Aberrant phosphorylation and subsequent aggregation of the trans-activation response (TAR) elemen... more Aberrant phosphorylation and subsequent aggregation of the trans-activation response (TAR) element DNA binding protein 43 (TDP-43) is a common feature of multiple neurodegenerative disorders and contributes to disease severity. Here, we investigated whether pathologic phosphorylation of TDP-43 (pTDP-43) is a hallmark of human immunodeficiency virus (HIV)- infected brains. We evaluated pTDP-43 immunoreactivity and TDP-43 kinases in HIV-infected (HIV + ) and seronegative post-mortem brain samples. We then used an inducible transgenic mouse model of the HIV-1 protein Tat and primary neuronal cultures, to decipher the underlying mechanism of the proteinopathy. Since opioid use disorder (OUD) can exaggerate HIV neuropathology, we explored interactions between HIV-1 Tat and morphine, a prototypical opioid, for all outcome measures. Cytoplasmic pTDP-43 and TDP-43 immunoreactivities were increased in neurons of the basal ganglia of post-mortem, HIV+ human tissues compared to seronegative controls. An evaluation of TDP-43 kinases revealed an increase in the levels of cytoplasmic casein kinase 2 (CK2) in HIV-positive human tissues but not CK1δ. There was a significant positive correlation between pTDP-43 and CK2 levels. Eight weeks of Tat induction and 2-week subcutaneous morphine exposure (10–40 mg/kg, increasing by 10 mg/kg/b.i.d.) independently produced similar outcomes for cytoplasmic pTDP-43 and CK2 levels in the mouse striatum. In primary, mouse striatal neuronal cultures, co-exposure to Tat and morphine for 24 h increased pTDP-43 levels and CK2 activity. Co-treatment with the CK2 antagonist CX-4945 prevented the Tat- and morphine-induced increases in pTDP-43 levels. Our results demonstrate that CK2 may be a viable therapeutic target for treating pTDP-43 proteinopathy in neuroHIV and OUD. Summary Statement HIV/HIV-1 Tat and morphine independently increase pathologic phosphorylation of TAR DNA binding protein 43 in the striatum. HIV- and opioid-induced pathologic phosphorylation of TAR DNA binding protein 43 may involve enhanced CK2 activity and protein levels.
The shared practice of pathology via the Internet holds great potential for pathologists in sub-S... more The shared practice of pathology via the Internet holds great potential for pathologists in sub-Saharan Africa (SSA) and their global partners. Application of the Internet is constrained by issues of bandwidth, cost, and power. The penetration of mobile telephony and the arrival of smartphones have changed the use of Internet and social media in Africa and therefore the work of the 4 African pathologists featured in this article. As pathology in SSA struggles for visibility and usefulness, the Internet and its electronic applications provide a critical infrastructure as well as a podium for pathologists across the continent.
bioRxiv (Cold Spring Harbor Laboratory), Mar 15, 2023
If we are to break new grounds in TB research, we need to have a complete understanding of what i... more If we are to break new grounds in TB research, we need to have a complete understanding of what is occurring at the site of infection in humans. Postmortem studies give us an opportunity to compare TB-involved and-uninvolved tissues, in both diseased and nondiseased individuals. We examined the feasibility of carrying out a postmortem study in Mulago and Kiruddu National Referral Hospitals in Uganda, to determine whether we could use immune cells collected postmortem for immunological studies. We report that we can consent the Next-of-Kin, perform postmortem procedures and process tissues within 8 hours of death, and that immune cells remain viable and functional up to 14 hours after death. We found subtle differences in T cell subsets within TB groups. We found a depletion of the CD4 CD69 + CD103 + T cell subset in the lungs and BAL, which was associated with HIV, and that the CD8 CD69 + CD103-T cell subset was depleted in BAL only, and was associated with TB. Our data show overall changes Tissue Resident Memory T cells within, and between, TB-infected and TB-uninfected human lungs.
Kaposi’s sarcoma (KS) is an endothelial cancer caused by the Kaposi’s sarcoma-associated herpesvi... more Kaposi’s sarcoma (KS) is an endothelial cancer caused by the Kaposi’s sarcoma-associated herpesvirus (KSHV) and is one of the most common cancers in sub-Saharan Africa. In limited-resource settings, traditional pathology infrastructure is often insufficient for timely diagnosis, leading to frequent diagnoses at advanced-stage disease where survival is poor. In this study, we investigate molecular diagnosis of KS performed in a point-of-care device to circumvent the limited infrastructure for traditional diagnosis. Using 506 mucocutaneous biopsies collected from patients at three HIV clinics in Uganda, we achieved 97% sensitivity, 92% specificity, and 96% accuracy compared to gold standard U.S.-based pathology. The results presented in this manuscript show that LAMP-based quantification of KSHV DNA extracted from KS-suspected biopsies has the potential to serve as a successful diagnostic for the disease and that diagnosis may be accurately achieved using a point-of-care device, reducing the barriers to obtaining KS diagnosis while increasing diagnostic accuracy.
Journal of cancer and tumor international, Apr 3, 2019
Aims: The aims of the current study were to determine the prevalence of epidermal growth factor (... more Aims: The aims of the current study were to determine the prevalence of epidermal growth factor (EGFR) receptor in patients diagnosed with esophageal squamous cell carcinoma (ESCC) as well as assessing the correlation of overexpression of EGFR with age, gender and tumor grades of the cases. Study Design: This was a cross-sectional analytical study.
IMPACT: This work represents a novel way in which genetic information can be used to improve clin... more IMPACT: This work represents a novel way in which genetic information can be used to improve clinical decision making as it pertains to both treatment and management of congenital heart disease. OBJECTIVES/GOALS: Our lab found that MYH6 variants are both enriched in hypoplastic left heart syndrome (HLHS) and associated with decreased cardiac transplant-free survival. To elucidate the mechanisms of MYH6 variant pathogenicity, we are assessing their impact on atrial function during HLHS development and progression. METHODS/STUDY POPULATION:We are using 2D speckle-based tracking to retrospectively evaluate echocardiograms (echos) from 51 HLHS patients, 17 with MYH6 variants and 34 matched controls. Atrial function will be assessed by myocardial strain and strain rate at seven time points, beginning at the time of the patients’ earliest prenatal echo, and ending with their last available echo before death or cardiac transplant. Early left atrial function will examine the role of MYH6 va...
Background Fine needle aspiration cytology of the thyroid gland has proven to be effective in the... more Background Fine needle aspiration cytology of the thyroid gland has proven to be effective in the categorization of thyroid lesions into benign, atypical, suspicious and malignant categories. However only scanty data has been available regarding the prevalence of thyroid lesions in Uganda. This study aimed at classifying thyroid lesions cytologically and to determine their correlation with the social demographic characteristics. Methodology: A laboratory based retrospective study involving a review of 170 cases was conducted at MAKCHS pathology department between 2012 and 2016. FNA results were independently categorized into 4 groups in accordance to Bethesda system for reporting thyroid cytopathology. The data collected was entered using Epidata software and exported to SPSS for analysis. Results Out of the 170 cases reviewed, 148 (87.1%) were benign, 3 (1.8%) were suspicious for Follicular Neoplasm, 5 (2.9%) were suspicious of malignancy & 14 (8.2%) were malignant. Colloidal nodul...
Background: haematoxylin and eosin staining has remained the standard diagnostic method for Burki... more Background: haematoxylin and eosin staining has remained the standard diagnostic method for Burkitt's lymphoma. ancillary tests including immunohistochemistry, not widely available in developing countries, are important tools in verifying the diagnosis of lymphomas with equivocal morphological findings. Objective: to evaluate the reliability of haematoxylin and eosin staining in the diagnosis of Burkitt's lymphoma using immunohistochemistry as the gold standard. Design: cross sectional study. Setting: department of pathology laboratory, makerere university medical school, uganda. Subjects: One hundred and thirty eight formalin fixed paraffin embedded biopsies of Burkitt's lymphoma diagnosed from January 2001 to december 2005. Results: of the 138 tumours, 88.4% were extra-nodal: jaw 36.2%, ovary 21%, gastrointestinal tract 12.3% other abdominal 11.6%. Males (55.8%) predominated. The sensitivity and specificity of haematoxylin and eosin were 93.2% and 50% respectively. the positive and negative predictive values were 91.7% and 55.6%. Conclusion: histology using haematoxylin and eosin staining is sensitive in the diagnosis of Burkitt's lymphoma but not very specific. Immunohistochemical staining with CD20, Ki-67 and bcl-2 is necessary for difficult cases.
Background Optimal penetration of anti-infectives in the female genital tract (FGT) is paramount ... more Background Optimal penetration of anti-infectives in the female genital tract (FGT) is paramount in the treatment and prevention of infectious diseases. While exposure of anti-infectives in lower FGT tissues (e.g. cervix, vagina) has been described, little data exist on upper genital tissues (e.g. ovary, uterus). Methods Autopsies were performed and post-mortem tissues were collected within 24 h of death for female participants with advanced HIV in Uganda (n = 27). Tenofovir, lamivudine, efavirenz and fluconazole concentrations were measured using LC-MS/MS in plasma, ovarian, uterine, cervical and vaginal tissues. Tissue penetration was calculated as tissue-to-plasma concentration ratios (TPRs). Results TPRs of tenofovir, lamivudine and fluconazole were highest in vaginal tissue (medians 1.86, 1.83 and 0.94, respectively), while the TPR of efavirenz was highest in ovarian tissue (median 0.65). With cervix as a reference compartment, vaginal TPRs were significantly higher than cervic...
BACKGROUND The burden of cancer in Africa is growing. Although cancer outcomes are understood as ... more BACKGROUND The burden of cancer in Africa is growing. Although cancer outcomes are understood as the product of influences at multiple socioecological levels, population-based studies of geographical factors and cancer outcomes in Africa are scarce. The objective of this study was to identify parish-level social factors associated with cervical cancer incidence in the Kampala Cancer Registry catchment area, using a novel linkage between population-based cancer registry data and small-area census data from Uganda. METHODS Kampala Cancer Registry cervical cancer records (2008-15) were augmented to add the parish of residence at diagnosis. Parish-level population and housing profile data (2014) were obtained from the Uganda Bureau of Statistics and linked to Kampala Cancer Registry records. Stepwise forward Poisson regression modelling was used to estimate incidence ratios (IR) assessing associations between social factors and incidence. Housing tenure, infrastructure, gender equality, economic status, and employment were examined, controlling for population density. The significance level was set at α=0·05. FINDINGS Factors related to higher incidence included a higher girl-to-boy ratio of 6-12-year-olds not attending school (IR 1·33 [95% CI 1·15-1·54]; p<0.001), a higher percentage of 10-17-year-olds ever married (IR 1·22 [95% CI 1·06-1·40]; p=0·006) and a higher percentage of households receiving remittances (IR 1·03 [95% CI 1·00-1·06]; p=0·026). Factors associated with lower incidence included a higher percentage of household owner occupancy (IR 0·95 [95% CI 0·92-0·98]; p=0·002) and a higher percentage of households with piped water (IR 0·97 [95% CI 0·95-0·99]; p=0·009). INTERPRETATION Parish-level social factors predict cervical cancer incidence in Uganda. Communities most at risk are characterised by inequity in educational access for girls, higher prevalence of child marriage, low home ownership, inadequate infrastructure, and financial dependence. These communities would benefit from HPV vaccination and screening campaigns to prevent and control cervical cancer. Investments should be made to enhance population-based cancer surveillance and census data collection in Africa to offer new strategies and targets for cancer prevention and control. FUNDING Medical College of Wisconsin Cancer Center and Medical College of Wisconsin Office of Global Health.
Fungal infections cause substantial morbidity and mortality. However, the burden of deep fungal i... more Fungal infections cause substantial morbidity and mortality. However, the burden of deep fungal infections is not well described in Uganda. We aimed to estimate the burden and etiology of histologically diagnosed deep fungal infections in Uganda. We retrospectively reviewed histology reports at the Pathology Reference Laboratory, Department of Pathology, Makerere University, Kampala, Uganda from January 1950 to September 2019 to identify any reports that had a fungal infection as the diagnosis. Over the study period, 697 cases of deep fungal infections were identified with an average incidence of 0.73/100,000 persons per decade. There was a general decline in the number of cases detected. Median age of the cases was 28 years (IQR: 11–40) and majority (59%) were male. The age group of 0–10 years were the most affected. The foot was the most affected part of the body (26%). Deep mycoses identified include eumycetoma (32%), subcutaneous phycomycosis (26%), histoplasmosis (9.2%), chromo...
The central nervous system (CNS) is a known HIV reservoir, yet little is known about drug exposur... more The central nervous system (CNS) is a known HIV reservoir, yet little is known about drug exposure in the brain. Our primary objective was to quantify exposure of three common antiretrovirals in brain tissue and compare exposures to plasma and cerebrospinal fluid (CSF). We also sought to identify pockets of brain most vulnerable to inadequate drug exposures and examine the role of meningitis in drug penetration into the CNS. Tenofovir, lamivudine, and efavirenz concentrations were measured using liquid chromatography and tandem mass spectrometry in plasma and CSF from 14 individuals with HIV, 7 with cryptococcal meningitis. In four individuals (three with meningitis) drug concentrations were also measured in 13 distinct brain tissue regions. In subjects with meningitis, geometric mean ratio (95% confidence interval) of tenofovir CSF to plasma was 66% (7-598%) and 14% (6-31%) in subjects without meningitis. Lamivudine CSF penetration was 100% (25-409%) in subjects with meningitis and 30% (24-37%) in subjects without meningitis. Tenofovir brain tissue concentrations were 36% (14-124%) of plasma and 49% (1-572%) of CSF. Lamivudine brain concentrations were 37% (23-64%) of plasma and 27% (1-104%) of CSF. Efavirenz brain tissue concentrations were 128% (108-179%) of plasma. Tissues collected postmortem provide a unique opportunity to assess drug distribution in tissues difficult to sample in living subjects. CSF is a poor surrogate for drug exposure throughout the CNS. Antiretrovirals differentially penetrate into the CNS and penetration may be enhanced by meningitis. Despite HIV therapy, persistent viral replication occurs in certain anatomic sites, and systemic viral rebound will occur within weeks of treatment interruption. 1 The central nervous system (CNS) is hypothesized to be a potential HIV reservoir, harboring virus that is either resistant or not exposed to adequate antiretroviral therapy. 2 CNS Penetration-Effectiveness scores, largely based on drug exposure in cerebrospinal fluid (CSF), were developed for antiretrovirals to provide a prescribers guidance on which regimens have greatest likelihood of achieving therapeutic CNS concentrations. 3 However, brain extracellular fluid can either be passively secreted to CSF or actively transported
Background There is a growing epidemic of cancer and other non-communicable diseases in sub-Sahar... more Background There is a growing epidemic of cancer and other non-communicable diseases in sub-Saharan Africa. Targeted, specific, cost-effective strategies are needed to manage the growing burden of cancer. In high-resource settings, geospatial analysis has transformed cancer control through geographic targeting of interventions and policies. A similar approach could improve cancer control in sub-Saharan Africa; however, georeferenced cancer data and increased geospatial research capacity are needed. Here, we aimed to assess the feasibility of geocoding and mapping small-area cancer data from a cancer registry in Uganda. Methods We established a partnership including the
The shared practice of pathology via the Internet holds great potential for pathologists in sub-S... more The shared practice of pathology via the Internet holds great potential for pathologists in sub-Saharan Africa (SSA) and their global partners. Application of the Internet is constrained by issues of bandwidth, cost, and power. The penetration of mobile telephony and the arrival of smartphones have changed the use of Internet and social media in Africa and therefore the work of the 4 African pathologists featured in this article. As pathology in SSA struggles for visibility and usefulness, the Internet and its electronic applications provide a critical infrastructure as well as a podium for pathologists across the continent.
<p>1. Renal TB 2. Passage of whole <i>Mycobacterium tuberculosis</i> through th... more <p>1. Renal TB 2. Passage of whole <i>Mycobacterium tuberculosis</i> through the glomerular basement membrane into the urine 3. Passage of LAM captured by immune-complexes through the glomerular basement membrane into the urine 4. Passage of uncomplexed LAM through the glomerular basement membrane into the urine 5. Impaired tubular resorption of uncomplexed LAM 6. Genital tract TB (Fig. based on [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0123323#pone.0123323.ref021" target="_blank">21</a>]). MTB: <i>Mycobacterium tuberculosis</i>; GBM: glomerular basement membrane; LAM: lipoarabinomannan.</p
Despite the persistently high HIV-related mortality in sub-Saharan Africa, limited information on... more Despite the persistently high HIV-related mortality in sub-Saharan Africa, limited information on the causes of death is available. Pathological autopsies are the gold standard to establish causes of death. In this review we describe the autopsy series performed among HIV-infected individuals in sub-Saharan Africa over the last two decades. We identified nine complete and 11 partial or minimally invasive autopsy series. Complete autopsies were performed in 593 HIV-positive adults and 177 HIV-positive children. Postmortem diagnoses were mainly infectious diseases. Tuberculosis was the most frequent, present in 21-54% of HIV-positive adults and was considered the cause of death in 32-45%. Overall, pulmonary infections accounted for approximately 66% of pathology and central nervous system infections for approximately 20%. A high discordance between clinical and postmortem diagnoses was observed. This review emphasizes the need for reliable information on causes of death in order to im...
Microglia provide protection against a range of brain infections, but how these glial cells respo... more Microglia provide protection against a range of brain infections, but how these glial cells respond to fungi is poorly understood. We investigated the role of microglia in the context of cryptococcal meningitis, the most common cause of fungal brain infections in humans. Using a series of transgenic- and chemical-based microglia depletion methods we found that, contrary to their protective role during other infections, microglia supported cryptococcal fungal brain infection. We show that microglia become hosts for intracellular fungal growth and are a site in which the fungus accesses the restricted micronutrient copper. We developed a reporter fungal strain to track copper starvation responses by the fungus and found that yeast were protected from copper starvation within microglia. Lastly, we show that stimulation of microglia with IFNγ causes restriction of phagosomal copper to intracellular fungi. These data provide a mechanistic explanation for why microglia depletion has a the...
Background The global burden of cervical cancer is concentrated in low-and middle-income countrie... more Background The global burden of cervical cancer is concentrated in low-and middle-income countries (LMICs), with the greatest burden in Africa. Targeting limited resources to populations with the greatest need to maximize impact is essential. The objectives of this study were to geocode cervical cancer data from a population-based cancer registry in Kampala, Uganda, to create high-resolution disease maps for cervical cancer prevention and control planning, and to share lessons learned to optimize efforts in other low-resource settings. Methods Kampala Cancer Registry records for cervical cancer diagnoses between 2008 and 2015 were updated to include geographies of residence at diagnosis. Population data by age and sex for 2014 was obtained from the Uganda Bureau of Statistics. Indirectly age-standardized incidence ratios were calculated for sub-counties and estimated continuously across the study area using parish level data. Results Overall, among 1873 records, 89.6% included a valid sub-county and 89.2% included a valid parish name. Maps revealed specific areas of high cervical cancer incidence in the region, with significant variation within sub-counties, highlighting the importance of high-resolution spatial detail. Conclusions Population-based cancer registry data and geospatial mapping can be used in low-resource settings to support cancer prevention and control efforts, and to create the potential for research examining geographic factors that influence cancer outcomes. It is essential to support LMIC cancer registries to maximize the benefits of limited cancer control resources.
Aberrant phosphorylation and subsequent aggregation of the trans-activation response (TAR) elemen... more Aberrant phosphorylation and subsequent aggregation of the trans-activation response (TAR) element DNA binding protein 43 (TDP-43) is a common feature of multiple neurodegenerative disorders and contributes to disease severity. Here, we investigated whether pathologic phosphorylation of TDP-43 (pTDP-43) is a hallmark of human immunodeficiency virus (HIV)- infected brains. We evaluated pTDP-43 immunoreactivity and TDP-43 kinases in HIV-infected (HIV + ) and seronegative post-mortem brain samples. We then used an inducible transgenic mouse model of the HIV-1 protein Tat and primary neuronal cultures, to decipher the underlying mechanism of the proteinopathy. Since opioid use disorder (OUD) can exaggerate HIV neuropathology, we explored interactions between HIV-1 Tat and morphine, a prototypical opioid, for all outcome measures. Cytoplasmic pTDP-43 and TDP-43 immunoreactivities were increased in neurons of the basal ganglia of post-mortem, HIV+ human tissues compared to seronegative controls. An evaluation of TDP-43 kinases revealed an increase in the levels of cytoplasmic casein kinase 2 (CK2) in HIV-positive human tissues but not CK1δ. There was a significant positive correlation between pTDP-43 and CK2 levels. Eight weeks of Tat induction and 2-week subcutaneous morphine exposure (10–40 mg/kg, increasing by 10 mg/kg/b.i.d.) independently produced similar outcomes for cytoplasmic pTDP-43 and CK2 levels in the mouse striatum. In primary, mouse striatal neuronal cultures, co-exposure to Tat and morphine for 24 h increased pTDP-43 levels and CK2 activity. Co-treatment with the CK2 antagonist CX-4945 prevented the Tat- and morphine-induced increases in pTDP-43 levels. Our results demonstrate that CK2 may be a viable therapeutic target for treating pTDP-43 proteinopathy in neuroHIV and OUD. Summary Statement HIV/HIV-1 Tat and morphine independently increase pathologic phosphorylation of TAR DNA binding protein 43 in the striatum. HIV- and opioid-induced pathologic phosphorylation of TAR DNA binding protein 43 may involve enhanced CK2 activity and protein levels.
The shared practice of pathology via the Internet holds great potential for pathologists in sub-S... more The shared practice of pathology via the Internet holds great potential for pathologists in sub-Saharan Africa (SSA) and their global partners. Application of the Internet is constrained by issues of bandwidth, cost, and power. The penetration of mobile telephony and the arrival of smartphones have changed the use of Internet and social media in Africa and therefore the work of the 4 African pathologists featured in this article. As pathology in SSA struggles for visibility and usefulness, the Internet and its electronic applications provide a critical infrastructure as well as a podium for pathologists across the continent.
bioRxiv (Cold Spring Harbor Laboratory), Mar 15, 2023
If we are to break new grounds in TB research, we need to have a complete understanding of what i... more If we are to break new grounds in TB research, we need to have a complete understanding of what is occurring at the site of infection in humans. Postmortem studies give us an opportunity to compare TB-involved and-uninvolved tissues, in both diseased and nondiseased individuals. We examined the feasibility of carrying out a postmortem study in Mulago and Kiruddu National Referral Hospitals in Uganda, to determine whether we could use immune cells collected postmortem for immunological studies. We report that we can consent the Next-of-Kin, perform postmortem procedures and process tissues within 8 hours of death, and that immune cells remain viable and functional up to 14 hours after death. We found subtle differences in T cell subsets within TB groups. We found a depletion of the CD4 CD69 + CD103 + T cell subset in the lungs and BAL, which was associated with HIV, and that the CD8 CD69 + CD103-T cell subset was depleted in BAL only, and was associated with TB. Our data show overall changes Tissue Resident Memory T cells within, and between, TB-infected and TB-uninfected human lungs.
Kaposi’s sarcoma (KS) is an endothelial cancer caused by the Kaposi’s sarcoma-associated herpesvi... more Kaposi’s sarcoma (KS) is an endothelial cancer caused by the Kaposi’s sarcoma-associated herpesvirus (KSHV) and is one of the most common cancers in sub-Saharan Africa. In limited-resource settings, traditional pathology infrastructure is often insufficient for timely diagnosis, leading to frequent diagnoses at advanced-stage disease where survival is poor. In this study, we investigate molecular diagnosis of KS performed in a point-of-care device to circumvent the limited infrastructure for traditional diagnosis. Using 506 mucocutaneous biopsies collected from patients at three HIV clinics in Uganda, we achieved 97% sensitivity, 92% specificity, and 96% accuracy compared to gold standard U.S.-based pathology. The results presented in this manuscript show that LAMP-based quantification of KSHV DNA extracted from KS-suspected biopsies has the potential to serve as a successful diagnostic for the disease and that diagnosis may be accurately achieved using a point-of-care device, reducing the barriers to obtaining KS diagnosis while increasing diagnostic accuracy.
Journal of cancer and tumor international, Apr 3, 2019
Aims: The aims of the current study were to determine the prevalence of epidermal growth factor (... more Aims: The aims of the current study were to determine the prevalence of epidermal growth factor (EGFR) receptor in patients diagnosed with esophageal squamous cell carcinoma (ESCC) as well as assessing the correlation of overexpression of EGFR with age, gender and tumor grades of the cases. Study Design: This was a cross-sectional analytical study.
IMPACT: This work represents a novel way in which genetic information can be used to improve clin... more IMPACT: This work represents a novel way in which genetic information can be used to improve clinical decision making as it pertains to both treatment and management of congenital heart disease. OBJECTIVES/GOALS: Our lab found that MYH6 variants are both enriched in hypoplastic left heart syndrome (HLHS) and associated with decreased cardiac transplant-free survival. To elucidate the mechanisms of MYH6 variant pathogenicity, we are assessing their impact on atrial function during HLHS development and progression. METHODS/STUDY POPULATION:We are using 2D speckle-based tracking to retrospectively evaluate echocardiograms (echos) from 51 HLHS patients, 17 with MYH6 variants and 34 matched controls. Atrial function will be assessed by myocardial strain and strain rate at seven time points, beginning at the time of the patients’ earliest prenatal echo, and ending with their last available echo before death or cardiac transplant. Early left atrial function will examine the role of MYH6 va...
Background Fine needle aspiration cytology of the thyroid gland has proven to be effective in the... more Background Fine needle aspiration cytology of the thyroid gland has proven to be effective in the categorization of thyroid lesions into benign, atypical, suspicious and malignant categories. However only scanty data has been available regarding the prevalence of thyroid lesions in Uganda. This study aimed at classifying thyroid lesions cytologically and to determine their correlation with the social demographic characteristics. Methodology: A laboratory based retrospective study involving a review of 170 cases was conducted at MAKCHS pathology department between 2012 and 2016. FNA results were independently categorized into 4 groups in accordance to Bethesda system for reporting thyroid cytopathology. The data collected was entered using Epidata software and exported to SPSS for analysis. Results Out of the 170 cases reviewed, 148 (87.1%) were benign, 3 (1.8%) were suspicious for Follicular Neoplasm, 5 (2.9%) were suspicious of malignancy & 14 (8.2%) were malignant. Colloidal nodul...
Background: haematoxylin and eosin staining has remained the standard diagnostic method for Burki... more Background: haematoxylin and eosin staining has remained the standard diagnostic method for Burkitt's lymphoma. ancillary tests including immunohistochemistry, not widely available in developing countries, are important tools in verifying the diagnosis of lymphomas with equivocal morphological findings. Objective: to evaluate the reliability of haematoxylin and eosin staining in the diagnosis of Burkitt's lymphoma using immunohistochemistry as the gold standard. Design: cross sectional study. Setting: department of pathology laboratory, makerere university medical school, uganda. Subjects: One hundred and thirty eight formalin fixed paraffin embedded biopsies of Burkitt's lymphoma diagnosed from January 2001 to december 2005. Results: of the 138 tumours, 88.4% were extra-nodal: jaw 36.2%, ovary 21%, gastrointestinal tract 12.3% other abdominal 11.6%. Males (55.8%) predominated. The sensitivity and specificity of haematoxylin and eosin were 93.2% and 50% respectively. the positive and negative predictive values were 91.7% and 55.6%. Conclusion: histology using haematoxylin and eosin staining is sensitive in the diagnosis of Burkitt's lymphoma but not very specific. Immunohistochemical staining with CD20, Ki-67 and bcl-2 is necessary for difficult cases.
Background Optimal penetration of anti-infectives in the female genital tract (FGT) is paramount ... more Background Optimal penetration of anti-infectives in the female genital tract (FGT) is paramount in the treatment and prevention of infectious diseases. While exposure of anti-infectives in lower FGT tissues (e.g. cervix, vagina) has been described, little data exist on upper genital tissues (e.g. ovary, uterus). Methods Autopsies were performed and post-mortem tissues were collected within 24 h of death for female participants with advanced HIV in Uganda (n = 27). Tenofovir, lamivudine, efavirenz and fluconazole concentrations were measured using LC-MS/MS in plasma, ovarian, uterine, cervical and vaginal tissues. Tissue penetration was calculated as tissue-to-plasma concentration ratios (TPRs). Results TPRs of tenofovir, lamivudine and fluconazole were highest in vaginal tissue (medians 1.86, 1.83 and 0.94, respectively), while the TPR of efavirenz was highest in ovarian tissue (median 0.65). With cervix as a reference compartment, vaginal TPRs were significantly higher than cervic...
BACKGROUND The burden of cancer in Africa is growing. Although cancer outcomes are understood as ... more BACKGROUND The burden of cancer in Africa is growing. Although cancer outcomes are understood as the product of influences at multiple socioecological levels, population-based studies of geographical factors and cancer outcomes in Africa are scarce. The objective of this study was to identify parish-level social factors associated with cervical cancer incidence in the Kampala Cancer Registry catchment area, using a novel linkage between population-based cancer registry data and small-area census data from Uganda. METHODS Kampala Cancer Registry cervical cancer records (2008-15) were augmented to add the parish of residence at diagnosis. Parish-level population and housing profile data (2014) were obtained from the Uganda Bureau of Statistics and linked to Kampala Cancer Registry records. Stepwise forward Poisson regression modelling was used to estimate incidence ratios (IR) assessing associations between social factors and incidence. Housing tenure, infrastructure, gender equality, economic status, and employment were examined, controlling for population density. The significance level was set at α=0·05. FINDINGS Factors related to higher incidence included a higher girl-to-boy ratio of 6-12-year-olds not attending school (IR 1·33 [95% CI 1·15-1·54]; p<0.001), a higher percentage of 10-17-year-olds ever married (IR 1·22 [95% CI 1·06-1·40]; p=0·006) and a higher percentage of households receiving remittances (IR 1·03 [95% CI 1·00-1·06]; p=0·026). Factors associated with lower incidence included a higher percentage of household owner occupancy (IR 0·95 [95% CI 0·92-0·98]; p=0·002) and a higher percentage of households with piped water (IR 0·97 [95% CI 0·95-0·99]; p=0·009). INTERPRETATION Parish-level social factors predict cervical cancer incidence in Uganda. Communities most at risk are characterised by inequity in educational access for girls, higher prevalence of child marriage, low home ownership, inadequate infrastructure, and financial dependence. These communities would benefit from HPV vaccination and screening campaigns to prevent and control cervical cancer. Investments should be made to enhance population-based cancer surveillance and census data collection in Africa to offer new strategies and targets for cancer prevention and control. FUNDING Medical College of Wisconsin Cancer Center and Medical College of Wisconsin Office of Global Health.
Fungal infections cause substantial morbidity and mortality. However, the burden of deep fungal i... more Fungal infections cause substantial morbidity and mortality. However, the burden of deep fungal infections is not well described in Uganda. We aimed to estimate the burden and etiology of histologically diagnosed deep fungal infections in Uganda. We retrospectively reviewed histology reports at the Pathology Reference Laboratory, Department of Pathology, Makerere University, Kampala, Uganda from January 1950 to September 2019 to identify any reports that had a fungal infection as the diagnosis. Over the study period, 697 cases of deep fungal infections were identified with an average incidence of 0.73/100,000 persons per decade. There was a general decline in the number of cases detected. Median age of the cases was 28 years (IQR: 11–40) and majority (59%) were male. The age group of 0–10 years were the most affected. The foot was the most affected part of the body (26%). Deep mycoses identified include eumycetoma (32%), subcutaneous phycomycosis (26%), histoplasmosis (9.2%), chromo...
The central nervous system (CNS) is a known HIV reservoir, yet little is known about drug exposur... more The central nervous system (CNS) is a known HIV reservoir, yet little is known about drug exposure in the brain. Our primary objective was to quantify exposure of three common antiretrovirals in brain tissue and compare exposures to plasma and cerebrospinal fluid (CSF). We also sought to identify pockets of brain most vulnerable to inadequate drug exposures and examine the role of meningitis in drug penetration into the CNS. Tenofovir, lamivudine, and efavirenz concentrations were measured using liquid chromatography and tandem mass spectrometry in plasma and CSF from 14 individuals with HIV, 7 with cryptococcal meningitis. In four individuals (three with meningitis) drug concentrations were also measured in 13 distinct brain tissue regions. In subjects with meningitis, geometric mean ratio (95% confidence interval) of tenofovir CSF to plasma was 66% (7-598%) and 14% (6-31%) in subjects without meningitis. Lamivudine CSF penetration was 100% (25-409%) in subjects with meningitis and 30% (24-37%) in subjects without meningitis. Tenofovir brain tissue concentrations were 36% (14-124%) of plasma and 49% (1-572%) of CSF. Lamivudine brain concentrations were 37% (23-64%) of plasma and 27% (1-104%) of CSF. Efavirenz brain tissue concentrations were 128% (108-179%) of plasma. Tissues collected postmortem provide a unique opportunity to assess drug distribution in tissues difficult to sample in living subjects. CSF is a poor surrogate for drug exposure throughout the CNS. Antiretrovirals differentially penetrate into the CNS and penetration may be enhanced by meningitis. Despite HIV therapy, persistent viral replication occurs in certain anatomic sites, and systemic viral rebound will occur within weeks of treatment interruption. 1 The central nervous system (CNS) is hypothesized to be a potential HIV reservoir, harboring virus that is either resistant or not exposed to adequate antiretroviral therapy. 2 CNS Penetration-Effectiveness scores, largely based on drug exposure in cerebrospinal fluid (CSF), were developed for antiretrovirals to provide a prescribers guidance on which regimens have greatest likelihood of achieving therapeutic CNS concentrations. 3 However, brain extracellular fluid can either be passively secreted to CSF or actively transported
Background There is a growing epidemic of cancer and other non-communicable diseases in sub-Sahar... more Background There is a growing epidemic of cancer and other non-communicable diseases in sub-Saharan Africa. Targeted, specific, cost-effective strategies are needed to manage the growing burden of cancer. In high-resource settings, geospatial analysis has transformed cancer control through geographic targeting of interventions and policies. A similar approach could improve cancer control in sub-Saharan Africa; however, georeferenced cancer data and increased geospatial research capacity are needed. Here, we aimed to assess the feasibility of geocoding and mapping small-area cancer data from a cancer registry in Uganda. Methods We established a partnership including the
The shared practice of pathology via the Internet holds great potential for pathologists in sub-S... more The shared practice of pathology via the Internet holds great potential for pathologists in sub-Saharan Africa (SSA) and their global partners. Application of the Internet is constrained by issues of bandwidth, cost, and power. The penetration of mobile telephony and the arrival of smartphones have changed the use of Internet and social media in Africa and therefore the work of the 4 African pathologists featured in this article. As pathology in SSA struggles for visibility and usefulness, the Internet and its electronic applications provide a critical infrastructure as well as a podium for pathologists across the continent.
<p>1. Renal TB 2. Passage of whole <i>Mycobacterium tuberculosis</i> through th... more <p>1. Renal TB 2. Passage of whole <i>Mycobacterium tuberculosis</i> through the glomerular basement membrane into the urine 3. Passage of LAM captured by immune-complexes through the glomerular basement membrane into the urine 4. Passage of uncomplexed LAM through the glomerular basement membrane into the urine 5. Impaired tubular resorption of uncomplexed LAM 6. Genital tract TB (Fig. based on [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0123323#pone.0123323.ref021" target="_blank">21</a>]). MTB: <i>Mycobacterium tuberculosis</i>; GBM: glomerular basement membrane; LAM: lipoarabinomannan.</p
Despite the persistently high HIV-related mortality in sub-Saharan Africa, limited information on... more Despite the persistently high HIV-related mortality in sub-Saharan Africa, limited information on the causes of death is available. Pathological autopsies are the gold standard to establish causes of death. In this review we describe the autopsy series performed among HIV-infected individuals in sub-Saharan Africa over the last two decades. We identified nine complete and 11 partial or minimally invasive autopsy series. Complete autopsies were performed in 593 HIV-positive adults and 177 HIV-positive children. Postmortem diagnoses were mainly infectious diseases. Tuberculosis was the most frequent, present in 21-54% of HIV-positive adults and was considered the cause of death in 32-45%. Overall, pulmonary infections accounted for approximately 66% of pathology and central nervous system infections for approximately 20%. A high discordance between clinical and postmortem diagnoses was observed. This review emphasizes the need for reliable information on causes of death in order to im...
Microglia provide protection against a range of brain infections, but how these glial cells respo... more Microglia provide protection against a range of brain infections, but how these glial cells respond to fungi is poorly understood. We investigated the role of microglia in the context of cryptococcal meningitis, the most common cause of fungal brain infections in humans. Using a series of transgenic- and chemical-based microglia depletion methods we found that, contrary to their protective role during other infections, microglia supported cryptococcal fungal brain infection. We show that microglia become hosts for intracellular fungal growth and are a site in which the fungus accesses the restricted micronutrient copper. We developed a reporter fungal strain to track copper starvation responses by the fungus and found that yeast were protected from copper starvation within microglia. Lastly, we show that stimulation of microglia with IFNγ causes restriction of phagosomal copper to intracellular fungi. These data provide a mechanistic explanation for why microglia depletion has a the...
Background The global burden of cervical cancer is concentrated in low-and middle-income countrie... more Background The global burden of cervical cancer is concentrated in low-and middle-income countries (LMICs), with the greatest burden in Africa. Targeting limited resources to populations with the greatest need to maximize impact is essential. The objectives of this study were to geocode cervical cancer data from a population-based cancer registry in Kampala, Uganda, to create high-resolution disease maps for cervical cancer prevention and control planning, and to share lessons learned to optimize efforts in other low-resource settings. Methods Kampala Cancer Registry records for cervical cancer diagnoses between 2008 and 2015 were updated to include geographies of residence at diagnosis. Population data by age and sex for 2014 was obtained from the Uganda Bureau of Statistics. Indirectly age-standardized incidence ratios were calculated for sub-counties and estimated continuously across the study area using parish level data. Results Overall, among 1873 records, 89.6% included a valid sub-county and 89.2% included a valid parish name. Maps revealed specific areas of high cervical cancer incidence in the region, with significant variation within sub-counties, highlighting the importance of high-resolution spatial detail. Conclusions Population-based cancer registry data and geospatial mapping can be used in low-resource settings to support cancer prevention and control efforts, and to create the potential for research examining geographic factors that influence cancer outcomes. It is essential to support LMIC cancer registries to maximize the benefits of limited cancer control resources.
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Papers by Robert Lukande