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Biomedical subjects

J Klein

Publications and source records attributed to J Klein.

At least 73 records · Page 4Linked to original sources
PubMed2003

Regulation of gonadotropin releasing hormone release by neuropeptide Y at the median eminence during the preovulatory period in ewes.

The median eminence (ME) of the hypothalamus is known to be an important brain site where hypophysiotropic release might be regulated by excitatory and inhibitory signals impinging on their neuronal terminals. Since a role for neuropeptide Y (NPY) on preovulatory luteinizing hormone (LH) release has been suggested, we hypothesized that NPY might act at the ME to control preovulatory gonadotropin-releasing hormone (GnRH) release and thus the onset of the preovulatory surge of LH. To examine this possibility, we used the ewe as an animal model to determine: (a) immunocytochemical distribution of GnRH and NPY in the ewe ME; (b) changes in in vivo release of NPY and GnRH using ME push-pull cannula (PPC) perfusate samples, as well as in plasma LH, during the luteal, follicular and preovulatory phases of a synchronized estrous cycle, and (c) effects of ME perfusion of NPY or a Y1-NPY antagonist, or an NPY antiserum on in vivo release of ME-GnRH and plasma LH during a synchronized follicular phase. Immunolocalization reveals a dense plexus of beaded GnRH-containing neurites in the arcuate nucleus and in its vicinity, the pituitary stalk and the palisade. In contrast, a dense plexus of NPY-containing neurites occurs in the internal layer, with occasional fibers found in the intermediate and lateral external zone of the ME. In the area between the lateral internal and lateral external layers, both NPY and GnRH-containing processes were found, thus providing opportunities for synaptic and/or paracrine interactions between NPY- and GnRH-containing neurons. Hormonal analysis indicated that a synchronized preovulatory surge of LH is elicited within a 2-hour window by the sequential implantation and removal of silastic-encased estradiol (E2) or progesterone (P4) implants. In this paradigm, there was a parallel increase in ME release of both NPY and GnRH preceding the synchronized LH surge. The onset of this synchronized LH surge was advanced by ME perfusion of exogenous NPY and was both delayed and blunted by ME perfusion with the NPY antagonist (both were perfused through the PPC probe for 2 h, starting 2-3 h before the expected onset of the LH surge). In addition, NPY perfusion in the ME increases, while perfusion of the Y1-NPY antagonist or of the NPY antiserum decreases ME-PPC GnRH content and plasma levels of LH in early follicular ewes. Finally, perfusion of NPY antiserum during an ongoing LH surge disrupted LH release. These results suggest that interactions between NPY and GnRH neurons are important in controlling the timing, magnitude and maintenance of the preovulatory LH surge.

Animals↗
PubMed2003

GH is a positive regulator of tumor necrosis factor alpha-induced adipose related protein in 3T3-L1 adipocytes.

Tumor necrosis factor (TNF) alpha-induced adipose-related protein (TIARP) has recently been cloned as a TNFalpha-stimulated protein expressed in adipocytes. Its expression is differentiation-dependent and potentially involved in mediating TNFalpha-induced insulin resistance. To further characterize regulation of TIARP gene expression, 3T3-L1 adipocytes were treated with key hormones modulating insulin sensitivity and influencing adipocyte metabolism, and TIARP gene expression was determined by quantitative real-time RT-PCR. Interestingly, TIARP mRNA expression was stimulated almost 9-fold after 500 ng/ml GH were added for 16 h whereas addition of 10 microM isoproterenol, 100 nM insulin and 100 nM dexamethasone for 16 h significantly decreased TIARP gene expression to between 35 and 50% of control levels. In contrast, angiotensin 2 (10 microM) and triiodothyronine (1 microM) did not have any effect. The stimulatory effect of GH was time- and dose-dependent with stimulation occurring as early as 1 h after effector addition and at concentrations as low as 5 ng/ml GH. Moreover, pharmacological inhibition of Janus kinase 2 and p42/44 mitogen-activated protein kinase reversed the stimulatory effect of GH, suggesting that both signaling molecules are involved in activation of TIARP gene expression by GH. Furthermore, an increase of TIARP mRNA could be completely reversed to control levels by withdrawal of GH for 24 h. Taken together, these results show that TIARP is not only responsive to TNFalpha but also to important other hormones influencing glucose homeostasis and adipocyte metabolism. Thus, this factor may play an integrative role in the pathogenesis of insulin resistance and its link to obesity.

3T3-L1 Cells↗
PubMed2003

Understanding the effects of age on female fertility.

This review focuses on age-related changes occurring in the female reproductive system and their effect on reproductive material. Aging significantly impact the female reproductive system but, despite this fact, there is nowadays a trend to postpone pregnancy to pursue educational and vocational goals. Therefore a remarkable effort is now being made by the medical establishment to understand the effects of aging in women. The assessment and treatment of this decline in fertility is discussed, as well as assisted reproductive techniques and new therapeutic options.

Adult↗
PubMed2003

[Importance of neoadjuvant chemotherapy in the treatment of advanced bronchogenic carcinoma].

BACKGROUND: The basic task of induction (neo-adjuvant) therapy is elimination of occult micrometastatic dissemination found in some cases already in localized stages of non-small cell pulmonary cancer (stage I-IIIA NSCLC). An equally important effect is also cytoreduction in primary tumours which have before the local intervention an intact vascular supply. A difficult problem remains the correct selection of patients who from the long-term aspect may profit from such a procedure. MATERIAL AND METHODS: The authors evaluated perspectively aspects of oncological treatment and circumstances of surgical intervention after induction chemotherapy in 81 and 87 patients resp. in stage IIIA NSCLC evaluated before initiated neo-adjuvant chemotherapy. RESULTS: Complete remission was recorded in 4.9%, partial remission in 50.6%, stabilized disease in 23.5% and progression in 21% patients. Down-staging was recorded in 26%, 70.3% patients were indicated for surgery. In the group of 87 patients operated after induction therapy pneumonectomies predominated--41 (46%), only one operated patient died within 30 days after surgery (1.1%), complications were neither frequent nor serious. The median of survival after radical resection is 26 months. CONCLUSIONS: Neo-adjuvant chemotherapy by modern cytostatics is usually well tolerated and creates satisfactory conditions for successful complete resection. The operation proper may be more difficult but need not be associated with serious complications. By this treatment it is probably possible to influence long-term results not only in stage IIIA but to reduce also the risk of a later more remote metastatic dissemination in some patients operated in lower stages of lung cancer. Our present aim is to test parameters which will be able to predict possible failure of induction therapy, and seek factors predicting risk behaviour of the tumour also in lower stages (stage I and II TNM classification).

Antineoplastic Combined Chemotherapy Protocols↗
PubMed2003

Cardiac risk and perioperative management.

Cardiovascular complications are the major cause of perioperative and late morbidity and mortality in patients undergoing major vascular surgery. This is related to the frequent presence of underlying coronary artery disease (CAD). CAD may be asymptomatic because of reduced exercise capacity due to pre-existing non-cardiac conditions like stroke or claudication. Careful preoperative evaluation of CAD and perioperative management with beta-blockers and statins may offer the physician a unique opportunity to improve patients' perioperative and long-term outcome.

Adrenergic beta-Antagonists↗
PubMed2002

Linkage relationships of genes coding for alpha2-macroglobulin, C3 and C4 in the zebrafish: implications for the evolution of the complement and Mhc systems.

The alpha2-macroglobulin (A2M) and the complement components C3 and C4 are related proteins derived from a common ancestor. Theoretically, this derivation could have occurred either by tandem duplications of their encoding genes or by polyploidization involving chromosomal segments, a chromosome or the whole genome. In tetrapods the A2M-, C3- and C4-encoding genes are generally each located on a different chromosome. This observation has been interpreted as supporting their origin by polyploidization. We identified and mapped (with the help of a radiation hybrid panel of cell lines) the A2M, C3 and C4 loci in the zebrafish, Danio rerio. Each of the three types of loci is present in the zebrafish in multiple copies, but all of the identified copies of a given type map to the same region in linkage groups 1 (C3) and 15 (A2M, C4). The A2M and C4 loci are mapped in the same region not linked to any of the class I or class II major histocompatibility complex (Mhc) loci. These observations are interpreted as supporting the origin of the A2M family of genes by tandem duplications, followed by the dispersal of the copies to different chromosomes. It is also argued that the association of C4 with the class I/II loci in tetrapods is accidental and without functional significance.

Animals↗
PubMed2002

Laparoscopic transhiatal resection of epiphrenic diverticulum.

Our experience with videolaparoscopic operations for hiatus hernia and achalasia, which have almost replaced classical procedures, enabled us to use the same technique for other interventions in the distal third of the thoracic esophagus. Thus, we were able to treat epiphrenic diverticulum using a minimally invasive approach. We report our experience with videolaparoscopic diverticulum resection. The procedure was performed in three patients, all of them elderly men with ventilation limitation and a history of a chest intervention. The procedure included myotomy and an antireflux procedure. No significant complications occurred during the operations or postoperative periods; a minor leak that was successfully managed conservatively occurred in one patient. We conclude that videolaparoscopy could be a possible alternative to the standard classical left-side thoracotomy approach for patients in whom a classical operation is not feasible.

Aged↗
PubMed2002

The polymer mat: arrested rebound of a compressed polymer layer.

Compression of an adsorbed polymer layer distorts its relaxed structure. Surface force measurements from different laboratories show that the return to this relaxed structure after the compression is released can require tens of minutes and that the recovery time can grow rapidly with molecular weight. We argue that the arrested state of the free layer before relaxation can be described as a Guiselin brush structure (O. Guiselin, Europhys. Lett. 17, 225 (1992)), in which the monomer density falls off only weakly with distance from the surface. This brush structure predicts an exponential falloff of the force at large distance with a decay length that varies as the initial compression distance to the 6/5 power. This exponential falloff is consistent with surface force measurements. We propose a relaxation mechanism that accounts for the increase in relaxation time with chain length.

Journal Article↗
PubMed2002

Direct effects of ciliary neurotrophic factor on brown adipocytes: evidence for a role in peripheral regulation of energy homeostasis.

Ciliary neurotrophic factor (CNTF) plays an important role in regulating neuronal growth. Recently, central anorexigenic effects of this cytokine have been characterized. However, peripheral effects on tissues that actively contribute to the regulation of energy homeostasis have not been described. Here, we report direct potent and selective effects of CNTF on growth factor and metabolic signalling intermediates in mouse brown adipocytes. CNTF stimulates STAT3, MAP kinase, Akt, and p70 S6 kinase. We find that, next to mediating Akt and p70 S6 kinase activation, both phosphatidylinositol 3-kinase and protein kinase C are separately acting, main intermediates for inducing mitogen-activated protein (MAP) kinase activation. On a functional level, CNTF enhances beta3-adrenergic induction of uncoupling protein-1. Thus, these results demonstrate direct effects of CNTF on adipose tissue signalling and metabolism and suggest a novel role for this cytokine in the peripheral regulation of energy homeostasis.

Adipocytes↗
PubMed2002

Leptin secretion and negative autocrine crosstalk with insulin in brown adipocytes.

Leptin is an important adipocytokine whose main regulative effects on energy metabolism are exerted via activation of signalling pathways in the central nervous system. Another important regulator of energy homeostasis is insulin. The role of direct autocrine leptin effects on adipose tissue and crosstalk with insulin, in particular in the thermogenically active brown adipose tissue, remains unclear. In the present study, we have investigated leptin secretion and interaction with insulin in highly insulin-responsive immortalised mouse brown adipocytes. Leptin was secreted in a differentiation-dependent manner, and acute leptin treatment of mature adipocytes dose- and time-dependently stimulated phosphorylation of STAT3 and MAP kinase. Interestingly, acute pretreatment of fully differentiated brown adipocytes with leptin (100 nM) significantly diminished insulin-induced glucose uptake by approximately 25%. This inhibitory effect was time-dependent and maximal after 60 min of leptin prestimulation. Furthermore, it correlated with a 35% reduction in insulin-stimulated insulin receptor kinase activity after acute leptin pretreatment. Insulin-induced insulin receptor substrate-1 tyrosine phosphorylation and binding to the regulatory subunit p85 of phosphatidylinositol 3-kinase (PI 3-kinase) were diminished by approximately 60% and 40%, respectively. Taken together, this study has demonstrated strong differentiation-dependent leptin secretion in brown adipocytes and PI 3-kinase-mediated negative autocrine effects of this hormone on insulin action. Direct peripheral leptin-insulin crosstalk may play an important role in the regulation of energy homeostasis.

Adipose Tissue, Brown↗
PubMed2002

Isoproterenol is a positive regulator of the suppressor of cytokine signaling-3 gene expression in 3T3-L1 adipocytes.

SOCS (suppressor of cytokine signaling)-3 has recently been shown to be an insulin- and tumor necrosis factor (TNF)-alpha-induced negative regulator of insulin signaling. To further clarify a potential involvement of SOCS-3 in the development of insulin resistance, we measured differentiation-dependent SOCS-3 mRNA expression in 3T3-L1 adipocytes and studied its regulation by various hormones known to impair insulin signaling using quantitative real-time RT-PCR. There was a differentiation-dependent downregulation of SOCS-3 mRNA by 50% over the 9 day adipocyte differentiation course. Interestingly, besides insulin and TNF-alpha, chronic treatment of differentiated 3T3-L1 cells with 10 microM isoproterenol for 16 h stimulated SOCS-3 gene expression by about 3.5-fold. Furthermore, isoproterenol stimulated SOCS-3 mRNA expression in a dose-dependent manner with significant activation detectable at concentrations as low as 10 nM isoproterenol. Moreover, a strong 27- and 47-fold activation of SOCS-3 mRNA expression could be seen after 1 h of isoproterenol and GH treatment respectively. The stimulatory effect of isoproterenol could be almost completely reversed by pretreatment of 3T3-L1 cells with the beta-adrenergic antagonist propranolol. Finally, isoproterenol's action could be mimicked by stimulation of G(S)-proteins with cholera toxin and of adenylyl cyclase with forskolin and dibutyryl cAMP. Taken together, our results demonstrate a differentiation-dependent downregulation of SOCS-3 in adipocytes and suggest that SOCS-3 gene expression is stimulated by beta-adrenergic agents via activation of a G(S)-protein-adenylyl cyclase-dependent pathway. As SOCS-3 is a novel inhibitor of insulin signaling, the data support a possible role of this protein as a selectively regulated mediator of catecholamine-induced insulin resistance.

3T3 Cells↗
PubMed2002

[Role of modern absorbable suture materials in decreasing the occurrence of early complications after laparotomy].

UNLABELLED: Uncomplicated healing of surgical wounds is one of the most important factors which contribute to the success of operations. In particular dehiscence of laparotomy is an important complication associated with considerable morbidity and lethality. A number of factors which contribute to the healing of the surgical wound at the time of operation cannot be influenced, it is however possible to influence the technique of wound closure and the material used. The authors compare in their study early postoperative and long-term results of closure of laparotomy in three groups of patients where for closure of laparatomy PDSII loop suture was used, continuous suture with Vicryl and classical suture by individual silone stitches. The investigation revealed a significantly lower incidence of early postoperative complications in patients where modern absorbable materials were used, in particular early infections. The incidence of dehiscence of laparotomy or hernia in the scar did not differ significantly although in the group with absorbable materials, contrary to the group with silone, there was no dehiscence of laparotomy without an infectious complication of wound healing. CONCLUSION: Empirical experience and the conclusions of some major investigations indicate that the best method of closure of laparotomy is continuous suture using absorbable material.

Absorption↗
PubMed2001

Evolution of Mhc class II B genes in Darwin's finches and their closest relatives: birth of a new gene.

The 15 extant species of Darwin's finches on the Galápagos and Cocos Islands are the products of an unfinished adaptive radiation from a founder flock of birds related to the South American species Tiaris obscura. Molecular characterization of their major histocompatibility complex ( Mhc) class II B genes has revealed the existence of several related groups of sequences (presumably encoded in distinct loci) from which one (group 5) stands out because of its low divergence over extended time periods. Analysis of group 5 exon 2 and intron 2 sequences has revealed that the encoding locus apparently arose 2-3 million years ago in the Tiaris group of South and Central American Thraupini. The locus shows no evidence of inactivation, but displays a very low degree of polymorphism, both in terms of number of alleles and genetic distances between alleles. Some of the polymorphism, however, appears to be trans-specific. All the observed intergenic differences can be explained by point mutations and most of the exon 2 changes represent non-synonymous substitutions, although the rate of non-synonymous and synonymous substitutions appears to be the same. The origin of the new locus is explained by the birth-and-death model of Mhc evolution with two important extensions. First, the ancestor of the group 5 genes may have arisen without new gene duplication and second, the birth of the new group may have been brought about by a switch from balancing to directional selection. The ancestor of the group 5 genes may have been a classical class II B allele (one of many) which directional selection fixed in the ancestral population and drove into the category of nonclassical genes.

Animals↗
PubMed2001

Tumor necrosis factor alpha is a negative regulator of resistin gene expression and secretion in 3T3-L1 adipocytes.

Resistin has recently been implicated as an adipocytokine leading to insulin resistance and, therefore, potentially linking obesity and diabetes. To further characterize the regulation of this fat-secreted protein by insulin sensitivity-modulating hormones, 3T3-L1 adipocytes were treated with tumor necrosis factor (TNF) alpha, angiotensin (AT) 2, as well as growth hormone (GH), and resistin gene expression and protein secretion were determined by quantitative real-time reverse transcription-polymerase chain reaction and Western blotting. Interestingly, both, resistin mRNA expression and protein secretion, were inhibited by 70-90% after TNFalpha-treatment whereas AT2 and GH did not have any effect. The inhibitory effect of TNFalpha was time- and dose-dependent with significant inhibition occurring as early as 4 h after effector addition and at concentrations as low as 1 ng/ml TNFalpha. Pharmacological inhibition of protein kinase A (PKA), p44/42, and p38 mitogen-activated protein (MAP) kinase did not reverse the inhibitory effect of TNFalpha suggesting that neither of these signaling molecules is involved in suppression of resistin gene expression by TNFalpha. Furthermore, suppression of resistin mRNA levels could be completely reversed to control levels by withdrawal of TNFalpha for 24 h. Taken together, these results suggest that TNFalpha is a pivotal negative regulator of resistin gene expression. This may have important implications for the pathogenesis of insulin resistance and its link to obesity.

3T3 Cells↗
PubMed2001

Mhc class I genes of swordtail fishes, Xiphophorus: variation in the number of loci and existence of ancient gene families.

Swordtail fishes and platies in the genus Xiphophorus (order Cyprinodontiformes, Teleostei) encompass 22 closely related species which are the products of a recent adaptive radiation in the streams of Central America. To investigate the evolution of the major histocompatibility complex (Mhc) genes in the period immediately following speciation, the class I genes from 20 of the 22 species were cloned and characterized by sequencing. The analysis revealed the existence of multiple loci (at least seven in some individuals) whose numbers vary among the different species and probably also among individuals of the same species. The variation does not seem to bear any relationship to the taxonomy of the genus. Genes at the different loci are distinguished by their intron sequences and by the presence of characteristic motifs in exons 2 and 3. The variation in copy number of loci may have been effected in part by unequal crossing over occurring between introns of misaligned closely related genes. The sequences of the genes fall into two groups, A and B, which represent ancient lineages. The groups define two families of loci, which diverged from each other an estimated 85 million years ago, before the separation of the Acanthopterygii from the Paracanthopterygii of the advanced bony fishes. Evolution of the genes within each family can be explained by the birth-and-death process driven by gene duplications and mutational differentiation.

Amino Acid Sequence↗
PubMed2001

Adiponectin gene expression is inhibited by beta-adrenergic stimulation via protein kinase A in 3T3-L1 adipocytes.

Recently, it has been shown that the fat-derived factor adiponectin is downregulated in insulin resistance and obesity and that replenishment of this adipocytokine reverses insulin resistance in mice. Growing evidence, on the other hand, suggests that raised levels of catecholamines due to increased activity of the sympathetic nervous system are an integral part in the development of insulin resistance. To clarify whether catecholamines might exert their insulin resistance-inducing effects at least partly via downregulation of adiponectin gene expression, 3T3-L1 adipocytes were treated with isoproterenol, and adiponectin mRNA was measured by quantitative real-time reverse transcription-polymerase chain reaction. In fact, isoproterenol treatment reduced the level of adiponectin mRNA by about 75% in a dose-dependent fashion with significant inhibition detectable at concentrations as low as 10 nM isoproterenol. Furthermore, the inhibitory effect of isoproterenol was almost completely reversed by pretreatment of 3T3-L1 cells with the beta-adrenergic antagonist propranolol and the protein kinase A (PKA) inhibitor H-89. Moreover, the effects of isoproterenol could be mimicked by stimulation of stimulatory guanine nucleotide-binding (G(S))-proteins with cholera toxin and adenylyl cyclase with forskolin. Thus, our results suggest that adiponectin gene expression is severely suppressed by beta-adrenergic agents via activation of a G(S)-protein-PKA-dependent pathway. The data support a possible role of adiponectin in catecholamine-induced insulin resistance.

3T3 Cells↗
PubMed2001

Neuroimmune alterations in the complex regional pain syndrome.

This review focuses on some clinical aspects of the complex regional pain syndrome, such as oedema, local temperature changes and chronic pain, as a result of supposed neurogenic inflammation. Involvement of the immune system could imply the subsequent release of neuropeptides, pro-inflammatory cytokines and eicosanoids, which in turn leads to a complex cross-talk of primary and secondary generated mediators of inflammation. The development and application of drugs that act through selective receptor antagonism or enzymatic synthesis inhibition to prevent further stimulation of this cascade that could inevitably lead to chronicity of this disease are extensively discussed.

Animals↗