Occurrence of Horizontal Gene Transfer of PIB-type ATPase Genes among Bacteria Isolated from the Uranium Rich Deposit of Domiasiat in North East India

Uranium (U) tolerant aerobic heterotrophs were isolated from the subsurface soils of one of the pre-mined U-rich deposits at Domiasiat located in the north-eastern part of India. On screening of genomic DNA from 62 isolates exhibiting superior U and heavy metal tolerance, 32 isolates were found to be positive for P_IB-type ATPase genes. Phylogenetic incongruence and anomalous DNA base compositions revealed the acquisition of P_IB-type ATPase genes by six isolates through horizontal gene transfer (HGT). Three of these instances of HGT appeared to have occurred at inter-phylum level and the other three instances indicated to have taken place at intra-phylum level. This study provides an insight into one of the possible survival strategies that bacteria might employ to adapt to environments rich in uranium and heavy metals.     

Spinal ascending projections to the nucleus tractus spinalis nervi trigemini of the dog

Seven dogs were subjected 30 min to ligation of the thoracic aorta and were then kept alive 6-7 days after the ligature had been removed. Their spinal cord and brain stem were treated by the Nauta-Gygax method and the extent and appearance of preterminal and terminal degeneration of certain ascending spinal systems were analysed. In the medulla oblongata region, marked degenerating fibres from the lower thoracic and lumbosacral cord segments were found in the nucleus tractus spinalis nervi trigemini. Preterminal and terminal degenerating fibres were visualized in the caudal part of the trigeminal nuclear complex. Comparison with the literature showed these to be previously unknown projections with a relationship to the nucleus tractus spinalis nervi trigemini.     

Visualization of multiple protein bands on the same nitrocellulose membrane by double immunoblotting

A method has been developed which allows the simultaneous immunodetection of more than one type of protein on the same nitrocellulose membrane. This procedure does not require special labeling of samples or elution of antibodies from the membrane as do the alternatives cited in the literature (1,2). Proteins are separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electrophoretically transferred to the membrane before specific immunostaining with either peroxidase/4-chloro-1-naphthol or immunogold/silver staining. Antigen identity is visually determined by the formation of different-colored precipitates on the membrane. This innovation in protein blotting offers a savings in time and reagents as well as permitting identification of closely spaced bands with certainty.     

Lipid-bound sugars in malignant human breast tumors

Microsomal preparations from malignant human breast tumors catalyzed the transfer of mannose and glucose from GDP-[14C]-Man and UDP-[14C]-Glc into lipid-linked sugars and glycoprotein-like substances. As judged by several criteria the obtained lipid-linked monosaccharides behaved as dolichyl phosphate mannose and dolichyl phosphate glucose whereas lipid-linked oligosaccharides behaved as polyprenyl diphosphate derivatives. The optimum conditions for mannosyl- and glucosyl-transfer reactions and the effect of dolichyl phosphate, detergent and EDTA on incubation mixture were described.     

Effect of the cumulus on in vitro fertilization of bovine matured oocytes

The effect of the cumulus on in vitro fertilization in bovines was examined. Follicular oocytes were cultured in medium 199 plus OCS and extra granulosa cells. Frozen-thawed bovine spermatozoa was separated by the swim-up technique, suspended in Talp medium and capacitated with heparin. Fresh sheep and goat semen was incubated for 4 h at room temperature, washed and spermatozoa were then suspended in Talp medium and capacitated by incubation at 38.5 °C and 5% CO₂ in air and heparin.

In experiment 1, cumulus-enclosed oocytes, denuded oocytes and denuded oocytes plus additional cumulus cells were incubated with a reduced concentration of bovine spermatozoa for 8 or 18 h. In Experiment 2, cumulus enclosed and denuded oocytes were incubated with bovine spermatozoa for 4, 6, 8 and 18 h using a sperm concentration adjusted to secure high fertilization rates. In Experiment 3, cumulus-enclosed and denuded bovine oocytes were incubated with either sheep or goat spermatozoa for 18 h. Fertilization rates were then calculated and compared statistically. The results showed that 1) the cumulus improved the fertilization rate only when cumulus cells were associated with the oocytes 2) the timing of sperm penetration was not modified by the cumulus and started at 4 h after sperm incubation and 3) the presence of the cumulus improved the heterologous fertilization rate only when sheep spermatozoa were used. The results suggest that the cumulus improves fertilization rate by providing a capacitation-inducing mechanism and by facilitating the interaction between capacitated spermatozoa and the zona pellucida surface.     

L cells expressing DQ molecules of the DR3 and DR4 haplotypes: reactivity patterns with mAbs

cDNAs coding for the HLA class II DR and DQ and chains of the diabetogenic haplotypes DR3 and DR4 were introduced into a mammalian expression vector and transfected into L-cell mouse fibroblasts to produce cells expressing individual human class II molecules. Stable L transfectants were generated expressing each of the DR or DQ isotypes of the cis-encoded and chains of the DR3 or DR4 haplotypes, as well as the trans-encoded and chains of the DQ molecules of the two haplotypes. However, isotype mismatched combinations (DR/DQ or DQ/DR) did not result in any stable transfectants. The stable DQ L-cell transfectants obtained, along with homozygous B-cell lines expressing the DQ2 and DQ8 specificities, were tested against a large panel of twentyone anti-HLA class II monoclonal antibodies (mAbs). Their unusual reactivity patterns are described including the failure of most pan-DQ mAbs to react with all DQ expressing L-cell transfectants. Interestingly, some mAbs react with certain heterodimers expressed on B-LCL but fail to recognize the same heterodimers expressed on the transfectants. This is suggestive of minor structural modifications that class II molecules undergo depending on the cells they are expressed on.The nucleotide sequence data reported in this paper have been submitted to the GenBank nucleotide sequence database and have been assigned the accession number U07848. The name DQB1 ^* 0202 was officially assigned by the WHO Nomenclature Committee in April 1994. This follows the agreed policy that, subject to the conditions stated in the most recent Nomenclature Report (Bodmer et al. 1992), names will be assigned to new sequences as they are identified. Lists of such new names will be published in the following WHO Nomenclature Report     

Habitat and host associations of the fish‐burrowing parasite Artystone minima (Cymothoidae: Isopoda) in eastern Amazonia

Cymothoid fish parasites settle on hosts in ways that may impact fish health and energetics. High abundances of Artystone minima observed in Nannostomus beckfordi from the Jeju River in eastern Amazonia were investigated to answer the following questions: (a) What factors are associated with the high prevalence at this locality?; (b) Is high abundance associated with co‐infestation of alternative hosts?; and (c) Is parasite presence associated with host species growth and/or reproduction? Fish assemblages were sampled quarterly (August 2017–May 2018) from five habitats along with environmental data. Parasitic indices were calculated, and parasite presence used to evaluate differences in growth of hosts using analysis of covariance considering host sex and sampling season (wet vs. dry). Parasites were only abundant in one of the habitats, a large, shallow backwater bay with macrophytes. Abiotic environmental factors (flow and depth) likely impact parasite transmission and are, therefore, particularly important in producing these local patterns. Two secondary hosts, Hyphessobrycon cf. rosaceus and Moenkhausia collettii, were found in the wet season. Based on host biology compared to other fish in the habitat, parasite infestation is inferred to be depth associated and long‐term infestation is apparently limited in alternative hosts. Parasite presence was significantly associated with reduced weight (standardized for length) of female Nannostomus beckfordi in the wet season. Furthermore, ovaries of non‐parasitized females from the wet season presented a range of maturation stages, while parasitized females were all immature, indicating a significant association of parasites with host reproductive capacity. Abstract in Portuguese is available with online material