Upregulation of estrogen receptor alpha (ERα) expression in transgenic mice expressing human CYP4Z1.

PURPOSE: CYP4Z1 is a human cytochrome P450 enzyme involved in breast cancer progression and prognosis, but its functional role in these processes is not understood. In order to gain more insight into CYP4Z1's properties it was recombinantly expressed in a host animal that does not have an endogenous homologue. METHODS: We generated a transgenic mouse model that specifically expresses human CYP4Z1 in breast tissue under the control of the whey acidic protein promoter. Complementary experiments were done using cell lines derived from human breast cell. RESULTS: Induction of CYP4Z1 expression led to reduction of body weight, activity, and birth rates. Histological analysis revealed no evidence for tumor formation. However, a strong increase in estrogen receptor alpha was observed by immunohistochemistry; weaker but significantly increased immunoreactivity was also detected for collagen I and fibronectin. Overexpression of CYP4Z1 in the human breast cancer cell line MCF7 also led to increased ERα expression. Moreover, increased expression of both CYP4Z1 and ERα was observed in MCF-10A normal breast cells upon cocultivation with MCF-7 cells (with or without overexpression of CYP4Z1). CONCLUSION: These data suggest that CYP4Z1 facilitates breast cancer development by induction of ERα expression via an as yet undefined mechanism.

Nutritional and social contribution of meat in diets: Interplays among young urban and rural men.

Considering the recent increase in the demand for meat and its subsequent implications for health and food security, there is an increasing need to explore its nutritional and social importance among young men in settings experiencing nutrition transition. A better understanding of meat in the diets of this group could contribute to the design of socio-culturally appropriate interventions to improve healthy eating, as these men are key decision makers in family food choices. This mixed-methods study aimed to assess the nutritional and social contribution of meat in the diet of young adult men in urban and rural Zambia. A food frequency questionnaire, multiple pass 24-h dietary recall, anthropometric measurements and a socio-demographic questionnaire were utilized while qualitative interviews explored the socio-cultural importance of meat consumption. Rural and urban participants had an isocaloric diet. All macronutrient intakes except carbohydrates were significantly higher in the urban population than the rural population (p < 0.01). Zinc intake was significantly greater in the urban than the rural sample (χ2 (39) = 40, p-value = 0.04). Except for vitamin A, calcium and folate, participants met the recommendations for all micronutrients. Regardless of being rural or urban, the higher the participant's level of education, the weaker the socio-cultural importance of meat. In both settings, increased consumption of meat was associated with prosperity, authority and respect within society. There are strong social and cultural beliefs among participants about meat consumption, reflecting the symbolic meaning in their customs. These findings could help improve the design and implementation of dietary interventions, incorporating specific cultural beliefs and socio-economic factors in the targeted population, to achieve healthy eating practices.

Combined chemotherapy manifest less severe immunopathology effects in helminth-protozoa comorbidity.

BACKGROUND: Co-infection with Leishmania major and Schistosoma mansoni may have significant consequences for disease progression, severity and subsequent transmission dynamics. Pentavalent antimonials and Praziquantel (PZQ) are used as first line of treatment for Leishmania and Schistosoma infections respectively. However, there is limited insight on how combined therapy with the standard drugs impacts the host in comorbidity. The study aimed to determine the efficacy of combined chemotherapy using Pentostam (P) and PZQ in murine model co-infected with L. major and S. mansoni. METHODS: A 3 × 4 factorial design with three parasite infection groups (Lm, Sm, Lm + Sm to represent L. major, S. mansoni and L. major + S. mansoni respectively) and four treatment regimens [P, PZQ, P + PZQ, and PBS designating Pentostam (GlaxoSmithKline UK), Praziquantel (Biltricide®, Bayer Ag. Leverkusen, Germany), Pentostam + Praziquantel and Phosphate buffered saline] as factors was applied. RESULTS: Significant changes were observed in the serum Interferon gamma (IFN-γ), and Macrophage inflammatory protein-one alpha (MIP-1α) levels among various treatment groups between week 8 and week 10 (p < 0.05). There was increased IFN-γ in the L. major infected mice subjected to PZQ and PBS, and in L. major + S. mansoni infected BALB/c mice treated with P + PZQ. Subsequently, MIP-1α levels increased significantly in both the L. major infected mice under PZQ and PBS and in L. major + S. mansoni infected BALB/c mice undergoing concurrent chemotherapy with P + PZQ between 8 and 10 weeks (p < 0.05). In the comorbidity, simultaneous chemotherapy resulted in less severe histopathological effects in the liver. CONCLUSION: It was evident, combined first line of treatment is a more effective strategy in managing co-infection of L. major and S. mansoni. The findings denote simultaneous chemotherapy compliments immunomodulation in the helminth-protozoa comorbidity hence, less severe pathological effects following the parasites infection. Recent cases of increased incidences of polyparasitism in vertebrates call for better ways to manage co-infections. The findings presented necessitate intrinsic biological interest on examining optimal combined chemotherapeutic agents strategies in helminth-protozoa concomitance and the related infections abatement trends vis-a-vis host-parasite relationships.

Determinants of breast cancer early detection for cues to expanded control and care: the lived experiences among women from Western Kenya.

BACKGROUND: Estimately, 70-80% of cancer cases are diagnosed in late stages in Kenya with breast cancer being a common cause of mortality among women where late diagnosis is the major ubiquitous concern. Numerous studies have focused on epidemiological and health policy dynamics essentially underestimating the determining factors that shape people's choices and cues to health care service uptake. The study sought to evaluate the knowledge, attitude and health seeking behavior towards breast cancer and its screening in a quest to explain why women present for prognosis and treatment when symptomatic pointers are in advanced stages, impeding primary prevention strategies. METHODS: Eight focus groups (6-10 members per group) and four key informant interviews were conducted among adult participants from rural and urban settings. Sessions were audio-recorded and transcribed. A thematic analysis of the data was based on the concepts of the health belief model. Data analysis was conducted using NVIVO10. RESULTS: Most women perceived breast cancer as a fatal disease and conveyed fear of having early screening. Rural women preferred self-prescribed medications and the use of alternative medicine for long periods before presenting for professional care on suspicion that the lump is cancerous. Accessibility to equipped health facilities, lack of information to establish effective follow-up treatment and low-income status were underscored as their major health seeking behavior barriers whereas, urban women identified marital status as their main barrier. Key informant interviews revealed that health communication programs emphasized more on communicable diseases. This could in part explain why there is a high rate of misconception and suspicion about breast cancer among rural and urban women in the study setting. CONCLUSIONS: Creating breast cancer awareness alongside clear guidelines on accessing screening and treatment infrastructure is critical. It was evident, a diagnosis of breast cancer or lump brings unexpected confrontation with mortality; fear, pain, cultural barriers, emotional and financial distress. Without clear referral channels to enable those with suspicious lumps or early stage disease to get prompt diagnosis and treatment, then well-meaning awareness will not necessarily contribute to reducing morbidity and mortality.

Policy environment for prevention, control and management of cardiovascular diseases in primary health care in Kenya.

BACKGROUND: In Kenya, cardiovascular diseases (CVDs) accounted for more than 10% of total deaths and 4% of total Disability-Adjusted Life Years (DALYs) in 2015 with a steady increase over the past decade. The main objective of this paper was to review the existing policies and their content in relation to prevention, control and management of CVDs at primary health care (PHC) level in Kenya. METHODS: A targeted document search in Google engine using keywords "Kenya national policy on cardiovascular diseases" and "Kenya national policy on non-communicable diseases (NCDs)" was conducted in addition to key informant interviews with Kenyan policy makers. Relevant regional and international policy documents were also included. The contents of documents identified were reviewed to assess how well they aligned with global health policies on CVD prevention, control and management. Thematic content analysis of the key informant interviews was also conducted to supplement the document reviews. RESULTS: A total of 17 documents were reviewed and three key informants interviewed. Besides the Tobacco Control Act (2007), all policy documents for CVD prevention, control and management were developed after 2013. The national policies were preceded by global initiatives and guidelines and were similar in content with the global policies. The Kenya health policy (2014-2030), The Kenya Health Sector Strategic and Investment Plan (2014-2018) and the Kenya National Strategy for the Prevention and Control of Non-communicable diseases (2015-2020) had strategies on NCDs including CVDs. Other policy documents for behavioral risk factors (The Tobacco Control Act 2007, Alcoholic Drinks Control (Licensing) Regulations (2010)) were available. The National Nutrition Action Plan (2012-2017) was available as a draft. Although Kenya has a tiered health care system comprising primary healthcare, integration of CVD prevention and control at PHC level was not explicitly mentioned in the policy documents. CONCLUSION: This review revealed important gaps in the policy environment for prevention, control and management of CVDs in PHC settings in Kenya. There is need to continuously engage the ministry of health and other sectors to prioritize inclusion of CVD services in PHC.

Challenges and opportunities of optimal breastfeeding in the context of HIV option B+ guidelines.

BACKGROUND: In 2013, the World Health Organization released a new set of guidelines widely known as Option B+. Prior to that there were guidelines released in 2010. Option B+ recommends lifelong antiretroviral treatment for all pregnant and breastfeeding women living with Human Immunodeficiency Virus. The study aimed at investigating challenges and opportunities in implementing Infant and Young Child Feeding in the context of Prevention of Mother To Child Transmission (PMTCT) guidelines among HIV positive mothers of children aged 0-24 months. The study also examined implications presented by implementing the 2013 PMTCT consolidated guidelines in the transition phase from the 2010 approach in Zambia. METHODS: A mixed methods approach was employed in the descriptive cross sectional study utilizing semi structured questionnaires and Focused Group Discussions. Further, data was captured from the Health Information Management System. RESULTS: During the PMTCT transition, associated needs and challenges in institutionalizing the enhanced guidelines from option A and B to option B+ were observed. Nonetheless, there was a decline in Mother to Child Transmission (MTCT) of HIV rates with an average of 4%. Mothers faced challenges in complying with optimal breastfeeding practices owing to lack of community support systems and breast infections due to poor breast feeding occasioned by infants' oral health challenges. Moreover, some mothers were hesitant of lifelong ARVs. Health workers faced programmatic and operational challenges such as compromised counseling services. CONCLUSION: Despite the ambitious timelines for PMTCT transition, the need to inculcate new knowledge and vary known practice among mothers and the shift in counseling content for health workers, the consolidated guidelines for PMTCT proved effective. Some mothers were hesitant of lifelong ARVs, rationalizing the debated paradigm that prolonged chemotherapy/polypharmacy may be a future challenge in the success of ART in PMTCT. Conflicting breast feeding practices was a common observation across mothers thus underpinning the need to strongly invigorate Infant and Young Child Feeding information sharing across the continuum of heath care from facility level to community and up to the family; for cultural norms, practices and attitudes enshrined within communities play a vital role in child care.

Vector competence of Aedes aegypti in transmitting Chikungunya virus: effects and implications of extrinsic incubation temperature on dissemination and infection rates.

BACKGROUND: Aedes aegypti is a competent arthropod vector of chikungunya virus (CHIKV). The rate at which the virus disseminate in the vector is limited by temperature of their environment which can be an important determinant of geographical and seasonal limits to transmission by the arthropods in the tropics. This study investigated the vector competence of Ae. aegypti for CHIKV at ambient temperature of 32 and 26 °C (Coastal and Western Kenya respectively) reared at Extrinsic Incubation Temperature (EIT) of 32 and 26 °C that resembles those in the two regions. METHODS: Ae. aegypti eggs were collected from coastal and Western Kenya, hatched in the insectary and reared to F1 generation. Four-day old mosquitoes were exposed to CHIKV through a membrane feeding. They were then incubated in temperatures mimicking the mean annual temperatures for Trans-Nzoia (26 °C) and Lamu (32 °C). After every 7, 10 and 13 days post infection (DPI); one third of exposed mosquitoes were sampled and assayed for virus infection and dissemination. RESULTS: The midgut infection rates (MIR) of Ae. aegypti sampled from Coastal Region was significantly (p < 0.05) higher than those sampled from Western Kenya, with no statistical differences observed for the coastal Ae. aegypti at EIT 26 and at 32 °C. The MIR of Ae. aegypti from the Western Region was significantly (p < 0.05) affected by the EIT, with mosquito reared at EIT 32 °C exhibiting higher MIR than those reared at EIT 26 °C. There was a significant (p < 0.05) interactive effects of the region, EIT and DPI on MIR. The disseminated infection rates for the CHIKV in Ae. aegypti in the legs (DIR-L) was higher in mosquitoes sampled from Coast regardless of the EIT while those from Western Kenya, dissemination rates were significantly higher at higher EIT of 32 °C. CONCLUSIONS: Vector competence was higher in mosquito populations reared under high temperatures which weakens the midgut infection barrier. Hence, suggesting Lamu population is more susceptible to CHIKV therefore having a weaker mid gut infection barrier than the Trans Nzoia population. These underscores importance of examining the course of infection at various ambient temperatures and EIT between regions mosquito populations.

Subcellular dissemination of prothymosin alpha at normal physiology: immunohistochemical vis-a-vis western blotting perspective.

BACKGROUND: The cell type, cell status and specific localization of Prothymosin α (PTMA) within cells seemingly determine its function. PTMA undergoes 2 types of protease proteolytic modifications that are useful in elucidating its interactions with other molecules; a factor that typifies its roles. Preferably a nuclear protein, PTMA has been shown to function in the cytoplasm and extracellularly with much evidence leaning on pathognomonic status. As such, determination of its cellular distribution under normal physiological context while utilizing varied techniques is key to illuminating prospective validation of its distinct functions in different tissues. Differential distribution insights at normal physiology would also portent better basis for further clarification of its interactions and proteolytic modifications under pathological conditions like numerous cancer, ischemic stroke and immunomodulation. We therefore raised an antibody against the C terminal of PTMA to use in tandem with available antibody against the N terminal in a murine model to explicate the differences in its distribution in brain cell types and major peripheral organs through western blotting and immunohistochemical approaches. RESULTS: The newly generated antibody was applied against the N-terminal antibody to distinguish truncated versions of PTMA or deduce possible masking of the protein by other interacting molecules. Western blot analysis indicated presence of a truncated form of the protein only in the thymus, while immunohistochemical analysis showed that in brain hippocampus the full-length PTMA was stained prominently in the nucleus whereas in the stomach full-length PTMA staining was not observed in the nucleus but in the cytoplasm. CONCLUSION: Truncated PTMA could not be detected by western blotting when both antibodies were applied in all tissues examined except the thymus. However, immunohistochemistry revealed differential staining by these antibodies suggesting possible masking of epitopes by interacting molecules. The differential localization patterns observed in the context of nucleic versus cytoplasmic presence as well as punctate versus diffuse pattern in tissues and cell types, warrant further investigations as to the forms of PTMA interacting partners.

Symptomatic malaria diagnosis overestimate malaria prevalence, but underestimate anaemia burdens in children: results of a follow up study in Kenya.

BACKGROUND: The commonly accepted gold standard diagnostic method for detecting malaria is a microscopic reading of Giemsa-stained blood films. However, symptomatic diagnosis remains the basis of therapeutic care for the majority of febrile patients in malaria endemic areas. This study aims to compare the discrepancy in malaria and anaemia burdens between symptomatic diagnosed patients with those diagnosed through the laboratory. METHODS: Data were collected from Western Kenya during a follow-up study of 887 children with suspected cases of malaria visiting the health facilities. In the laboratory, blood samples were analysed for malaria parasite and haemoglobin levels. Differences in malaria prevalence between symptomatic diagnosis and laboratory diagnosis were analysed by Chi-square test. Bayesian probabilities were used for the approximation of the malaria and anaemia burdens. Regression analysis was applied to: (1) determine the relationships between haemoglobin levels, and malaria parasite density and (2) relate the prevalence of anaemia and the prevalence of malaria. RESULTS: The prevalence of malaria and anaemia ranged from 10% to 34%, being highest during the rainy seasons. The predominant malaria parasite was P. falciparum (92.3%), which occurred in higher density in children aged 2‒5 years. Fever, high temperature, sweating, shivering, vomiting and severe headache symptoms were associated with malaria during presumptive diagnosis. After conducting laboratory diagnosis, lower malaria prevalence was reported among the presumptively diagnosed patients. Surprisingly, there were no attempts to detect anaemia in the same cohort. There was a significant negative correlation between Hb levels and parasite density. We also found a positive correlation between the prevalence of anaemia and the prevalence of malaria after laboratory diagnosis indicating possible co-occurrence of malaria and anaemia. CONCLUSION: Symptomatic diagnosis of malaria overestimates malaria prevalence, but underestimates the anaemia burden in children. Good clinical practice dictates that a laboratory should confirm the presence of parasites for all suspected cases of malaria.

Monitoring of autoantibodies against CYP4Z1 in patients with colon, ovarian, or prostate cancer.

We have previously monitored the detection of autoantibodies (aAbs) directed against CYP4Z1 in the sera of breast and lung cancer patients. In the present study, the occurence of anti-CYP4Z1 aAbs in patients suffering from colon (n = 100), ovarian (n = 72), or prostate (n = 85) cancer was examined. Determination of aAbs was done using our previously established ELISA method. On average, the levels of anti-CYP4Z1 aAbs detected in sera from all cancer patients were not significantly higher than controls. No correlations were found with respect to gender or tumor stage. However, a subgroup of colon cancer patients with increased anti-CYP4Z1 aAb titers exhibited positive fecal occult blood test (FOBT) results and higher levels of both carbohydrate antigen 19-9 (CA19-9) and carcinoembryonic antigen (CEA). These results do not suggest that anti-CYP4Z1 aAbs have value as an independent biomarker for the detection of either colon, ovarian, or prostate cancer. However, they might be useful in combination with other biomarkers for the identification of a subset of colon cancers. Investigations involving a more powered sample size of this subgroup are needed to support this notion.