Zero malaria: a mirage or reality for populations of sub-Saharan Africa in health transition.

The global burden of malaria continues to be a significant public health concern. Despite advances made in therapeutics for malaria, there continues to be high morbidity and mortality associated with this infectious disease. Sub-Saharan Africa continues to be the most affected by the disease, but unfortunately the region is burdened with indigent health systems. With the recent increase in lifestyle diseases, the region is currently in a health transition, complicating the situation by posing a double challenge to the already ailing health sector. In answer to the continuous challenge of malaria, the African Union has started a "zero malaria starts with me" campaign that seeks to personalize malaria prevention and bring it down to the grass-root level. This review discusses the contribution of sub-Saharan Africa, whose population is in a health transition, to malaria elimination. In addition, the review explores the challenges that health systems in these countries face, that may hinder the attainment of a zero-malaria goal.

Immunochromatographic and microscopic detection of Plasmodium falciparum in recipients of P. falciparum-infected donor blood.

Blood transfusion practice is an essential medical intervention; however, it poses problems of transmissibility of infectious diseases including malaria. This study was designed to determine the potential of transfusion-transmitted malaria (TTM) by detecting malaria antigens and parasites in recipients of infected donor blood. After successful blood transfusion, remnants of transfused blood were screened for Plasmodium falciparum HRP2 antigen and parasitemia using CareStart malaria RDT and 10% Giemsa stain microscopy respectively according to established protocols. Recipients of microscopy detectable P. falciparum in infected blood who tested negative for malaria by both microscopy and mRDT prior to receiving infected donor blood were followed up weekly for 35 days. Donor P. falciparum antigenemia and parasitemia were 12.1% and 8.4%, respectively, while the prevalence of blood recipient parasitemia was 3.2%. Blood stored for 2-5 days recorded mean parasitemia higher than those stored for a day and after 5 days. Additionally, parasitemia was observed in all follow-up days with marginally high frequencies in days 7, 14, and 35. There was no association between the attributes (storage days, blood group, and parasite count range) of the infected donor blood units and the characteristics of blood recipients with post-transfusion parasitemia. This study provides baseline data on TTM in Ghana. However, further studies should establish the genetic relatedness of the implicated parasites since new infections and/or recrudescence of previous infections could account for this observation.

Prevalence and risk factors of human Balantidium coli infection and its association with haematological and biochemical parameters in Ga West Municipality, Ghana.

BACKGROUND: In Ghana, Balantidium coli (B. coli) has been identified in vegetables and in pigs, although there is a paucity of data regarding human balantidiosis. This study sought to assess human B. coli infection in Ghana, factors associated with the infection as well as its association with haematological and biochemical parameters. METHODS: Two pig rearing communities in the Ga West Municipality, Ghana, were involved in this study. Stool and blood samples were collected from pig farmers and their exposed household members as well as relevant information on potential associated factors. Eosin-saline wet preparation was done on the same day of stool samples were collected while formol ether concentration technique was performed later. Haematological, biochemical parameters and serum electrolytes were determined using Celltac MEK-6500 K, PKL-125 biochemical analyser, and FT-320 electrolyte analyser, respectively. RESULTS: The overall prevalence of balantidiosis was 10.4 %, significantly higher among farmers (21.7 %) than in exposed household members (5.8 %) (x2 = 17.8, p = 0.000025). Of the 43 infected individuals, 20.9 % were co-infected with either Entamoeba histolytica, Giardia lamblia, or Schistosoma mansoni. In B. coli infection, mild to moderate anaemia together with a reduction in levels of platelet, albumin and, sodium, chloride, and bicarbonate ions were observed. However, white blood cells were significantly elevated in infected states. Poor farming practices such as free-range systems, improper disposal of pig faeces, lack of use of protective farming clothing, and unavailability of dedicated farming clothing were found to be associated with B. coli infection status. Finally, frequent diarrhea (OR = 12.30, p = 0.006) with occult blood (OR = 25.94, p < 0.0001) were found to be predictors of B. coli infection. CONCLUSIONS: Human balantidiosis is endemic in Ga West Municipality, Ghana. Individuals living closed to pig rearing communities presenting with frequent diarrhea with occult blood in stool should be screened and treated for balantidiosis to mitigate the clinical consequences of the infection.

Evaluating the potential of using urine and saliva specimens for malaria diagnosis in suspected patients in Ghana.

BACKGROUND: This study aimed at detecting PfHRP2 and pLDH malaria antigens in urine and salivary specimens of suspected malaria patients using RDT kits, and identifying factors influencing the detection of these antigens. METHODS: Malaria rapid test kit (SD Bioline RDT kit) was used to detect malaria antigens, PfHRP2 and pLDH, in blood, urine and saliva samples received from patients suspected of malaria. Subsequently, malaria parasitaemia was determined. From the same patients, body temperature readings and haemoglobin concentrations were recorded. Also, micro-haematuria and saliva occult blood were determined. Relative to blood, the sensitivities and the performance of urine and saliva as alternative samples were evaluated. RESULTS: A total of 706 suspected malaria patients provided all three specimens. Prevalence of malaria by microscopy and RDT was 44.2% and 53.9%, respectively. Compared to blood, the sensitivities of urine and saliva were 35.2% and 57.0% respectively. Haemoglobin concentration < 9.9 g/dL, body temperature > 38.7 °C and occult blood influenced the detection of malaria antigens in both urine and saliva. Furthermore, the antigens were not detected in urine and saliva when parasitaemia was < 60,000 parasites/µL and < 40,000 parasites/µL, respectively. CONCLUSION: Saliva, with or without blood contamination, was found to be more efficient that urine samples. Therefore these non-blood specimens have the potential to be used as non-invasive samples for malaria diagnosis. However, this approach is useful in severe to moderate anaemia, hyperthermia, parasitaemia > 60,000 parasites/µL and samples contaminated with blood.

Characterization of putative drug resistant biomarkers in Plasmodium falciparum isolated from Ghanaian blood donors.

BACKGROUND: Plasmodium falciparum parasites, which could harbour anti-malaria drug resistance genes, are commonly detected in blood donors in malaria-endemic areas. Notwithstanding, anti-malaria drug resistant biomarkers have not been characterized in blood donors with asymptomatic P. falciparum infection. METHODS: A total of 771 blood donors were selected from five districts in the Greater Accra Region, Ghana. Each donor sample was screened with malaria rapid diagnostic test (RDT) kit and parasitaemia quantified microscopically. Dried blood spots from malaria positive samples were genotyped for P. falciparum chloroquine resistance transporter (Pfcrt), P. falciparum multi-drug resistance (Pfmdr1), P. falciparum dihydropteroate-synthetase (Pfdhps), P. falciparum dihydrofolate-reductase (Pfdhfr) and Kelch 13 propeller domain on chromosome 13 (Kelch 13) genes. RESULTS: Of the 771 blood donors, 91 (11.8%) were positive by RDT. Analysis of sequence reads indicated successful genotyping of Pfcrt, Pfmdr1, Pfdhfr, Pfdhps and Kelch 13 genes in 84.6, 81.3, 86.8, 86.9 and 92.3% of the isolates respectively. Overall, 21 different mutant haplotypes were identified in 69 isolates (75.8%). In Pfcrt, CVIET haplotype was observed in 11.6% samples while in Pfmdr1, triple mutation (resulting in YFN haplotype) was detected in 8.1% of isolates. In Pfdhfr gene, triple mutation resulting in IRNI haplotype and in Pfdhps gene, quintuple mutation resulting in AGESS haplotype was identified in 17.7% parasite isolates. Finally, five non-synonymous Kelch 13 alleles were detected; C580Y (3.6%), P615L (4.8%), A578S (4.8%), I543V (2.4%) and A676S (1.2%) were detected. CONCLUSION: Results obtained in this study indicated various frequencies of mutant alleles in Pfcrt, Pfmdr1, Pfdhfr, Pfdhps and Kelch 13 genes from P. falciparum infected blood donors. These alleles could reduce the efficacy of standard malaria treatment in transfusion-transmitted malaria cases. Incorporating malaria screening into donor screening protocol to defer infected donors is therefore recommended.

Evaluating 18s-rRNA LAMP and selective whole genome amplification (sWGA) assay in detecting asymptomatic Plasmodium falciparum infections in blood donors.

BACKGROUND: Undesirable consequences of donor Plasmodium falciparum parasitaemia on stored donor blood have been reported. Therefore, it is imperative that all prospective blood donors are screened for P. falciparum infections using sensitive techniques. In this study, the sensitivities of microscopy, rapid diagnostic test (RDT), loop-mediated isothermal amplification (LAMP) assay and selective whole genome amplification (sWGA) technique in detecting P. falciparum infections in blood donors was assessed. METHODS: Randomly selected blood donors from 5 districts in Greater Accra Region of Ghana were screened for asymptomatic P. falciparum infections. Each donor sample was screened with SD Bioline RDT kit for P. falciparum histidine rich protein 2 and Plasmodium lactate dehydrogenase antigens, sWGA and 18s-rRNA LAMP. Crude DNA LAMP (crDNA-LAMP) was compared to purified DNA LAMP (pDNA-LAMP). RESULTS: A total of 771 blood donors were screened. The respective overall prevalence of P. falciparum in Ghana by microscopy, RDT, crDNA-LAMP, pDNA-LAMP and sWGA was 7.4%, 11.8%, 16.9%, 17.5% and 18.0%. Using sWGA as the reference test, the sensitivities of microscopy, RDT, crDNA-LAMP and pDNA-LAMP were 41.0% (95% CI 32.7-49.7), 65.5% (95% CI 56.9-73.3), 82.6% (95% CI 75.8-88.3) and 95.7% (95% CI 90.1-98.4), respectively. There was near perfect agreement between LAMP and sWGA (sWGA vs. crDNA-LAMP, κ = 0.87; sWGA vs. pDNA-LAMP, κ = 0.96), while crDNA-LAMP and pDNA-LAMP agreed perfectly (κ = 0.91). Goodness of fit test indicated non-significant difference between the performance of LAMP and sWGA (crDNA-LAMP vs. sWGA: x2 = 0.71, p = 0.399 and pDNA-LAMP vs. sWGA: x2 = 0.14, p = 0.707). Finally, compared to sWGA, the performance of LAMP did not differ in detecting sub-microscopic parasitaemia (sWGA vs. crDNA-LAMP: x2 = 1.12, p = 0.290 and sWGA vs. pDNA-LAMP: x2 = 0.22, p = 0.638). CONCLUSIONS: LAMP assay agreed near perfectly with sWGA with non-significant differences in their ability to detect asymptomatic P. falciparum parasitaemia in blood donors. Therefore, it is recommended that LAMP based assays are employed to detect P. falciparum infections in blood donors due to its high sensitivity, simplicity, cost-effectiveness and user-friendliness.

Generation and characterization of a novel human IgG1 antibody against vascular endothelial growth factor receptor 2.

VEGF and its receptors, especially VEGFR2 (KDR), are known to play a critical role in angiogenesis under both physiological and pathological conditions, including cancer and angiogenic retinopathies. This study was aimed at developing a fully human IgG1 antibody (mAb-04) constructed from a phage-derived scFv, targeting the VEGF/VEGFR2 pathway. Firstly, an innovative transfection system, containing two recombinant expression vectors (pMH3 and pCApuro), were introduced into CHO-s cells and clones with higher yield selected accordingly. After an optimal fermentation condition was determined, fed-batch fermentation was performed in 5-L bioreactor with a final yield up to 60 mg/L. Further, cell proliferation, wound healing, transwell invasion, tube formation and chick embryo chorioallantoic membrane assays showed significant anti-angiogenic activity of mAb-04 in vitro and in vivo. In addition, the results of Western blotting indicated the ability of mAb-04 to inhibit VEGF-induced VEGFR2 signaling pathway. Finally, ADCC assay demonstrated that mAb-04 is capable of mediating tumor cell killing in presence of effector cells. This study has therefore proved that the full-length antibody targeting human VEGFR2 has potential clinical applications in the treatment of cancer and other diseases where pathological angiogenesis is involved.

Molecular speciation of Plasmodium and multiplicity of P. falciparum infection in the Central region of Ghana.

Malaria is endemic in the Central region of Ghana, however, the ecological and the seasonal variations of Plasmodium population structure and the intensity of malaria transmission in multiple sites in the region have not been explored. In this cross-sectional study, five districts in the region were involved. The districts were Agona Swedru, Assin Central and Gomoa East (representing the forest zone) and Abura-Asebu-Kwamankese and Cape Coast representing the coastal zone. Systematically, blood samples were collected from patients with malaria. The malaria status was screened with a rapid diagnostic test (RDT) kit (CareStart manufactured by Access Bio in Somerset, USA) and the positive ones confirmed microscopically. Approximately, 200 µL of blood was used to prepare four dried blood spots of 50µL from each microscopy positive sample. The Plasmodium genome was sequenced at the Malaria Genome Laboratory (MGL) of Wellcome Sanger Institute (WSI), Hinxton, UK. The single nucleotide polymorphisms (SNPs) in the parasite mitochondria (PfMIT:270) core genome aided the species identification of Plasmodium. Subsequently, the complexity of infection (COI) was determined using the complexity of infection likelihood (COIL) computational analysis. In all, 566 microscopy positive samples were sequenced. Of this number, Plasmodium genome was detected in 522 (92.2%). However, whole genome sequencing was successful in 409/522 (72.3%) samples. In total, 516/522 (98.8%) of the samples contained P. falciparum mono-infection while the rest (1.2%) were either P. falciparum/P. ovale (Pf/Po) (n = 4, 0.8%) or P. falciparum/P. malariae/P. vivax (Pf/Pm/Pv) mixed-infection (n = 2, 0.4%). All the four Pf/Po infections were identified in samples from the Assin Central municipality whilst the two Pf/Pm/Pv triple infections were identified in Abura-Asebu-Kwamankese district and Cape Coast metropolis. Analysis of the 409 successfully sequenced genome yielded between 1-6 P. falciparum clones per individual infection. The overall mean COI was 1.78±0.92 (95% CI: 1.55-2.00). Among the study districts, the differences in the mean COI between ecological zones (p = 0.0681) and seasons (p = 0.8034) were not significant. However, regression analysis indicated that the transmission of malaria was more than twice among study participants aged 15-19 years (OR = 2.16, p = 0.017) and almost twice among participants aged over 60 years (OR = 1.91, p = 0.021) compared to participants between 20-59 years. Between genders, mean COI was similar except in Gomoa East where females recorded higher values. In conclusion, the study reported, for the first time, P. vivax in Ghana. Additionally, intense malaria transmission was found to be higher in the 15-19 and > 60 years, compared to other age groups. Therefore, active surveillance for P. vivax in Ghana and enhanced malaria control measures in the 15-19 year group years and those over 60 years are recommended.

Point-of-care circulating cathodic antigen positivity and associated factors in school children one year after mass praziquantel administration in an endemic district in Ghana.

Background: Mass drug administration of praziquantel is expected to reduce Schistosome carriage in treated children in endemic communities. However, the effectiveness of this annual exercise has not been assessed in Ghana. Therefore, this study aimed to detect viable Schistosoma mansoni infection using point-of-care circulating cathodic antigen (POC-CCA) positivity as proxy and associated factors in children previously treated with praziquantel in an endemic municipality in Ghana. Materials and methods: This cross-sectional study was done in the Assin Central municipality in the Central Region of Ghana. School children, less than 16 years of age, treated with 40 mg/kg of praziquantel (treatment period: February-March 2019), provided early morning urine (∼40 mL) and stool (∼4 g) samples. Immediately, POC-CCA (ICT International, South Africa) was done, while S. mansoni ova were detected in formalin fixed samples using microscopy later. Additionally, participant's socio-demographic information and factors associated with S, mansoni infection transmission were collected from each child. Results: A total of 520 children participated in the study (males-51.9%, majority age range [9-11 years, 34.4%]). Overall, 244 (46.9%) were positive for urinary CCA with no S. mansoni detected by microscopy. POC-CCA positivity was higher in females (48.4%), children with 2-3 siblings (49.3%), children aged 6-8-year range (55.4%) and residents of Brofoyedur (52%). However, age (x2 = 16.1, p = 0.0003) and town of residence (x2 = 11.7, p = 0.019) associated with CCA positivity. Further, location of water body (x2 = 16.4, p = 0.008), frequency of water contact (x2 = 12.3, p = 0.015) and handling of the Biomphalaria intermediate host (x2 = 5.1, p = 0.024) associated with POC-CCA outcome. Conclusion: About 47% of the school children were positive for CCA, one year after mass praziquantel administration in the Assin Central municipality. Varied factors associated with the post-praziquantel administration POC-CCA positivity. This study should be replicated in other endemic areas to identify groups at risk of parasite persistence or reinfection to inform modification of control and preventive measures.

Periplasmic expression optimization of VEGFR2 D3 adopting response surface methodology: antiangiogenic activity study.

Vascular endothelial growth factor (VEGF) is one of the most significant mediators of angiogenesis, which interacts with a specific membrane receptor: VEGF receptor 2 (VEGFR2). Studies elsewhere have shown that, a VEGF-blocker can regulate several vital processes of tumor promotion. However, there is no literature evidence of investigation on antiangiogenic ability of single domain 3 of VEGFR-2 (VEGFR2 D3), as the key domain in signal transduction of VEGF. In this article, we aimed at developing an efficient method for producing soluble form of this receptor as therapeutic applications. The optimization of the production of soluble VEGFR2 D3 in Escherichia coli was firstly done by testing the periplasmic expression in different expression systems using three osmotic shock methods. To enhance the yield, vital factors were selected from nine factors by Plackett-Burman design and the level of each viral factor was optimized via a response surface methodology based central composite design. After purification and identification of the protein, the bioactivity assays: quantitative ELISA, VEGF-induced proliferation and in vivo chick chorioallantoic membrane assay were employed in our study. The outcome showed that, E. coli Rosetta-gami (DE3)/pET22b-VEGFR2 D3 was the most effective expression system. Furthermore, the inducing time, peptone and glycerol concentration affected the periplasmic expression of VEGFR2 D3 significantly. The corresponding level was also optimized. The bioactivity assay studies showed VEGFR2 D3 could suppress both VEGF stimulated cell proliferation in vitro and neovascularization in vivo. We have therefore provided a novel antiangiogenic drug candidate relating to VEGF-VEGFR2 pathway.

Pf7: an open dataset of Plasmodium falciparum genome variation in 20,000 worldwide samples.

We describe the MalariaGEN Pf7 data resource, the seventh release of Plasmodium falciparum genome variation data from the MalariaGEN network.  It comprises over 20,000 samples from 82 partner studies in 33 countries, including several malaria endemic regions that were previously underrepresented.  For the first time we include dried blood spot samples that were sequenced after selective whole genome amplification, necessitating new methods to genotype copy number variations.  We identify a large number of newly emerging crt mutations in parts of Southeast Asia, and show examples of heterogeneities in patterns of drug resistance within Africa and within the Indian subcontinent.  We describe the profile of variations in the C-terminal of the csp gene and relate this to the sequence used in the RTS,S and R21 malaria vaccines.  Pf7 provides high-quality data on genotype calls for 6 million SNPs and short indels, analysis of large deletions that cause failure of rapid diagnostic tests, and systematic characterisation of six major drug resistance loci, all of which can be freely downloaded from the MalariaGEN website.

Oxidative stress and associated clinical manifestations in malaria and sickle cell (HbSS) comorbidity.

In Ghana, uncomplicated malaria and sickle cell disease (SCD) is common, hence comorbidity is not farfetched. However, the extent of oxidative stress and the array of clinical manifestations in this comorbidity (presence of both malaria and SCD) has not been fully explored. This study highlights the impact of uncomplicated malaria on SCD. The level of isoprostane, 8-iso-prostaglandin F2α (8-iso-PGF2α) was used to assess oxidative stress while plasma biochemistry and urinalysis was used to assess renal function. Hematological profiling was also done to assess the impact of comorbidity on the hematological cell lines. Of the 411 study participants with malaria, 45 (11%) had SCD. Mean body temperature was significantly higher in comorbidity compared to malaria and SCD cohorts, while a lower parasite density range was obtained in comorbidity compared to malaria cohorts. Furthermore, in comorbidity, the 8-iso-PGF2α oxidative stress biomarker was significantly elevated in all ages, parasite density ranges and gender groups. Comorbidity affected both leukocytic and erythrocytic cell lines with significant eosinophilia and monocytosis coexisting with erythrocytic parameters consistent with severe anemia. Biochemically, while plasma creatinine and bilirubin were significantly elevated in comorbidity, spot urinary creatinine was significantly reduced. Additionally, urine samples in the comorbid state were slightly acidic and hypersthenuric with significant hematuria, proteinuria, and bilirubinemia. Finally, 80% or more malaria-SCD presented with chills, fever, anorexia, headache, joint pains, lethargy, and vomiting. In conclusion, malaria could induce vaso-occlusive crisis in sickle cell disease, therefore, prompt management will alleviate the severity of this comorbidity.

Ecological and seasonal variations and other factors associated with clinical malaria in the Central Region of Ghana: A cross-sectional study.

BACKGROUND: This study investigated malaria transmission under various contrasting settings in the Central Region, a malaria endemic region in Ghana. METHODS: This cross-sectional study was carried out in five randomly selected districts in the Central Region of Ghana. Three of the districts were forested, while the rest was coastal. Study participants were selected to coincide with either the regular rainy or dry season. From each study site, hospital attendees were randomly selected with prior consent. Consciously, study participants were selected in both rainy (September and October, 2020) and dry (November and December, 2020) seasons. Clinical data for each patient was checked for clinical malaria suspicion and microscopic confirmation of malaria. Using SPSS Version 24 (Chicago, IL, USA), bivariate analysis was done to determine the association of independent variables (ecological and seasonal variations) with malaria status. When the overall analysis did not yield significant association, further statistical analysis was performed after stratification of variables (into age and gender) to determine whether any or both of them would significantly associate with the dependent variable. RESULTS: Of the 3993 study participants, 62.5% were suspected of malaria whereas 38.2% were confirmed to have clinical falciparum malaria. Data analysis revealed that in both rainy and dry seasons, malaria cases were significantly higher in forested districts ) than coastal districts (x2 = 217.9 vs x2 = 50.9; p < 0.001). Taken together, the risk of malaria was significantly higher in the dry season (COR = 1.471, p < 0.001) and lower in coastal zones (COR = 0.826, p = 0.007). There was significant reduced risk of participants aged over 39 years of malaria (COR=0.657, p < 0.001). Whereas, in general patients between 10 and 19 years were insignificantly less likely to have malaria (COR = 0.911, p = 0.518) compared to participants aged less than< 10 years, the reverse was observed in coastal districts where patients less than 10 years of age in coastal districts were less likely to have malaria (COR=2.440, p = 0.003). In general, gender did not associate with malaria, but when stratified by study district, the risk of female gender to malaria was significantly higher in Agona Swedru (COR = 5.605, p < 0.001), Assin central (COR = 2.172, p < 0.001), Awutu Senya (COR = 2.410, p < 0.001) and Cape Coast (COR = 3.939, p < 0.001) compared to Abura-Asebu-Kwamankese. CONCLUSION: This study demonstrated that the predictors of malaria differ from one endemic area to another. Therefore, malaria control interventions such as distribution of long-lasting insecticide treated bed nets, residual spraying with insecticide and mass distribution of antimalaria prophylaxis must be intensified in forested districts in all seasons with particular attention on females.

The Use of Screening Algorithm to Defer Blood Donors with Subclinical Malaria.

Plasmodium falciparum infection in blood donors is common in malaria endemic countries, including Ghana. To date, there are no established exclusion criteria to defer a donor carrying malaria parasites. Therefore, based on significant independent variables identified in this study, donor malaria screening algorithm was developed to be used by blood banks to screen blood donors for subclinical malaria. Each significant variable was weighted one (1) point and its alternative response was weighted negative one (-1) point. Accumulation of the points determines the risk level of the donor. These weighted points were used to categorize infected donors as having negligible (<2 points), tolerable (3-4 points), undesirable (5-8 points), or intolerable (>9 points) risk. Based on accumulated weight of ≥5 points, the algorithm was 94.7% (54/57) sensitive but 82% (298/364) specific. With this level of specificity, 18% of the blood donors without malaria would have been deferred. Therefore, it is imperative that all donors with accumulated risk ≥5 be screened for malaria using either malaria rapid test kit or microscopy.

Prospect of reprogramming replication licensing for cancer drug development.

Eukaryotic chromosomal DNA replication is preceded by replication licensing which involves the identification of the origin of replication by origin recognition complex (ORC). The ORC loads pre-replication complexes (pre-RCs) through a series of tightly regulated mechanisms where the ORC interacts with Cdc6 to recruit cdt1-MCM2-7 complexes to the origin of replication. In eukaryotes, adherence to regulatory mechanisms of the replication program is required to ensure that all daughter cells carry the exact copy of genetic material as the parent cell. Failure of which leads to the development of genome instability syndromes like cancer, diabetes, etc. In an event of such occurrence, preventing cells from carrying the defaulted genetic material and passing it to other cells hinges on the regulation of chromosomal DNA replication. Thus, understanding the mechanisms underpinning chromosomal DNA replication and particularly replication licensing can expose druggable enzymes, effector molecules, and secondary messengers that can be targeted for diagnosis and therapeutic purposes. Effectively drugging these molecular markers to reprogram pre-replication events can be used to control the fate of chromosomal DNA replication for the treatment of genome instability disorders and in this case, cancer. This review discusses available knowledge of replication licensing in the contest of molecular drug discovery for the treatment of cancer.

Sterilizing Immunity against COVID-19: Developing Helper T cells I and II activating vaccines is imperative.

Six months after the publication of the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) sequence, a record number of vaccine candidates were listed, and quite a number of them have since been approved for emergency use against the novel coronavirus disease 2019 (COVID-19). This unprecedented pharmaceutical feat did not only show commitment, creativity and collaboration of the scientific community, but also provided a swift solution that prevented global healthcare system breakdown. Notwithstanding, the available data show that most of the approved COVID-19 vaccines protect only a proportion of recipients against severe disease but do not prevent clinical manifestation of COVID-19. There is therefore the need to probe further to establish whether these vaccines can induce sterilizing immunity, otherwise, COVID-19 vaccination would have to become a regular phenomenon. The emergence of SARS-CoV-2 variants could further affect the capability of the available COVID-19 vaccines to prevent infection and protect recipients from a severe form of the disease. These notwithstanding, data about which vaccine(s), if any, can confer sterilizing immunity are unavailable. Here, we discuss the immune responses to viral infection with emphasis on COVID-19, and the specific adaptive immune response to SARS-CoV-2 and how it can be harnessed to develop COVID-19 vaccines capable of conferring sterilizing immunity. We further propose factors that could be considered in the development of COVID-19 vaccines capable of stimulating sterilizing immunity. Also, an old, but effective vaccine development technology that can be applied in the development of COVID-19 vaccines with sterilizing immunity potential is reviewed.

Abrus precatorius Leaf Extract Reverses Alloxan/Nicotinamide-Induced Diabetes Mellitus in Rats through Hormonal (Insulin, GLP-1, and Glucagon) and Enzymatic (α-Amylase/α-Glucosidase) Modulation.

BACKGROUND: Abrus precatorius is used in folk medicine across Afro-Asian regions of the world. Earlier, glucose lowering and pancreato-protective effects of Abrus precatorius leaf extract (APLE) was confirmed experimentally in STZ/nicotinamide-induced diabetic rats; however, the underlying mechanism of antidiabetic effect and pancreato-protection remained unknown. OBJECTIVE: This study elucidated antidiabetic mechanisms and pancreato-protective effects of APLE in diabetic rats. MATERIALS AND METHODS: APLE was prepared by ethanol/Soxhlet extraction method. Total phenols and flavonoids were quantified calorimetrically after initial phytochemical screening. Diabetes mellitus (DM) was established in adult Sprague-Dawley rats (weighing 120-180 g) of both sexes by daily sequential injection of nicotinamide (48 mg/kg; ip) and Alloxan (120 mg/kg; ip) over a period of 7 days. Except control rats which had fasting blood glucose (FBG) of 4.60 mmol/L, rats having stable FBG (16-21 mmol/L) 7 days post-nicotinamide/Alloxan injection were considered diabetic and were randomly reassigned to one of the following groups (model, APLE (100, 200, and 400 mg/kg, respectively; po) and metformin (300 mg/kg; po)) and treated daily for 18 days. Bodyweight and FBG were measured every 72 hours for 18 days. On day 18, rats were sacrificed under deep anesthesia; organs (kidney, liver, pancreas, and spleen) were isolated and weighed. Blood was collected for estimation of serum insulin, glucagon, and GLP-1 using a rat-specific ELISA kit. The pancreas was processed, sectioned, and H&E-stained for histological examination. Effect of APLE on enzymatic activity of alpha (α)-amylase and α-glucosidase was assessed. Antioxidant and free radical scavenging properties of APLE were assessed using standard methods. RESULTS: APLE dose-dependently decreased the initial FBG by 68.67%, 31.07%, and 4.39% compared to model (4.34%) and metformin (43.63%). APLE (100 mg/kg) treatment restored weight loss relative to model. APLE increased serum insulin and GLP-1 but decreased serum glucagon relative to model. APLE increased both the number and median crosssectional area (×106 µm2) of pancreatic islets compared to that of model. APLE produced concentration-dependent inhibition of α-amylase and α-glucosidase relative to acarbose. APLE concentration dependently scavenged DPPH and nitric oxide (NO) radicals and demonstrated increased ferric reducing antioxidant capacity (FRAC) relative to standards. CONCLUSION: Antidiabetic effect of APLE is mediated through modulation of insulin and GLP-1 inversely with glucagon, noncompetitive inhibition of α-amylase and α-glucosidase, free radical scavenging, and recovery of damaged/necro-apoptosized pancreatic ß-cells.

Immunological, haematological, and clinical attributes of rural and urban malaria: a case-control study in Ghana.

To compare clinical presentations, haematological and immunological parameters in urban and rural malaria patients. Clinically suspected malaria patients, resident in either rural or urban communities, were selected from seven health facilities in the Greater Accra region of Ghana. For each suspected malaria patient, parasites were detected microscopically and quantified subsequently. In each study site, an equal number of cases and age-matched controls were selected. In both cases and controls, clinical presentations, nutritional status, haematological, and immunological parameters were profiled. A total of 149 malaria patients and 149 nonmalaria controls were selected. Compared to rural dwellers with malaria, parasitaemia was significantly higher in both males and females and in the various age groups in urban dwellers with malaria. Additionally, mean lymphocytes, haemoglobin, haematocrit, mean cell haemoglobin, platelets, and mean platelet volume levels were significantly lower in urban dwellers with malaria. However, TNF-α, IL-6, and IL-12 levels in urban dwellers with malaria were significantly higher, while IL-10, CD4+, CD3+, CD8+ T-cells levels and CD4+/ CD3+ ratio were significantly lower in urban dwellers with malaria. Furthermore, chills, diarrhoea, fever, and pallor were significantly associated with urban dwellers with malaria. This study concluded that urban dwellers are more prone to severe malaria while rural dwellers tend to have more measured immune response against malaria infection, and therefore experienced better controlled inflammatory processes associated with mild form of the disease.

Effect of Asymptomatic Plasmodium falciparum Parasitaemia on Platelets Thrombogenicity in Blood Donors.

Currently, blood donors in Ghana are not screened for malaria parasites. Therefore, this study assessed platelet thrombogenicity in blood donors infected asymptomatically with Plasmodium falciparum and the relationship between tumour necrosis factor alpha (TNF-α), 8-iso-prostaglandin F2α oxidative stress biomarker (8-iso-PG2α), C-reactive protein (hs-CRP) and D-dimer, and platelet thrombogenes levels. Haematology analyser was used to enumerate platelet count and platelet indices in 80 P. falciparum infected blood donors and 160 matched non-infected controls. Replicate serum levels of von Willebrand Factor (vWF), platelet factor 4 (PF4), P-selectin thrombogenic factors as well as TNF-α and 8-iso-PG2α were determined using enzyme immuno-assay while high sensitive hs-CRP and D-dimer concentrations were determined by fluorescent immunoassay. The geometric mean of parasite density in malaria infected donors was 1784 parasites/µL (505-2478 parasites/µL). This led to significant increase in the mean levels of 8-iso-PG2α, hs-CRP, TNF-α and D-dimer. However, PF4, P-selectin were significantly lower in infected donors while vWF levels did not differ significantly among the groups even though lower levels were observed in the infected donors. Significant direct relationship existed between both P-selectin and PF4 and platelet count, and plateletcrit and platelet large cell ratio whereas these thrombogenic factors varied inversely to 8-iso-PG2α, TNF-α and hs-CRP. Relative thrombocytopaenia was associated with significant reduction in P-selectin and platelet factor 4 levels together with increased 8-iso-PG2α, hs-CRP, TNF-α and D-dimer levels. Taken together, it is recommended that all P. falciparum infected blood donors should be deferred.

Zanthoxylum zanthoxyloides Alkaloidal Extract Improves CCl4-Induced Hepatocellular Carcinoma-Like Phenotypes in Rats.

BACKGROUND: Despite the enrollment of new small molecules such as Sorafenib for the treatment of hepatocellular carcinoma (HCC), HCC still remains a significant contributor to cancer-related mortality and morbidity globally. Zanthoxylum zanthoxyloides is long suspected of possessing anticancer bioactive compounds that may hold the prospect of adjunctive therapy against inflammation-related cancers such as HCC. OBJECTIVE: This study assessed the effects of an alkaloidal extract of the leaves of Zanthoxylum zanthoxyloides on CCl4/olive oil (1 : 1 v/v)-induced HCC-like phenotypes in rats. MATERIALS AND METHODS: Zanthoxylum zanthoxyloides alkaloidal extract (ZZAE) was prepared using Soxhlet and liquid-liquid extraction methods. Subsequently, ZZAE was characterized phytochemically. In the curative method, experimental HCC was established in adult (8-10 weeks old) male Sprague-Dawley rats weighing 150-300 g by twice-daily administration of CCl4/olive oil (1 : 1 v/v) (2 mL/kg ip). After confirmation of experimental HCC in rats, the rats were randomly reassigned into seven (7) groups of seven (7) rats each and treated daily for 12 weeks as follows: control (normal saline, 5 ml/kg po), model (CCl4, 5 ml/kg, ip), ZZAE (50, 100, and 200 mg/kg po), carvedilol (6.25 mg/kg po), and 20% Tween20 (1 mL/rat, po). To assess whether ZZAE has a prophylactic (preventive) effect, rats were first treated with ZZAE and later exposed to CCl4 reconstituted in olive oil. RESULTS: ZZAE (100 and 200 mg/kg) and carvedilol decreased tumor incidence compared to that of control. Compared to control, ZZAE (100 and 200 mg/kg) significantly (P < 0.05) improved serum GGT. Compared to control, ZZAE improved hepatohistological distortions induced by CCl4/olive oil and also improved liver/body weight ratio. Compared to water, ZZAE arrested mitosis in the Allium cepa assay. CONCLUSION: ZZAE ameliorated CCl4/olive oil-induced HCC-like phenotype in rats and demonstrated general hepatoprotective effects by improving liver and kidney function markers. This finding rationalizes the need for further studies on ZZAE as a potential source of bioactive anti-HCC compounds.