Prolactin,hypercalcemia and corpuscles of stannius in seawater eels

In intact eels in sea water (SW), ovine prolactin (PRL) treatment induces hypercalcemia, but its mechanism of action, which is discussed, remains to be defined. Corpuscles of Stannius (CSt) are modified simultaneously: two cell categories then become evident. The first cell type (type 1) predominates; it has an oval shape and large granules, it shows a nuclear and nucleolar hypertrophy and a mitotic activity, and appears greatly stimulated by PRL; it may elaborate a hypocalcemic factor (hypocalcin) which would compensate for the PRL-induced hypercalcemia. A similar effect, although slightly less intense, is detected in hypophysectomized-PRL treated eels in SW. A second cell type (type 2), is more elongated, smaller in size, and has an oval nucleus and fine granules. Scarcely less active in SW, it is significantly stimulated by PRL despite an increased blood sodium and potassium level. This experiment does not help to clarify its function.     

Lactose repressor hinge domain independently binds DNA

The short 8–10 amino acid “hinge” sequence in lactose repressor (LacI), present in other LacI/GalR family members, links DNA and inducer‐binding domains. Structural studies of full‐length or truncated LacI‐operator DNA complexes demonstrate insertion of the dimeric helical “hinge” structure at the center of the operator sequence. This association bends the DNA ～40° and aligns flanking semi‐symmetric DNA sites for optimal contact by the N‐terminal helix‐turn‐helix (HtH) sequences within each dimer. In contrast, the hinge region remains unfolded when bound to nonspecific DNA sequences. To determine ability of the hinge helix alone to mediate DNA binding, we examined (i) binding of LacI variants with deletion of residues 1–50 to remove the HtH DNA binding domain or residues 1–58 to remove both HtH and hinge domains and (ii) binding of a synthetic peptide corresponding to the hinge sequence with a Val52Cys substitution that allows reversible dimer formation via a disulfide linkage. Binding affinity for DNA is orders of magnitude lower in the absence of the helix‐turn‐helix domain with its highly positive charge. LacI missing residues 1–50 binds to DNA with ～4‐fold greater affinity for operator than for nonspecific sequences with minimal impact of inducer presence; in contrast, LacI missing residues 1–58 exhibits no detectable affinity for DNA. In oxidized form, the dimeric hinge peptide alone binds to O1 and nonspecific DNA with similarly small difference in affinity; reduction to monomer diminished binding to both O1 and nonspecific targets. These results comport with recent reports regarding LacI hinge interaction with DNA sequences.     

A Single Administration of CRISPR/Cas9 Lipid Nanoparticles Achieves Robust and Persistent In Vivo Genome Editing

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Differential Effect of Sodium and Magnesium on the Pituitary of Goldfish Adapted to Calcium-Free Environments

Adaptation to deionized water (DW) affects several cell types in the goldfish. The pars intermedia PAS-positive cells are highly stimulated. Their low response or the absence of changes in goldfish kept in 1/3 Ca-free seawater (SW) and in Ca-free SW-adapted eels, respectively, suggest that sodium and/or magnesium are interfering. To test this hypothesis, young goldfish were adapted to DW supplemented or not with sodium (50 and 140 mM) for 8 and 16 days or with magnesium (16.5 and 50 mM) for 16 and 30 days. Cytological and morphometric studies of the pituitary showed that prolactin (PRL) cell activity was reduced by sodium. Thyrotropic (TSH) cells were stimulated. The activity of melanocyte-stimulating (MSH) cells increased in DW + Na+. Stimulation of the pars intermedia PAS+ cells in DW was partly inhibited by adding sodium; the cellular and nuclear areas increased only moderately, the endoplasmic reticulum (ER) was not conspicuous and mitotic activity disappeared. In DW + Mg²⁺ the activity of PRL, TSH and MSH cells tended to be lower after a long-term adaptation. The response of the PAS+ cells was as high as that noted in DW; complete degranulation, enlargement of the ER and important mitotic activity. Differential responses to Na+ and Mg²⁺ are not due to pH differences in the solutions. External sodium is able then to reduce the response of the PAS+ cells in a Ca-free environment, while magnesium is not inhibitory. Other cell types are also affected by high levels of Na+ and Mg²⁺.     

Effect of 17 α-Methyltestosterone on the Cytology of the Pituitary and the Liver,and on Plasma Electrolytes in Male Silver Eels

Male silver eels kept in fresh water were injected with 17α-methyltestosterone (MT). They received 5, 10, 15 or 20 injections (8 μg/g every other day) or 6 or 9 injections (4 μg/g or 2.5 μg/g once a week). Compared to solvent-injected controls, the treated eels showed changes in the pituitary: prolactin, corticotropic, thyrotropic and melanotropic cells were stimulated. Gonadotropic cells had enlarged nuclei, but synthesis and storage of granules and large globules were not evident, despite the progressive testicular maturation. High doses of MT induced a liver hypertrophy, a drop in plasma osmolarity and sodium level. An increase in plasma calcium level is possibly involved in the reduced activity of PAS-positive Ca-sensitive cells of the pars intermedia. These responses, previously observed in estradiol (E2) treated eels, may suggest that MT is aromatized in the brain and/or the pituitary. However, the considerable stimulation of GTH cells which always occurs in E2-treated eels is absent in MT-treated eels. With lower doses of MT, changes in the pituitary and the testis were similar to those produced by high doses, although the Ca-sensitive cells were more active. The decrease in plasma osmolarity and sodium level was less marked, plasma calcium was slightly reduced and the liver was not modified. There is no clear evidence in favour of E2 production. These data suggest that MT may stimulate the gonad directly, in agreement with biochemical data by Dufour et al. (1983).     

Gender effects on trapezius surface EMG during delayed onset muscle soreness due to eccentric shoulder exercise.

The purpose of this study was to investigate gender-specific motor control strategies during eccentric exercise and delayed onset muscle soreness (DOMS) in the shoulder region. Twelve healthy males and females participated in the study. Eccentric shoulder exercises were conducted on the dominant shoulder while the other side served as control. The exerted force, range of shoulder elevation, rating of perceived exertion, pain intensity, and surface electromyography (EMG) from the trapezius muscles were recorded and analyzed. A significant decrease in exerted force during exercise was only found in males despite similar rating of perceived exertion among genders. During eccentric exercise: males showed increasing root mean square (RMS) of the EMG while a decrease occurred for females, no difference between genders in mean power frequency of the EMG were seen. During static and dynamic contractions: no differences between genders in pain intensity or RMS were observed; RMS of the exercised side were lower than that of the control side (P<0.05) at 24 h after exercise. The results indicated a more prominent muscle fatigue resistance in females compared with males and mobilization of different muscle activation strategies during eccentric exercise. A protective adaptation to DOMS, i.e. decrease in RMS values was found with no gender differences.     

Evaluation of Micro Complement Fixation Tests for Antibodies Against Group A Streptococcal M and M-Associated Antigens in Rabbit and Human Sera

The microslide gel-diffusion and macro-tube agglutination techniques to detect Brucella canis antibodies in dogs were compared. Sera from dogs experimentally infected with B. canis and a random sample of dog sera with unknown histories of exposure to this organism were examined. The results of the gel-diffusion method employing specific rough Brucella saline-extract antigens of B. canis and Brucella ovis were comparable to those obtained by the tube agglutination test. The easily prepared, stable R antigen in the freeze-dried form offers a convenient, simple, and reliable diagnostic method for the serological detection of canine brucellosis by the gel-diffusion test.     

Sporulation and Enterotoxin Production by Mutants of Clostridium perfringens

The ability of Clostridium perfringens type A to produce an enterotoxin active in human food poisoning has been shown to be directly related to the ability of the organism to sporulate. Enterotoxin was produced only in a sporulation medium and not in a growth medium in which sporulation was repressed. Mutants with an altered ability to sporulate were isolated from an sp(+) ent(+) strain either as spontaneous mutants or after mutagenesis with acridine orange or nitrosoguanidine. All sp(0) (-) mutants were ent(-). Except for one isolate, these mutants were not disturbed in other toxic functions characteristic of the wild type and unrelated to sporulation. A total of four of seven osp(0) mutants retained the ability to produce detectable levels of enterotoxin. None of the ent(-) mutants produced gene products serologically homologous to enterotoxin. A total of three sp(-) mutants, blocked at intermediate stages of sporulation, produced enterotoxin. Of these mutants, one was blocked at stage III, one probably at late stage IV, and one probably at stage V. A total of three sp(+) revertants isolated from an sp(-) ent(-) mutant regained not only the ability to sporulate but also the ability to produce enterotoxin. The enterotoxin appears to be a sporulation-specific gene product; however, the function of the enterotoxin in sporulation is unknown.