Co-infection and co-localization of Kaposi sarcoma-associated herpesvirus and Epstein-Barr virus in HIV-associated Kaposi sarcoma: a case report.

Kaposi sarcoma (KS), a multifocal vascular neoplasm frequently observed in HIV-positive individuals, primarily affects the skin, mucous membranes, visceral organs, and lymph nodes. KS is associated primarily with Kaposi sarcoma-associated herpesvirus (KSHV) infection. In this case report, we present a rare occurrence of co-infection and co-localization of KSHV and Epstein-Barr virus (EBV) in KS arising from the conjunctiva, which, to our knowledge, has not been reported previously. Immunohistochemistry (IHC), DNA polymerase chain reaction (PCR), and EBV-encoded RNA in situ hybridization (EBER-ISH) were utilized to demonstrate the presence of KSHV and EBV infection in the ocular KS lesion. Nearly all KSHV-positive cells displayed co-infection with EBV. In addition, the KS lesion revealed co-localization of KSHV Latency-Associated Nuclear Antigen (LANA) and EBV Epstein Barr virus Nuclear Antigen-1 (EBNA1) by multi-colored immunofluorescence staining with different anti-EBNA1 antibodies, indicating the possibility of interactions between these two gamma herpesviruses within the same lesion. Additional study is needed to determine whether EBV co-infection in KS is a common or an opportunistic event that might contribute to KS development and progression.

Yes-associated protein-1 overexpression in ocular surface squamous neoplasia; a potential diagnostic marker and therapeutic target.

Yes-associated protein-1 (YAP-1) is a Hippo system transcription factor, which serves as an oncogene in squamous cell carcinoma, and several solid tumors when the Hippo pathway is dysregulated. Yet, the activity of YAP-1 in ocular surface squamous neoplasia (OSSN) has not been determined. Here, we investigate the relationship between YAP-1 overexpression and OSSN. Using a cross-sectional study design, we recruited 227 OSSN patients from the University Teaching Hospitals in Lusaka, Zambia. Immunohistochemistry was used to assess YAP-1 protein overexpression in tumor tissue relative to surrounding benign squamous epithelium. OSSN patient samples (preinvasive, n = 62, 27% and invasive, n = 165, 73%) were studied. One hundred forty-nine invasive tumors contained adjacent preinvasive tissue, bringing the total number of preinvasive lesions examined to 211 (62 + 149). There was adjacent benign squamous epithelium in 50.2% (114/227) of OSSN samples. Nuclear YAP- 1 was significantly overexpressed in preinvasive (Fisher's (F): p <.0001, Monte Carlo (MC): p <.0001) and invasive (F: p <.0001, MC: p <.0001) OSSN in comparison to adjacent benign squamous epithelium when analyzed for basal keratinocyte positive count, staining intensity, expression pattern, and Immunostaining intensity-distribution index. YAP-1 expression did not differ between preinvasive and invasive OSSN (p >.05), keratinizing and non- keratinizing cancer (p >.05), or between T1/T2 and T3/T4 stages in invasive tumors (p >.05). However, grade 2 and 3 tumors had significantly stronger nucleus YAP-1 overexpression intensity than grade 1 tumors (F: p = .0078, MC: p = .0489). By immunohistochemistry, we identified significant overexpression (upregulation of YAP-1 protein expression) in preinvasive and invasive OSSN lesions compared to neighboring benign squamous epithelium. YAP-1 expression was significantly higher in poorly and moderately differentiated invasive squamous cancer than in well-differentiated carcinomas. Overexpression of YAP-1 within the margin of preinvasive and invasive OSSN, but not in the neighboring normal epithelium, indicates that it plays a role in the development and progression of OSSN.

High-risk human papillomavirus-associated vulvar neoplasia among women living with human immunodeficiency virus in Zambia.

Background: Globally, women living with HIV have a higher risk of vulvar neoplasia than HIV-negative women. Vulvar neoplasia among women living with HIV has not previously been characterised in Zambia. Objective: This study determined the clinical and pathologic features of vulvar neoplasia among women living with HIV at the University Teaching Hospital, Lusaka, Zambia. Methods: We conducted a cross-sectional study of vulvar lesions among 53 women living with HIV who presented with vulvar lesions between July 2017 and February 2018. The study assessed clinical and histological characteristics and prevalence of high-risk human papillomavirus (HRHPV). Results: Twenty-one patients were diagnosed with vulvar squamous cell carcinoma (VSCC), 20 with usual vulvar intraepithelial neoplasm (uVIN), and the rest with either benign lesions or non-neoplastic lesions (NNL). Participants' mean age was 40 years. Patients with VSCC were significantly older than those with NNL (mean (s.d.): 43 (21) vs 33 (10), p = 0.004). The prevalence of HRHPV was 88.9% in VSCC patients and 100.0% in high-grade squamous intraepithelial lesion patients. HPV16 was the most common (52.6%) genotype. The clinical features of neoplasia were similar to those of NNL. Conclusion: VSCC was significantly more common among women aged ≥ 40 years. HRHPV in VSCC and high-grade squamous intraepithelial lesions was high. Women with vulvar lesions, especially those aged > 40 years, should be evaluated for vulvar cancer. Young girls should be vaccinated to prevent vulvar cancer.

Epstein-Barr Virus, But Not Human Papillomavirus, Is Associated With Preinvasive and Invasive Ocular Surface Squamous Neoplasias in Zambian Patients.

Background: The etiopathogenesis of ocular surface squamous neoplasia (OSSN) is not fully understood. We assessed the frequency of oncogenic viruses in OSSN by immunohistochemistry (IHC) and polymerase chain reaction (PCR) for human papillomavirus (HPV), Epstein-Barr virus (EBV), Merkel cell polyomavirus (MCPyV), Kaposi sarcoma virus, and adenovirus. Cases from Zambia were prospectively enrolled using a cross-sectional study design between November 2017 and March 2020. Methods: Demographic and clinical data [age, sex, HIV status, antiretroviral therapy (ART) history, CD4 count, plasma viral load] and tumor biopsies were collected from 243 consenting patients. Tumor samples were bisected, and half was used for DNA isolation, while the other half was formalin fixed and paraffin embedded (FFPE) for histopathology analysis. The expressions of latent EBV nuclear antigen 1 (EBNA1), CDKN2A/p16INK4A (p16), and MCPyV large T-antigen (LT) were tested by IHC. Multiplex PCR was used to detect 16 HPV genotypes and four other DNA tumor viruses [Kaposi's sarcoma-associated herpesvirus (KSHV), EBV, MCPyV, and adenovirus]. Relationships between HIV status, viral DNA and protein expression, and tumor grades were determined by statistical analysis. Results: OSSN tumors from patients were 29.6% preinvasive and 70.4% invasive. Patients presented with unilateral tumors that were 70.4% late stage (T3/T4). OSSN patients were HIV positive (72.8%). IHC on 243 FFPE biopsies resulted in the detection of EBNA1 (EBV), p16 high-risk HPV (HR-HPV), and MCPyV LT expression in 89.0%, 4.9%, and 0.0%, respectively. EBNA1 was expressed in all grades of preinvasive [cornea-conjunctiva intraepithelial neoplasia (CIN)1, 100%; CIN2, 85.7%; CIN3, 95.8%; and carcinoma in situ (CIS), 83.8%] and in invasive (89.2%) OSSN. PCR on 178 samples detected EBV, HR-HPV, and MCPyV in 80.3%, 9.0%, and 13.5% of tumors, respectively. EBV was detected in all grades of preinvasive and invasive OSSN. EBV detection was associated with high HIV viral loads (p = 0.022). HR-HPV was detected in 0.0% CIN1, 0.0% CIN2, 5.6% CIN3, 13.0% CIS, and 7.0% invasive OSSN. Conclusions: Our findings of EBV DNA and EBNA1 protein in all the grades of preinvasive and especially invasive OSSN are consistent with a potential causal role for EBV in OSSN. A role of HPV in OSSN was not clearly established in this study.

Contextualisation of Early and Enhanced Clinical Exposure Model through Development of Curricula for Advanced Practice Nursing and Midwifery.

Background: An Advanced Practice Nurse is a generalist or specialized nurse who has acquired thorough graduate education a minimum of a master's degree. The need for Advanced Practice Nurses is increasingly recognized globally. This paper describes the process, which was undertaken by School of Nursing Sciences, University of Zambia in reviewing and developing advanced practice nursing and midwifery curricula which will be implemented using the Early and Enhanced Clinical Exposure model (EECE). Materials and Methods: The curricula development/review process utilized a modified Taba's Model which followed a step-by-step approach including: 1) desk review, 2) diagnosis of needs (needs assessment), 3) stakeholder consultations, 4) content development, 5) validations and approval from which several lessons were learnt and recommendations made. Findings and recommendations from different stages were used as a basis for reviewing and developing advanced practice nursing and midwifery curricula. Results: Desk review needs assessment and stakeholder consultations identified both strengths and weaknesses in the existing curricula. Major strengths were duration and core courses which met the minimum requirement for postgraduate nursing and midwifery training. Major weaknesses/gaps included some content that was too basic for the master's level and the delayed exposure to practicum sites which limited the development of advanced practice skills. Others were inadequate competence for advanced practice, inadequate research methodology course, lack of content to foster development of personal soft skills and predominant use of traditional teaching methods. Stakeholders recommended implementing advanced, clinical and hands-on Masters of Nursing and Midwifery programmes which resulted in the review of four existing and development of five demand-driven curricula. Conclusion: The reviewed and developed curricula were strengthened to close the identified gaps. Both the reviewed and developed curricula have been implemented using the Early and Enhanced Clinical Exposure Model with a view to producing Advanced Practice Nurses and Midwives who are competent to meet diverse health care needs and contribute to improving patient outcomes.

Application of Interprofessional Education Model to University Pre-Licensure Health Students in the Management of Chronic Care Conditions in Zambia.

Background: There is evidence that multidisciplinary healthcare teams can provide better quality of care and treatment outcomes compared to that delivered by individuals from a single health discipline. The project on which this article is based applied the interprofessional education model to university pre-licensure health students in the management of chronic care conditions in Zambia. Methods: Four distinct but interrelated approaches, namely desk review; module development workshops; review and validation of modules by experts; piloting and review of the training modules were employed. Results: Several models of interprofessional education currently in existence and used successfully by higher education institutions in other settings were identified. While several models of Interprofessional Education were identified, our project adapted the "didactic program, community-based experience, and interprofessional-simulation experience" models. To apply the models, modules of seven chronic care conditions were developed and piloted. The extent to which the module activities promoted interprofessional education were rated between 74 - 87% (agree or strongly agree) by the students. Conclusion: Three models of Interprofessional Education were identified and adapted in the project, and seven modules were developed and administered to the students. The process was effective for putting forth an interprofessional training program at the undergraduate level, with the potential to improve quality of care for patients.

von Willebrand factor activity and activated partial thromboplastin time as proxy biomarkers for coagulopathies in women with menorrhagia in Zambia: a case-control study.

INTRODUCTION: von Willebrand Disease (vWD) is the most prevalent bleeding disorder. Women are more likely to manifest abnormal bleeding symptoms due to physiologic events and menorrhagia is the most common presenting symptom. METHODS: this case-control study included 168 women aged between 18 and 45. The cases had menorrhagia whilst the controls did not. Blood grouping, activated partial thromboplastin time and von Willebrand factor activity tests were performed on samples collected from consenting study participants. RESULTS: the mean age was 29.96 ± 7.37. Mean vWF activity of cases was 66.6% and of controls 97.8%. The mean activated Partial ThromboplastinTime (aPTT) of cases was 31.09s and of controls was 30.40s. There was no difference in the vWF activity between blood group O (86.3%) and non-blood group O (88.0%) participants. Eight women were diagnosed with von Willebrand disease, 6 cases and 2 controls. Higher odds of von Willebrand disease were seen in the cases (OR = 6.6). Epistaxis, von Willebrand and factor activity levels and family history of menorrhagia were associated with an increased risk for menorrhagia. CONCLUSION: von Willebrand factor activity levels were associated with menorrhagia while activated partial thromboplastin time was not. vWF activity levels did not depend on any specific blood group. The prevalence of von Willebrand disease was significantly higher in participants with menorrhagia and repeated epistaxis and family history of menorrhagia pointed to a higher risk of menorrhagia.

Clinical and Pathologic Presentation of Primary Ocular Surface Tumors among Zambians.

AIM: This study aimed to characterize the clinical and pathologic presentation of ocular surface tumors (OSTs) and to more precisely differentiate the grades of ocular surface squamous neoplasia (OSSN) and benign lesions among Zambians. METHODS: Two-hundred sixty-five Zambian patients presenting with ocular surface growths, suspicious for OSSN, were recruited between November 2017 and November 2019 to a cross-sectional study to investigate their lesions. Sociodemographic data were collected, HIV infection status and vision tests were performed, and lesions were measured and documented. Lesions >2 mm in diameter were excised and sent for pathology analysis. In addition to the biopsies, tears, blood, and buccal swabs were collected. CD4+ T-cell counts were measured by flow cytometry. Lesions were classified according to the WHO guidelines. χ2 and bivariate correlations were used to analyze variable associations and strengths with phi/Cramer's V and correlation coefficients, respectively. Binary logistics was used to adjust for covariance. RESULTS: In this study, 68.3% of the participants were found to be HIV positive. The most frequent diagnoses were invasive OSSN (45.3%), preinvasive OSSN (29.1%), and pterygium (22.6%). Invasive OSSN comprised keratinizing squamous cell carcinoma (SCC) (87.5%), basaloid SCC (3.3%), and spindle cell carcinoma (3.3%). Unusual carcinomas, not described previously, included hybrid SCC (5.0%) and acantholytic SCC (0.8%). Invasive OSSN had advanced tumor (T3/T4) staging (93.3%) at diagnosis. Lymphadenopathy was rare (2.3%), and metastasis was absent. Patients were mostly female (59.2%). Median age was 36 (interquartile ranges 33-41) years (ranges 18-81). Patients with invasive OSSN were more likely to present with pain (p = 0.007), redness (p = 0.034), excessive tearing (p = 0.0001), discharge (p = 0.011), bleeding (p = 0.007), reduced vision (p = 0.0001), fungating lesion (p = 0.001), and blindness (p = 0.005); location at temporal limbus (p = 0.0001), inferior limbus (p = 0.0001), or circumlimbal (p = 0.001); and extension to cornea (p = 0.006) and forniceal palpebral conjunctiva (p = 0.001). Invasive OSSN was associated with any smoking habit and alcohol consumption (p = 0.04 and 0.03, respectively). HIV positivity was strongly associated with OSSN (74.6% OSSN vs. 49.3% benign lesions; p = 0.0001; phi: 0.237 [p = 0.0001]). CONCLUSION: OSTs are very common in Zambia and are strongly associated with HIV coinfection. Patients with OSSN were more likely to be HIV positive than those with pterygia. Despite the commonality of OSTs in sub-Saharan Africa, these cancers have historically been poorly characterized.

Characterization of Mycobacterium tuberculosis genotypes and their correlation to multidrug resistance in Lusaka, Zambia.

OBJECTIVES: The burden of multidrug-resistant tuberculosis (MDR-TB) has been reported to be increasing in Zambia. The reasons for the increase are still unclear. This study determined the diversity of Mycobacterium tuberculosis genotypes among isolates in Lusaka, the capital city, and investigated their association with MDR-TB. METHODS: Spoligotyping, large sequence polymorphism (LSP) analysis, and sequencing of MDR associated genes were performed on a total of 274 M. tuberculosis clinical isolates stored at the University Teaching Hospital from 2013 to 2017. Of these, 134 were MDR-TB while 126 were pan-susceptible. RESULTS: Spoligotyping showed the LAM family as the most predominant genotype (149/274, 54.4%) followed by the CAS family (44/274, 16.1%), T family (39/274, 14.2%), and minor proportions of X, S, Harleem, EAI and Beijing spoligofamilies were identified. Three M. bovis isolates were also observed. Among those, CAS1-Kili (SIT 21) and LAM1 (SIT 20) subfamilies showed a propensity for MDR-TB with p = 0.0001 and p = 0.001, respectively. CONCLUSIONS: This phenomenon might explain the future increase in the MDR-TB burden caused by specific lineages in Zambia. Therefore, it is recommended that the National TB control program in the country complements conventional control strategies with molecular analysis for monitoring and surveillance of MDR-TB epidemiology.

Prevalence of iodine deficiency among pregnant women in Gwembe and Sinazongwe districts of Southern Province, Zambia: a cross-sectional study.

BACKGROUND: Maternal iodine deficiency is one of the common causes of morbidity and mortality during pregnancy. Maternal iodine deficiency during pregnancy is associated with a number of adverse outcomes such as abortion, stillbirth, congenital anomalies, perinatal mortality and irreversible mental retardation. A study conducted in Zambia among pregnant women in 2013 on the prevalence of iodine deficiency showed that iodine deficiency was not a public health concern. The previous study used Urine Iodine concentration (UIC) as a marker of iodine deficiency among the pregnant women. Our study was conducted to assess the prevalence of iodine deficiency among pregnant women in Gwembe and Sinazongwe districts of Southern Province, Zambia, using urine iodine concentration and goitre presence by manual palpation. METHODS: We carried out a community based, cross sectional study in rural areas of Gwembe and Sinazongwe districts between April 2016 to March 2018. Data were collected from 412 pregnant women by a multistage cluster sampling technique. The presence of a goitre was examined by manual palpation and urinary iodine concentration was determined by the Ultra Violet Method using PerkinElmer Labda UV Spectrometer equipment made in Jena Germany (Model 107,745). As part of the existing baseline data, we used results of a 2013 countrywide study (n = 489) for household salt iodine content which showed a greater than 40 ppm at 76.2%, between 15 and 40 ppm at 19.21% and less than 15 ppm at 4.59%. Statistical analysis was done using Stata version 14.0. The outputs of analysis are presented as median and Interquartile range (IQR) as the urine data were not normally distributed. Further, the categorical and independent variables were presented as proportions (percentages) to describe the distribution and trends in the target sample population. RESULTS: The median Urine Iodine concentration (UIC) of the pregnant women was 150 µg/L (Interquartile Range (IQR): 100-200 µg/L). Based on the UIC, There were 49% pregnant women who had inadequate iodine intake with urine iodine concentration of less than 150 µg/L, 34.0% had UIC of 150-249 µg/L indicating adequate iodine intake, 13.0% with UIC of 250-499 µg/L indicating more than adequate iodine intake, and 5.0% with UIC of above 500 µg/L indicating excessive iodine intake. To determine whether the women had access to iodized salt, we used baseline data from 2013 Zambia national survey for iodine concentration in household salt samples as being an average of 40 ppm, which also showed that 95.41% households consumed adequately iodized salt (≥15 ppm). The prevalence of goitre in our study was very low at 0.02% among the pregnant women of all ages who participated in the study (18-49 years). CONCLUSION: Iodine deficiency was still not a public health concern among the pregnant women of Gwembe and Sinazongwe districts of Southern Province in Zambia. Goitre prevalence has remained very low in this study area. The UIC and goitre observations were consistent with the Zambia National Food and Nutrition Commission findings in 2013 report. However, our study showed more pregnant women with insufficient than adequate iodine status indicating the risk of developing IDD is still high in this region. It also reinforces the argument that strengthening of the existing salt iodization program is needed in order to make a homogenous iodated salt available to the communities. The National Food and Nutrition Commission of Zambia needs to find innovative ways of sensitizing people about the adverse effects of IDDs and how these could be prevented. It is recommended that iodine supplementation be introduced as part of the package of Antenatal clinic care for all pregnant women.

Mutations in rpoB and katG genes and the inhA operon in multidrug-resistant Mycobacterium tuberculosis isolates from Zambia.

OBJECTIVES: It is established that resistance to rifampicin (RIF) in 90% of RIF-resistant Mycobacterium tuberculosis isolates is attributable to point mutations in the rpoB gene, whilst 50-95% of M. tuberculosis resistance to isoniazid (INH) is caused by mutations in the katG gene. However, the patterns and frequencies of mutations vary by geographical region. In Zambia, the genetic mechanisms of resistance of M. tuberculosis to RIF and INH were unreported before this study. METHODS: Using gene sequencing, the rpoB, katG and inhA genes of 99 multidrug-resistant M. tuberculosis (MDR-TB) and 49 pan-susceptible M. tuberculosis isolates stored at a tuberculosis reference laboratory from 2013 to 2016 were analysed and were compared with published profiles from other African countries. RESULTS: Of the 99 MDR-TB isolates, 95 (96.0%) carried mutations in both rpoB and katG. No mutations were detected among the pan-susceptible isolates. The most common mutations among RIF- and INH-resistant isolates were in codon 531 of the rpoB gene (55.6%; 55/99) and codon 315 of the katG gene (94.9%; 94/99), respectively. Distinctly, katG mutations were predominantly high among Zambian isolates (96.0%) compared with other countries in the region. CONCLUSION: Resistance-associated mutations to RIF and INH circulating in Zambia are similar to those reported globally, therefore these data validate the applicability of molecular diagnostic tools in Zambia. However, katG mutations were predominantly high among M. tuberculosis isolates in this study compared with other regional countries and might distinguish cross-boundary transmission of MDR-TB from other African nations.

Evidence of EBV infection in lymphomas diagnosed in Lusaka, Zambia.

INTRODUCTION: Epstein-Barr virus (EBV) is a ubiquitous virus that infects more than 90% of the world's population, and is implicated in lymphoma pathogenesis. However, in Zambia during the diagnosis of these lymphomas, the association of the virus with the lymphomas is not established. Since most patients with lymphomas have poor prognosis, the identification of the virus within the lymphoma lesion will allow for more targeted therapy. The aim of this study was to provide evidence of the presence of the EBV in lymphomas diagnosed at the University Teaching Hospital (UTH) in Lusaka, Zambia. METHODS: One hundred and fifty archival formalin-fixed paraffin embedded suspected lymphoma tissues stored over a 4-year period in the Histopathology Laboratory at the UTH in Lusaka, Zambia, were analysed. Histological methods were used to identify the lymphomas, and the virus was detected using Polymerase Chain Reaction (PCR). Subtyping of the virus was achieved through DNA sequencing of the EBNA-2 region of the viral genome. Chi square or fisher's exact test was used to evaluate the association between EBV status, type of lymphoma and gender. RESULTS: The majority of the lymphomas identified were non-Hodgkin's lymphoma (NHL) (80%) followed by Hodgkin's lymphoma (HL) (20%). EBV was detected in 51.8% of the cases, 54.5% of which were associated with NHL cases, while 40.9% associated with HL cases. The predominant subtype of the virus in both types of lymphomas was subtype 1. One of the lymphoma cases harboured both subtype 1 and 2 of the virus. CONCLUSION: This study showed that EBV is closely associated with lymphomas. Therefore, providing evidence of the presence of the virus in lymphoma tissues will aid in targeted therapy. To our knowledge this is the first time such data has been generated in Zambia.

Seroprevalence of parvovirus B19 in blood donors: the risks and challenges of blood transfusion in Zambia in the era of HIV/AIDS at the Kitwe Central Hospital, blood bank.

BACKGROUND: Human Parvovirus (B19V) is a small, single-stranded, non-enveloped DNA virus which is pathogenic to humans causing a wide array of clinical complications which include erythema infectiosum, aplastic crisis and hydrops foetalis. It is generally harmless in healthy individuals but may be life threatening in immunocompromised individuals such as patients with sickle cell disease, cancer, HIV and pregnant women. It has been shown to be transmissible by blood transfusion but donor screening for the virus is not yet mandatory in most sub-Saharan African countries including Zambia. MATERIALS AND METHODS: This was a cross-sectional study undertaken at the Kitwe Central Hospital, blood bank and Tropical Diseases Research Centre at Ndola Central Hospital. A total of 192 blood samples were screened for Ig M antibodies against parvovirus B19 by ELISA. OBJECTIVES: The general objective of the study was to determine the seroprevalence of parvovirus B19 infections among healthy blood donors at the Kitwe Central Hospital blood bank. Specific Objectives were to detect parvovirus B19 Ig M antibodies in donor blood using serology and to analyse the age and sex distribution of parvovirus among blood donors. RESULTS: The prevalence of parvovirus B19 Ig M in this study was 15.6%. The majority of the positive cases were in the age group 15-22 years (17.8%) but there was no statistical significance between occurrence of parvovirus and age ( p value=0.703). Prevalence in males was higher than in females that is 16.4% and 13.8%, respectively. The relationship between gender and parvovirus B19 occurrence was however not significant either (p value=0.516). CONCLUSION: This study showed a 15.6% prevalence rate of acute Parvovirus B19 infections in blood donors at the Kitwe Central Hospital, blood bank. Studies with larger sample sizes are needed to validate these results.

Detection of Human Herpes Virus 8 in Kaposi's sarcoma tissues at the University Teaching Hospital, Lusaka, Zambia.

INTRODUCTION: Human herpes virus-8, a γ2-herpes virus, is the aetiological agent of Kaposi sarcoma. Recently, Kaposi's sarcoma cases have increased in Zambia. However, the diagnosis of this disease is based on morphological appearance of affected tissues using histological techniques, and the association with its causative agent, Human Herpes virus 8 is not sought. This means poor prognosis for affected patients since the causative agent is not targeted during diagnosis and KS lesions may be mistaken for other reactive and neoplastic vascular proliferations when only histological techniques are used. Therefore, this study was aimed at providing evidence of Human Herpes virus 8 infection in Kaposi's sarcoma tissues at the University Teaching Hospital in Lusaka, Zambia. METHODS: One hundred and twenty suspected Kaposi's sarcoma archival formalin-fixed paraffin-wax embedded tissues stored from January 2013 to December 2014 in the Histopathology Laboratory at the University Teaching Hospital, Lusaka, Zambia were analysed using histology and Polymerase Chain Reaction targeting the ORF26 gene of Human Herpes virus 8. RESULTS: The predominant histological type of Kaposi's sarcoma detected was the Nodular type (60.7%) followed by the plaque type (22.6%) and patch type (16.7%). The nodular lesion was identified mostly in males (40.5%, 34/84) than females (20.2%, 17/84) (p=0.041). Human Herpes virus 8 DNA was detected in 53.6% (45/84) and mostly in the nodular KS lesions (60%, 27/84) (p=0.035). CONCLUSION: The findings in this study show that the Human Herpes virus-8 is detectable in Kaposi's sarcoma tissues, and, as previously reported in other settings, is closely associated with Kaposi's sarcoma. The study has provided important baseline data for use in the diagnosis of this disease and the identification of the virus in the tissues will aid in targeted therapy.

Performance Characteristics of Kidney Injury Molecule-1 In Relation to Creatinine, Urea, and Microalbuminuria in the Diagnosis of Kidney Disease.

CONTEXT: The diagnosis and evaluation of impaired renal function remains a challenge owing to lack of reliable biomarker for assessment of kidney function. The existing panel of biomarkers currently displays several limitations, and recently kidney injury molecule-1 (KIM-1) has been suggested as a sensitive biomarker of renal function and proposed to enter clinical practice. AIMS: This study was conducted to determine the diagnostic value of serum creatinine, urea, and microalbuminuria (MAU) in relation to the novel biomarker, KIM-1. MATERIALS AND METHODS: Serum creatinine, urea, MAU, and KIM-1 were measured in forty individuals with and forty without kidney disease. Data were analyzed using multivariate methods of assessing diagnostic efficiency, test agreement, condition effects, and variability. RESULTS: The area under the receiver-operator characteristic curve revealed a diagnostic advantage of creatinine (0.924 ± 0.0066) and urea (0.925 ± 0.0068) over MAU (0.880 ± 0.078) and KIM-1 (0.35 ± 0.124). Overall diagnostic efficiency was higher for creatinine and urea (89.5% and 90.9%, respectively), followed by MAU (85.7%) and then KIM-1 (56.3%). Logistic regression analysis showed that creatinine and urea (R2 = 0.75 and R2 = 0.72, respectively, P < 0.001 for both) were better predictors of kidney disease than MAU (R2 = 0.64, P < 0.001) and KIM-1 (R2 = 0.046, P = 0.116). Further analysis of agreement showed that urea had an excellent agreement with creatinine (kappa r = 0.835, P < 0.001), with KIM-1 (kappa r = -0.198, P = 0.087) showing a poor agreement with creatinine. CONCLUSION: Our results indicate that elevated serum creatinine and urea above specific cutoff points reliably identifies patients with acute kidney injury or chronic kidney disease. However, more researches are warranted to further validate the diagnostic efficiency and application of MAU and for KIM-1 before its implementation in clinical practice.

Kidney injury molecule-1 and microalbuminuria levels in Zambian population: biomarkers of kidney injury.

INTRODUCTION: Kidney injury affects renal excretion of plasma analytes and metabolic waste products with grave pathologic consequences. Early detection, thus of kidney injury is essential for injury specific intervention that may avert permanent renal damage and delay progression of kidney injury. We aimed to evaluate Kidney Injury Molecule-1 (KIM-1) and Microalbuminuria (MAU), as biomarkers of kidney injury, in comparison with creatinine. METHODS: We compared the levels of urine MAU, urine KIM-1 and other plasma biochemical tests in specimens from 80 individuals with and without kidney disease. RESULTS: We found no difference in KIM-1 levels between the kidney disease group (2.82± 1.36ng/mL) and controls (3.29 ± 1.14ng/mL), p = 0.122. MAU was higher in participants with kidney disease (130.809± 84.744 µg/mL) than the controls (15.983± 20.442µg/mL), p ?0.001. KIM-1 showed a weak negative correlation with creatinine (r = -0.279, p = 0.09), whereas MAU was positively correlated with creatinine in participants with kidney disease with statistical significance (r = 0.556, p = 0.001). CONCLUSION: The study demonstrated that in Zambian setting MAU and creatinine are sensitive biomarkers in the diagnosis of kidney damage. We moreover propose further evaluation of KIM-1 as a biomarker of kidney injury.

Molecular identification of non-tuberculous mycobacteria isolated from clinical specimens in Zambia.

BACKGROUND: The emergence of Acquired Immunodeficiency Syndrome has highlighted the increased incidence and importance of the disease caused by Non-tuberculous Mycobacteria (NTM). While disease due to M. avium-intracellulare complex is apparently common throughout the world, other Non-tuberculous mycobacterial species have been isolated from both immunocompromised and immunocompetent individuals. The increasing number of infections caused by these organisms has made it clinically important to quickly identify mycobacterial species. The diagnosis of a pathogenic versus a non-pathogenic species not only has epidemiological implications but is also relevant to the demands of patient management. Since antibiotic treatment varies according to the species encountered, species identification would reduce the burden of some of these emerging opportunistic pathogens especially in immunocompromised patients and improve their quality of life. FINDINGS: A total of 91 NTM suspected isolates from four regions of Zambia were included in the study. These isolates were identified using the sequence analysis of the 16S-23S rRNA intergenic transcribed spacer (ITS) region of Mycobacteria. Fifty-four of the 91 (59%) isolates were identified as NTM and these included M. intracellulare (27.8%), M. lentiflavum (16.7%), M. avium (14.8%), M. fortuitum (7.4%), M. gordonae (7.4%), M. kumamotonense (3.7%), M. indicus pranii (3.7%), M. peregrinum (3.7%), M. elephantis (1.85%), M. flavescens (1.85%), M. asiaticum (1.85%), M. bouchedurhonense (1.85%), M. chimaera (1.85%), M. europaeum (1.85%), M. neourum (1.85%), M. nonchromogenicum (1.5%). CONCLUSION: The study has shown that DNA sequencing of the ITS region may be useful in the preliminary identification of NTM species. All species identified in this study were potentially pathogenic.

Neuropsychiatric and socioeconomic status impact antiretroviral adherence and mortality in rural Zambia.

We conducted a prospective cohort study of 496 adults starting antiretroviral treatment (ART) to determine the impact of neuropsychiatric symptoms and socioeconomic status on adherence and mortality. Almost 60% had good adherence based upon pharmacy records. Poor adherence was associated with being divorced, poorer, food insecure, and less educated. Longer travel time to clinic, concealing one's human immunodeficiency virus (HIV) status, and experiencing side effects predicted poor adherence. Over a third of the patients had cognitive impairment and poorer cognitive function was also associated with poor adherence. During follow-up (mean 275 days), 20% died-usually within 90 days of starting ART. Neuropsychiatric symptoms, advanced HIV, peripheral neuropathy symptoms, food insecurity, and poverty were associated with death. Neuropsychiatric symptoms, advanced HIV, and poverty remained significant independent predictors of death in a multivariate model adjusting for other significant factors. Social, economic, cognitive, and psychiatric problems impact adherence and survival for people receiving ART in rural Zambia.

Antiretroviral adherence in rural Zambia: the first year of treatment availability.

We conducted a retrospective chart review of antiretroviral therapy (ART) clinic patients treated during the first 12 months after clinics opened in rural Zambia and assessed adherence based on clinic attendance, patient report, and staff assessment. We identified 255 eligible patients (mean age, 39.7 years; 44.3% male; 56.5% married; and 45.5% with only primary school education). Twenty percent had partners known to be HIV positive. Twenty percent were widowed. Thirty-seven percent had disclosed their HIV status to their spouse. Disclosure was less likely among women (27.5% versus 49.6%, P = 0.0005); 36.5% had "clinic buddies" to provide adherence support. Adherence rates were good for 59.2%. Disclosure of HIV status to ones' spouse (P = 0.047), knowing spouses' HIV status (P = 0.02), and having a clinic buddy (P = 0.01) were associated with good adherence. Social support is a key patient-level resource impacting ART adherence in rural Zambia. Limited spousal disclosure affects women more than men. Clinic buddies are associated with better adherence.

Inappropriately low aldosterone concentrations in adults with AIDS-related diarrhoea in Zambia: a study of response to fluid challenge.

BACKGROUND: Chronic diarrhoea is one of the most debilitating consequences of HIV infection in sub-Saharan Africa and it carries a high mortality rate. We report unexpectedly low concentrations of circulating aldosterone in 12 patients (6 men, 6 women) in the University Teaching Hospital, Lusaka, who all had diarrhoea for over one month. Changes in serum electrolytes, blood pressure, Karnofsky score and serum aldosterone concentration were being monitored during a short study of responses to saline infusion (3 litres/24 h) over 72 hours. FINDINGS: At baseline, 9/12 (75%) of the patients were hyponatraemic, 10/11 (91%) were hypokalaemic, and 6/12 (50%) had undetectable aldosterone concentrations. Blood pressure and Karnofsky score rose and creatinine concentration fell in response to the infusion. CONCLUSION: Circulating aldosterone concentrations were inappropriately low and complicate the profound electrolyte deficiencies resulting from chronic diarrhoea. Management of these deficiencies needs to be more aggressive than is currently practised and consideration should be given to a formal clinical trial of mineralocorticoid replacement in these severely ill patients. If the inappropriately low aldosterone reflects a general adrenal failure, it may explain a considerable proportion of the high mortality seen both before and after initiation of anti-retroviral therapy.