Identification of heat shock protein gene expression in hair follicles as a novel indicator of heat stress in beef calves.

Heat shock proteins (HSPs) consist of highly preserved stress proteins that are expressed in response to stress. Two studies were carried out to investigate whether HSP genes in hair follicles from beef calves can be suggested as indicators of heat stress (HS). In study 1, hair follicles were harvested from three male Hanwoo calves (aged 172.2 ± 7.20 days) on six dates over the period of 10 April to 9 August 2017. These days provided varying temperature-humidity indices (THIs). In study 2, 16 Hanwoo male calves (aged 169.6 ± 4.60 days, with a BW of 136.9 ± 6.23 kg) were maintained (4 calves per experiment) in environmentally controlled chambers. A completely randomized design with a 2 × 4 factorial arrangement involving two periods (thermoneutral: TN; HS) and four THI treatment groups (threshold: THI = 68 to 70; mild: THI = 74 to 76; moderate THI = 81 to 83; severe: THI = 88 to 90). The calves in the different group were subjected to ambient temperature (22°C) for 7 days (TN) and subsequently to the temperature and humidity corresponding to the target THI level for 21 days (HS). Every three days (at 1400 h) during both the TN and HS periods, the heart rate (HR) and rectal temperature (RT) of each individual were measured, and hair follicles were subsequently collected from the tails of each individual. In study 1, the high variation (P < 0.0001) in THI indicated that the external environment influenced the HS to different extents. The expression levels of the HSP70 and HSP90 genes at the high-THI level were higher (P = 0.0120, P = 0.0002) than those at the low-THI level. In study 2, no differences in the THI (P = 0.2638), HR (P = 0.2181) or RT (P = 0.3846) were found among the groups during the TN period, whereas differences in these indices (P < 0.0001, P < 0.0001 and P < 0.0001, respectively) were observed during the HS period. The expression levels of the HSP70 (P = 0.0010, moderate; P = 0.0065, severe) and HSP90 (P = 0.0040, severe) genes were increased after rapid exposure to heat-stress conditions (moderate and severe levels). We conclude that HSP gene expression in hair follicles provides precise and accurate data for evaluating HS and can be considered a novel indicator of HS in Hanwoo calves maintained in both external and climatic chambers.

Effects of dietary mixture of garlic (Allium sativum), coriander (Coriandrum sativum) and probiotics on immune responses and caecal counts in young laying hens.

This study was conducted to evaluate the effects of a combined mixture of phytogenic extracts (garlic and coriander) and probiotics on growth performance and immune responses in laying hens based on the results of in vitro studies to screen for immunomodulatory potency of each ingredient. Several parameters of immunomodulatory potency were estimated using lamina propria leucocytes (LPLs) isolated from rat intestinal mucosa tissue. Results show that the combined mixture enhanced LPLs proliferation, increased LPL-mediated cytotoxicity against YAC-1 tumour cells, and decreased lipopolysaccharide (LPS)-induced cytokine production including tumour necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) in LPLs. For in vivo study, laying hens (n = 50/each diet group) were fed with control diet, a diet containing antibiotics (0.01% per kg feed) or the combined mixture (0.02% per kg feed) for 21 days. The dietary combined mixture improved egg production (p < 0.05) but not growth performance and carcass traits. Interestingly, the patterns of suppressing plasma IFN-γ productions during inflammation by LPS injection and decreasing caecal E. coli counts in the combined mixture group were comparable to those in the antibiotics group. Taken together, our results suggested that the 0.02% of combined mixture of phytogenic extracts and probiotics as ingredients has potential immunomodulatory effects in laying hens.

Proteomic Analysis Reveals PGAM1 Altering cis-9, trans-11 Conjugated Linoleic Acid Synthesis in Bovine Mammary Gland.

cis-9, trans-11 Conjugated linoleic acid (CLA) is one of the most extensively studied CLA isomers due to its multiple isomer-specific effects. However, the molecular mechanisms of cis-9,trans-11 CLA synthesis in ruminant mammary gland are still not clearly understood. This process may be mediated, to a certain extent, by trans-11 C18:1 regulated by stearoyl-CoA desaturase-1 (SCD1) and/or its syntrophic proteins. This study aimed to investigate the effects of TVA on SCD1-mediated cis-9,trans-11 CLA synthesis in MAC-T cells and its potential molecular mechanism. Results showed that trans-11 C18:1 was continually taken up and converted into cis-9,trans-11 CLA in MAC-T cells during the 4-h incubation of 50 µM trans-11 C18:1. SCD1 protein expression increased more than twofold at 2 h (P < 0.01) and 2.5 h (P < 0.05) before decreasing to less than half of the normal level at 4 h (P < 0.05). One up-regulated (RAS guanyl releasing protein 4 isoform 1 [RASGRP4]) and six down-regulated proteins (glucosamine-6-phosphate deaminase 1 [GNPDA1], triosephosphate isomerase [TPI1], phosphoglycerate mutase 1 [PGAM1], heat shock protein beta-1 [HSPB1], annexin A3 [ANXA3], thiopurine S-methyltransferase [TPMT]) were found in MAC-T cells treated with trans-11 C18:1. Of these seven identified proteins, the presence of GNPDA1 and PGAM1 was verified in several models. More trans-11 C18:1 was taken up after PGAM1 knockdown by small interfering RNA (siRNA). In conclusion, our data suggested that PGAM1 may have a negative relationship with SCD1 and seemed to be involved in cis-9, trans-11 CLA synthesis by facilitating the absorption of trans-11 C18:1 in the bovine mammary gland.

Decreased prevalence of high-risk human papillomavirus infection is associated with obesity.

PURPOSE OF INVESTIGATION: Obesity is correlated with low education, low economic status, and lower rates of Pap smears, which are known as socio-demographic risk factors for cervical cancer. However, the association between obesity and high-risk human papillomavirus (HR-HPV) infection, the necessary cause of cervical cancer, and its related precursors, is not established. MATERIALS AND METHODS: The authors examined the association between obesity and HR-HPV infection in 6,868 patients, who participated in annual health examinations at the Kangbuk Samsung Hospital in Seoul, Korea, from January through December 2007. RESULTS: The prevalence of HR-HPV infection was 14.8%. Women infected with HR-HPV had a lower body mass index (BMI), when compared with non-infected women. After adjustment for alcohol intake, cigarette smoking, and marital status, HR-HPV infection was found to be negatively associated with BMI. When the analysis was stratified according to BMI, the risk of HR-HPV infection was significantly lower among those who were overweight (OR = 0.817, 95% CI = 0.680-0.982), or obese (OR = 0.688, 95% CI = 0.556-0.851), when compared with women with normal weight. CONCLUSION: HR-HPV infection was associated with obesity defined by BMI, with a lower prevalence of infection observed in obese women.

Accessory polar renal artery encountered in transperitoneal systemic laparoscopic paraaortic lymphadenectomy.

OBJECTIVE: To increase vigilance among gynecological surgeons for the presence of accessory polar renal artery (APRA) encountered with transperitoneal systemic laparoscopic paraaortic lymphadenectomy (LPAL). METHODS: A retrospective review was conducted on 156 women who underwent LPAL for various gynecologic malignancies between November 2003 and December 2009. RESULTS: The median age, parity, body mass index, and number of previous abdominal surgeries, respectively, of the women were 52 years (range, 23-82 years), two (range, 0-7), 24.1 kg/m2 (range, 17.4-35.0 kg/m2), and 0 (range, 0-3). During the study period, we found four women with APRA. There were three cases of right lower APRAs arising from the abdominal aorta, caudal to the inferior mesenteric artery (IMA), terminating at the parenchyma of the lower pole of the right kidney. In the other case, the APRA arose from the abdominal aorta superior to the IMA. There were no vascular complications, such as transection or ligation of the APRA. CONCLUSION: It is important for the gynecological oncologic surgeon to have knowledge of retroperitoneal vascular anatomy, experience in laparoscopic surgery, and an accurate surgical technique to avoid vascular injury during LPAL.

Genome-wide analysis of gene expression regulated by the yeast cell wall integrity signalling pathway.

The cell integrity pathway of Saccharomyces cerevisiae monitors cell wall remodelling during growth and differentiation. Additionally, this pathway responds to environmental stresses that challenge the integrity of the cell wall. We conducted a genome-wide survey of genes whose expression was altered in response to activation of Mpk1/Slt2, the MAP kinase, under the control of cell integrity signalling. We identified 25 genes whose regulation was altered by Mpk1 activity. Among these, 20 were positively regulated by Mpk1, and five were negatively regulated. Most of the genes identified encode either known or suspected cell wall proteins or enzymes involved in cell wall biogenesis. These include glycosyl-phosphatidylinositol (GPI) proteins, members of the Pir family of cell wall proteins, Mpk1 itself and others. All of the regulation detected was mediated by the Rlm1 transcription factor, a MADS-box protein that is phosphorylated and activated by Mpk1, but for which no transcriptional targets had been identified. A similar pattern of regulation was observed when cell integrity signalling was induced by environmental stress (i.e. temperature upshift).

Temperature-induced expression of yeast FKS2 is under the dual control of protein kinase C and calcineurin.

FKS1 and FKS2 are alternative subunits of the glucan synthase complex, which is responsible for synthesizing 1,3-beta-glucan chains, the major structural polymer of the Saccharomyces cerevisiae cell wall. Expression of FKS1 predominates during growth under optimal conditions. In contrast, FKS2 expression is induced by mating pheromone, high extracellular [Ca2+], growth on poor carbon sources, or in an fks1 mutant. Induction of FKS2 expression in response to pheromone, CaCl2, or loss of FKS1 function requires the Ca2+/calmodulin-dependent protein phosphatase calcineurin. Therefore, a double mutant in calcineurin (CNB1) and FKS1 is inviable due to a deficiency in FKS2 expression. To identify novel regulators of FKS2 expression, we isolated genes whose overexpression obviates the calcineurin requirement for viability of an fks1 mutant. Two components of the cell integrity signaling pathway controlled by the RHO1 G protein (MKK1 and RLM1) were identified through this screen. This signaling pathway is activated during growth at moderately high temperatures. We demonstrate that calcineurin and the cell integrity pathway function in parallel, through separable promoter elements, to induce FKS2 expression during growth at 39 degrees C. Because RHO1 also serves as a regulatory subunit of the glucan synthase, our results define a regulatory circuit through which RHO1 controls both the activity of this enzyme complex and the expression of at least one of its components. We show also that FKS2 induction during growth on poor carbon sources is a response to glucose depletion and is under the control of the SNF1 protein kinase and the MIG1 transcriptional repressor. Finally, we show that FKS2 expression is induced as cells enter stationary phase through a SNF1-, calcineurin-, and cell integrity signaling-independent pathway.

The protein kinase C-activated MAP kinase pathway of Saccharomyces cerevisiae mediates a novel aspect of the heat shock response.

The PKC1 gene of budding yeast encodes a homolog of the alpha, beta, and gamma isoforms of mammalian PKC that is proposed to regulate a MAPK-activation pathway. Mutants in this pathway undergo cell lysis resulting from a deficiency in cell wall construction when they attempt to grow at elevated temperatures. We show that the PKC1-regulated pathway is important for induced thermotolerance and that the MPK1 protein kinase (the MAPK of this pathway) is strongly activated by mild heat shock. This activation is sustained during growth at high temperature and is dependent on the function of pathway components proposed to function upstream of MPK1, including PKC1. Expression of genes under the control of known heat shock-inducible promoter elements (HSEs and STREs) was not compromised in PKC1 pathway mutants, indicating that this pathway mediates a novel aspect of the yeast heat shock response. We propose that the heat-induced signal for pathway activation is generated in response to weakness in the cell wall created during growth under thermal stress, perhaps as a result of increased membrane fluidity. Evidence is presented that the mechanism by which the cell detects this weakness is by measuring stretch of the plasma membrane.