Mechanism of the enhancing effect of sorbitol on ileal Ca uptake in rat enterocytes

The effect of sorbitol on Ca uptake by isolated ileal epithelial cells was investigated. Intestinal cells were isolated from rat ileum by mechanical vibration.⁴⁵Ca uptake was approximately 2 times higher in cells exposed to 200 mM sorbitol ofd-alanine than in control cells. This enhancing effect of sorbitol on percentage Ca uptake decreased with increasing Ca concentrations in the incubation medium suggesting an effect on Ca entry velocity. The addition of 10 M nifedipine or 200 M verapamil to the incubation medium was devoid of any effect on Ca uptake in ileal cells, whereas 100 M trifluoperazine or chlorpromazine abolished the stimulatory effect of sorbitol. Finally, the effect of sorbitol on isolated cells was independent of a measurable change of cellular ATP content. In conclusion, the stimulatory effect of sorbitol on ileal Ca uptake is probably exerted through mechanisms other than an increase in intracellular ATP concentration. Sorbitol may enhance enterocyte Ca transport via a direct interaction with calmodulin and/or the Ca pump. It may also exert its effect through an inhibition of the basolateral Na Ca exchanger.     

An outbreak of trichinellosis in farmed wild boar in Finland

Nine farmed wild boar out of 25 slaughtered from a single farm were condemned at meat inspection because of trichinellosis. With RAPD-PCR, Trichinella spiralis was identified in all positive wild boar. Out of the available serum samples (n=7), all wild boar which had failed the meat inspection showed seroconversion in ELISA and Western blotting, as did one additional animal which had passed the inspection. The animals became infected during an invasion of rats from an improperly closed dump near the farm. Unfortunately, by the time trichinellosis was discovered in the wild boar, the invasion had already been brought under control; thus, no samples from rats were available. However, having lived through the rat invasion was shown to be a risk factor for trichinellosis in wild boar (relative risk, RR=6.3). In wildlife samples from surrounding areas, sylvatic trichinellosis was found to be very common (74%; n= 19 red foxes). Intriguingly, the prevalent species in trichinella-positive foxes differed from that in wild boar, Trichinella nativa and T. spiralis being found in 12 foxes and in one fox, respectively.     

Systemic and mucosal antibody responses specific to SARS-CoV-2 during mild versus severe COVID-19

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The linear plasmid pSA3239 is essential for the replication of the Streptomyces lavendulae subsp. lavendulae CCM 3239 chromosome

We previously reported the complete genome of Streptomyces lavendulae subsp. lavendulae CCM 3239, containing the linear chromosome and the large linear plasmid pSA3239. Although the chromosome exhibited replication features characteristic for the archetypal end-patching replication, it lacked the tap/tpg gene pair for two proteins essential for this process. However, this archetypal tpgSa-tapSa operon is present in pSA3239. Complete genomic sequence of the S. lavendulae Del-LP strain lacking this plasmid revealed the circularization of its chromosome with a large deletion of both arms. These results suggest an essential role of pSA3239-encoded TapSa/TpgSa in the end-patching replication of the chromosome.     

Pyrford钢丝环扎加张力带内固定治疗髌骨骨折的生物力学分析

目的　测试Pyrford钢丝环扎加张力带内固定治疗髌骨骨折的生物力学特性 ,为临床治疗提供理论依据。方法　采集新鲜牛尸体膝关节标本 6具 ,制成髌骨骨折模型 ,用Pyrford钢丝环扎加张力带固定 ,并与克氏针张力带、单纯钢丝环扎进行对照比较。结果　Pyrford钢丝环扎加张力带内固定完全符合髌骨的生物力学性能 ,与克氏针张力带相当 ,能满足膝关节的强度 ,符合人体正常生理功能要求。结论　Pyrford钢丝环扎加张力带固定治疗髌骨骨折 ,具有优越的生物力学性能 ,且能达到解剖复位 ,操作简便 ,固定牢固 ,适应早期功能锻炼的目的     

Pathologic features of the femoral head in Mseleni disease

Severe but atypical osteoarthritic deformities were found in each of 12 femoral heads removed during total hip arthroplasty for Mseleni disease. Although degenerative and regenerative changes were present throughout the cartilage, there was a paucity of eburnation. Histomorphometric analysis of bone at the line of excision indicated that mild osteomalacia was present in four of the 12 specimens. The percentage of the endosteum occupied by osteoid was 9.7 +/- 7.96 (SD) in the patients with Mseleni disease, compared with 5.6 +/- 4.33 in seven African black and 4.1 +/- 2.13 in 13 American white control subjects. The mean thickness of the osteoid seams was not increased. The findings suggest that osteomalacia is not a major pathogenetic factor in Mseleni disease.     

Leydig cell types in primary testicular disorders

Ultrastructural study of testicular biopsy specimens from 67 adults with primary testicular disorders (Klinefelter's syndrome, XX male syndrome, Del Castillo's syndrome, and cryptorchidism) revealed the following four Leydig cell types: 1) normal or nearly normal Leydig cells with abundant smooth endoplasmic reticulum, mitochondria with tubular cristae, lipid droplets, and Reinke's crystals; 2) abnormally differentiated Leydig cells without either lipid droplets or Reinke's crystals but with altered mitochondria, concentric unfenestrated cisternae of smooth endoplasmic reticulum, and both paracrystalline and filamentous inclusions; 3) multivacuolated Leydig cells containing abundant lipid droplets; and 4) immature Leydig cells with scarce development of the smooth endoplasmic reticulum and mitochondria, and numerous cytoplasmic microfilaments. Abnormally differentiated Leydig cells might represent dysgenetic cells, whereas immature, normal, and vacuolated Leydig cells might represent three progressive stages in the Leydig cell cycle (undifferentiated, mature, and old involuting Leydig cells). An inverse correlation between the proportion of abnormal Leydig cells and testosterone levels was observed in each of these testicular disorders.     

The 49 K subunit of NADH: ubiquinone reductase (complex I) from Neurospora crassa mitochondria: primary structure of the gene and the protein

Summary The primary structure of the 49 K subunit of the respiratory chain NADH:ubiquinone reductase (complex I) from Neurospora crassa was determined by sequencing cDNA, genomic DNA and the N-terminus of the mature protein. The sequence lengths correlate to a molecular mass of 54002 daltons for the preprotein and 49239 daltons for the mature protein. The presequence consists of 42 amino acids of typical composition for sequences which target nuclear-encoded proteins into mitochondria. The mature protein consists of 436 amino acids and shows 64% similarity to a 49 K subunit of bovine heart NADH:ubiquinone reductase and 33% to a predicted translation product of an open reading frame in the chloroplast DNAs of Marchantia polymorpha and Nicotiana tabacum. Evidence for an iron-sulfur cluster in the subunit is discussed.     

Functional characterization of human natural killer cells responding to Mycobacterium bovis bacille Calmette-Guérin

The kinetics of activation and induction of several effector functions of human natural killer (NK) cells in response to Mycobacterium bovis bacille Calmette-Guérin (BCG) were investigated. Owing to the central role of monocytes/macrophages (MM) in the initiation and maintenance of the immune response to pathogens, two different experimental culture conditions were analysed. In the first, monocyte-depleted nylon wool non-adherent (NW) cells from healthy donors were stimulated with autologous MM preinfected with BCG (intracellular BCG). In the second, the NW cells were directly incubated with BCG, which was therefore extracellular. In the presence of MM, CD4+ T lymphocytes were the cell subset mainly expressing the activation marker, CD25, and proliferating with a peak after 7 days of culture. In contrast, in response to extracellular BCG, the peak of the proliferative response was observed after 6 days of stimulation, and CD56+ CD3- cells (NK cells) were the cell subset preferentially involved. Such proliferation of NK cells did not require a prior sensitization to mycobacterial antigens, and appeared to be dependent upon contact between cell populations and bacteria. Following stimulation with extracellular BCG, the majority of interferon-gamma (IFN-gamma)-producing cells were NK cells, with a peak IFN-gamma production at 24-30 hr. Interleukin (IL)-2 and IL-4 were not detectable in NK cells or in CD3+ T lymphocytes at any time tested. IL-12 was not detectable in the culture supernatant of NW cells stimulated with extracellular BCG. Compared to the non-stimulated NW cells, the NW cells incubated for 16-20 hr with BCG induced the highest levels of expression of apoptotic/death marker on the NK-sensitive K562 cell line. BCG also induced expression of the activation marker, CD25, and proliferation, IFN-gamma production and cytotoxic activity, on negatively selected CD56+ CD3- cells. Altogether, the results of this study demonstrate that extracellular mycobacteria activate several NK-cell functions and suggest a possible alternative mechanism of NK-cell activation as the first line of defence against mycobacterial infections.     

DNA vaccine encoding human immunodeficiency virus-1 Gag, targeted to the major histocompatibility complex II compartment by lysosomal-associated membrane protein, elicits enhanced long-term memory response

Antigen presentation by major histocompatibility complex type II (MHC II) molecules and activation of CD4+ helper T cells are critical for the generation of immunological memory. We previously described a DNA vaccine encoding human immunodeficiency virus-1 p55Gag as a chimera with the lysosome-associated membrane protein (LAMP/gag). The LAMP/gag chimera protein traffics to the MHC II compartment of transfected cells and elicits enhanced immune responses as compared to a DNA vaccine encoding native gag not targeted to the MHC II compartment. We have now investigated the long-term responses of immunized mice and show that the LAMP/gag DNA vaccine promotes long-lasting B cell- and CD4+ and CD8+ T-cell memory responses induced by DNA encoding non-targeted Gag decay rapidly and elicit very low or undetectable levels of gag DNA is sufficient to generate T-cell memory. Following this initial priming immunization with LAMP/gag DNA, booster immunizations with native gag DNA or the LAMP/gag chimera are equally efficient in eliciting B- and T-cell secondary responses, results in accordance with observations that secondary expansion of CD8+ cells in the boost phase does not require additional CD4+ help. These findings underscore the significance of targeting DNA-encoded vaccine antigens to the MHC II processing compartments for induction of long-term immunological memory.