Causes and effects of conflict at social security institution: a case study of National Pension Scheme Authority (NAPSA) in Zambia.

ThesisThe purpose of this study is to investigate the Causes and Effects of Conflict at National Pension Scheme Authority in Zambia. The study employed descriptive research design and used questionnaire to collect data from sampling frame of 200 of which 20 respondents participated in this research in at NAPSA in Lusaka District. The research revealed that limited resources was the major cause of conflict and that conflicts had both negative and positive effects on institution when managed properly, the positive effects could be used to encourage organizational innovativeness and build cooperation among the employees. It also revealed that Management could also set standards that were specific, measurable, achievable, and realistic and should indeed have time limit as Strategies. The findings revealed that the way committee handle conflicts was effective in organization at NAPSA and that all issues of grievance were reviewed and resolve by the committee, by convening a meeting with affected parties and a mediator to preside, by interviewing employees involved and come up with a solution. Disputes were submitted to the committee and the aggrieved parties submitted their case and holding negotiable meetings. Conflict was an inseparable aspect of people as well as organizations' life. The result also show that conflicts occurred because of competition for supremacy, leadership style and scarcity of common resources. If a conflict was not well and timely managed, it could lead to low productivity or service delivery. The results show that conflict could sometimes produce positive results, if well managed. Thus, not all conflict situations were bad. Efforts should always be made to ensure that the causes of conflicts were addressed as soon as they were noticed. The research concluded that early recognition and paying attention to the conflicting parties and negotiation between parties involved in the conflict should be adopted in resolving conflicts while force or intimidation should never be used to resolve conflicting parties. Force and intimidation could only be counterproductive

Evidence for Placental HPV Infection in Both HIV Positive and Negative Women

Human papillomaviruses (HPVs) have previously been reported to infect epithelial trophoblast cells of the placenta. To investigate this possibility, 200 placental samples from Zambian women were separated into HIV+ and HIV− groups and tested for HPV by redundant primer PCR, using GP5+/GP6+ and CPI/CPII primer sets. Three HPV genotypes (HPV6, 16 and 90) were detected in placental samples. Whereas, 20 different HPV genotypes were detected in vaginal sampling of the same patients, suggesting that compartment specific sub-populations of HPV may exist. The incidence of HPV16 in placental samples was almost 2-fold greater in HIV+ women compared to HIV− (p = 0.0241). HPV16 L1 expression, detected by immunochemistry, was significantly higher in HIV+ than HIV− samples (p = 0.0231). HPV16 DNA was detected in the nuclei of trophoblast cells by in situ hybridization. Overall, these results suggest that HPVs infect the placenta and that HIV significantly influences these infections

Evidence for Placental HPV Infection in Both HIV Positive and Negative Women

Human papillomaviruses (HPVs) have previously been reported to infect epithelial trophoblast cells of the placenta. To investigate this possibility, 200 placental samples from Zambian women were separated into HIV+ and HIV− groups and tested for HPV by redundant primer PCR, using GP5+/GP6+ and CPI/CPII primer sets. Three HPV genotypes (HPV6, 16 and 90) were detected in placental samples. Whereas, 20 different HPV genotypes were detected in vaginal sampling of the same patients, suggesting that compartment specific sub-populations of HPV may exist. The incidence of HPV16 in placental samples was almost 2-fold greater in HIV+ women compared to HIV− (p = 0.0241). HPV16 L1 expression, detected by immunochemistry, was significantly higher in HIV+ than HIV− samples (p = 0.0231). HPV16 DNA was detected in the nuclei of trophoblast cells by in situ hybridization. Overall, these results suggest that HPVs infect the placenta and that HIV significantly influences these infections

Evidence for Placental HPV Infection in Both HIV Positive and Negative Women

Abstract Human papillomaviruses (HPVs) have previously been reported to infect epithelial trophoblast cells of the placenta. To investigate this possibility, 200 placental samples from Zambian women were separated into HIV+ and HIV− groups and tested for HPV by redundant primer PCR, using GP5+/GP6+ and CPI/CPII primer sets. Three HPV genotypes (HPV6, 16 and 90) were detected in placental samples. Whereas, 20 different HPV genotypes were detected in vaginal sampling of the same patients, suggesting that compartment specific sub-populations of HPV may exist. The incidence of HPV16 in placental samples was almost 2-fold greater in HIV+ women compared to HIV− (p = 0.0241). HPV16 L1 expression, detected by immunochemistry, was significantly higher in HIV+ than HIV− samples (p = 0.0231). HPV16 DNA was detected in the nuclei of trophoblast cells by in situ hybridization. Overall, these results suggest that HPVs infect the placenta and that HIV significantly influences these infections

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

Placental HPV Infection in HIV Positive and HIV Negative Zambian Women

Human papillomaviruses (HPVs) have been reported to infect epithelial trophoblastic cells of the placenta, induce cell death and even cause placental malfunction associated with spontaneous preterm delivery. To date, no study has been conducted to determine the role of HIV on HPV genotype distribution and pathogenesis in the placental compartment. This is despite the evidence that the human immunodeficiency virus (HIV) can decrease the cellular immune response and increase the incidence of malignant cancers in HPV patients. Therefore, in this study, we analyzed 200 genomic DNA (gDNA) samples extracted from paraffin embedded placental tissues of HIV positive and HIV negative Zambian women. The gDNA was PCR amplified using GP5+/GP6+ and CPI/CPII primers targeted to the L1 and E1regions of the HPV genome, respectively. We found the overall prevalence of HPV to be 85.0%. The prevalence of HPV in the HIV+ tissues was 84(80.8%), while that of the HIV- tissues was 86(89.6%). This difference in HPV prevalence between the HIV+ and HIV- placental tissues was not significant (p\u3e0.05; p=0.112). Direct sequencing of the PCR products revealed 3 HPV genotypes namely: HPV6, 16 and 90.We observed a significant difference (p To the best of our knowledge, we are the first group to study HPV in the context of HIV within the placental compartment. Advisor: Peter C. Angelett

Inhibition of the p38 MAPK pathway as a strategy to overcome tumour immune suppression in CD1c+ dendritic cells

Introduction: Harnessing the potential of the immune system to destroy and eliminate cancer cells has been a long-term dream for many researchers in the field of cancer immunotherapy. The licencing of immune checkpoint inhibitors (ICI) and targeted therapies for malignancies such as malignant melanoma confirms the value of immune responses with important advances in survival being achieved despite some notable toxicity. Despite these major advances, there remains a keen interest in new treatments with improved efficacy and safety profiles and the exploration of adoptive immunotherapy with dendritic cells (DC) remains topical. Dendritic cells belong to a heterogeneous population of professional antigen presenting cells (APC) and reside in peripheral tissues and lymphoid organs. Previously, our group described loss of function of myeloid CD1c+ DC in patients with advanced melanoma and showed promising data to support the concept of restoring immune function by inhibiting mitogen activated protein kinase (MAPK) signalling pathways in these cells. Notably, inhibition of p38 MAPK (p38i) with small molecule research inhibitors was shown to increase the balance of IL-12:IL-10 secretion to favour the differentiation of Th1 responses. This thesis explored the impact of inhibiting p38 MAPK with candidate small molecules suitable for clinical use on the chemotaxis of CD1c+DC. Methods: Health donor CD1c+ DC were isolated and cultured in the presence of highly selective clinical p38 inhibitors, AZD6703 and AZD7624, (AstraZeneca Ltd) and assayed for CCR7 and CD86 expression, IL-10 and IL-12p70 production. Experiments were designed to investigate how these compounds could be translated from the laboratory to a clinical manufacturing setting to align with the use of Miltenyi Biotech’s Prodigy platform for advanced cell manufacturer. RNA sequencing (RNASeq) was performed on mature dendritic cells (mDC) versus p38i+mDC to identify differentially expressed genes (DEG) that drive DC chemotaxis and motility. ViewRNA assay was also performed in partnership with AstraZeneca Ltd to understand the potential roles played by CD5, CD209 and Semaphorin 7A (Sema7A) in DC chemotaxis during reduced CCR7 expression. Results: The two clinical p38i had comparable effect to the research inhibitor (BIRB0796 1µM) previously used but were 100-fold more efficacious as they were active at 10nM. As anticipated, inhibition of p38 MAPK with these compounds enhanced the secretion of IL-12 but inhibited IL-10 release by activated DC. Furthermore, p38i resulted in a marked increase in CD86 expression beyond that achieved with maturation alone. Importantly, treatment of mature DC with the clinical p38i led to increased chemotaxis to CCR7 ligand (CCL19) despite reduced display of CCR7 per se. To explore the wider alterations in gene expression following inhibition of p38 MAPK, RNASeq analysis on CD1c+ DC was undertaken. Transcriptomics analysis revealed differentially expressed genes (DEG) which upon Kyoto encyclopaedia genes and genomes (KEGG) pathway analysis with WEB-based Gene Set Analysis Toolkit (WebGestalt) identified the chemokine signalling pathway as being crucial for driving DC migration and motility. Central to this pathway was the Wiskott-Aldrich syndrome protein (WASP). Importantly, the study showed concordance between cell culture results and transcriptomics data for CCR7 and CD86 expression as well as IL-10 and IL-12p70 production all of which served as important makers for Th1 polarisation. Surprisingly, no significant differences were recorded in the ViewRNA expression profiles of the target molecules. To develop a process for future clinical manufacture of CD1c+ DC, we explored the scheduling of p38 MAPK inhibition. Importantly, it was possible to inhibit the p38 MAPK pathway in CD1c+ DC in the presence of other cells prior to their isolation and yet retain the functional impact of inhibiting p38 MAPK. However, in the presence of other cells a 100-fold increase in p38i was required to elicit functional efficacy comparable to MAPK inhibition of already isolated cells. Conclusions: This study identified a suitable clinical inhibitor, AZD7624, that will now be used in an advanced cell therapy manufacturing for melanoma patients. Additionally, RNASeq analysis revealed that the chemokine signalling pathway was crucial for mediating DC migration and motility. The presence of WASP in this pathway has important implications for DC cytoskeleton rearrangement and immunological synapse formation with naïve T cells for potent adaptive immune responses. We will shortly initiate a first-in-man clinical trial (Innovate UK funded) of p38i CD1c+ DC plus plasmacytoid DC for patients with advanced malignant melanoma who have failed immune checkpoint blockade

Inhibition of the p38 MAPK pathway as a strategy to overcome tumour immune suppression in CD1c+ dendritic cells

Introduction: Harnessing the potential of the immune system to destroy and eliminate cancer cells has been a long-term dream for many researchers in the field of cancer immunotherapy. The licencing of immune checkpoint inhibitors (ICI) and targeted therapies for malignancies such as malignant melanoma confirms the value of immune responses with important advances in survival being achieved despite some notable toxicity. Despite these major advances, there remains a keen interest in new treatments with improved efficacy and safety profiles and the exploration of adoptive immunotherapy with dendritic cells (DC) remains topical. Dendritic cells belong to a heterogeneous population of professional antigen presenting cells (APC) and reside in peripheral tissues and lymphoid organs. Previously, our group described loss of function of myeloid CD1c+ DC in patients with advanced melanoma and showed promising data to support the concept of restoring immune function by inhibiting mitogen activated protein kinase (MAPK) signalling pathways in these cells. Notably, inhibition of p38 MAPK (p38i) with small molecule research inhibitors was shown to increase the balance of IL-12:IL-10 secretion to favour the differentiation of Th1 responses. This thesis explored the impact of inhibiting p38 MAPK with candidate small molecules suitable for clinical use on the chemotaxis of CD1c+DC. Methods: Health donor CD1c+ DC were isolated and cultured in the presence of highly selective clinical p38 inhibitors, AZD6703 and AZD7624, (AstraZeneca Ltd) and assayed for CCR7 and CD86 expression, IL-10 and IL-12p70 production. Experiments were designed to investigate how these compounds could be translated from the laboratory to a clinical manufacturing setting to align with the use of Miltenyi Biotech’s Prodigy platform for advanced cell manufacturer. RNA sequencing (RNASeq) was performed on mature dendritic cells (mDC) versus p38i+mDC to identify differentially expressed genes (DEG) that drive DC chemotaxis and motility. ViewRNA assay was also performed in partnership with AstraZeneca Ltd to understand the potential roles played by CD5, CD209 and Semaphorin 7A (Sema7A) in DC chemotaxis during reduced CCR7 expression. Results: The two clinical p38i had comparable effect to the research inhibitor (BIRB0796 1µM) previously used but were 100-fold more efficacious as they were active at 10nM. As anticipated, inhibition of p38 MAPK with these compounds enhanced the secretion of IL-12 but inhibited IL-10 release by activated DC. Furthermore, p38i resulted in a marked increase in CD86 expression beyond that achieved with maturation alone. Importantly, treatment of mature DC with the clinical p38i led to increased chemotaxis to CCR7 ligand (CCL19) despite reduced display of CCR7 per se. To explore the wider alterations in gene expression following inhibition of p38 MAPK, RNASeq analysis on CD1c+ DC was undertaken. Transcriptomics analysis revealed differentially expressed genes (DEG) which upon Kyoto encyclopaedia genes and genomes (KEGG) pathway analysis with WEB-based Gene Set Analysis Toolkit (WebGestalt) identified the chemokine signalling pathway as being crucial for driving DC migration and motility. Central to this pathway was the Wiskott-Aldrich syndrome protein (WASP). Importantly, the study showed concordance between cell culture results and transcriptomics data for CCR7 and CD86 expression as well as IL-10 and IL-12p70 production all of which served as important makers for Th1 polarisation. Surprisingly, no significant differences were recorded in the ViewRNA expression profiles of the target molecules. To develop a process for future clinical manufacture of CD1c+ DC, we explored the scheduling of p38 MAPK inhibition. Importantly, it was possible to inhibit the p38 MAPK pathway in CD1c+ DC in the presence of other cells prior to their isolation and yet retain the functional impact of inhibiting p38 MAPK. However, in the presence of other cells a 100-fold increase in p38i was required to elicit functional efficacy comparable to MAPK inhibition of already isolated cells. Conclusions: This study identified a suitable clinical inhibitor, AZD7624, that will now be used in an advanced cell therapy manufacturing for melanoma patients. Additionally, RNASeq analysis revealed that the chemokine signalling pathway was crucial for mediating DC migration and motility. The presence of WASP in this pathway has important implications for DC cytoskeleton rearrangement and immunological synapse formation with naïve T cells for potent adaptive immune responses. We will shortly initiate a first-in-man clinical trial (Innovate UK funded) of p38i CD1c+ DC plus plasmacytoid DC for patients with advanced malignant melanoma who have failed immune checkpoint blockade

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 formalinfixed 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.Keywords: Human Herpes Virus 8, Kaposi´s sarcoma, histological type