Assessment of Diagnostics Tools for Sarcopenia Severity Using the Item Response Theory (IRT).

OBJECTIVE: To use the item response theory (IRT) methods to examine the degree to which the four selected tools reflect sarcopenia and to arrange them according to their ability to estimate sarcopenia severity. DESIGN: A cross-sectional study aimed at verifying the possibilities of using diagnostic tools for sarcopenia. SETTING AND PARTICIPANTS: The study included residents living in an assisted living unit at the Senior Centre in Blansko (South Moravia, Czech Republic) (n=77). Sarcopenia was estimated according to the proposals of the European Working Group on Sarcopenia in Older People (EWGSOP) using calf circumference, the EWGSOP algorithm, hand grip strength, and the Short Physical Performance Battery (SPPB). RESULTS: The results from the IRT model showed that these four methods indicate strong unidimensionality so that they measure the same latent variable. The methods ranked according to the discrimination level ranging from high to low discrimination where the calf circumference was the most discriminatory (Hi = 0.86) and the SPPB together with hand grip strength were the least discriminatory (both Hi = 0.44). CONCLUSION: We are recommending to identify mild sarcopenia by SPPB or hand grip strength, moderate sarcopenia by the EWGSOP algorithm and severe sarcopenia by the calf circumference.

Relation between cigarette smoking and sarcopenia: meta-analysis.

Cigarette smoking is a risk factor for many diseases. It could be associated with sarcopenia. The aim of this meta-analysis was to determine whether smoking is an isolated risk factor for sarcopenia. We searched PubMed, Web of Science, EBSCO, and Science Direct for articles addressing the relationship between cigarette smoking and sarcopenia. A total of 12 studies containing information on 22,515 participants were included in this meta-analysis. Odds ratio (OR) was calculated for each study group and for all studies together. An OR was also calculated separately for each sex. We used a fixed-effect model in overall estimation and in males, because results of small studies were significantly different from the results of large studies in those cases and in females where the estimation showed only moderate heterogeneity we used a random-effect model. According to proposes of the Cochrane Handbook for Systematic Reviews. The resulting OR in the fixed-effect model was 1.12 (95 % CI 1.03-1.21), OR for each sex was in the fixed-effect model 1.20 (95 % CI 1.06-1.35) in males and in the random-effect model 1.21 (95 % CI 0.92-1.59) in females. The results of this meta-analysis indicate that cigarette smoking as an isolated factor may contribute to the development of sarcopenia. However, the results of the individual studies were largely inconsistent due to different approaches of measuring the main variables which affected the results.

[Subclinical alveolar echinococcosis in a dog initially showing clinical features of leptospirosis]. / Subklinische alveoläre Echinokokkose bei einem Hund mit anfänglicher Symptomatik einer Leptospireninfektion.

A 3.5-year-old male Labrador retriever dog showed a short history of illness characterized by vomiting, apathy, and fever. Ultrasonographically, large nodular liver masses of high echogenicity were noted in both left and right liver lobes. Cytological and bacteriological examinations revealed a neutrophilic hepatitis without detectable agents. During treatment with doxycycline a four-fold decrease of serum titers to Leptospira (L.) icterohaemorrhagiae and L. sejroe was detected in paired serum samples by use of the complement-fixation test. The dog remained without clinical signs and no significant biochemical changes were recorded. However, ultrasonsographic examinations showed a progression of the hepatic lesions, presenting now as nodular parts with high echogenicity and cavernous parts with lower echogenicity. Diagnostic laparotomy was performed and the dog was euthanized due to severity of hepatic lesions. Histopathologically, a severe chronic granulomatous hepatitis with numerous parasitic structures was diagnosed. Morphology of the parasitic structures was comparable to the metacestode stage of Echinococcus multilocularis.

Effect of the MTHFR 677C/T polymorphism on homocysteinemia in response to creatine supplementation: a case study.

Creatine (Cr) is recommended as a dietary supplement especially for athletes but its therapeutic potential is also discussed. It is assumed that human body uses Cr for the formation of phosphocreatine, which is necessary for muscular work as a source of energy. Production of Cr in a body is closely connected to methionine cycle where guanidinoacetate (GAA) is in a final step methylated from S-adenosylmethionine (SAM). Increased availability of SAM for phosphatidylcholine (PC) and sarcosine synthesis can potentially stimulate endogenous production of betaine a thus methylation of homocysteine (HCy) to form methionine. Our subject who was methylenetetrahydrofolate reductase (MTHFR) 677TT homozygote lowered plasma HCy from 33.3 micromol/l to 17.1 micromol/l following one-month Cr supplementation (5 g/day) opposite to 677CC and CT genotypes whose HCy levels tended to increase (but still in normal ranges). We suppose that Cr supplementation stimulates pathways leading to production of sarcosine which can serve to regenerate tetrahydrofolate (THF) to form 5,10-methylene-THF. This could potentially increase MTHFR enzyme activity which may later result in increased HCy methylation. Cr supplementation significantly effects metabolism of one carbon unit and potentially lower body´s demands for methyl groups. This could be beneficial as in the case of reduced enzyme activity such as MTHFR 677C/T polymorphism.

Estrous cycle-dependent activity of neutrophils in the porcine endometrium: possible involvement of heat shock protein 27 and lactoferrin.

Neutrophil infiltration into the porcine endometrium is thought to be a specific feature during the follicular phase of the estrous cycle. To specify the localization and distribution of neutrophil granulocytes at different stages of the estrous cycle, porcine uterine samples were evaluated by immunohistochemical methods using anti-bovine lactoferrin (LF) antibody. Additionally, blood samples were collected from 30 pigs at different stages of the estrous cycle with a special focus on peri-estrous phase. Manual 100-cell differential counts were performed on routinely stained blood smears and evaluated statistically. Finally, the expression of granulocyte-colony-stimulating factor (G-CSF) and heat shock protein 27 (HSP 27), which are known to influence activation of the neutrophilic granulocytic lineage, was analyzed in porcine uteri using immunohistochemistry. Results show that LF is expressed regularly in the cytoplasm of neutrophil granulocytes. An increasing infiltration of subepithelial neutrophils was detected in the follicular phase. The highest number of intra- and subepithelial LF-positive cells was found on d 2 of the estrous cycle. Maximum level was followed by a strong decrease on d 3. Blood analysis revealed that the percentage of neutrophil granulocytes was significantly lower at d 2 (26.2+/-11.1%) than d 1 (42.1+/-11%) of the estrous cycle. HSP 27 staining was predominantly localized to luminal epithelium (LE) and glandular epithelium (GE) depending on stage of the estrous cycle. Strong immunostaining of HSP 27 is only found in LE during estrus. At d 2 of the estrous cycle, HSP 27 immunoreactivity in LE and superficial GE is reduced but moderate staining is found in deep GE. G-CSF immunostaining is uniformly not detected in endometrial cells of cyclic pigs. In conclusion, there is a clinically relevant relationship between neutrophil count in the blood and neutrophil infiltrate in the endometrium of the pig during the estrous cycle. This association may reflect the possibility of translocation of neutrophils from the blood to the endometrium up to d 2 of the estrous cycle. Additionally, HSP 27 could be a good candidate involved in migration and/or function of neutrophils within the porcine endometrium.

Localization of transforming growth factor beta3 in squamous metaplasia of the porcine oviduct: a case report.

Squamous metaplasia of the oviduct epithelium is a rare disorder of reproductive organs. We noted squamous metaplasia of the oviduct epithelium in a sow routinely slaughtered at day 2 of the oestrous cycle. Expression of transforming growth factor beta3 (TGF beta3) in the metaplastic epithelia was evaluated by immunohistochemistry, because TGF beta3 appears to play a key role as regulator of a variety of tissue remodelling events. Our results show that TGF beta3 immunostaining was specifically localized to foci of squamous metaplasia of the epithelial linings. Non-metaplastic epithelial cells of the oviduct were not immunostained with anti-TGF beta3 antibody. At the subcellular level, TGF beta3-labelled cells occasionally showed signs of apoptotic cell death. It is concluded that signals produced by TGF beta3 in metaplastic lesions of the oviduct are potentially involved in pathophysiological processes.

Expression of transforming growth factor-beta3 (TGF-beta3) in the porcine ovary during the oestrus cycle.

Transforming growth factor-beta (TGF-beta) proteins are growth factors that have been shown to be involved in regulation of ovarian follicular development. Ovarian expression, activity and functional significance of TGF-beta1 and TGF-beta2 isoforms were extensively studied in most species. However, little is known about the biological role of TGF-beta3 previously shown to be expressed independently of the other two isoforms. Therefore, expression of TGF-beta3 mRNA and protein was evaluated by RT-PCR and immunohistochemistry, respectively, in porcine ovaries collected during different phases of the oestrus cycle. Results of RT-PCR analysis showed that TGF-beta3 mRNA is expressed throughout the oestrus cycle. The level of TGF-beta3 mRNA expression was found to be higher at metoestrus and dioestrus. Weak TGF-beta3 immunoreactivity was present in follicular epithelial cells and oocytes of preantral follicles in all stages examined. TGF-beta3 protein expression was exclusively present in theca interna cell layer of antral follicles, and was particularly prominent in large antral follicles. Immediately after ovulation, almost all theca cells outside of the granulosa cell layer were intensively stained with anti-TGF-beta3. Immunostaining of TGF-beta3 in theca lutein cells rapidly decreased during corpus luteum development. It is suggested that TGF-beta3 may play an important role in modulating theca cell function of pre- and postovulatory follicles of the pig.

Review of apoptotic and non-apoptotic events in non-ciliated cells of the mammalian oviduct.

Reproductive organs are known to undergo dynamic changes during the oestrus cycle and pregnancy. Cell growth and regeneration of the reproductive tissues are closely correlated with ovarian steroid hormone levels. This review focuses on apoptotic and non-apoptotic degenerative events within oviduct epithelium that occur in a species-, cycle-, and segment-specific manner. Epithelial extrusion of larger cell fragments including nuclei and whole cells is the characteristic feature of non-apoptotic cell loss of non-ciliated cells in large (pig, sheep, goat, cattle) and small animals (dog). This mechanism of epithelial cell loss is most frequently observed in the luteal phase of the oestrus cycle and after progesterone treatment, respectively. Using light- and electron-microscopic techniques, typical apoptotic epithelial cells characterized by extensive nuclear and cytoplasmic fragmentation are found very sporadically in most species. In contrast, oviduct epithelial cells of subhuman primates and cats in part show marked signs of apoptosis, which could be explained by their respective cycle-specific characteristics. Recent investigations using histochemical markers of apoptosis and our own findings in the porcine oviduct suggest that the degenerative process in the mammalian oviduct includes the death of numerous epithelial cells by apoptosis. Advancement in the knowledge of elimination of oviduct epithelial cells is necessary to understand the physiological process of epithelial renewal and pathological processes caused by imbalances between cell renewal and elimination.

Clusterin expression in porcine endocrine cells during islet development.

Clusterin is a well-known glycoprotein expressed by many cell types involved in multiple physiological functions. In rat pancreatic tissue it is expressed along with islet cell development and found to be involved in regeneration of pancreatic endocrine cells after various types of tissue injuries. These results led us to propose that clusterin might play a crucial role in organization and assembling processes of islet cells during pre- and postnatal development. Therefore, the aim of this study was to find out whether and in which cell type clusterin is expressed during islet cell organization in the porcine species which could play a future role in the field of xenotransplantation. For this purpose we examined the expression pattern of clusterin at different developing stages in the porcine pancreas by double-immunostaining with antibodies against chromogranin A and clusterin, and clarified whether distinct islet hormones were coexpressed with clusterin. Further, we checked by RT-PCR whether clusterin was locally expressed or possibly locally bound to the corresponding receptor. In newborn and up to 3-month-old animals clusterin was found to be expressed in a special cell type which is closely associated and intermingled with other endocrine cells. In fully developed adult islets clusterin-cells then reorganize and were found to be mainly localized in the mantle area of Langerhans islets. Double-immunostaining with antibodies against clusterin and different islet hormones such as insulin, glucagon, and somatostatin clearly demonstrate that clusterin expression was found in an own special cell type and it is also present in a subset of glucagon producing A-cells. Taken together, our results show that clusterin expression in porcine species is found in an own, as yet unidentified cell type during postnatal developmental stages, and probably labels immature precursor cells in adult animals, which finally have the potential to differentiate into glucagon-expressing cells.

Expression and localisation of extracellular matrix degrading proteases and their inhibitors during the oestrous cycle and after induced luteolysis in the bovine corpus luteum.

The corpus luteum (CL) offers the opportunity to study high proliferative processes during its development and degradation processes during its regression. We examined the mRNA expression of matrix metalloproteases (MMP)-1, MMP-2, MMP-9, MMP-14, MMP-19, tissue inhibitor of MMP (TIMP)-1, TIMP-2, tissue plasminogen activator (tPA), urokinase plasminogen activator (uPA), uPA-receptor (uPAR), PA-inhibitors (PAI)-1, PAI-2 in follicles 20 h after GnRH application, CLs during days 1-2, 3-4, 5-7 and 8-12 of the oestrous cycle as well as after induced luteolysis. Cows in the mid-luteal phase were injected with Cloprostenol and the CLs were collected at 0.5, 2, 4, 12, 24, 48 and 64 h after PGF2alpha injection. Real-time RT-PCR determined mRNA expressions. Expression from 20 h after GnRH to day 12: MMP-1, MMP-2, MMP-14 and tPA showed a clear expression, but no regulation. TIMP-1 and uPAR mRNA increased when compared with the follicular phase. TIMP-2, MMP-9, MMP-19 and uPA increased from the follicular phase to days 8-12. PAI-1 and PAI-2 expression increased from days 1-7 and decreased to days 8-12. Induced luteolysis: MMP-1, MMP-2, MMP-9, MMP-14, MMP-19 and TIMP-1 all increased at different time points and intensities, whereas TIMP-2 was constantly decreased from 24 to 64 h. The plasminogen activator system and their inhibitors were up-regulated from 2 to 64 h, tPA was already increased after 0.5 h. Immunohistochemistry for MMP-1, MMP-2, MMP-14: an increased staining for MMP-1 and MMP-14 was seen in large luteal cells beginning 24 h after PGF2alpha application. MMP-2 showed a strong increase in staining in endothelial cells at 48 h.

Non-neuronal acetylcholine and choline acetyltransferase in oviductal epithelial cells of cyclic and pregnant pigs.

Certain female reproductive tissues are known to express the non-neuronal cholinergic system. Using different experimental approaches, we tested the hypothesis that acetylcholine (ACh) in the porcine oviduct may also be derived from non-neuronal structures. Immunohistochemistry was performed to detect acetylcholine synthesizing enzyme choline acetyltransferase (ChAT) in different segments of the oviduct of cyclic and pregnant sows. Immunohistochemical experiments revealed strong immunoexpression of ChAT in the entire oviductal epithelium at metoestrus. Thereby, a particular pronounced staining was found in the supranuclear region of almost all epithelial cells. Immunostaining of ChAT decreased markedly during dioestrus and prooestrus stages, respectively. At prooestrus, ChAT immunoreactivity was confined to ciliated cells. Furthermore, we found elevated level of staining intensity of ChAT in the pregnant oviduct at day 13. Using the same ChAT antibody for Western blot analyses, we detected immunoreactive bands of MW 69,000 and 46,000 mainly in ampulla, while MW 58,000 and 30,000 forms were present mainly in infundibulum and isthmus. Furthermore ACh was detected by HPLC and fluorimetric methods in oviductal epithelium. In conclusion, we show expression of ChAT in oviductal epithelial cells at different stages of the oestrus cycle and pregnancy, indicating that these cells can synthesize ACh in a cycle-dependent manner. These results suggest as yet unexplored roles of epithelial ACh in the oviduct.

Prion protein expression in bovine podocytes and extraglomerular mesangial cells.

The cellular form of the prion protein (PrP(c)) is thought to be a substrate for an abnormal isoform of the prion protein (PrP(sc)). One emerging hypothesis is that the proposed conversion phenomenon takes place at the site at which the infectious agent meets PrP(c). PrP(c) is abundant in the central nervous system, but little is known about the cell-type-specific distribution of PrP(c) in non-neuronal tissues of cattle. We have studied whether PrP(c), a protein found predominantly in neurons, also exists in bovine podocytes, since neurons and podocytes share a large number of similarities. We have therefore examined the expression of PrP(c) by immunohistochemistry, reverse transcription/polymerase chain reaction and enzyme-linked immunosorbent analysis. Immunostained serial sections and specific antibodies against PrP(c) have revealed that PrP(c) is selectively localized in podocytes and is particularly strongly expressed in extraglomerular mesangial cells but not in endothelial or intraglomerular mesangial cells. The selective expression of PrP(c) in podocytes is of special importance, as it suggests that these cells represent possible targets for peripheral infection with prions and demonstrates that PrP(c) can be added to the list of neuronal factors expressed in mammalian podocytes.

Colocalization of chromogranin A and inositol 1,4,5-trisphosphate receptor in ciliated cells of the bovine oviduct.

Previous investigations of the expression of chromogranin A (CgA) have been performed primarily in neuroendocrine tissues containing amine and peptide secretory vesicles. More recently it has been shown that CgA, as a high capacity Ca2+ storage protein, interacts with the inositol 1,4,5-trisphosphate receptor/Ca2+ channel (InsP3R) which has been found to be selectively localized in oviductal cells of the mouse. To examine a possible role of this coupling in the Ca2+-dependent ciliary movement, we investigated the topographical and cellular distribution of cells positive for CgA and inositol 1,4,5-trisphosphate receptor type 2 (InsP3R2) in the bovine oviduct at different stages of the oestrous cycle. Using immunohistochemical techniques on paraffin-embedded tissue we have successfully shown that CgA is selectively expressed in ciliated cells of the bovine oviduct. The labelled cells show intense positive staining in the apical surface area in close vicinity to the ciliary apparatus. CgA-positive ciliated cells are most frequently observed at dioestrous while a lower number appears at oestrous. Additionally, secretory and intraepithelial neuroendocrine cells consistently do not stain with the CgA-antiserum. We then investigated whether the reported expression of the InsP3R in oviductal cells of the mouse corresponds to the expression of the InsP3R in bovine oviductal cells. Using a polyclonal antibody to the type 2 InsP3R, we found that the receptor is also selectively expressed in a similar matter to CgA in the apical cytoplasm of ciliated cells. This is the first morphological demonstration of the colocalization of CgA and InsP3R in epithelial ciliated cells of the bovine oviduct. Our results suggest that CgA and InsP3R could be involved in controlling the ciliary activity of oviductal epithelial cells.

Expression of cytokeratin 18 during pre- and post-natal porcine lung development.

The expression pattern of the intermediate filament protein cytokeratin 18 (CK 18) is described during pre- and post-natal development of the porcine lung using a monoclonal antibody against human CK 18. Lungs from 16 foetuses in pseudoglandular, canalicular, saccular and alveolar stages of lung development and lungs from 12 pigs ranging in age from birth to 49 days after birth were studied by immunohistochemistry. In the early pseudoglandular stage of development (day 70 of gestation) all the columnar epithelial cells lining the tubular endbuds strongly expressed CK 18 predominantly in the apical cell compartment. A modest staining was found in the more cuboidal cells of the canalicular stage (day 80 of gestation) where the labelling occurred as a distinct positive rim at the apical cell membrane in most of the cells lining the canaliculi. In 96- and 100-day-old foetuses, parts of the gas exchanging area were formed as terminal sacs by extreme attenuation of the epithelium. In this stage, CK 18 was clearly detectable in the flat type I as well as in the cuboidal type II alveolar epithelial cells. A marked change of the CK 18 expression pattern occurred during formation of the alveoli by septal outgrowth and maturation of the epithelium in 105- and 111-day-old foetuses. Differentiated type I cells no longer expressed CK 18, whereas type II cells were still labelled. Moreover, a specific change in the subcellular distribution pattern from the luminal periphery in immature porcine type II cells to a cytoplasmic localization in differentiated type II cells could be observed. Our investigation additionally demonstrated that the epithelium of bronchi, bronchioli and terminal bronchioli expressed CK 18 in all pre- and post-natal developmental stages. From the 96 days of gestation onwards the epithelial cells of developing bronchial glands were also labelled. Our results clearly show that during porcine lung development profound changes in the cellular expression pattern of CK 18 occur and that CK 18 can be regarded as a selective marker for differentiated porcine alveolar type II cells from the 105th day of gestation onwards. We also assume that the intermediate filament CK 18 could be of significance in the maturation process of the type II alveolar cells.

[Not Available]. / Wirkung von Deoxynivalenol (DON) auf den Magen-Darm-Trakt männlicher Läuferschweine.

Following an experiment feeding Deoxynivalenol to growing pigs histological studies have been carried out to evaluate tissue damaging effects of DON. Pigs were fed with naturally contaminated wheat containing 4000 and 6000 µg DON/kg feed. A number of two animals was chosen from control and high exposed group, respectively. Samples of the gastrointestinal tract, liver and kidney were examined. Although, clear effects in feed consumption and weight gain could be realized, only one animal from the high exposed group showed focal alterations in tissues of the gastrointestinal tract.

[Surgical treatment of Zenker's diverticulum]. / Chirurgická lécba Zenkerova divertiklu.

The authors present an account of diagnostic methods, endoscopic problems and surgical treatment of Zenker's diverticula of the oesophagus. They analyze possibilities of X-ray and endoscopic diagnosis; a frequent problem is the insertion of an oesophageal probe before operation; tamponade of the diverticula before operation is useful. Surgical treatment is indicated in major diverticula, small flat ones are followed up. The surgical approach is from the left side of the neck. In the authors' department always total resection of the dicerticulum was performed in major diverticule, invagination could be used in minor diverticula. Early and late complications of operations are discussed.