ALS2CL, the novel protein highly homologous to the carboxy-terminal half of ALS2, binds to Rab5 and modulates endosome dynamics

ALS2, the causative gene product for juvenile recessive amyotrophic lateral sclerosis (ALS2), is a guanine-nucleotide exchange factor for the small GTPase Rab5. Here, we report a novel ALS2 homologous gene, ALS2 C-terminal like (ALS2CL), which encodes a 108-kD ALS2CL protein. ALS2CL exhibited a specific but a relatively weak Rab5-GEF activity with accompanying rather strong Rab5-binding properties. In HeLa cells, co-expression of ALS2CL and Rab5A resulted in a unique tubulation phenotype of endosome compartments with significant colocalization of ALS2CL and Rab5A. These results suggest that ALS2CL is a novel factor modulating the Rab5-mediated endosome dynamics in the cells.     

Gastrointestinal ischemia-reperfusion injury is lectin complement pathway dependent without involving C1q

Complement activation plays an important role in local and remote tissue injury associated with gastrointestinal ischemia-reperfusion (GI/R). The role of the classical and lectin complement pathways in GI/R injury was evaluated using C1q-deficient (C1q KO), MBL-A/C-deficient (MBL-null), complement factor 2- and factor B-deficient (C2/fB KO), and wild-type (WT) mice. Gastrointestinal ischemia (20 min), followed by 3-h reperfusion, induced intestinal and lung injury in C1q KO and WT mice, but not in C2/fB KO mice. Addition of human C2 to C2/fB KO mice significantly restored GI/R injury, demonstrating that GI/R injury is mediated via the lectin and/or classical pathway. Tissue C3 deposition in C1q KO and WT, but not C2/fB KO, mice after GI/R demonstrated that complement was activated in C1q KO mice. GI/R significantly increased serum alanine aminotransferase, gastrointestinal barrier dysfunction, and neutrophil infiltration into the lung and gut in C1q KO and WT, but not C2/fB KO, mice. MBL-null mice displayed little gut injury after GI/R, but lung injury was present. Addition of recombinant human MBL (rhuMBL) to MBL-null mice significantly increased injury compared with MBL-null mice after GI/R and was reversed by anti-MBL mAb treatment. However, MBL-null mice were not protected from secondary lung injury after GI/R. These data demonstrate that C2 and MBL, but not C1q, are necessary for gut injury after GI/R. Lung injury in mice after GI/R is MBL and C1q independent, but C2 dependent, suggesting a potential role for ficolins in this model.     

Role of Breast Milk in Acquisition of Cytomegalovirus Infection

The prevalence of IgG antibody against cytomegalovirus (CMV) was compared between the age-matched (0 month to 2 years of age) groups of 212 breast-fed children and 223 bottle-fed children to examine the role of breast milk for acquisition of CMV. Mothers of both groups of children were also examined for CMV IgG antibodies. Both the breast-fed and bottle-fed children groups showed high seropositivity for CMV at 0 to 2 months of age, which gradually decreased and bottomed at 6 to 8 months of age. Thereafter, in the breast-fed children group, the seropositivity rate increased up to 70% by 1 year of age. In contrast, in the bottle-fed children group, the seropositivity rate remained at the bottom level of lower than 30%, without showing any apparent increases. The serological data of the children whose mothers were confirmed to be seropositive, revealed that mother-to-child transmission of CMV occurred in 11 of 17 (64.7%) of the breast-fed children and in 24 of 87 (27.6%) of the bottle-fed children. All the bottle-fed children born to seronegative mothers remained seronegative for CMV up to 1 year of age. The bottle-fed children showed significantly lower seropositivity than the breast-fed children, although most of both groups of children were born to seropositive mothers. The results strongly suggested that about 40% of the breast-fed children acquire CMV via breast milk and breast-feeding has certain protective effects on congenital CMV disease in the offspring.     

Seroepidemiology of human parechovirus types 1, 3, and 6 in Yamagata,Japan, in 2014

To clarify the seroepidemiology of human parechovirus type 1 (HPeV1), 3 and 6, neutralizing antibodies (NT Abs) were measured in 214 serum specimens collected in 2014 in Yamagata, Japan. The seroprevalence against HPeV1 was 100% in all age groups, while that against HPeV3 and HPeV6 was 79.4% and 66.8%, respectively, overall. The geometric mean titers of NT Abs against HPeV1, 3 and 6 were 755.2, 255.0 and 55.9, respectively, overall. Our findings indicate that HPeV1 is the most prevalent HPeV circulating in Yamagata, followed by HPeV3 and HPeV6.     

Substantial Downregulation of Myogenic Transcripts in Skeletal Muscle of Atlantic Cod during the Spawning Period

Gonadal maturation is an extremely energy consuming process for batch spawners and it is associated with a significant decrease in growth and seasonal deterioration in flesh quality. Our knowledge about the molecular mechanisms linking sexual maturation and muscle growth is still limited. In the present study, we performed RNA-Seq using 454 GS-FLX pyrosequencing in fast skeletal muscle sampled from two-year-old Atlantic cod (Gadus morhua) at representative time points throughout the reproductive cycle (August, March and May). In total, 126,937 good quality reads were obtained, with 546 nucleotide length and 52% GC content on average. RNA-Seq analysis using the CLC Genomics Workbench with the Atlantic cod reference UniGene cDNA data revealed 59,581 (46.9%) uniquely annotated reads. Pairwise comparison for expression levels identified 153 differentially expressed UniGenes between time points. Notably, we found a significant suppression of myh13 and myofibrillar gene isoforms in fast skeletal muscle during the spawning season. This study uncovered a large number of differentially expressed genes that may be influenced by gonadal maturation, thus representing a significant contribution to our limited understanding of the molecular mechanisms regulating muscle wasting and regeneration in batch spawners during their reproductive cycle.     

Optimal conditions of in vitro mycelial growth of <Emphasis Type="Italic">Lentinus strigosus</Emphasis>, an edible mushroom isolated in the Brazilian Amazon

The protocols of in vitro cultivation described in the literature for mushrooms are usually correlated with temperate climate habitat, but it is necessary to study protocols for species of tropical climates. In this article, we collected, isolated, and evaluated the conditions of in vitro mycelial growth of Lentinus strigosus and correlated these with the characteristics of its habitat. These results indicate, as optimal conditions of in vitro mycelial growth for L. strigosus, the use of 35°C for incubation, initial pH from 5 to 7, without illumination, Sabouraud dextrose agar medium, and agitation for culture in liquid medium.     

Computer simulation of Gumboro disease outbreak. III. Construction model G-4

Following the simulation mode., G-3, of Gumboro disease outbreak, Model G-4 was constructed. The algorithm for computer simulation is shown in a flow chart. The postulates added to those for Models G-1 and G-2 are as follows: (1) The source of contamination is the virus remaining in the house and declining gradually in value with the lapse of time. (2) Any diseased bird excretes the virus during a certain period, so that the virus may be added to the source of contamination. (3) The morbid status of the diseased bird becomes worse in process of time, but the infection remains subclinical until a threshold value is reached. Beyond this value the bird becomes clinically diseased. In this model, more than 20 parameters are involved, and random numbers used for expressing the individual differences in the four variables, viz., the level of innate resistance, parentally conferred immunity, virus-intake, and threshold of clinical manifestation.