Organization of the ribosomal ribonucleic acid genes in various wild-type strains and wild-collected strains of Neurospora

Summary The organization of the ribosomal DNA (rDNA) repcat unit in the standard wild-type strain of Neurospora crassa, 74-OR23-1A, and in 30 other wild-type strains and wild-collected strains of N. crassa, N. tetrasperma, N. sitophila, N. intermedia, and N. discreta isolated from nature, was investigated by restriction enzyme digestion of genomic DNA, and probing of the Southern-blotted DNA fragments with specific cloned pieces of the rDNA unit from 74-OR23-1A. The size of the rDNA unit in 74-OR23-1A was shown to be 9.20 kilobase pairs (kb) from blotting data, and the average for all strains was 9.11+0.21 kb; standard error=0.038; coefficient of variation (C.V.)=2.34%. These data indicate that the rDNA repeat unit size has been highly conserved among the Neurospora strains investigated. However, while all strains have a conserved HindIII site near the 5 end of the 25 S rDNA coding sequence, a polymorphism in the number and/or position of HindIII sites in the nontranscribed spacer region was found between strains. The 74-OR23-1A strain has two HindIII sites in the spacer, while others have from 0 to at least 3. This restriction site polymorphism is strain-specific and not species-specific. It was confirmed for some strains by restriction analysis of clones containing most of the rDNA repeat unit. The current restriction map of the 74-OR23-1A rDNA repeat unit is presented.     

Mechanisms of genetic control of immune responses

The mechanisms underlying Ir gene control of CMI were addressed by examining the DTH and Tprlf responses specific for the synthetic polymers GT, GAT, and GA. We show that BALB/c mice (GAT/GA responders, GT nonresponders) primed with GT fail to develop DTH and Tprlf responses specific for GT, GAT, or GA. GAT immunization resulted in DTH responses that could be elicited not only with GAT and GA but also with GT, demonstrating that GT-specific T_DH are present in nonresponder mice. GT-specific DTH was transferred with Thy-1⁺ Lyt-1⁺2^–, H-2 Irestricted, nylon wool nonadherent cells. GA-primed BALB/c mice developed GAT- and GA-, but not GT-apecific DTH responses, indicating that GA and GT do not cross-react at the T-cell level. The ability of GAT [but not a mixture of GA plus GT, or GT electrostatically complexed to the immunogenic carrier MBSA (GT-MBSA)] to induce GT-specific DTH suggested a requirement for covalent linkage of stimulatory GA and nonstimulatory GT determinants present on the GAT molecule. Similarly, GT-specific in vitro Tprlf responses could be demonstrated in GAT-primed mice exhibiting significant levels of GT-specific DTH but not in GT- or GT-MBSA-primed mice. Tolerization experiments also suggested that GT-specific Th were involved in the development of GT-specific DTH in GAT-primed mice. The GT nonresponsiveness of BALB/c mice for DTH and Tprlf responses could not be reversed by treatments designed to abrogate Ts activity (priming with GT-MBSA and CY injection), nor could GT-primed cells be shown to inhibit the development or elicitation of GT-specific CMI in GAT-primed mice during the afferent and/or efferent stages of DTH. Our results suggest that GT nonresponsiveness does not result from the absence of GT-specific T cells or preferential induction of Ts. The results are discussed in the context of hole-in-the-repertoire and antigen presentation (determinant selection) models of Ir gene control.Abbreviations used in this paper APC antigen-presenting cells - BSA bovine serum albumin - BSS Mishell-Dutton balanced salt solution - CFA complete Freund's adjuvant - CMI cell-mediated immunity - CY cyclophosphamide - DTH delayed-type hypersensitivity - GA poly(Glu⁶⁰Ala⁴⁰) - GAT poly (Glu⁶⁰Ala³⁰Tyr¹⁰) - GT poly(Glu⁵⁰Tyr⁵⁰) - GT-MBSA GT complexed to methylated bovine serum albumin - It immune response - LN lymph node - PPD purified protein derivative of tuberculin - TDH DTH T cells - Th helper T cells - Tprlf T-cell proliferation - Ts suppressor T cells - TsF T-cell suppressor factor(s)     

Shape, size and density of daytime surface swarms of the euphausiid Meganyctiphanes norvegica in the Bay of Fundy

Daytime surface swarms of ^{Meganyctiphanes norvegica} in the Bayof Fundy were examined using a variety of techniques to providemeasurements of their shapes, sizes and densities. Shapes andsizes were determined from two aerial photographs: swarms werespherical, ribbon-like or amorphous. were up to 28 6 m longand ranged in area from 0.4–111 7 m². Densities were measuredby a bag-sampling device which gave figures of up to 41 000animals m^–3 by photographic methods which gave figuresof up to 770 000 animals m^–3 and by a plankton net whichgave maximum values of six animals m^–3 Using the photographicmethod the maximum euphausud biomass was estimated to be 154kg m^–3 within swarms and the largest swarm measured wasestimated to contain up to 2 1 tonnes of M. norvegica. Meanpatch biomass estimates for the two aerial photographs rangedfrom 77.8–778 g m^–3 and 15 6–155.6 g m^–3which are similar to figures obtained by other authors usingintegrating sampling techniques at depth.     

Thermoregulatory physiology of the carpenter bee,Xylocopa varipuncta

Summary The carpenter beesXylocopa varipuncta maintain thoracic temperatures of 33.0°C to 46.5°C during continuous free flight from 12°C to 40°C. Since the thoracic temperature excess is not constant (decreasing from 24°C at low air temperatures to 6°C at high) the bees are thermoregulating. We document physiological transfer of relatively large amounts of heat to the abdomen and to the head during pre-flight warm-up and during artificial thoracic heating. Most of the temperature increase of the head is due to passive conduction, while that of the abdomen is due to active physiological heat transfer despite a series of convolutions of the aorta in the petiole that anatomically conform to a counter-current heat exchanger. Although the thermoregulatory mechanisms during flight are far from clarified, our data suggest that thermoregulation involves a strong reliance on active convective cooling through increased flight speed.     

Dopamine- and octopamine-sensitive adenylate cyclase in the brain of adultCulex pipiens mosquitoes

The effects of dopamine and octopamine on adenylate cyclase activity were studied on the head homogenate of adult Culex pipiens mosquitoes in vitro. Both dopamine and octopamine were shown to increase the cyclic AMP content in the homogenate. The antagonist haloperidol blocked the production of cyclic AMP induced from dopamine but had no effect on the production of cyclic AMP induced by octopamine at the concentrations tested. The opiate agonist etorphine was ineffective at reducing cyclic AMP levels induced by either dopamine or octopamine at the concentrations tested.     

Heliobacterium chlorum: cell organization and structure

The basic cellular organization of Heliobacterium chlorum is described using the freeze-etching technique. Internal cell membranes have not been observed in most cells, leading to the conclusion that the photosynthetic apparatus of these organisms must be localized in the cell membrane of the bacterium. The two fracture faces of the cell membrane are markedly different. The cytoplasmic (PF) face is covered with densely packed particles averaging 8 nm in diameter, while the exoplasmic (EF) face contains far fewer particles, averaging approximately 10 nm in diameter. Although a few differentiated regions were noted within these fracture faces, the overall appearance of the cell membrane was remarkably uniform. The Heliobacterium chlorum cell wall is a strikingly regular structure, composed of repeating subunits arranged in a rectangular pattern at a spacing of 11 nm in either direction. We have isolated cell wall fragments by brief sonication in distilled water, and visualized the cell wall structure by negative staining as well as deep-etching.Abbreviations PF protoplasmic fracture face - EF exoplasmic fracture face     

Dinitrogen fixation by obligate and facultative anaerobic bacteria in association with cellulolytic fungi

The anaerobic bacteriumClostridium butyricum is the major contributor to nitrogen gains by a cellulolytic/nitrogen-fixing population isolated from straw. Growth of the anaerobe is supported by the products of fungal cellulases. The facultative anaerobeEnterobacter cloacae does not make a significant direct contribution to nitrogen fixation but in association withC. butyricum allows the anaerobe to grow under aerobic conditions. The major function ofE. cloacae is though to be provision of oxygen-depleted microsites.     

Transformation ofBacillus spp.: An examination of the transformation ofBacillus protoplasts by plasmids pUB110 and pHV33

The protoplasting and transformation techniques described by Chang and Cohen [5] have been modified by the inclusion of mutanolysin and these techniques have been used to prepare protoplasts of a number ofBacillus spp. Cells of some, however, remained resistant to cell wall hydrolysis by both mutanolysin and lysozyme. Protoplasts were prepared from sixBacillus species, including a strain ofB. subtilis, and transformed with the plasmid pUB110. Transformation with the shuttle vector pHV33 was, however, less successful and antibiotic-resistant protoplasts, although detected, either failed to regenerate their osmotic stability or rapidly lost their antibiotic resistance.     

Population dynamics,production and angling catch of brown trout,Salmo trutta,in a mature upland reservoir in mid-Wales

Synopsis The brown trout in Llyn Frongoch, a mature upland reservoir, and its nursery stream was sampled during 1983. The stream stock consisted largely of the 1983 and 1982 year classes, with fish reaching mean lengths of 7.0 and 11.6 cm at one and two years of age. The size and biomass of the stream stock at the beginning of 1983 and 1984 were estimated to be 120 and 125 (1.20 and 1.25 fish m^–2) and 1.41 and 0.69 kg (14.1 g m^–2 and 6.9 g m^–2) respectively. Annual stream production ranged from an estimated minimum of 2.49 kg (24.9 g m^–2) to an estimated maximum of 4.59 kg (45.9 g m^–2). Both downstream and upstream movements of 0+ juveniles were recorded. The adult spawning stock was estimated at 79 males and 32 females, a sex ratio of 2.5:1, with most spawners belonging to the 1980 yearclass. The average size of the lake stock over the year was estimated to be 1 650 (229 fish ha^–1) or 250.8 kg (34.8 kg ha^–1). The 1980 yearclass was predominant; there were few fish older than five years. Seasonal variations in netting catches suggested movements to and from the littoral region. Growth in the lake was moderately fast, with fish reaching mean lengths of 21.7 and 27.2 cm by three and four years of age. Fish entering the lake after one year appeared to grow faster than fish which remained in the stream for two years. Annual production in the lake was estimated at 136.7 kg (19.0 kg ha^–1). The total angling catch for the season was estimated to be 62.6 kg (8.7 kg ha^–1).     

The effect of the tumor promoter, phorbol myristate acetate (PMA), on hyaluronic acid (HA) synthesis by chicken embryo fibroblasts

Treatment of chicken embryo frbroblasts (CEF) with the tumor promoter, phorbol myristate accetate (PMA), resulted in a rapid increase in their ability to synthesize the glycosaminoglycan, hyaluronic acid (HA), whereas the parent compound, phorbol, had no effect. CEF cultures incubated with PMA (100 ng/ml) for 6 h resulted in a 15-fold increase in HA synthetase activity compared with phorbol-treated control cultures. The incorporation of [³H]acetate into HA and chemical determination of this polymer also demonstrated increased synthesis of HA by cells treated with PMA. We have previously shown that CEF infected with a temperature-sensitive mutant of Rous sarcoma virus, LA24, exhibit increased synthesis of HA upon transformation. PMA treatment of cells transformed with RSV-LA24 results in a further 1.5-fold stimulation of HA synthesis. These data indicate that PMA causes an increased synthesis of HA in CEF which is analogous to the increased synthesis of HA found in virally transformed CEF.