T-cell responsiveness in Mycobacterium lepraemurium infections in a "resistant" (CBA) and a "susceptible" (BALB/c) mouse strain

Antigen-specific and mitogen-nonspecific T-lymphocyte proliferation and lymphokine release (interleukin 2 and macrophage activation factor) were studied in BALB/c and CBA mice infected intravenously with 10(8) Mycobacterium lepraemurium organisms. The responsiveness of spleen cells from infected animals to Con A and specific MLM antigen declined as the infection progressed. Thus, the decreased responsiveness appeared earlier and was more profound in the relatively susceptible BALB/c strain than in the relatively resistant CBA strain. Nylon-wool-purified, T-cell-enriched spleen cells from both strains, however, responded to both M. lepraemurium antigen and Con A until the later stages of infection (17 weeks postinfection). The relevance of nonspecific immunodepression mediated by nylon-wool-adherent spleen cells to the progressive nature of this infection is discussed.     

The metabolic clearance of progesterone in the pregnant rat: absence of a physiological role for the lung

The metabolic clearance rate (MCR) of progesterone is among the highest for all steroid hormones studied, yet it is difficult to apportion this high MCR to specific organ contributions. The isolated lung has been shown to metabolize progesterone, and since this tissue receives the entire cardiac output, potentially it could make a major contribution to the overall MCR. This possibility was examined in the present study by measuring lung extraction of [3H]progesterone under steady-state conditions in the intact pregnant rat. Anesthetized rats (n = 6) were infused with [3H]progesterone via a femoral vein for 100 min on Day 16 of pregnancy. After the onset of steady state (40 min), four blood samples were obtained at 20-min intervals from the right ventricle and from the aorta, and the concentrations of [3H]progesterone and its metabolites were determined. Throughout the sampling period, mean arterial pressure and heart rate remained stable (two-way analysis of variance), as did the production rate (3.76 +/- 0.35 mg/day; mean +/- SEM) and the MCR (34.8 +/- 3.5 ml/min) of progesterone. Despite this high rate of clearance, there was no difference between the concentration of [3H]progesterone in arterial and right ventricular blood, indicating no net extraction of progesterone during passage through the lung. Furthermore, there was no change in the concentration of either lipid-soluble or aqueous-soluble [3H]progesterone metabolites during trans-lung passage. These observations demonstrate that the lung does not contribute to the MCR of progesterone when measured under physiological and steady-state conditions. Therefore, the relationship, MCR (ml/min) = whole-body extraction (%) x cardiac output (ml/min), is upheld for progesterone in the rat.     

Evidence for regulation of actin synthesis in cytochalasin D-treated HEp-2 cells

In HEp-2 cells treated with 0.2 or 2.0 μM cytochalasin D (CD), the relative rate of actin synthesis increased for about 12 h and then reached a plateau; this increase was suppressed by actinomycin D (AD). When CD was washed from cells which had been treated for 20 h, the elevated rate of actin synthesis declined to the control value within ca 4 h, as the actin-containing cytoskeletal components rearranged by CD recovered their normal morphology. Subsequently, actin synthesis was depressed below control values for a prolonged period; during recovery from 2 h treatment with CD, this depression was of much shorter duration. Re-addition of CD to cells after a 3 h recovery period again induced the cytoskeletal alterations characteristic of CD treatment but did not reverse the prior decline in the rate of actin synthesis. In HEp-2 cells treated with cycloheximide during exposure to CD for 20 h, the relative rate of actin synthesis measured after removal of cycloheximide was twofold higher than with CD alone and such cells exhibited a twofold slower decline in the rate of actin synthesis during recovery from CD in the continued presence of cycloheximide. These effects of cycloheximide, which resemble observations on “super-induction”, suggest that actin synthesis in CD-treated and recovering HEp-2 cells may be regulated by a repressor protein. The possibility that the proposed repressor protein is actin and that actin may thus be a feedback inhibitor of its own synthesis is discussed.     

Localization of the circadian pacemakers of Hemideina thoracica (Orthoptera; Stenopelmatidae)

The location of the circadian pacemakers of the orthopteran Hemideina thoracica (White) has been investigated through observation of the effects of surgical removal of brain tissues (principally optic lobes and tracts) on free-running and entrained locomotor rhythms. Bilobectomy and severance of optic tracts invariably resulted in arrhythmicity, whereas rhythmicity was sustained following unilateral lobectomy, generally with increases in the free-running period (FRP) and decreases in both the active-phase lengths and activity-to-rest ratios of the rhythm. Bilobectomized subjects could be entrained by temperature cycles, but exhibited no transients or residual rhythmicity, indicating that temperature brought about a direct response or masking effect. These results support the hypothesis that the circadian locomotor pacemakers of Hemideina are located within each optic lobe, and that there are no extraoptic centers for the control of the timing of locomotor activity. Although confirmation of the pacemaker role of the optic lobes requires transplantation of the tissues, the conclusion may be drawn by inference from other studies (e.g., Leucophaea maderae--Page, 1983; Gryllus bimaculatus--Tomioka and Chiba, 1986). Light entrainment continued after surgical binding and blackening of the compound eyes and ocelli, supporting the view that direct illumination of neural tissue through the cuticle may be one possible pathway for light entrainment.     

Phenotypic and karyotypic transitions in the spontaneous transformation of a rat cell line

After 20-50 transfers, a rat myofibroblast line, Hmf-n, 'spontaneously' transforms to an established (immortalized) line of smaller, rapidly cycling fibroblastoid cells (tHmf-f). From these 1 degree transformants, colonies of larger, slower growing anchorage-independent (tHmf-e) cells of epithelioid phenotype emerge. Both transformants grow in low serum and low calcium media, but the tHmf-f cells are highly tumorigenic in nude mice, have diminished substrate adhesivity, and limited anchorage independence, whereas tHmf-e are less tumorigenic, firmly substrate adherent, and markedly anchorage independent. Most tHmf-f are trisomic; most tHmf-e transformants are hypodiploid, a third are tetraploid, and all have chromosomal abnormalities, but no trisomy. Hmf-n cells have polar stress fiber arrays terminating in vinculin adhesion plaques, colinear extracellular fibronectin matrices, and linear non-coincident deposits of fodrin. Microtubules (mt) and vimentin-intermediate filaments (IF) parallel the actin cables. Stress fibers of the tHmf-f are moderately reduced, their vinculin adhesion plaques and fibronectin matrices intact; fodrin is diffuse. Mts and IFs are normal and axial. Most epithelioid tHmf-e have no stress fibers, adhesion plaques, or extracellular fibronectin; instead, dense actin microfilament meshworks are attached to plasma membrane, as is fodrin. Mt and IF are radial. Both transformed phenotypes are stable over greater than 300 continuous passages. The differentiation-inducing agents DMSO, cyclic AMP, 5-azacytidine, and mezerein, were ineffective in normalizing shape or cytoskeleton of transformed Hmf, and butyrate was selectively toxic to 50% of tHmf-e. But hydrocortisone induced striking polarization, and increase in number, and alignment of stress fibers of both tHmf-f and tHmf-e. Growth, anchorage, cytoskeletal arrangements, and tumorigenic potential are not closely correlated in these stable, spontaneously transformed lines of distinct pheno- and karyotype originating from the same normal parental cell, suggesting independent acquisition of properties associated with transformation.     

Frostbite and the significance of a temperature-sensation scale in field and laboratory conditions

The literature indicates that rapid thawing in water between 42 and 45C provides the best prognosis for frostbitten limbs.A temperature:sensation scale for the normal hand was derived and tested in laboratory and field conditions. The scale permitted the preparation of water suitable for frostbite treatment by the majority of subjects without access to a thermometer. The maximum observed inaccuracy was 1.2C outside the preferred range but was well withing the safe range for treatment.

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Zusammenfassung Die beste Prognose zur Behandlung von Erfrierungen bietet nach den Erfahrungen in der Literatur das schnelle Auftauen von erfrorenen Gliedern bei Wassertemperaturen von 42–45C. Für die gesunde Hand wurde eine Temperatur-Wahrnehmungsskala aufgestellt und unter verschiedenen Bedingungen im Laboratorium und im Freien geprüft. Ohne Zugang zu einem Thermometer gestattete die Skala in der Mehrzahl der FÄlle die ErwÄrmung von Wasser auf Temperaturen, die für die Behandlung von Erfrierungen geeignet sind. Die grösste Ungenauigkeit zwischen SchÄtzwert und Thermometerwert war 1.2C ausserhalb des gewünschten aber noch innerhalb des für die Behandlung zulÄssigen Bereiches.

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Résumé D'après les indications de plusieurs auteurs, le meilleur traitement de membres gelés consiste à les dégeler dans de l'eau à une temperature entre 42 et 45C. Une échelle température-sensation pour la main normale a été mise au point et vérifiée au laboratoire ainsi qu'en plain air.Cette méthode a permis la préparation d'eau en vue d'un traitement de gelures sans faire usage d'un thermomètre dans la plupart des cas. L'erreur maximum observée était de 1,2 C sans que les limites de l'intervalle propice à un traitement ne soient jamais dépassées.

     

Lipid composition and food quality of some freshwater phytoplankton for cladoceran zooplankters

The nutritional value of several planktonic algae was testedby means of feeding trials with three cladoceran zooplankters.The algae were monocultures and included two blue-greens, fourgreens and four flagellates with a size range of 5–48µm. The specific growth rates of the zooplankters werechosen as the measure of the nutritional value of the algae.The three cladocerans showed large differences in growth ratein the different algae, but the two cryptomonads were withoutdoubt best suited as food for all. The fatty acid compositionfor the cryptomonads were different from the other algae. Theycontained high percentages of the polyunsaturated fatty acids20:5æ3 (EPA) and 22:6æ3 (DHA), which also are commonin fish. It is suggested that the lipid composition is a probablefactor determining the nutritional quality of the algae.     

Genetic variation,breeding system evolution,and conservation of the narrow sand dune endemic Stellaria arenicola and the widespread S. longipes (Caryophyllaceae)

Genetic variation was examined by electrophoresis in 14 populations of Stellaria arenicola, an endemic of the Athabasca sand dunes in northern Saskatchewan, Canada, and seven populations of S. longipes, its progenitor. Three of the 5. longipes populations were sympatric with the endemic. Populations of the endemic were found to have fewer alleles per polymorphic locus (2.21 vs. 2.37), fewer polymorphic loci (29.9 vs. 33.8), and lower genetic diversity (0.087 vs. 0.107) than populations of the progenitor. Genetic identities for all pairs of populations were high (0.932 to 1.000). The endemic had one novel allele and shared ten alleles with progenitor populations from the sand dunes that were not found in other populations of S. longipes. Populations of both species were found to partition most of their genetic variation within populations. An investigation of the multilocus outcrossing rates revealed that S. arenicola had higher rates of selling and biparental inbreeding than S. longipes. This study suggests that partial genetic isolation through a shift in the breeding system, in addition to previously reported strong directional selection, has been important in the sympatric evolution of the endemic S. arenicola. The close genetic relationship between populations of S. arenicola and S. longipes found on the Athabasca sand dunes supports the suggestion that the endemic evolved while sympatric to the gene pool of the progenitor species that is found presently in the region.