Non-pathogenic Forms Research Articles

Intermittent Detections of ISAV-HPR0 in a Salmon Recirculating Aquaculture System, and Implications for Sampling

Infectious salmon anemia virus (ISAV) is an important pathogen in global Atlantic salmon (Salmo salar L.) aquaculture. The existence of both non-pathogenic (ISAV-HPR0) and pathogenic (HPR-deleted ISAV) forms of the virus impacts hatchery management. In November 2016, fish tested positive for ISAV-HPR0 at the National Cold Water Marine Aquaculture Center in Maine. A cohort exposed to the fish testing positive for ISAV were lethally sampled over a 7-month period (February–August 2017). No positive samples were detected during this time. Additional testing aimed to determine the extent of the ISAV infections in the facility’s fish and to investigate the water sources as potential virus entry points. Fish testing was designed to detect 2% pathogen prevalence with 95% confidence (assuming diagnostic sensitivity of 85%). Over a three-year period, ISAV-HPR0 was detected in spawning fish annually and once in smolts. Repeat testing of smolts from the affected tank three weeks later failed to detect ISAV-HPR0. Over a one-year period of weekly or biweekly evaluation of the incoming water sources, ISAV was never detected. These findings suggest that ISAV-HPR0 infections in monitored hatchery populations can evade detection and that episodes of high prevalence of ISAV-HPR0 associated with spawning can be highly transient. In both cases, conventional surveillance based on recurrent testing of healthy populations may provide only a very limited indication of the HPR0 status. Instead, targeting surveillance to periods of physiological stress, such as spawning and smoltification, and adjusting the sample sizes to account for a related surge in prevalence, should enhance the detection capacity in hatchery settings while also reducing testing costs.

The polyglutamine domain is the primary driver of seeding in huntingtin aggregation.

Huntington's Disease (HD) is a fatal, neurodegenerative disease caused by aggregation of the huntingtin protein (htt) with an expanded polyglutamine (polyQ) domain into amyloid fibrils. Htt aggregation is modified by flanking sequences surrounding the polyQ domain as well as the binding of htt to lipid membranes. Upon fibrillization, htt fibrils are able to template the aggregation of monomers into fibrils in a phenomenon known as seeding, and this process appears to play a critical role in cell-to-cell spread of HD. Here, exposure of C. elegans expressing a nonpathogenic N-terminal htt fragment (15-repeat glutamine residues) to preformed htt-exon1 fibrils induced inclusion formation and resulted in decreased viability in a dose dependent manner, demonstrating that seeding can induce toxic aggregation of nonpathogenic forms of htt. To better understand this seeding process, the impact of flanking sequences adjacent to the polyQ stretch, polyQ length, and the presence of model lipid membranes on htt seeding was investigated. Htt seeding readily occurred across polyQ lengths and was independent of flanking sequence, suggesting that the structured polyQ domain within fibrils is the key contributor to the seeding phenomenon. However, the addition of lipid vesicles modified seeding efficiency in a manner suggesting that seeding primarily occurs in bulk solution and not at the membrane interface. In addition, fibrils formed in the presence of lipid membranes displayed similar seeding efficiencies. Collectively, this suggests that the polyQ domain that forms the amyloid fibril core is the main driver of seeding in htt aggregation.

Collection of Bacterial Community Associated with Size Fractionated Aerosols from Kuwait

Airborne particles play a significant role in the spread of bacterial communities. The prevalence of both pathogenic and non-pathogenic forms in the inhalable fractions of aerosols is known. The abundance of microorganisms in the aerosols heightens the likely health hazards due to inhalation since they serve as carriers for pathogens and allergens, often acting as a vector for pulmonary/respiratory infections. Not much information is available on the occurrence and prevalence of bacterial communities in different size-fractionated aerosols in Kuwait. A high-volume air sampler with a six-stage cascade impactor was deployed for sample collection at two sites representing a remote and an urban site. A total volume of 815 ± 5 m3 of air was passed through the filters to trap the particulate matter ranging from 0.39 to >10.2 μm in size (Stage 1 to Stage 5 and base filter). Aeromonas dominated all the stages at the urban site and Stage 5 at the remote site, whereas Sphingobium was prevalent at Stages, 2, 3 and 4 at the remote site. Brevundimonas were found at Stages 1 and 5, and the base filter at the remote site. These results show that the bacterial community is altered in different size fractions of aerosols. Stages 1–4 form the respirable fraction, whereas Stage 5 and particles on the base filter are the inhalable fractions. Many species of Aeromonas cause disease, and hence their presence in inhalable fractions is a health concern, meaning that species-level identification is warranted.

Random Lasing Detection of Mutant Huntingtin Expression in Cells.

Huntington’s disease (HD) is an autosomal dominant, incurable neurodegenerative disease caused by mutation in the huntingtin gene (HTT). HTT mutation leads to protein misfolding and aggregation, which affect cells’ functions and structural features. Because these changes might modify the scattering strength of affected cells, we propose that random lasing (RL) is an appropriate technique for detecting cells that express mutated HTT. To explore this hypothesis, we used a cell model of HD based on the expression of two different forms—pathogenic and non-pathogenic—of HTT. The RL signals from both cell profiles were compared. A multivariate statistical analysis of the RL signals based on the principal component analysis (PCA) and linear discriminant analysis (LDA) techniques revealed substantial differences between cells that expressed the pathogenic and the non-pathogenic forms of HTT.

PROSPECTS FOR RATIONAL USE OF MINERAL RESOURCES OF THE DZHAKSY-KLYCH DEPOSIT, THE ARAL REGION

Rational use of mineral resources is becoming an important task for the development and economic growth of Kazakhstan. The purpose of the research was to study the state of salt-containing raw materials to determine the prospects for rational use of mineral resources of the Dzhaksy-Klych Deposit, one of the salty halite lakes of the Aral region. Exploration studies have shown that the Deposit is layered, where the halite layer is underlain by a sulfate layer: astrakhanite, mirabilite, tenardite, located on a layer of silt, the bed of salt deposits is dark brown clays, less often - clay Sands. Mineralogically, the halite formation is represented by, %: halite – 90-96, epsomite -1,2-2,6, mirabilite -0,2-1,9, gypsum-0,2-1,4. The production of table salt results in waste containing 65.5% chlorides, 24.5% sulfates, 6.5% sodium carbonates, and 3.5% sodium silicates, which can be used for pharmaceutical and medical purposes. Bottom silty mud by its origin and chemical composition belongs to the mainland silt-sulfide type and can be used for Spa and balneological treatment. The reserves of sulphate salts in the field are significant, with the average thickness of the sulphate reservoir in the southern basin being 0.87 m, and in the Northern basin 0.91 m. Intergranular and surface brine containing sodium and magnesium chloride-sulfate salts is of particular interest. Analysis of the state of mineral resources of the Dzhaksy-Klych Deposit revealed prerequisites for expanding the range of opportunities for using salt-containing raw materials. A promising direction for the development of the mineral resource base is the production of soda based on sodium chloride, as well as the production of a commercial product based on sodium sulfate and magnesium chloride. Microbiological examination showed the presence of non-pathogenic forms of halophilic bacteria in the salt-containing raw materials, which indicates safety for use in pharmaceutical and medical practice. In the Aral sea region, there is every reason for the development of the cosmetology industry, where a wide range of cosmetology and pharmaceutical products can be produced based on a combination of salt-containing and local vegetable raw materials.

The ILE56 mutation on different genetic backgrounds of alanine:glyoxylate aminotransferase: Clinical features and biochemical characterization

Primary Hyperoxaluria type I (PH1) is a rare disease caused by mutations in the AGXT gene encoding alanine:glyoxylate aminotransferase (AGT), a liver enzyme involved in the detoxification of glyoxylate, the failure of which results in accumulation of oxalate and kidney stones formation. The role of protein misfolding in the AGT deficit caused by most PH1-causing mutations is increasingly being recognized. In addition, the genetic background in which a mutation occurs is emerging as a critical risk factor for disease onset and/or severity. Based on these premises, in this study we have analyzed the clinical, biochemical and cellular effects of the p.Ile56Asn mutation, recently described in a PH1 patient, as a function of the residue at position 11, a hot-spot for both polymorphic (p.Pro11Leu) and pathogenic (p.Pro11Arg) mutations. We have found that the p.Ile56Asn mutation induces a structural defect mostly related to the apo-form of AGT. The effects are more pronounced when the substitution of Ile56 is combined with the p.Pro11Leu and, at higher degree, the p.Pro11Arg mutation. As compared with the non-pathogenic forms, AGT variants display reduced expression and activity in mammalian cells. Vitamin B6, a currently approved treatment for PH1, can overcome the effects of the p.Ile56Asn mutation only when it is associated with Pro at position 11. Our results provide a first proof that the genetic background influences the effects of PH1-causing mutations and the responsiveness to treatment and suggest that molecular and cellular studies can integrate clinical data to identify the best therapeutic strategy for PH1 patients.

Recommended Vaccination During Pregnancy: A Short Commentary

Vaccination is the process of stimulating a protective adaptive immune response against microbes by exposure to non-pathogenic forms or microbial components. Live vaccines killed vaccines and pathogen derived purified macromolecules are the three types of vaccines. Maternal vaccination provide protection to both the mother and fetus from the morbidity of certain serious infectious diseases which can otherwise be prevented by vaccination. Influenza and Pertussis vaccines are routinely recommended during pregnancy. Vaccines can also result in adverse reactions both in the mother and child. This review focuses on guidelines and recommendations for vaccination during pregnancy.

Novel Rickettsia genotypes in ticks in French Guiana, South America

Rickettsia are obligate intracellular bacteria often associated with ticks and best known for causing human diseases (rickettsiosis), including typhus fever and sporadic cases of serious infection. In this study, we conducted a large survey of ticks in French Guiana to understand the overall diversity of Rickettsia in this remote area largely covered by dense rainforests. Out of 819 individuals (22 tick species in six genera), 252 (30.8%) samples were positive for Rickettsia infection. Multilocus typing and phylogenetic analysis identified 19 Rickettsia genotypes, but none was 100% identical to already known Rickettsia species or strains. Among these 19 genotypes, we identified two validated Rickettsia species, Rickettsia amblyommatis (spotted fever group) and Rickettsia bellii (bellii group), and characterized a novel and divergent Rickettsia phylogenetic group, the guiana group. While some tick hosts of these Rickettsia genotypes are among the most common ticks to bite humans in French Guiana, their potential pathogenicity remains entirely unknown. However, we found a strong association between Rickettsia genotypes and their host tick species, suggesting that most of these Rickettsia genotypes may be nonpathogenic forms maintained through transovarial transmission.

Natural history of GM1 gangliosidosis mouse models generated by The Jackson Laboratory Rare and Orphan Disease Center

Primary Hyperoxaluria type I (PH1) is a rare disease caused by mutations in the AGXT gene encoding alanine:glyoxylate aminotransferase (AGT), a liver enzyme involved in the detoxification of glyoxylate, the failure of which results in accumulation of oxalate and kidney stones formation. The role of protein misfolding in the AGT deficit caused by most PH1-causing mutations is increasingly being recognized. In addition, the genetic background in which a mutation occurs is emerging as a critical risk factor for disease onset and/or severity. Based on these premises, in this study we have analyzed the clinical, biochemical and cellular effects of the p.Ile56Asn mutation, recently described in a PH1 patient, as a function of the residue at position 11, a hot-spot for both polymorphic (p.Pro11Leu) and pathogenic (p.Pro11Arg) mutations. We have found that the p.Ile56Asn mutation induces a structural defect mostly related to the apo-form of AGT. The effects are more pronounced when the substitution of Ile56 is combined with the p.Pro11Leu and, at higher degree, the p.Pro11Arg mutation. As compared with the non-pathogenic forms, AGT variants display reduced expression and activity in mammalian cells. Vitamin B6, a currently approved treatment for PH1, can overcome the effects of the p.Ile56Asn mutation only when it is associated with Pro at position 11. Our results provide a first proof that the genetic background influences the effects of PH1-causing mutations and the responsiveness to treatment and suggest that molecular and cellular studies can integrate clinical data to identify the best therapeutic strategy for PH1 patients.

Gene therapy PR006 increased progranulin levels and improved lysosomal related phenotypes in model systems

Primary Hyperoxaluria type I (PH1) is a rare disease caused by mutations in the AGXT gene encoding alanine:glyoxylate aminotransferase (AGT), a liver enzyme involved in the detoxification of glyoxylate, the failure of which results in accumulation of oxalate and kidney stones formation. The role of protein misfolding in the AGT deficit caused by most PH1-causing mutations is increasingly being recognized. In addition, the genetic background in which a mutation occurs is emerging as a critical risk factor for disease onset and/or severity. Based on these premises, in this study we have analyzed the clinical, biochemical and cellular effects of the p.Ile56Asn mutation, recently described in a PH1 patient, as a function of the residue at position 11, a hot-spot for both polymorphic (p.Pro11Leu) and pathogenic (p.Pro11Arg) mutations. We have found that the p.Ile56Asn mutation induces a structural defect mostly related to the apo-form of AGT. The effects are more pronounced when the substitution of Ile56 is combined with the p.Pro11Leu and, at higher degree, the p.Pro11Arg mutation. As compared with the non-pathogenic forms, AGT variants display reduced expression and activity in mammalian cells. Vitamin B6, a currently approved treatment for PH1, can overcome the effects of the p.Ile56Asn mutation only when it is associated with Pro at position 11. Our results provide a first proof that the genetic background influences the effects of PH1-causing mutations and the responsiveness to treatment and suggest that molecular and cellular studies can integrate clinical data to identify the best therapeutic strategy for PH1 patients.

Glucocerebrosidase Activity Modulates Neuronal Susceptibility to Pathological α-Synuclein Insult

Mutations in the GBA1 gene are the most common genetic risk factor for Parkinson's disease (PD) and dementia with Lewy bodies (DLB). GBA1 encodes the lysosomal lipid hydrolase glucocerebrosidase (GCase), and its activity has been linked to accumulation of α-synuclein. The current study systematically examines the relationship between GCase activity and both pathogenic and non-pathogenic forms of α-synuclein in primary hippocampal, cortical, and midbrain neuron and astrocyte cultures, as well as in transgenic mice and a non-transgenic mouse model of PD. We find that reduced GCase activity does not result in aggregation of α-synuclein. However, in the context of extant misfolded α-synuclein, GCase activity modulates neuronal susceptibility to pathology. Furthermore, this modulation does not depend on neuron type but rather is driven by the level of pathological α-synuclein seeds. This study has implications for understanding how GBA1 mutations influence PD pathogenesis and provides a platform for testing novel therapeutics.

Isolation and screening of biofilm forming Vibrio spp. from fish sample around south east region of Tamil Nadu and Puducherry

Abstract Biofilm are group of microbial community enclosed inside a well-organized complex structure made up of extracellular polymeric substances. Biofilm forming bacteria are the growing concern of morbidity and mortality in aquatic population. Among the different biofilm forming bacteria, Vibrio spp. are pathogenic, diverse and abundant group of aquatic organism. Aquatic animals like fish, mollusks and shrimp harbors pathogenic and non-pathogenic forms of Vibrio spp. In the present study, Vibrio strains were isolated from various region of Tamil Nadu (Chennai, Cuddalore and Tiruchirappalli) and Puducherry. A total of 155 strains were isolated from various sampling sites, among them 84 (54.7%) were confirmed as Vibrio spp. through biochemical characterization and Vibrio specific 16S rRNA gene amplification. Among the 84 strains 50 strains were confirmed as biofilm formers and they were categorized as strong (10), moderate (17) and weak (23), based on their biofilm forming ability. Further the pathogenicity of all the Vibrio strains were tested through lytic enzyme production (protease, gelatinase and hemolysin) and swarming motility. After 16S rRNA gene sequencing, the 10 strong biofilm forming strains were Vibrio alginolyticus (TFM17, TFM30, and UFM24), Vibrio parahaemolyticus (TFM1), Vibrio harveyi (CUS2), Vibrio spp. (CUS4) and Gammaproteobacteria (TFM4, TFM19, TFM61, CUS5). The present study is a first report dealing with the biofilm forming Vibrio spp. isolation from south east coastal regions of Tamil Nadu and Puducherry. Further the informations in the present study will help in finding novel strategies to combat multi drug resistance biofilm formers in aquaculture industries.

Functional and biological studies of α-galactosidase A variants with uncertain significance from newborn screening in Taiwan.

Fabry disease is an X-linked disorder resulted from deficiency of α-galactosidase A (GLA) activity. In Taiwan, a total of 792,247 newborns were screened from 2008 to 2014 in two newborn screening centers, and 13 variants of uncertain significance (VOUS) in the GLA gene were identified. To determine whether these variants were pathogenic or not, functional, biochemical, clinical and pedigree analyses were performed. In vitro functional assay was established through site-directed mutagenesis, and four in silico tools were used to predict pathogenesis. The enzyme activity of dried blood spots and plasma metabolite lyso-Gb3 level from subjects with the variants were measured. Additionally, clinical manifestations were evaluated extensively from the subjects and their relatives. Our results revealed that p.G104V, p.I232T, p.D322H, and p.G360C all exhibited relatively low residual enzyme activities and elevated plasma lyso-Gb3 level. These data strongly suggest that these Fabry mutations may cause classical or later-onset phenotypes. In contrast, neither significantly clinical symptoms nor elevated lyso-Gb3 level was found in cases with p.P60S, p.A108T, p.S304T, p.R356Q, and p.P362T variants, which may be non-pathogenic or milder forms of Fabry variants. More data need to be included for the patients with p.N53D, p.P210S, p.M296L, and p.K391T variants. The established system provides us more information to classify these GLA variants.

Microbial transcriptome profiling of black band disease in a Faviid coral during a seasonal disease peak.

The etiology of black band disease (BBD), a persistent, globally distributed coral disease characterized by a dark microbial mat, is still unclear. A metatranscriptomics approach was used to unravel the roles of the major mat constituents in the disease process. By comparing the transcriptomes of the mat constituents with those of the surface microbiota of diseased and healthy corals, we showed a shift in bacterial composition and function in BBD-affected corals. mRNA reads of Cyanobacteria, Bacteroidetes and Firmicutes phyla were prominent in the BBD mat. Cyanobacterial adenosylhomocysteinase, involved in cyanotoxin production, was the most transcribed gene in the band consortium. Pathogenic and non-pathogenic forms of Vibrio spp., mainly transcribing the thiamine ABC transporter, were abundant and highly active in both the band and surface tissues. Desulfovibrio desulfuricans was the primary producer of sulfide in the band. Members of the Bacilli class expressed high levels of rhodanese, an enzyme responsible for cyanide and sulfide detoxification. These results offer a first look at the varied functions of the microbiota in the disease mat and surrounding coral surface and enabled us to develop an improved functional model for this disease.

Diversity of endophytic fungi in Glycine max

Endophytic fungi are microorganisms that live within plant tissues without causing disease during part of their life cycle. With the isolation and identification of these fungi, new species are being discovered, and ecological relationships with their hosts have also been studied. In Glycine max, limited studies have investigated the isolation and distribution of endophytic fungi throughout leaves and roots. The distribution of these fungi in various plant organs differs in diversity and abundance, even when analyzed using molecular techniques that can evaluate fungal communities in different parts of the plants, such as denaturing gradient gel electrophoresis (DGGE). Our results show there is greater species richness of culturable endophytic filamentous fungi in the leaves G. max as compared to roots. Additionally, the leaves had high values for diversity indices, i.e. Simpsons, Shannon and Equitability. Conversely, dominance index was higher in roots as compared to leaves. The fungi Ampelomyces sp., Cladosporium cladosporioides, Colletotrichum gloeosporioides, Diaporthe helianthi, Guignardia mangiferae and Phoma sp. were more frequently isolated from the leaves, whereas the fungi Fusarium oxysporum, Fusarium solani and Fusarium sp. were prevalent in the roots. However, by evaluating the two communities by DGGE, we concluded that the species richness was higher in the roots than in the leaves. UPGMA analysis showed consistent clustering of isolates; however, the fungus Leptospora rubella, which belongs to the order Dothideales, was grouped among species of the order Pleosporales. The presence of endophytic Fusarium species in G. max roots is unsurprising, since Fusarium spp. isolates have been previously described as endophyte in other reports. However, it remains to be determined whether the G. max Fusarium endophytes are latent pathogens or non-pathogenic forms that benefit the plant. This study provides a broader knowledge of the distribution of the fungal community in G. max leaves and roots, and identifies the genetic relationships among the isolated species.

The Uptake of GABA in Trypanosoma cruzi.

Gamma aminobutyric acid (GABA) is widely known as a neurotransmitter and signal transduction molecule found in vertebrates, plants, and some protozoan organisms. However, the presence of GABA and its role in trypanosomatids is unknown. Here, we report the presence of intracellular GABA and the biochemical characterization of its uptake in Trypanosoma cruzi, the etiological agent of Chagas' disease. Kinetic parameters indicated that GABA is taken up by a single transport system in pathogenic and nonpathogenic forms. Temperature dependence assays showed a profile similar to glutamate transport, but the effect of extracellular cations Na(+) , K(+) , and H(+) on GABA uptake differed, suggesting a different uptake mechanism. In contrast to reports for other amino acid transporters in T. cruzi, GABA uptake was Na(+) dependent and increased with pH, with a maximum activity at pH 8.5. The sensitivity to oligomycin showed that GABA uptake is dependent on ATP synthesis. These data point to a secondary active Na(+) /GABA symporter energized by Na(+) -exporting ATPase. Finally, we show that GABA occurs in the parasite's cytoplasm under normal culture conditions, indicating that it is regularly taken up from the culture medium or synthesized through an still undescribed metabolic pathway.

Invasion ability and disease dynamics of environmentally growing opportunistic pathogens under outside-host competition.

Most theories of the evolution of virulence concentrate on obligatory host-pathogen relationship. Yet, many pathogens replicate in the environment outside-host where they compete with non-pathogenic forms. Thus, replication and competition in the outside-host environment may have profound influence on the evolution of virulence and disease dynamics. These environmentally growing opportunistic pathogens are also a logical step towards obligatory pathogenicity. Efficient treatment methods against these diseases, such as columnaris disease in fishes, are lacking because of their opportunist nature. We present a novel epidemiological model in which replication and competition in the outside-host environment influences the invasion ability of a novel pathogen. We also analyze the long-term host-pathogen dynamics. Model parameterization is based on the columnaris disease, a bacterial fresh water fish disease that causes major losses in fish farms worldwide. Our model demonstrates that strong competition in the outside-host environment can prevent the invasion of a new environmentally growing opportunist pathogen and long-term disease outbreaks.

146 Amyloid fibril polymorphism probed by advanced vibrational spectroscopy

Amyloid fibrils are associated with many neurodegenerative diseases. All known amyloids including pathogenic and nonpathogenic forms display functional and structural heterogeneity (polymorphism) which determines the level of their toxicity. Despite a significant biological and biomedical importance, the nature of the amyloid fibril polymorphism remains elusive. We utilized for the first time three most advanced vibrational techniques to probe the core, the surface, and supramolecular chirality of fibril polymorphs. A new type of folding, aggregation phenomenon, spontaneous refolding from one polymorph to another, was discovered (Kurouski, Lauro et al., 2010). Hydrogen–deuterium exchange deep UV resonance Raman spectroscopy (Oladepo, Xiong et al., 2012) combined with advanced statistical analysis (Shashilov & Lednev, 2010) allowed for structural characterization of the highly ordered cross-β core of amyloid fibrils. We reported several examples showing significant variations in the core structure for fibril polymorphs. Amyloid fibrils are generally composed of several protofibrils and may adopt variable morphologies, such as twisted ribbons or flat-like sheets. We discovered the existence of another level of amyloid polymorphism, namely, that associated with fibril supramolecular chirality. Two chiral polymorphs of insulin, which can be controllably grown by means of small pH variations, exhibit opposite signs of vibrational circular dichroism (VCD) spectra (Kurouski, Dukor et al. 2012). VCD supramolecular chirality is correlated not only by the apparent fibril handedness but also by the sense of supramolecular chirality from a deeper level of chiral organization at the protofilament level of fibril structure. A small pH change initiates spontaneous transformation of insulin fibrils from one polymorph to another. As a result, fibril supramolecular chirality overturns both accompanying morphological and structural changes (Kurouski, Dukor et al. 2012). No conventional methods could probe the fibril surface despite its significant role in the biological activity. We utilized tip-enhanced Raman spectroscopy (TERS) to characterize the surface structure of an individual fibril due to a high depth and lateral spatial resolution of the method in the nanometer range (Kurouski, Deckert-Gaudig et al. 2012). It was found that the surface is strongly heterogeneous and consists of clusters with various protein conformations and amino acid composition.

PCR Detection of <i>Entamoeba histolytica</i> in Microscopically Positive Stool Samples of Hospital Patients in Soroti, Eastern Uganda

Amoebiasis is an infection caused by water borne protozoan parasite Entamoeba histolytica. In Uganda where sanitation infrastructure and health education was not adequate, amoebiasis was thought to be still an important health problem. However there was little or no data on prevalence of this very important protozoan infection. In addition, microscopy remained the main method for the diagnosis of amoebiasis but could not differentiate between Entamoeba dispar/moshkovskii and Entamoeba histolytica infections. This made determination of true prevalence of Entamoeba histolytica infections difficult. It was against this background that this study was designed to carry out species specific diagnosis of Entamoeba histolytica and Entamoeba dispar/moshkovskii in Uganda where these species had been reported to be endemic. This study used microscopy and polymerase chain reaction amplification of Serine-rich Entamoeba histolytica (SREHP) gene. It was shown that 36.7% (n=22) of the samples initially diagnosed as positive by microscopy were positive by PCR. The true prevalence of E. histolytica and E.dispar/ moshkovskii was found to be 7.31% and 12.6% respectively. It was concluded that Entamoeba infection in Soroti, Eastern Uganda is more frequently due to E. dispar /moshkovskii (13.3%) the non-pathogenic forms than to E. histolytica, the pathogen (7.31%).Key words: Entamoeba histolytica, Microscopy, Polymerase chain reaction, Prevalence.

In vivo P2X7 inhibition reduces amyloid plaques in Alzheimer's disease through GSK3β and secretases

β-amyloid (Aβ) peptide production from amyloid precursor protein (APP) is essential in the formation of the β-amyloid plaques characteristic of Alzheimer's disease. However, the extracellular signals that maintain the balance between nonpathogenic and pathologic forms of APP processing, mediated by α-secretase and β-secretase respectively, remain poorly understood. In the present work, we describe regulation of the processing of APP via the adenosine triphosphate (ATP) receptor P2X7R. In 2 different cellular lines, the inhibition of either native or overexpressed P2X7R increased α-secretase activity through inhibition of glycogen synthase kinase 3 (GSK-3). In vivo inhibition of the P2X7R in J20 mice, transgenic for mutant human APP, induced a significant decrease in the number of hippocampal amyloid plaques. This reduction correlated with a decrease in glycogen synthase kinase 3 activity in J20 mice, increasing the proteolytic processing of APP through an increase in α-secretase activity. The in vivo findings presented here demonstrate for the first time the therapeutic potential of P2X7R antagonism in the treatment of familiar Alzheimer's disease (FAD).