Molecular characterization of full-length genomic segment 2 of a bovine picobirnavirus (PBV) strain: evidence for high genetic diversity with genogroup I PBVs.
We report here the molecular characterization of a bovine genogroup I picobirnavirus strain RUBV-P detected from a 1-month-old diarrhoeic calf in eastern India. Sequence comparisons and phylogenetic analysis of a short stretch of gene segment 2 of RUBV-P revealed low nucleotide identities (51.2-64.9%) with and distant genetic relatedness to other genogroup I picobirnaviruses. The complete gene segment 2 sequence of RUBV-P was obtained by the single primer amplification method with modifications. Gene segment 2 of RUBV-P was 1758 bp long, encoded a predicted protein of 554 aa and exhibited low nucleotide (58.1-58.8%) and amino acid (51.3-55.4%) identities with genogroup I human strains Hy005102 and 1-CHN-97. The 5'- and 3'-end nucleotide sequences, and the three motifs of RNA-dependent RNA polymerases of double-stranded RNA viruses, were conserved among these strains. Our findings suggested that bovine strain RUBV-P might be distinct from genogroup I picobirnaviruses of humans and other animals.

title: Full genomic analyses of two human G2P[4] rotavirus strains detected in 2005: identification of a caprine-like VP3 gene.
abstract: Although G2P[4] rotaviruses are common causes of infantile diarrhoea, to date only the full genomes of the prototype (strain DS-1) and another old strain, TB-Chen, have been analysed. We report here the full genomic analyses of two Bangladeshi G2P[4] strains, MMC6 and MMC88, detected in 2005. Both the strains exhibited a DS-1-like genotype constellation. Excluding the VP4 and VP7 genes, and except for VP3 of MMC88, the MMC strains were genetically more closely related to the contemporary G2P[4] and several non-G2P[4] human strains than the prototype G2P[4] strain. However, by phylogenetic analyses, the VP2, VP3 (except MMC88), NSP1 and NSP3-5 genes of these strains appeared to share a common origin with those of the prototype strain, whilst their VP1, VP6 and NSP2 genes clustered near a caprine strain. The VP3 gene of MMC88 exhibited maximum relatedness to a local caprine strain, representing the first reported human G2P[4] strain with a gene of animal origin.

title: Whole genomic analysis reveals the porcine origin of human G9P[19] rotavirus strains Mc323 and Mc345.
abstract: The group A rotavirus (RVA) P[19] is a rare P-genotype of the RVA VP4 gene, reported so far in humans and pigs. Whole genomic analyses of P[19] strains are essential to study their origin and evolutionary patterns. To date, all the 11 genes of only two P[19] strains, RVA/Human-wt/IND/RMC321/1990/G9P[19] and RVA/Human-wt/IND/mani-97/2006/G9P[19], have been analyzed, providing evidence for their porcine origin. In the present study, the whole genomes of the first reported human P[19] strains, RVA/Human-tc/THA/Mc323/1989/G9P[19] and RVA/Human-tc/THA/Mc345/1989/G9P[19], were analyzed. Strains Mc323 and Mc345 exhibited a G9-P[19]-I5-R1-C1-M1-A8-N1-T1-E1-H1 genotype constellation. With the exception of the NSP5 gene, both the strains were closely related to each other. Most of the genes of Mc323 (VP2-4, VP6-7, NSP1-4 genes) and Mc345 (VP2-4, VP6-7 and NSP1-5 genes) appeared to be of porcine origin, whilst the exact origin of VP1 and NSP5 genes of Mc323 and VP1 gene of Mc345 could not be ascertained. Therefore, strains Mc323 and Mc345 were found to have a porcine RVA genetic backbone, and are likely of porcine origin. Taken together, our observations corroborated the hypothesis that P[19] strains might be derived from porcine RVAs, providing important insights into the origin of P[19] strains, and on interspecies transmission of RVAs.

title: The origin of two rare human P[10] rotavirus strains.
abstract: The Group A rotavirus (RVA) P[10] is a rare genotype of the RVA VP4 gene. To date, the whole genome sequence of only a single P[10] RVA strain, RVA/Human-tc/IDN/69M/1980/G8P4[10], has been determined, revealing a DS-1-like genotype constellation. Whole genomic analyses of P[10] RVA strains with other VP7 genotypes are essential to obtain conclusive data on the origin and genetic diversity of the P10] RVAs. In the present study, the whole genome of a human G4P[10] RVA strain, RVA/Human-tc/IDN/57M/1980/G4P[10], was analyzed. Strain 57M exhibited an unusual G4-P[10]-I1-R1-C1-M1-A1-N1-T2-E1-H2 genotype constellation, and was found to originate from intergenogroup reassortment events involving acquisition of RVA strain 69M-like VP4, NSP3 and NSP5 genes by a co-circulating Wa-like human G4 RVA strain. Although the reference P[10] strain, 69M, exhibits a DS-1-like genotype constellation, the exact origin of this RVA remains to be elucidated. By detailed phylogenetic analyses, we found that the VP1-VP3, VP6, NSP2 and NSP4 genes of 69M originated from artiodactyl and/or artiodactyl-like human P[14] strains, whilst its NSP1, NSP3 and NSP5 genes were more related to those of typical human DS-1-like strains than those of other RVAs. On the other hand, the origin of the VP4 gene of 69M could not be established. Nevertheless, these observations clearly indicated that strain 69M might have originated from reassortment events involving at least the artiodactyl or artiodactyl-like human RVAs and the typical human DS-1-like strains. The present study provided rare evidence for intergenogroup reassortment events involving co-circulating typical human Wa-like RVAs and unusual RVAs of the DS-1-like genogroup, and revealed the presence of artiodactyl-like genes in a human P[10] strain, highlighting the complex evolutionary patterns of the P[10] RVAs.

title: Whole-genomic analysis of a human G1P[9] rotavirus strain reveals intergenogroup-reassortment events.
abstract: Group A rotavirus (RVA) strain K8 (RVA/Human-tc/JPN/K8/1977/G1P[9]) was found to have Wa-like VP7 and NSP1 genes and AU-1-like VP4 and NSP5 genes. To determine the exact origin and overall genetic makeup of this unusual RVA strain, the remaining genes (VP1-VP3, VP6 and NSP2-NSP4) of K8 were analysed in this study. Strain K8 exhibited a G1-P[9]-I1-R3-C3-M3-A1-N1-T3-E3-H3 genotype constellation, not reported previously. The VP6 and NSP2 genes of strain K8 were related closely to those of common human Wa-like G1P[8] and/or G3P[8] strains, whilst its VP1-VP3, NSP3 and NSP4 genes were related more closely to those of AU-1-like RVAs and/or AU-1-like genes of multi-reassortant strains than to those of other RVAs. Therefore, strain K8 might have originated from intergenogroup-reassortment events involving acquisition of four Wa-like genes, possibly from G1P[8] RVAs, by an AU-1-like P[9] strain. Whole-genomic analysis of strain K8 has provided important insights into the complex genetic diversity of RVAs.

title: Full genomic analysis of a simian SA11-like G3P[2] rotavirus strain isolated from an asymptomatic infant: identification of novel VP1, VP6 and NSP4 genotypes.
abstract: We report here the full genomic analysis of a simian SA11-like G3P[2] group A rotavirus (GAR) strain, B10, isolated from an asymptomatic infant in Kenya in 1987. By nucleotide sequence identities and phylogenetic analyses, the VP7-VP4-VP2-VP3-NSP1-NSP2-NSP3-NSP5 genes of strain B10 exhibited maximum genetic relatedness to those of the different isolates of simian strain SA11, and were assigned to the G3-P[2]-C5-M5-A5-N5-T5-H5 genotypes, respectively. On the other hand, the VP1, VP6 and NSP4 genes of strain B10 did not belong to any of the established GAR genotypes, and therefore, were assigned to new genotype numbers R8, I16 and E13, respectively, by the Rotavirus Classification Working Group. These observations suggested that strain B10 might have originated from reassortment event/s involving simian SA11-like strains and GAR strains from unknown animal host species (possibly other wild animals) preceding transmission to humans. Alternatively, considering the lack of data on simian GARs, it might be also possible that the VP1, VP6 and NSP4 genes of strain B10 are those of unknown simian strains, and that strain B10 might be a typical simian strain that was directly transmitted to humans. Therefore, either hypothesis pointed towards a rare instance of possible direct transmission of GARs from an animal host (possibly a monkey or some other wild animal) to humans. This was corroborated by the presence of different species of wild animals including non-human primates, and unhygienic conditions at the sampling site. To our knowledge, the present study is the first report on the detection of a simian SA11-like G3P[2] GAR strain in humans.

title: Idiopathic intracranial hypertension in a child after hemodialysis.
abstract: Idiopathic intracranial hypertension remains a disease of unknown etiology. Epidemiology, associations, features, and prognosis in children have changed over the years. The main clinical complaint of pediatric idiopathic intracranial hypertension is headache with visual disturbance. We report on a patient with childhood idiopathic intracranial hypertension, who presented without headache after hemodialysis for acute renal failure. Idiopathic intracranial hypertension without headache is believed to have a poor prognosis. However, this child demonstrated a favorable outcome with medical therapy. The child was followed with serial optical coherence tomography of retinal nerve fiber layer thickness around the optic nerve head. In the absence of a reliable procedure to evaluate the therapeutic efficacy in the follow-up of patients with idiopathic intracranial hypertension, the importance of optical coherence tomography, a noninvasive, objective, and reproducible procedure, is highlighted as a useful adjunct in the management of idiopathic intracranial hypertension.

title: Comparison of the cortical connections of areas 4 gamma and 4 delta in the cat cerebral cortex.
abstract: Area 4 of the cat cerebral cortex has been subdivided into several regions based on cytoarchitectonic studies: areas 4 gamma, 4 delta, 4sfu, and 4fu (Hassler and Muhs-Clement [1964] J. Hirnforsch. 6:377-420). In a previous study, we found separate representations of contralateral limb movements in areas 4 gamma and 4 delta (Ghosh [1997] ). To investigate the relationship between these representations, the ipsilateral cortical connections of area 4 gamma and 4 delta were compared by the use of the retrograde neural tracers. After intracortical microstimulation of area 4, tracer was injected into one or two of the following regions: the forelimb regions of the rostral and caudal subdivisions of areas 4 gamma and 4 delta (r4 gamma, r4 delta, c4 gamma, c4 delta, separated by the cruciate sulcus) and the hindlimb regions of c4 gamma and c4 delta. Retrogradely labeled neural profiles were counted in every fourth section of the coronal series and located in cytoarchitectonic areas of the ipsilateral cortex. We found topographically organized reciprocal connections between areas 4 gamma and 4 delta; these connections were part of a rich network of interconnections between the cytoarchitectonic subdivisions of area 4. The forelimb regions of c4 gamma and c4 delta, of r4 gamma and c4 delta, and of r4 gamma and r4 delta were interconnected. These findings support the location of a secondary motor area in 4 delta. No interconnections between the forelimb regions of r4 gamma and c4 gamma, and of r4 delta and c4 gamma, could be found. Area 6 (particularly area 6a gamma) was found to project strongly to the forelimb regions of r4 gamma and r4 delta and relatively weakly to the forelimb region of c4 delta. Retrogradely labeled neurons were also detected in areas 3a, 3b, 1, 2pri, 5, 7, and insula after tracer injections in area 4.

title: Application of rbcL based molecular diversity analysis to algae in wastewater treatment plants.
abstract: The molecular diversity of algae in the final clarifier or denitrification filter outfall from three wastewater treatment plants (WWTPs) with activated sludge based treatment was analyzed using the rbcL gene as a phylogenetic marker. The rbcL gene encodes the large subunit of the CO(2) fixing enzyme RuBisCO. Among algae identified at the WWTPs were diatoms, green algae, cyanobacteria, Eustigmatophyceae, and unknown heterokonts. A high level of diversity was observed within WWTPs with 19-24 unique rbcL sequences detected at each plant. Algae composition also varied between treatment plants. Our results show that the rbcL gene can be used as a phylogenetic marker for algae diversity analysis in a wastewater treatment context.

title: Molecular characterization of rare bovine group A rotavirus G15P[11] and G15P[21] strains from eastern India: identification of simian SA11-like VP6 genes in G15P[21] strains
abstract: During a surveillance study (November 2001–March 2005), one rare G15P[11] and two rare G15P[21] bovine group A rotavirus strains were detected in diarrhoeic calves in Eastern India. Sequence analysis of the VP8*, VP6, NSP4 and NSP5 genes of the G15P[11] strain confirmed its bovine origin. Although the NSP4 and NSP5 genes of the two G15P[21] strains were of bovine origin, their VP6 genes shared higher nucleotide and amino acid identities with simian strain SA11 (92.5–93.1% and 98.5–98.7%) than bovine strains (88.5–88.9% and 97–97.2%), and by phylogenetic analysis, exhibited clustering with SA11, distantly related to bovine strains. All these pointed towards a possible reassortment event of VP6 gene between bovine and simian (SA11-like) strains. Therefore, the present study provided molecular evidence for bovine origin of G15 strains and revealed a rare instance of genetic diversity in the bovine VP6 gene, otherwise conserved in group A rotavirus strains from cattle.

title: Macrophage cholesteryl ester mobilization and atherosclerosis.
abstract: Accumulation of cholesteryl esters (CE) stored as cytoplasmic lipid droplets is the main characteristic of macrophage foam cells that are central to the development of atherosclerotic plaques. Since only unesterified or free cholesterol (FC) can be effluxed from the cells to extracellular cholesterol acceptors, hydrolysis of CE is the obligatory first step in CE mobilization from macrophages. This reaction, catalyzed by neutral cholesteryl ester hydrolase (CEH), is increasingly being recognized as the rate-limiting step in FC efflux. CEH, therefore, regulates the process of reverse cholesterol transport and ultimate elimination of cholesterol from the body. In this review, we summarize the earlier controversies surrounding the identity of CEH in macrophages, discuss the characteristics of the various candidates recognized to date and examine their role in mobilizing cellular CE and thus regulating atherogenesis. In addition, physiological requirements to hydrolyze lipid droplet-associated substrate and complexities of interfacial catalysis are also discussed to emphasize the importance of evaluating the biochemical characteristics of candidate enzymes that may be targeted in the future to attenuate atherosclerosis.