Expression of surface membrane IgG on pokeweed mitogen-reactive anti-tetanus toxoid antibody-producing cells

Anti-tetanus toxoid antibody-producing cells, differentially expressing surface membrane IgM, were analyzed for the additional expression of surface membrane IgG. ⁺ and ^– cells were rosetted with anti--ox red blood cells and separated by density centrifugation into fractions enriched or depleted or ⁺ cells. These B-cell subsets were assayed for the production of IgM and IgG anti-tetanus toxoid antibody and total IgM and IgG. The results indicated that the majority of anti-tetanus toxoid antibody synthesis in the ^– fraction was by ⁺ cells. In the ⁺ fraction, however, both IgM and IgG anti-tetanus toxoid antibody production was detected in the ^+– and ⁺⁺ fraction. The inclusion of isotype-specific antisera during the first 2 days of culture further established that was expressed on the surface of the majority of the precursors for IgG anti-tetanus antibody productionin vitro. Studies performed to determine the culture requirements of ^– and ⁺ cells revealed that production of IgG anti-tetanus toxoid antibody by both cell subsets was dependent on T cells and pokeweed mitogen. However, some ^– cells could produce IgG in the presence of T cells alone.     

Ribonuclease activity of the “myofibrillary” fraction of normal and autolyzed muscle tissue

The separation in a sucrose gradient of the myofibrillary fraction of normal and autolyzed muscle tissue gave 4 components. During post-mortem destruction of the tissue there was observed a slight decrease of the myofibrillary fraction yield and also certain changes in the distribution of protein between different components. Under the selected conditions RNase activity was found in all 4 components. During the course of autolysis enzymatic activity increased in the whole myofibrillary fraction, as well as in the lysosomal-mitochondrial components of myofibrils.Research Laboratory, Ministry of Health of the USSR, Moscow. Translated from Buylleten' Éksperimental'noi Biologii i Meditsiny, Vol. 85, No. 5, pp. 533–537, May, 1978.     

Recent progress in chromosome painting of Arabidopsis and related species

This paper reports on state-of-the-art achievements of chromosome painting in Arabidopsis thaliana (2n=10). Arabidopsis chromosomes 1, 2 and 4 were painted using chromosome-specific BAC contigs. We consider technical aspects of the painting approach and document major applications, such as the tracing of Arabidopsis chromosomes as interphase chromosome territories and during mitotic and meiotic cell cycles as well as comparative chromosome painting in related species. This is the first report of successful interspecific chromosome painting in plants. The evolutionary history of chromosomes homeologous to Arabidopsis chromosome 4 was reconstructed by hybridization of chromosome-4-specific painting probes to karyotypes of Brassicaceae species with x=8 chromosomes. Future perspectives of chromosome painting in A. thaliana and its wild relatives are outlined.     

Genome Organization and Expression of the Penicillium stoloniferum Virus S

The complete sequences of two double-stranded RNAs (dsRNAs) (referred to S1 and S2) of Penicillium stoloniferum virus S (PsV-S) were established. The S1 dsRNA was 1,690bp in length, and it contained a unique open reading frame (ORF) of 539 amino acids (molecular weight of 62kDa, referred to P62). The S2 dsRNA was 1,523bp in length, and also it contained one ORF of 434 amino acids (molecular weight of 47kDa, referred to P47). Both S1 and S2 ORFs were identified only on the positive strand of each dsRNA segment. A sequence motif of (5-CUG-3) was found at the 3-termini of the positive strands of PsV-S1 and S2 dsRNAs. The predicted amino acid sequences of S1 dsRNA showed high sequence homology with the putative RNA-dependent RNA polymerases of RNA viruses. Near full-length and positive-sense single-stranded RNAs derived from the S1 and S2 dsRNAs were detected from the PsV-infected host cell. The expressed proteins of P62 and P47 showed a positive reaction against PsV-S antiserum in Western blot analysis. Phylogenetic analysis using the RDRP sequences and the capsid proteins of the various partitiviruses revealed that PsV-S is a definite member of the partitivirus, the family Partitiviridae, and especially clusters well along with D. destructiva virus 1 and 2.     

Photobehavioral differentiation in natural populations ofDrosophila pseudoobscura andDrosophila persimilis

The photoresponses of natural populations ofD. pseudoobscura andD. persimilis, occurring sympatrically, are measured in two environmental conditions (at rest and disturbed). Comparisons of the responses, intraspecifically and interspecifically, lead to the following conclusions. These must be considered within the confines of the operational nature of the measurement of laboratory photoresponses. (1) Within each species population, significant nonenvironmental differentiation has been allowed or produced by selection in the at rest photoresponse. No significant nonenvironmental differentiation is found in the photoresponse measured in a disturbed condition. (2) Within each species population, a higher mean disturbed photoresponse has been favored. The intensities or patterns of selection acting on these two photoresponses have differed such that more intrapopulation differentiation has been allowed or produced in the at rest photoresponse. (3) A higher mean photoresponse has been favored inD. persimilis for both conditions. The intensities or patterns of selection acting between these two species populations on the at rest photoresponse have differed such that more intrapopulation differentiation has been allowed or produced inD. persimilis. (4) Comparisons of this study with one on intraspecific and interspecific differentiaiton in wing length lead to the conclusion that the selective differences inferred above have acted at a level more specifically attuned to photobehavior.     

Direct observation of sexual competition inDrosophila melanogaster: The mutantWhite in competition with other genotypes

When two types ofDrosophila are in competition, the frequency dependence of mating success routinely is measured in our laboratory by direct observation of mating pairs in Elens-Wattiaux observation chambers. The present experiments concern white mutants (used here as a reference standard) in competition with three other genotypes: wild-type Canton S, giantgt w ^a , and giantgt ^13z /w ⁺ Y/C(1)Dx, y f. From the present observations, the frequency dependence of mating success seems a very common phenomenon: a rare-type sexual advantage exists for females and for males. Sexual activity of male and femalewhite mutants is significantly higher when food is present in the mating chamber. Males of the other strains also are more active in the presence of food. Homogamic matings are more frequent amonggt ^13z /w ⁺ Y/C(1)Dx, y f flies.     

Altered synthesis of interferon-γ and expression of interferon-γ receptor by peripheral blood mononuclear cells from patients with IgA nephropathy and non-IgA proliferative glomerulonephritis

Previously we reported disease-specific interaction between interferon- (IFN-) and interleukin-4 (IL-4) in patients with IgA nephropathy (IgAN), suggesting the existence of unusual T cell behavior in this disease. In the present study, we investigated characteristic synthesis of interferon- (IFN-) and expression of IFN- receptor (IFN-R) in the peripheral blood mononuclear cells (PBMC) from patients with IgAN and other chronic proliferative glomerulonephritis (PGN). Heparinized peripheral blood samples were obtained from 38 patients with chronic mesangial proliferative glomerulonephritis (CGN; including 24 with IgA nephropathy) and 20 healthy controls. PBMC were isolated by gradient centrifugation and fragments were cultured in Iscove's modified Dulbecco's medium (IMDM) supplemented with 10% fetal calf serum (FCS) for 72 hr. IFN- concentrations in supernatants were evaluated by the enzyme-linked immunosorbent assay (ELISA). Other parts of PBMC pellets were reacted with anti-human IFN-R monoclonal antibody and FITC-labeled anti-mouse second antibody for analysis of IFN-R expression on these cells by FACScan. The remaining PBMC were fractionated into CD4⁺ T cells, CD8⁺ T cells, B cells, NK, cells and macrophages using the MACS cell sorting system. The isolated cells were evaluated for IFN- or IFN-R mRNA expression by the semiquantitative RT-PCR method.In vitro IFN- synthesis was enhanced in patients with CGN, and NK cells were revealed to be responsible for such enhancement. On the other hand, the expression of IFN-R on macrophages was suppressed in CGN patients. These results suggest that impairment of regulation of the IFN- system might be involved in the development of CGN.     

Nachweis von 6 neuen Steroiden im Plasma von menschlichem Placentablut (Nabelsehnurblut)

Zusammenfassung 16-Hydroxyprogesteron, 17-Hydroxyprogesteron, ⁴-Pregnen-20-ol-3-on, ⁴-Pregnen-17, 20-diol-3-on, Adrenosteron und 11-Hydroxyandrostendion wurden in Extrakten von Plasma des menschlichen Placentablutes (Nabelschnurblut) nachgewiesen.     

Mating-type suppression of the DNA-repair defect of the yeast rad6Δ mutation requires the activity of genes in the RAD52 epistasis group

The RAD6 gene of Saccharomyces cerevisiae is required for post-replication repair of UV-damaged DNA, UV mutagenesis, and sporulation. Here, we show that the radiation sensitivity of a MAT a rad6 strain can be suppressed by the MAT2 gene carried on a multicopy plasmid. The a1-2 suppression is specific to the RAD6 pathway, as mutations in genes required for nucleotide excision repair or for recombinational repair do not show such mating-type suppression. The a1-2 suppression of the rad6 mutation requires the activity of the RAD52 group of genes, suggesting that suppression occurs by channelling of post-replication gaps present in the rad6 mutant into the RAD52 recombinational repair pathway. The a1-2 repressor could mediate this suppression via an enhancement in the expression, or the activity, of recombination genes.     

Effect of cyclic nucleotides and of isopropylnoradrenalin on the partial pressure and consumption of oxygen

Isopropylnoradrenalin, cyclic AMP, and dibutyryl-3,5-AMP (DB) lower the partial pressure of oxygen (pO₂) in the liver and spleen and increase the oxygen consumption of the body. The time course of these two effects for isopropylnoradrenalin and cyclic AMP correlates closely. The increase in heat production was not accompanied by any significant change in the respiratory quotient. The effects of cyclic AMP were not abolished by pempidine or by dihydroergotamine and were only reduced to some degree by propranolol. The effect of catecholamines on pO₂ is evidently mediated through an increase in the O₂ consumption as a result of the effect of cyclic AMP; the action of cyclic AMP is basically direct, but in vivo it cannot take place through the adrenoreceptor; the compound 2,3-AMP reduces pO₂ and also the O₂ consumption.Department of Biochemistry, Krasnoyarsk Medical Institute. (Presented by Academician of the Academy of Medical Sciences of the USSR S. S. Debov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 83, No. 2, pp. 177–180, February, 1977.     

Pattern of antinuclear autoantibodies in diseases with systemic connective tissue lesions

Several types of antinuclear antibodies (ANA) were discovered by the fluorescent antibody method in diseases accompanied by systemic lesions of connective tissue and also in certain other diseases and in clinically healthy blood donors, depending on the character of fluorescence in the nuclei. Diffuse fluorescence of nuclei, diffuse fluorescence without fluorescence of the nucleoli, annular fluorescence, fluorescence in the form of granules, selective fluorescence of nucleoli, and fluorescnece in the form of long, thin, interweaving bands with simultaneous fluorescence in the region of the nuclear membrane were distinguished. The last type of ANA was observed only in various forms of lupus erythematosus, and the character of fluorescence in the nuclei differed from the reticular and filamentous types of fluorescence described previously.Department of Pathological Anatomy and Department of Skin Diseases, N. I. Pirogov Second Moscow Medical Institute. (Presented by Academician of the Academy of Medical Sciences of the USSR A. P. Avtsyn.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 84, No. 7, pp. 75–78, July, 1977.     

Foraging strategies ofDrosophila melanogaster: A chromosomal analysis

Two larval foraging strategies inDrosophila melanogaster were identified, rover and sitter. Rovers traverse a large area while feeding whereas sitters cover a small area. The difference between rovers and sitters was analyzed genetically by chromosomal substitutions between isogenic stocks. Differences in larval locomotor behavior (crawling behavior) can be attributed to the second chromosome, the rover strategy being dominant over the sitter strategy. Differences in feeding rate (shoveling behavior) are affected additively by both the second and third chromosomes. Natural populations ofDrosophila larvae were sampled three times over a 2-month period; rovers and sitters were at constant frequencies in these populations. The two foraging strategies are discussed in the light of resource utilization in environments where food is distributed continuously or discontinuously.     

Activation of atrial muscarinic K+ channels by low concentrations ofβγ subunits of rat brain G protein

Effects of G protein subunits from rat brain on cardiac K⁺ channel was examined in single atrial cells of guinea-pig, using patch clamp techniques. We found that 10 pM concentration of rat brain subunits preparation could activate the atrial muscarine receptor-gated K⁺ channel (I_K.ACh). Neither the detergent, CHAPS, used to suspend nor the boiled preparation activated I_K.ACh. Furthermore, preincubation of subunits preparation in Mg²⁺-free solution, which easily inactivated -GTP-S, did not affect -activation of I_K.ACh. We concluded, therefore, that subunits themselves can activate I_K.ACh.Supported by the grants from the Ministry of Education, Culture and Science of Japan and from the Calcium Signal Workshop on Cardiovascular Systems     

Synthesis of Seoul virus RNA and structural proteins in cultured cells

Summary. Seoul virus is a hantavirus that causes hemorrhagic fever with renal syndrome (HFRS). The virion has a tripartite (S, M, and L) negative-stranded RNA genome, which is characteristic of the family Bunyaviridae. However, the molecular basis of virus replication is not well known. We established a Northern blot hybridization (NB) procedure using digoxygenin-labeled RNA probes, to quantitate the hantaviral plus- and minus-strand RNAs separately. Virus RNA replication was analyzed in infected Vero E6 cells. When the Vero E6 cells were infected with Seoul virus strain KI-83-262 (KI) at m.o.i.=0.25, the plus-strand RNA was detected within 1h post-infection (hpi), and the minus-strand RNA was detected subsequently. Using laser confocal microscopy, the nucleocapsid protein (NP) was detected within 2hpi, and accumulated as scattered granules in the cytoplasm until 24hpi. In contrast, the G2 protein first appeared at 8hpi, was immediately transported to the Golgi, and accumulated in the Golgi until 24hpi. Infectious virus particles were released into the medium at 24hhpi. These findings indicate that hantavirus RNA replication starts with the appearance of NP at 2hpi, glycoproteins then accumulate gradually in the Golgi, and virion formation is initiated once the viral RNAs and proteins have accumulated.Received October 10, 2002; accepted April 25, 2003 Published online July 17, 2003     

Molecular Analysis of Fiji Disease Fijivirus Genome Segments 1 and 3

Fiji disease fijivirus (FDV) genomic segments 1 (S1) and 3 (S3) were completely sequenced. FDV S1 comprised 4532nt and was predicted to encode a 170.6kDa protein. FDV S3 comprised 3623nt and was predicted to encode a 135.5kDa protein. The terminal sequences of S1 and S3 were 5 AAGUUUUU......CAGCUAGCGUC 3 and 5 AAGUUUUU......CAGCAGAUGUC 3, respectively, and located immediately adjacent to these sequences were 12bp imperfect inverted repeats. The predicted translation product of FDV S1 showed highest similarity to Rice black-streaked dwarf virus (RBSDV) S1 and is thought to encode the viral RNA-dependent RNA polymerase (RdRp). The predicted translation product of FDV S3 was found to be most similar to RBSDV S4 which is thought to encode the 'B-spike' protein. The FDV sequence contained an ATP/GTP binding motif and a leucine zipper motif, but these motifs were not found in the RBSDV sequence. Phylogenetic analysis based on the amino acid sequences of the RdRp of FDV S1 and other reoviruses revealed that the fijiviruses form a cluster close to the oryzaviruses. The RdRp sequences were grouped into genera that were consistent with the current reovirus classification scheme that is based on physico-chemical and biological properties.     

A novel H101Q mutation causes PKCγ loss in spinocerebellar ataxia type 14

Spinocerebellar ataxia type 14 (SCA14) is an autosomal dominant neurodegenerative disorder, first described in a Japanese family, showing linkage to chromosome 19q13.4-qter. Recently, mutations have been identified in the PRKCG gene in families with SCA14. The PRKCG gene encodes the protein kinase C (PKC), a member of a serine/threonine kinase family involved in signal transduction important for several cellular processes, including cell proliferation and synaptic transmission. To identify the disease-causing mutation in a large group of ataxia patients, we searched for mutations in the PRKCG gene. We ascertained 366 unrelated patients with spinocerebellar ataxia, either pure or with associated features such as epilepsy, mental retardation, seizures, paraplegia, and tremor. A C-to-G transversion in exon 4, resulting in a histidine-to-glutamine change at codon 101 of the PKC protein, was identified in patients from a family with slowly progressive pure cerebellar ataxia. Functional studies performed in HEK293 cells transfected with normal or mutant construct showed that this mutation affects PKC stability or solubility, verified by time-dependent decreased protein levels in cell culture. In conclusion, the H101Q mutation causes slowly progressive uncomplicated ataxia by interfering with PKC stability or solubility, which consequently may cause in either case a decrease in the overall PKC-dependent phosphorylation.     

Mother-Baby psychiatric units (MBUs): National data collection in France and in Belgium (1999–2000)

Summary The French version of the Marcé checklist was used to collect data for 176 joint admissions to 11 psychiatric mother-baby units in 1999 and 2000. Mean age of the babies at admission ranged from 4 to 16 weeks. Two units also admitted older children. Mothers admitted were diagnosed with schizophrenia or chronic delusional disorders (n=44), acute transitory psychosis Bouffée délirante (n=20), bipolar disorders (n=20), depressive illness (n=38), personality disorders or intellectual disability (n=39), and other disorders (n=15). The mean duration of hospitalisation was 11 weeks. Units that also offered day-care admission in the same or a near-by unit had shorter mean admissions. More than half the womens partners (or babies fathers) had mental health problems. Women with schizophrenia or chronic delusional disorders and personality disorders or intellectual disability remained hospitalised longer, improved less, and were more often separated from their babies, or discharged with supervision, than women admitted with other diagnoses.     

Molecular organisation of an A mating type factor of the basidiomycete fungus Coprinus cinereus

Summary The A3 and A3 genes, which together constitute the A42 mating type factor of Coprinus cinereus, were isolated from a cosmid genomic library by walking 50 kb, a map distance of 0.5 units, from the closely linked metabolic gene pab-1. Cosmid clones having A gene function were identified by transformation into compatible A6 (22) and A5 (11) host cells where either 3 or 3 was expected to elicit the A factor — regulated development of unfused clamp cells. DNAs were digested with various enzymes before transformation in order to identify the smallest fragments containing an active 3 or 3 gene. Two non-overlapping fragments were identified as containing the 3 and 3 genes respectively. Southern hybridisation analyses showed that these two cloned genes had no detectable sequence homology, and that there was little or no hybridisation to the and gene alleles that constitute the A5 and A6 factors. 3 and 3 were shown to be less than 2.0 kb apart and embedded in a DNA sequence extending over 9.0 kb which was unique to our A42 strain and may contain a third A factor gene.     

A noninvasive membrane filter method for in vivo determination of leukocyte migration in man

A new in vivo method for measuring leukocyte migration in man involves the insertion of platelets prepared from membrane filters into the lower conjunctival sac. The platelets contain casein and are coated unilaterally with a foil, permitting the casein to diffuse in one direction only. A chemotactic gradient is thereby formed, in which neutrophil granulocytes originating from the conjunctival mucosa migrate. The total amount of immigrant cells, as well as their distribution within the filter platelets are evaluated. The method yields information as to the speed of migration of the cells and the intensity of egress from the vessels, A few cases of diseases are demonstrated where the total amount of cells and their distribution patterns differ from the standard values.     

Interspecific genetic complementation analysis of human and sheep fibroblasts withβ-galactosidase deficiency

Interspecific somatic cell hybrids were analyzed by genetic complementation to determine if a lysosomal storage disease in sheep associated with deficiencies of -galactosidase and -neuraminidase was homologous with any of four -galactosidase-deficient human diseases. Fibroblasts from -glactosidase-deficient sheep, cats, and human patients were fused and assayed histochemically for -galactosidase, with 5-bromo-4-chloro-3-indolyl -d-galactoside. We observed complementation in heterokaryons consisting of fibroblasts from -galactosidase-deficient sheep and fibroblasts from patients with galactosialidosis or mucolipidosis type II, but no complementation in heterokaryons consisting of fibroblasts from -galactosidase-deficient sheep and fibroblasts from human or feline GM₁, gangliosidosis (type I) or from human mucopolysaccharidosis type IVB fibroblasts. We conclude that the ovine disease is due to a mutation at the genetic locus homologous with that of GM₁, gangliosidosis and mucopolysaccharidosis type IVB, suggesting that the primary defect in the ovine disease is a mutation of the -galactosidase structural gene.