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Biomedical subjects

S Klaus

Publications and source records attributed to S Klaus.

At least 19 recordsLinked to original sources
PubMed1992

Hibernoma formation in transgenic mice and isolation of a brown adipocyte cell line expressing the uncoupling protein gene.

Transgenic mice were produced containing the adipocyte-specific regulatory region from the adipocyte P2 (aP2) gene linked to the simian virus 40 transforming genes. Most of the transgenic mice developed brown fat tumors (hibernomas) in their interscapular brown adipose tissue. Hibernoma formation was noticeable in some of the mice as early as 1 day after birth and most of the mice developed very large tumors by 1 month of age. All of the tumor tissue expressed the brown fat-specific uncoupling protein (UCP) gene as well as the aP2 gene. Several of the tumors have been used to establish cultured cell lines and at least one of these lines can be induced to differentiate into brown adipocytes. The cultured adipocytes express mRNA for UCP upon stimulation with N6,O2'-dibutyryladenosine 3',5'-cyclic monophosphate, norepinephrine, isoproterenol or D7114, a beta 3 adrenergic agonist. Thus, regulation of the key thermogenic gene UCP can now be studied in an established cell line.

Adipose Tissue, Brown↗
PubMed1992

Antibodies selected from whole antiserum by fusion proteins as tools for the study of the topology of mitochondrial membrane proteins. Evidence that the N-terminal extremity of the sixth alpha-helix of the uncoupling protein is facing the matrix.

The reactivity to freeze-thawed mitochondria or submitochondrial particles of a whole antiserum raised against the uncoupling protein has been investigated. Incubation with freeze-thawed brown adipose tissue mitochondria trapped antibodies reactive toward accessible parts of the uncoupling protein. One-third to one-half of antibodies against uncoupling protein which were present in the serum remained free. These antibodies were highly reactive with the vesicles obtained by sonication of mitochondria, in which the matricial side of the inner membrane was made accessible. To define epitopes recognized by the antiserum, different fusion proteins made up of MalE protein and uncoupling protein fragments were used. Immunoaffinity chromatography, using an immobilized purified fusion protein containing amino acids 253 to 290 of uncoupling protein, selected antibodies specifically directed against this part of the protein. A more precise localization of the main epitope recognized by these antibodies is proposed. These purified antibodies reacted with the protein only in submitochondrial particles, indicating a matricial orientation of this epitope. This result, associated with other data concerning uncoupling protein or related mitochondrial carriers such as the ADP/ATP translocator and the phosphate carrier, allowed us to determine the orientation of the sixth alpha-helix of the uncoupling protein.

Amino Acid Sequence↗
PubMed1992

Human cutaneous leishmaniasis: in-vitro parasite--mononuclear cell interactions in immune and naive individuals.

The aim of this study was to compare the ability of Leishmania parasites to survive in mononuclear cells from immune individuals with their ability to survive in cells from naive individuals. For this purpose we established an in vitro system based on the co-culture in suspension of human peripheral blood leukocytes derived from immune and naive subjects and L. major promastigotes. The proportion of monocytes containing intracellular parasites and the number of amastigotes per 100 infected monocytes (parasite burden) were determined 24 and 72 h after in-vitro infection. The proportion of infected cells from naive individuals did not change, and the number of amastigotes either did not change or increased by 1.2 to 1.7-fold between 24 and 72 h incubation. In contrast, in the immune subjects, the proportion of infected monocytes 24 h after infection was lower than in the naive individuals, and a 30-90% decrease in both the proportion of infected monocytes and the parasite burden was observed after 72 h incubation. Based on these results, three characteristics of leishmanicidal activity of mononuclear cells from immune individuals were determined: (a) the proportion of infected monocytes 24 h after infection was lower than 22%; (b) there was a decrease of more than 30% in the proportion of infected monocytes between 24 and 72 h after infection; and (c) there was a significant decrease in the number of amastigotes between 24 and 72 h after infection. The results of this study demonstrate an enhanced leishmanicidal activity of mononuclear cells from immune individuals.

Humans↗
PubMed1991

Clinical observations related to head lice infestation.

Living lice and eggs were found in 19% of 2643 children examined in Israel. Boys and girls were equally infested. Bite reactions, pruritus, excoriations, lymphadenopathy, and conjunctivitis were the most common signs and symptoms. Bite reactions, pruritus, excoriations, and conjunctivitis were found more frequently in infested children than in noninfested children. Lymphadenopathy, eczema, influenza, herpes, crusts behind ears, blepharitis, seborrhea, psoriasis, erythema, vitiligo, secondary infections, and scars were equally common in both groups. The sequence of the skin reactions of a volunteer to continuous exposure to lice bites was as follow: phase I, no clinical symptoms; phase II, papules accompanied by pruritus of medium intensity; phase III, wheal formation immediately after the bite, followed by a delayed papular reaction and intense itching; phase IV, papular reaction with diminished reactivity of the skin and mild pruritus. Healed bite reactions reappeared when other parts of the skin were again exposed to the lice.

Animals↗
PubMed1991

Production of interferon gamma in cultures of whole blood obtained in the course of and after healing of cutaneous leishmaniasis.

Whole-blood cultures from individuals with cutaneous leishmaniasis due to Leishmania major, tested under field conditions, showed a good correlation between the production of interferon-gamma and total blood lymphocyte proliferation (TLP), assayed by response to specific leishmania antigen. A group of patients, re-tested on several occasions, converted to positive in both parameters of cell mediated immunity (CMI) at the same time. It is suggested that the TLP assay and the production of interferon-gamma detect similar CMI activity in human cutaneous leishmaniasis. The measurement of interferon-gamma production has the advantage over the TLP assay that the supernatants of the cultures can be frozen and stored for prolonged periods before the assay is performed. For this reason, this could present a suitable method for field studies.

Antigens, Protozoan↗
PubMed1991

Development of Phodopus sungorus brown preadipocytes in primary cell culture: effect of an atypical beta-adrenergic agonist, insulin, and triiodothyronine on differentiation, mitochondrial development, and expression of the uncoupling protein UCP.

A new cellular model for the study of brown adipocyte development and differentiation in vitro is presented. Preadipocytes isolated from brown adipose tissue (BAT) of the djungarian dwarf hamster Phodopus sungorus are able to proliferate and differentiate in vitro into true brown adipocytes able to express the BAT marker protein the uncoupling protein (UCP). Whereas basal UCP expression is very low, its mRNA levels as well as the UCP detected by immunoblotting are highly increased by beta-adrenergic stimulation. The novel, atypical beta-adrenergic compound D7114 (ICI Pharmaceuticals, Macclesfield, Cheshire, England) was found to increase the number of adipocytes as well as UCP mRNA and UCP content of mitochondria, indicating the involvement of an atypical or beta 3 receptor. Insulin was found to play an important role in brown adipocyte differentiation and mitochondrial development, whereas T3 seemed to be implicated more directly in UCP expression. In a defined, serum-free medium a synergistic stimulatory action of insulin and T3 on UCP expression was found, which seems to involve a pathway different from that of beta-adrenergic UCP stimulation.

Adipose Tissue, Brown↗
PubMed1990

The mRNA of protein disulfide isomerase and its homologue the thyroid hormone binding protein is strongly expressed in adipose tissue.

The thyroid hormone 3,3',5-triiodothyronine (T3) plays an important role in the differentiation of adipocytes, as well as in the thermogenic activity of brown adipose tissue. Recently a T3 binding protein (T3BP), which is associated with plasma membranes, has been isolated and cloned from liver. It proved to be homologous to the multifunctional enzyme protein disulfide isomerase (PDI), which is involved in posttranslational modifications of secretory proteins. In this study we investigated the T3BP/PDI mRNA expression in white and brown adipose tissue of rat and bovine, as well as in several rodent adipose cell lines at various states of differentiation. T3BP/PDI mRNA expression was found in white and brown adipose tissue, as well as in preadipocytes and adipocytes at all states of differentiation. Comparison to other tissues (liver, kidney, heart, brain) revealed that its expression was highest in white fat. No modulation of T3BP/PDI mRNA corresponding to different adaptational or developmental situations could be detected in adipose tissue.

Adipose Tissue↗
PubMed1990

Expression of the brown fat mitochondria uncoupling protein in Xenopus oocytes and important into mitochondrial membrane.

Non shivering thermogenesis of brown adipose tissue is due to the uncoupling protein (UCP), located in the inner mitochondrial membrane, which functions as a proton translocator and can thus uncouple mitochondrial respiration. We describe here the expression of UCP in Xenopus laevis oocytes after injection of UCP mRNA, which was transcribed in vitro. UCP seems to be correctly transported into mitochondria and integrated into the membrane, but we were not able to establish definitely the functionality of this UCP. We conclude that this expression system could be suitable for the study of the mitochondrial import mechanism but not for the examination of physiological properties of UCP.

Adipose Tissue, Brown↗
PubMed1990

Stable expression of functional mitochondrial uncoupling protein in Chinese hamster ovary cells.

The mitochondrial uncoupling protein (UCP) is a membranous proton carrier exclusively synthesized in brown adipocytes. The cDNA for the rat UCP was placed in an expression vector and transfected into mammalian cells. Its expression was tested in transiently transfected CHO cells. In these cells the UCP was detected in mitochondria by using antibodies. Permanent expression of the UCP was achieved in stable transformed CHO cell lines. In these cells the UCP was characterized in mitochondrial membranes, by using antibodies and hydroxyapatite purification. The protein expressed in CHO cells displayed the functional characteristics of brown adipocyte UCP. It induced the uncoupling of respiration in isolated CHO mitochondria. The membrane potential of transformed mitochondria was also significantly lowered, as a result of the proton translocating activity of the UCP. GDP is known to inhibit the proton pathway in brown fat mitochondria. Addition of GDP to CHO mitochondria containing UCP resulted in a recoupling of respiration and an increase in membrane potential. Thus we conclude that functional UCP is expressed in CHO cells and that the insertion of the UCP alone in any mitochondria is sufficient to induce the uncoupling of respiration. This approach should allow studies on the structure-function relationship of the UCP and of several other related mitochondrial carriers.

Adipose Tissue, Brown↗
PubMed1990

Epidemiological studies on head lice infestation in Israel. I. Parasitological examination of children.

Of 3,079 children in a medium-sized community near Jerusalem, 11.2% were found to be infested with living lice and eggs and another 23.4% had only nits. Boys and girls were equally infested. The incidence of infestation was highest among children 4-11 years of age. About 78% of the infested children had from one to ten lice. Approximately 80% of the infested children had nits that were 2-5 cm away from the scalp. Boys with medium length hair and girls with short hair showed the highest incidence of lice infestation. Children with brown and red hair were more infested than those with black and blond hair. Boys and girls with straight and wavy hair were more infested than those with curly and frizzy hair.

Adolescent↗
PubMed1990

Increased nonshivering thermogenesis, brown fat cytochrome-c oxidase activity, GDP binding, and uncoupling protein mRNA levels after short daily cold exposure of Phodopus sungorus.

In their natural environment, burrowing rodents experience rather fluctuating ambient temperatures and are acutely cold exposed only for short periods outside their burrows. The effect of short daily cold exposure on basal metabolic rate, nonshivering thermogenesis, brown fat thermogenesis, and uncoupling protein mRNA was studied in the Djungarian hamster, Phodopus sungorus. They were kept at 23 degrees C and exposed to 5 degrees C daily either for one 4-h period or twice for 2 h (in 12-h intervals). At the same time control hamsters were kept continuously either at thermoneutrality (23 degrees C) or at 5 degrees C. Two 2-h cold exposures daily were sufficient to increase basal metabolic rate and nonshivering thermogenesis to the same level as continuous cold exposure, whereas one 4-h cold period per day did not result in a significant increase of both parameters. Brown fat thermogenesis (as measured by cytochrome-c oxidase activity and GDP binding to the mitochondrial uncoupling protein) increased to the same extent by both treatments with short daily cold exposure. However, this increase was less than in the chronically cold-exposed hamsters. A similar result was found for uncoupling protein mRNA: both short-term cold-exposed hamsters increased uncoupling protein mRNA levels to a similar extent, but less than after chronic cold treatment. It is concluded that short daily cold exposures are sufficient to cause adaptive increases of the capacity of metabolic heat production as well as brown fat thermogenic properties.

Adipose Tissue↗
PubMed1990

Inhibition of 5'-deiodination of thyroxine suppresses the cold-induced increase in brown adipose tissue messenger ribonucleic acid for mitochondrial uncoupling protein without influencing lipoprotein lipase activity.

Young adult male and female Djungarian hamsters were exposed to ambient temperatures of 23 or 0 C for 12 h; half of the animals in each group were treated with iopanoic acid to suppress the peripheral conversion of T4 to the thermotropically active thyroid hormone T3 by the enzyme 5'-deiodinase (5'D). Brown adipose tissue (BAT) mRNA for uncoupling protein (UCP), BAT lipoprotein lipase (LPL) activity, and 5'D activity were measured at the conclusion of the study. A temperature of 0 C produced large rises in 5'D and LPL activities and a similar large increase in UCP mRNA within the 12-h exposure period. When 5'D activity was inhibited with iopanoic acid, mRNA for UCP was reduced, while LPL activity was unaffected. The results show that the optimal production of mRNA for BAT UCP depends on the availability of T3; however, T3 is not required for the cold-induced activation of LPL activity in BAT.

Adipose Tissue, Brown↗
PubMed1989

Penile basal cell carcinoma. Report of a case and review of the literature.

We describe a 69-year-old man with penile basal cell carcinoma and review the literature on this condition. Basal cell carcinomas of the penis are rare, and there are only 26 other reported cases. There is no documented case of metastatic penile basal cell carcinoma. The one reported case of metastatic penile basal cell carcinoma actually represents urethral transitional cell carcinoma. Therefore conservative therapy with local removal is adequate treatment, and more extensive procedures are unnecessary. Ultraviolet light exposure may be an important etiologic factor for basal cell carcinomas located on sun-exposed areas. The occurrence of basal cell carcinomas on non-sun-exposed areas raises the possibility of other, not yet defined, etiologic agents.

Aged↗
PubMed1989

Short photoperiod and cold activate brown fat lipoprotein lipase in the Djungarian hamster.

Activation of lipoprotein lipase (LPL) in brown fat of the Djungarian hamster ensures substrate supply for nonshivering thermogenesis. Cold acclimation [5 degrees C ambient temperature (Ta)] in long photoperiod [light-dark (LD) 16:8] increased brown fat LPL activity from 4.7 to 22.7 nmol oleate.mg-1.min-1 within 1 day. Slight reduction of this high LPL activity was observed during prolonged cold exposure for up to 4 wk. Deacclimation (transfer from 5 to 23 degrees C Ta) caused total inactivation of brown fat LPL within 4 days. Short photoperiod (LD 8:16) also stimulates brown fat LPL activity. Its effect can be summarized by three different observations. At thermoneutrality short photoperiod elevated LPL activity to 7.1 instead of 4.7 nmol.mg-1.min-1 observed in long photoperiod. Second, at low ambient temperature short photoperiod reinforces the effect of cold acclimation. Maximum LPL activity of cold-exposed Djungarian hamsters averaged 23.7 nmol.mg-1.min-1 in long photoperiod and was elevated to 35.5 nmol.mg-1.min-1 in short photoperiod. Third, inactivation of LPL during deacclimation was delayed in short photoperiod. These results demonstrate that photoperiod as well as ambient temperature may be cooperatively used as environmental cues for seasonal acclimation of brown fat substrate supply for nonshivering thermogenesis.

Acclimatization↗
PubMed1988

Identification by heteroduplex analysis of an invertible element (Min) common among IncM group plasmids.

The analysis of heteroduplexes between EcoRI digested DNA derived from IncM plasmids in the transfer-off and in the transfer-on state revealed an internal loop of about 1 kbp on a distinct large EcoRI fragment. The presence of an invertible element (Min) is suggested which enables the formation of conjugative pili at 30 degrees C, but switches off the pilus formation at 37 degrees C incubation temperature.

DNA, Bacterial↗
PubMed1988

Characterization of a thermosensitive mutant of Streptomyces hygroscopicus defective in both DNA and RNA syntheses.

A stable temperature sensitive mutant of Streptomyces hygroscopicus JA6599 defective in both DNA and RNA syntheses is described. The mutant ts35 is characterized by an immediate stop of DNA synthesis and continued protein synthesis after transfer to restrictive temperature. The reinitiation of DNA synthesis begins immediately after a return to the permissive temperature. This kinetics of macromolecular synthesis at restrictive temperature appears to share similarities with a defect in the DNA elongation process, as described for Escherichia coli (Carl 1970, Hanna and Carl 1975). The simultaneous stop of both DNA and RNA syntheses may be caused by an additional mutational event affecting also the RNA synthesis. The data were discussed with respect to similar results in E. coli.

DNA, Bacterial↗