and selected (70) cases for expression of pan-leucocytic (CD45), B-cell (CD20), T-cell (CD3), Hodgkin/RS cell (CD30), histiocyte (CD68) and proliferation (Ki-67) antigen markers. Corresponding clinical records were also evaluated. Available sera from 38 ML patients were screened (ELISA) for HIV antibodies. The proportion of ML out of all diagnosed tumors at MNH during the 6 year period was 4.2% (176/4200) comprising 77.84% non-Hodgkin (NHL) including 19.32% Burkitt's (BL) and 22.16% Hodgkin's disease (HD). The ML tumors frequency increased from 0.42% (1997) to 0.70% (2001) and 23.7% of tested sera from these patients were HIV positive. The mean age for all ML was 30, age-range 3-91 and peak age was 1-20 years. The male:female ratio was 1.8:1. Supra-diaphragmatic presentation was commonest and histological sub-types were mostly aggressive B-cell lymphomas however, no clear cases of primary effusion lymphoma (PEL) and primary central nervous system lymphoma (PCNSL) were diagnosed. Malignant lymphomas apparently, increased significantly among diagnosed tumors at MNH between 1996 and 2001, predominantly among the young, HIV infected and AIDS patients. The frequent aggressive clinical and histological presentation as well as the dominant B-immunophenotype and the HIV serology indicate a pathogenic association with AIDS. Therefore, routine HIV screening of all malignant lymphoma patients at MNH is necessary to enable comprehensive ARL diagnosis and formulation of preventive and therapeutic protocols.\u

Cervical cytological changes in HIV-infected patients attending care and treatment clinic at Muhimbili National Hospital, Dar es Salaam, Tanzania

\ud Tanzania is among Sub-Saharan countries mostly affected by the HIV and AIDS pandemic, females being more vulnerable than males. HIV infected women appear to have a higher rate of persistent infection by high risk types of human papillomavirus (HPV) strongly associated with high-grade squamous intraepithelial lesions (HSIL) and invasive cervical carcinoma. Furthermore, although HIV infection and cervical cancer are major public health problems, the frequency and HIV/HPV association of cervical cancer and HSIL is not well documented in Tanzania, thus limiting the development of preventive and therapeutic strategies. A prospective unmatched, case-control study of HIV-seropositive, ≥ 18 years of age and consenting non-pregnant patients attending the care and treatment center (CTC) at Muhimbili National Hoospital (MNH) as cases was done between 2005 and 2006. HIV seronegative, non-pregnant and consenting women recruited from the Cervical Cancer Screening unit (CCSU) at ORCI were used as controls while those who did not consent to study participation and/or individuals under < 18 years were excluded. Pap smears were collected for routine cytodiagnosis and P53 immunohistochemistry (IHC). Cervical lesions were classified according to the Modified Bethesda System. A total of 170 participants from the two centers were recruited including 50 HIV-seronegative controls were from the CCSU. Ages ranged from 20-66 years (mean 40.5 years) for cases and 20-69 years (mean 41.6 years) for controls. The age group 36-45 years was the most affected by HIV (39.2%, n = 47). Cervicitis, squamous intraepithelial lesions (SIL) and carcinoma constituted 28.3% (n = 34), 38.3% (n = 46) and 5.8% (n = 7) respectively among cases, and 28% (n = 14), 34% (n = 17) and 2% (n = 1) for controls, although this was not statistically significant (P-value = 0.61). IHC showed that p53 was not detectable in HPV + Pap smears and cell blocks indicating possible degradation. The frequency of SIL and carcinoma appeared to be higher among HIV-infected women on HAART compared to seronegative controls and as expected increased with age. HIV seropositive patients appeared to present earlier with SIL compared to those HIV seronegative suggesting a role of HIV in altering the natural history of HPV infection and cervical lesions. The absence of p53 immunoreactivity in HPV + lesions is indicative of the ability of HPV E6 proteins to interact with the tumor suppressor gene and pave way for viral-induced oncogenesis in the studied Tanzanian women.\u

Kaposi's sarcoma and malignant lymphomas in Tanzania during the AIDS epidemic

Kaposi s sarcoma (KS) studies Kaposi´s sarcoma (KS) is the most frequent AIDS tumor (AKS) and associated with HHV-8, but controversial whether a clonal tumor or mostly a reactive proliferation of endothelial spindle cells (SC). In the present studies AKS and endemic KS (EKS) biopsies were compared for expression of HHV-8-LANA, the lymphatic endothelial (LEC) markers LYVE-1 and Ki-67 (proliferation). The study material is archival, formalin-fixed and paraffin-embedded (FFPE) surgical KS biopsies and sera from the Muhimbili National Hospital (MNH, Dar es Salaam, Tanzania) at early and late tumor stages of African AKS and EKS cases with cutaneous and oral (OKS) lesions from male, female, adult and juvenile KS patients. KS histopathological and immunohistochemical (IHC) studies (papers I-III) by single, double and/or triple antibody immunofluorescence (IFA) showed that: a) LANA+/CD34- cells were more frequent in early as compared to late KS and most of them expressed LEC markers LYVE-1, D2-40 and VEGFR-3, suggesting that the resident LECs represent an early target of primary HHV-8 infection; b) LANA+/CD34-/LYVE-1+ cells decreased from early (25%) to late (4%) KS while LANA+/CD34+ SC increased suggesting a phenotype switch from LEC to VEC, c) LANA appeared better correlated to LYVE-1 (LEC) than to CD34 (VEC) expression suggesting heterogeneous SC permissiveness to HHV-8 (paper I), d) the number of HHV-8 infected (LANA+) SC (CD34+) cells as well as LANA+ granules per SC nucleus was significantly higher in male vs. female, juvenile vs. adult and oral AKS [OAKS] vs. cutaneous KS lesions; e) similarly, tumor cell proliferation (Ki-67 immunoreactivity) in adult OAKS was significantly higher for male vs. female, juvenile vs. adult and OAKS vs. cutaneous KS lesions respectively; f) a positive correlation was apparently found between LANA+ SC and Ki-67 immunoreactivity (paper II). Together these findings suggest that tumor proliferation correlates with increasing HHV-8 LANA expression supporting the notion of viral (LANA)-driven cell proliferation during KS oncogenesis (paper II); g) a significantly higher viral load was also seen by quantitative RT-PCR in late nodular OAKS compared with cutaneous AKS lesions corroborating IHC results and indicating that the oral cavity is important for entry and/or reservoir for HHV-8 infection (papers II & III); h) the frequency of LANA+ SC in the tumor increases during the evolution of both cutaneous and oral KS lesions, suggesting a correlation between tumor growth and viral replication in the lesions, however, by RT-PCR, we observed that serum viral loads appeared to decrease in the corresponding patients whereas (paper III) all sera from early stage (patch-plaque) KS were positive for anti-HHV-8 antibodies while all HHV-8 negative sera were from late-stage nodular KS patients (paper IV). This supports our novel notion of a stage-dependent tissue-serum discrepancy in viral loads and HHV-8 antigen expression probably due to virus tissue retention, immune-segregation and/or selective clearance. Thus serum HHV-8 viral loads may not be sufficient for clinical and therapeutic prognostication (papers III & IV); i) the frequency of HHV-8 infection was high among the studied MNH cases and as expected, HHV-8 and HIV infection was more associated with KS compared to non-KS tumors and non-neoplastic conditions; j) the findings by IFA and ELISA had a high HHV-8 diagnostic concordance although ELISA seemed to have higher positive predictive value (PPV), specificity as well as a high sensitivity allowing more affordable HHV-8 screening in a resource-constrained country like Tanzania (paper IV). These findings also indicate the importance of routine HHV-8 screening among blood/organ donors to prevent transmission as well as in prospective transplant recipients to predict and possibly prevent iatrogenic KS (IKS) development (paper IV). Malignant lymphoma (ML) studies ML are second to KS as AIDS-related malignancies and have been reported to increase steadily with the HIV pandemic particularly in sub-Saharan Africa including Tanzania. The WHO classification, HIV and EBV association as well as ploidy and heterogeneity of diffuse large B-cell lymphoma (DLBCL) in Tanzania is not well documented. Our current study (paper V) has established that the WHO classification of lymphoid neoplasms can apparently be applied for the diagnosis of Tanzanian ML. Extranodal presentation of ML was frequent particularly for T-cell lymphomas (TCL). Diffuse large B-cell lymphoma (DLBCL) phenotype heterogeneity and frequency at MNH (Tanzania) was similar to that observed in Western countries suggesting applicability of similar, diagnostic, prognostic and therapeutic approaches (paper V). The attended ML at MNH had frequent aneuploidy, EBV infection as well as high DNA indices and cell proliferation (Ki-67). HIV infection was apparently associated with increased ML cell proliferation (paper V)