Liver biopsy specimens were obtained from 50 patients with chronic hepatitis C (CHC) and without detectable HCC before starting antiviral therapy. Patients’ characteristics are given in Table 1A. F3-F4 of histological stage was denoted as “advanced fibrosis.” Another 60 biopsies and 4 operative specimens were obtained from HCC tissues of patients with chronic hepatitis B (n = 6), C (n = 52), B and C coinfection (n = 2), or other causes Selleckchem DAPT (alcoholic: n = 1; primary biliary cirrhosis: n = 1; Wilson’s

disease: n = 1; unknown: n = 1; Table 1B). Four of the HCC samples and clinical data used in the current study were from a previous publication.[29] Tumor node metastasis (TNM) stage of HCC was determined according to the criteria of the International Union against Cancer and the American Joint Committee on Cancer,[30] and histological grading was performed according to the criteria of an International Working Party.[31] Tissues had been frozen for western blotting immediately at −80°C or formalin-fixed. Informed consent was obtained from each patient before study participation. Female 8-week-old Balb/c nude (nu/nu) mice were purchased from The Jackson Laboratory (Bar Harbor, ME). Animal studies were approved by the Institutional Animal Care and Use Committee of the Beth Israel Deaconess Medical Center (Boston, MA). HEPG2 cells were maintained at

37˚C for 3 days in vitro after a 10-minute exposure to 37˚C (controls), 45˚C, 48˚C, or 50˚C, followed HDAC inhibitor mechanism by harvest with trypsin/ethylenediaminetetraacetic

acid and resuspension in 50% growth-factor–reduced Matrigel (BD Biosciences) in PBS to a final cell count of 2.5 MCE公司 × 107 cells/mL. A volume of 0.2 mL of the cell suspensions was injected subcutaneously (SC) in the right flank of each mouse (6 mice per group), as described before.[32] Estimated tumor weight (ETW) was calculated every 2 days after injection using the following formula: ETW (mg) = Length (mm) × (Width (mm))2/2).[32] Fifteen days after tumor cell injection, animals were sacrificed and tumors were harvested and immediately stored in −80˚C for further analysis. Data are expressed as means ± standard error of the mean (SEM). Statistical analyses were performed using Microsoft Excel (Microsoft Corp., Redmond, WA) and GraphPad Prism (version 5.00; GraphPad Software Inc., San Diego, CA). Multiple comparisons were performed by one-way analysis of variance. Two planned comparisons were performed to each of the control groups using Dunnett’s post-test. The OS curve of patients with positive Shc-labeling indices (LI; %) was plotted using Kaplan-Meier’s method, and differences were analyzed statistically by the log-rank test. Differences among selected experimental groups with P values <0.05 were considered significant. Correlation coefficients were calculated by GraphPad Prism (version 5.00; GraphPad Software Inc.). A two-tailed P value was selected and confidence intervals were set to 95%. R > 0.

Previous studies have been hampered by problems with case ascertainment, definition, and have generally had limited numbers and/or follow-up, which could potentially lead to inaccurate estimates of disease burden.10-12 It is well established that cirrhotic patients presenting with overt synthetic liver dysfunction are more likely to develop liver-related complications and have a high overall mortality. However, some important aspects of the prognosis of patients with NAFLD still remain unclear. First, it is unclear how the long-term prognosis of patients with NAFLD compares with patients with liver disease of other etiologies, such as chronic hepatitis C virus (HCV)

infection. Second, what are the risks of liver-related complications, mTOR inhibitor including HCC, in patients with NAFLD with advanced fibrosis or cirrhosis and no overt synthetic dysfunction (i.e., Child-Pugh class A)? Third, the effect of NAFLD on non-liver-related sequelae, such as vascular outcomes (e.g., myocardial infarction, strokes, and vascular deaths), remains poorly described.13 Finally, it is unclear which, if any, risk factors can independently predict liver, vascular, and overall morbidity and mortality. To answer these questions, we carried out an international, multicenter prospective study to assess the natural history and outcomes of liver biopsy-confirmed NAFLD high throughput screening with advanced fibrosis

or cirrhosis from four medical centers. We sought to assess complications that occurred in these patients and identify the predictors of such events; we also compared their long-term morbidity and mortality to a group of patients with histologically confirmed chronic HCV infection and advanced fibrosis or cirrhosis. ALT,

alanine aminotransferase; AST, aspartate aminotransferase; BMI, body mass index; CI, confidence interval; HCC, hepatocellular cancer; HCV, hepatitis C virus; HDL, high-density lipoprotein; MELD, Model for End-Stage 上海皓元 Liver Disease; NAFLD, nonalcoholic fatty liver disease; NASH, nonalcoholic steatohepatitis; SD, standard deviation. A total of 247 Child-Pugh class A patients with biopsy-confirmed NAFLD and advanced fibrosis or cirrhosis comprised the NAFLD cohort. This cohort was recruited from 1984 to 2006. Patients were previously untreated and consecutively biopsied at four centers: Mayo Clinic (Rochester, MN) (n = 105); Newcastle Hospitals National Health Service Foundation Trust (Newcastle-upon-Tyne, UK) (n = 57); Westmead Hospital (Sydney, Australia) (n = 51); and University of Turin (Turin, Italy (n = 34). The comparator cohort consisted of 264 patients diagnosed with HCV infection and advanced fibrosis or cirrhosis, who were also Child-Pugh class A, enrolled from 1987 to 2005. HCV infection was confirmed by a positive polymerase chain reaction at baseline in all patients. HCV subjects were seen and consecutively biopsied at Westmead Hospital (n = 209) and University of Turin (n = 55).

Previous studies have been hampered by problems with case ascertainment, definition, and have generally had limited numbers and/or follow-up, which could potentially lead to inaccurate estimates of disease burden.10-12 It is well established that cirrhotic patients presenting with overt synthetic liver dysfunction are more likely to develop liver-related complications and have a high overall mortality. However, some important aspects of the prognosis of patients with NAFLD still remain unclear. First, it is unclear how the long-term prognosis of patients with NAFLD compares with patients with liver disease of other etiologies, such as chronic hepatitis C virus (HCV)

infection. Second, what are the risks of liver-related complications, p38 MAPK apoptosis including HCC, in patients with NAFLD with advanced fibrosis or cirrhosis and no overt synthetic dysfunction (i.e., Child-Pugh class A)? Third, the effect of NAFLD on non-liver-related sequelae, such as vascular outcomes (e.g., myocardial infarction, strokes, and vascular deaths), remains poorly described.13 Finally, it is unclear which, if any, risk factors can independently predict liver, vascular, and overall morbidity and mortality. To answer these questions, we carried out an international, multicenter prospective study to assess the natural history and outcomes of liver biopsy-confirmed NAFLD selleck screening library with advanced fibrosis

or cirrhosis from four medical centers. We sought to assess complications that occurred in these patients and identify the predictors of such events; we also compared their long-term morbidity and mortality to a group of patients with histologically confirmed chronic HCV infection and advanced fibrosis or cirrhosis. ALT,

alanine aminotransferase; AST, aspartate aminotransferase; BMI, body mass index; CI, confidence interval; HCC, hepatocellular cancer; HCV, hepatitis C virus; HDL, high-density lipoprotein; MELD, Model for End-Stage MCE公司 Liver Disease; NAFLD, nonalcoholic fatty liver disease; NASH, nonalcoholic steatohepatitis; SD, standard deviation. A total of 247 Child-Pugh class A patients with biopsy-confirmed NAFLD and advanced fibrosis or cirrhosis comprised the NAFLD cohort. This cohort was recruited from 1984 to 2006. Patients were previously untreated and consecutively biopsied at four centers: Mayo Clinic (Rochester, MN) (n = 105); Newcastle Hospitals National Health Service Foundation Trust (Newcastle-upon-Tyne, UK) (n = 57); Westmead Hospital (Sydney, Australia) (n = 51); and University of Turin (Turin, Italy (n = 34). The comparator cohort consisted of 264 patients diagnosed with HCV infection and advanced fibrosis or cirrhosis, who were also Child-Pugh class A, enrolled from 1987 to 2005. HCV infection was confirmed by a positive polymerase chain reaction at baseline in all patients. HCV subjects were seen and consecutively biopsied at Westmead Hospital (n = 209) and University of Turin (n = 55).

8, 9 Conversely, conventional histopathology quickly provides a wealth of irreplaceable data about structural integrity, spatial and temporal relationships, and rare events/cells. Only a tiny fraction of information is being harvested from tissue slides primarily because data extraction is dependent on a restricted staining repertoire and manual observation.

The challenge, therefore, is to develop a HM781-36B molecular weight modern replacement to the traditional histopathologic approach. Dramatic advances in robotics, digital imaging, and computing have spawned the “-omics” revolution that challenges routine histopathology for improved tissue utilization. Comprehensive examination of mRNA, protein, and metabolite expression data can be used as powerful screening tools

to query disease susceptibility, pathophysiology, and prognosis. These same innovations, however, are also revolutionizing histopathology. High-resolution whole-slide image (WSI) scanners now enable pathologists to view routinely prepared and stained slides on a computer screen instead of a microscope. Pathologists can then team with hardware and software engineers, mathematicians, and image analysis experts to greatly increase the value equation for histopathology in an era when there is increasing pressure to diagnose and monitor liver diseases noninvasively.8 We recently reported on the power of combining WSI, multiplex nanoparticle quantum dot http://www.selleckchem.com/products/Everolimus(RAD001).html staining, and automated 上海皓元医药股份有限公司 image analysis

to envisage and analyze multiple protein labels on a single slide to reveal biological mechanisms.7, 10-16 We use this novel approach to study liver epithelial diversity in formalin-fixed, paraffin-embedded, normal human liver tissue. In this report, automated quantitative data collection that described cell numbers, types, and nuclear/cytoplasmic analyte expression was spatially tethered to the tissue architecture. This enabled us to illustrate and locate preexisting diversity within BECs and hepatocytes in normal human liver, including the transition zone between these cell types in the canals of Hering (CH). This might lead to a better understanding of the considerable diversity that quickly appears during disease states.5 BEC, biliary epithelial cell; CD31, cluster of differentiation 31; CH, canal of Hering; EC, endothelial cell; HNF, hepatocyte nuclear factor; HPC, hepatic progenitor cell; ROI, region of interest; SMA, smooth muscle actin; SMC, smooth muscle cell; WSI, whole slide image. Four-μm sections from eight formalin-fixed paraffin-embedded and one frozen liver tissues were used for the analyses (Supporting Table 1; Institutional Review Board protocol 0404010, University of Pittsburgh). Before study inclusion, hematoxylin and eosin (H&E) slides from each case were reviewed. Tissues were prepared and stained as described.

006, Fig. 2A). The copy numbers for FGF3/FGF4 were 10.2 ± 0.8/6.7 ± 0.8, 26.7 ± 0.4/35.1 ± 3.1, and 162.5 ± 9.0/165.0 ± 12.5 copies in the amplified samples, whereas the copy numbers of FGF3 for all the other samples were below 5 copies. The correlation between the FGF3 locus and the FGF4 locus copy numbers was very high (R

= 0.998), indicating that the DNA copy number assay for FGF3/FGF4 was a sensitive and reproducible method. We examined the messenger RNA (mRNA) expression levels of FGF3/FGF4 in nine HCC samples that were available as frozen samples among the Ceritinib 48 sorafenib-treated samples, as shown in Fig. 2A. One amplified sample expressed extremely high mRNA levels of FGF3/FGF4 compared with nonamplified samples (Fig. 2B). The results demonstrated that FGF3/FGF4 gene amplification mediates the overexpression of FGF3/FGF4 mRNAs and proteins (Figs. 2B and 1D). We used FISH analysis to examine FGF3/FGF4 amplification and to verify the results of the above-described PCR-based DNA copy number assay. All FGF3/FGF4-amplified clinical samples were confirmed as exhibiting high-level FGF3 amplification using FISH analysis (Fig. 3). One patient showed multiple scattered signals, whereas two patients showed large clustered signals. Nonamplified HCC

yielded a negative result for gene amplification. These results clearly demonstrate the presence of FGF3/FGF4-amplified HCC among the clinical samples, and the FISH

analysis results were consistent with those for the copy number assay. To determine the frequency HSP inhibitor review of FGF3/FGF4 gene amplification in HCC, we performed a copy number assay for HCC samples without sorafenib treatment in a series of surgical specimens. Two of the 82 (2.4%) HCC samples exhibited FGF3/FGF4 gene amplification, with copy numbers 上海皓元医药股份有限公司 of 10.7/15.3 and 133.3/112.7 copies, respectively (Fig. 4). One amplified HCC was a poorly differentiated tumor, whereas the other was a moderately differentiated tumor. The clinico-pathological features of the sorafenib responders are shown in Table 1. A comparison of clinical factors (age, sex, viral status, alpha-fetoprotein level, PIVKA-II, clinical stage, primary tumor size, metastatic status, histological type, and tumor response between responders and nonresponders) is given in Table 2. Notably, multiple lung metastases over five nodules was significantly higher among responders to sorafenib (responders, 5/13 [38%]; nonresponders, 2/42 [5%]; P = 0.006). Although the difference was not significant, poorly differentiated HCC tended to be more common among responders to sorafenib (responders, 5/13 [38%]; nonresponders, 6/42 [14%]; P = 0.13). These results suggest that multiple lung metastases and a poorly differentiated histology may be clinical biomarkers for sorafenib treatment in patients with HCC.

Key Word(s): 1. Portal hypertensive gastropathy; 2. gastritis; 3. children; 4. varices Presenting Author: AI FUJIMOTO Additional Authors: OSAMU GOTO, YASUTOSHI OCHIAI, JYOICHIRO HORII, KOJI TAKAHASHI, KAORU TAKABAYASHI, MOTOKI SASAKI, RIEKO NAKAMURA, TOSHIHIRO NISHIZAWA, TADATERU MAEHATA, SEIJI SAGARA, SATOSHI KINOSHITA, TEPPEI AKIMOTO, TOSHIO URAOKA, NAOHISA YAHAGI Corresponding Author: AI FUJIMOTO Affiliations: Keio University, Keio University Hospital, Fukuyama Medical Center, Tokushima

Prefectural Central Hospital, Tokyo Medical Center, Keio University, Keio University, Keio University, Keio University,Keio University, Keio University, Keio University, Tokyo Medical Center, Keio University Objective: Indication of endoscopic submucosal EPZ-6438 cost dissection (ESD) has

been expanding due to endoscopic technique and device improvement. Recently, we sometimes performed ESD for total pathological this website diagnosis when preoperative diagnosis was unconfirmed. We examined treatment outcomes and adverse events of ESD in excluded indication criteria which were performed for total pathological diagnosis. Methods: We conducted a retrospective analysis for consecutive 28 early gastric cancers (EGC) in excluded indication criteria in 28 patients who were performed ESD between June 2010 and May 2014. We examined average of longer axis for lesions, procedure time, en bloc resection (ER) rate, en bloc complete resection with margin negative (ECR) rate, curative resection (CR) rate as treatment outcomes, and perforation rate, severe bleeding rate during ESD procedure, delayed bleeding rate, incidence of severe stenosis, incidence of severe aspiration pneumonia, incidence of disease-related death and emergency

surgery as adverse events. Results: The patients characteristics of 28 EGC in 28 patients were as follows: man : female 27:1, average age 68.5 ± 13.1. Treatment outcomes were as follows: average of longer axis for lesion 26.5 ± 13.2 mm, procedure time 75.7 ± 44.1 minutes, ER rate 28/28(100.0%), ECR rate 19/28(67.8%), CR rate 7/28(25.0%). Adverse 上海皓元医药股份有限公司 events were as follows: perforation 1/28 (3.5%), delayed bleeding 2/28 (7.1%). there were no cases of severe bleeding during ESD procedure, severe stenosis, aspiration pneumonia, emergency surgery and disease-related death. Conclusion: ESD for total pathological diagnosis in excluded indication criteria has significance because ESD is safety and diagnosis of EGC has limitations. Key Word(s): 1. ESD included indication criteria Presenting Author: SHAHRIYAR GHAZANFAR Additional Authors: SAJIDA QURESHI, SAAD KHALID NIAZ Corresponding Author: SHAHRIYAR GHAZANFAR Affiliations: Dow University of Health Sciences, Dow University of Health Sciences Objective: To evaluate the success and complications of endoscopic balloon dilatation in patients with Achalasia Cardia, in a tertiary care setup.

Key Word(s): 1. Portal hypertensive gastropathy; 2. gastritis; 3. children; 4. varices Presenting Author: AI FUJIMOTO Additional Authors: OSAMU GOTO, YASUTOSHI OCHIAI, JYOICHIRO HORII, KOJI TAKAHASHI, KAORU TAKABAYASHI, MOTOKI SASAKI, RIEKO NAKAMURA, TOSHIHIRO NISHIZAWA, TADATERU MAEHATA, SEIJI SAGARA, SATOSHI KINOSHITA, TEPPEI AKIMOTO, TOSHIO URAOKA, NAOHISA YAHAGI Corresponding Author: AI FUJIMOTO Affiliations: Keio University, Keio University Hospital, Fukuyama Medical Center, Tokushima

Prefectural Central Hospital, Tokyo Medical Center, Keio University, Keio University, Keio University, Keio University,Keio University, Keio University, Keio University, Tokyo Medical Center, Keio University Objective: Indication of endoscopic submucosal click here dissection (ESD) has

been expanding due to endoscopic technique and device improvement. Recently, we sometimes performed ESD for total pathological find more diagnosis when preoperative diagnosis was unconfirmed. We examined treatment outcomes and adverse events of ESD in excluded indication criteria which were performed for total pathological diagnosis. Methods: We conducted a retrospective analysis for consecutive 28 early gastric cancers (EGC) in excluded indication criteria in 28 patients who were performed ESD between June 2010 and May 2014. We examined average of longer axis for lesions, procedure time, en bloc resection (ER) rate, en bloc complete resection with margin negative (ECR) rate, curative resection (CR) rate as treatment outcomes, and perforation rate, severe bleeding rate during ESD procedure, delayed bleeding rate, incidence of severe stenosis, incidence of severe aspiration pneumonia, incidence of disease-related death and emergency

surgery as adverse events. Results: The patients characteristics of 28 EGC in 28 patients were as follows: man : female 27:1, average age 68.5 ± 13.1. Treatment outcomes were as follows: average of longer axis for lesion 26.5 ± 13.2 mm, procedure time 75.7 ± 44.1 minutes, ER rate 28/28(100.0%), ECR rate 19/28(67.8%), CR rate 7/28(25.0%). Adverse MCE events were as follows: perforation 1/28 (3.5%), delayed bleeding 2/28 (7.1%). there were no cases of severe bleeding during ESD procedure, severe stenosis, aspiration pneumonia, emergency surgery and disease-related death. Conclusion: ESD for total pathological diagnosis in excluded indication criteria has significance because ESD is safety and diagnosis of EGC has limitations. Key Word(s): 1. ESD included indication criteria Presenting Author: SHAHRIYAR GHAZANFAR Additional Authors: SAJIDA QURESHI, SAAD KHALID NIAZ Corresponding Author: SHAHRIYAR GHAZANFAR Affiliations: Dow University of Health Sciences, Dow University of Health Sciences Objective: To evaluate the success and complications of endoscopic balloon dilatation in patients with Achalasia Cardia, in a tertiary care setup.

Key Word(s): 1. Portal hypertensive gastropathy; 2. gastritis; 3. children; 4. varices Presenting Author: AI FUJIMOTO Additional Authors: OSAMU GOTO, YASUTOSHI OCHIAI, JYOICHIRO HORII, KOJI TAKAHASHI, KAORU TAKABAYASHI, MOTOKI SASAKI, RIEKO NAKAMURA, TOSHIHIRO NISHIZAWA, TADATERU MAEHATA, SEIJI SAGARA, SATOSHI KINOSHITA, TEPPEI AKIMOTO, TOSHIO URAOKA, NAOHISA YAHAGI Corresponding Author: AI FUJIMOTO Affiliations: Keio University, Keio University Hospital, Fukuyama Medical Center, Tokushima

Prefectural Central Hospital, Tokyo Medical Center, Keio University, Keio University, Keio University, Keio University,Keio University, Keio University, Keio University, Tokyo Medical Center, Keio University Objective: Indication of endoscopic submucosal Bortezomib cost dissection (ESD) has

been expanding due to endoscopic technique and device improvement. Recently, we sometimes performed ESD for total pathological 3-Methyladenine mouse diagnosis when preoperative diagnosis was unconfirmed. We examined treatment outcomes and adverse events of ESD in excluded indication criteria which were performed for total pathological diagnosis. Methods: We conducted a retrospective analysis for consecutive 28 early gastric cancers (EGC) in excluded indication criteria in 28 patients who were performed ESD between June 2010 and May 2014. We examined average of longer axis for lesions, procedure time, en bloc resection (ER) rate, en bloc complete resection with margin negative (ECR) rate, curative resection (CR) rate as treatment outcomes, and perforation rate, severe bleeding rate during ESD procedure, delayed bleeding rate, incidence of severe stenosis, incidence of severe aspiration pneumonia, incidence of disease-related death and emergency

surgery as adverse events. Results: The patients characteristics of 28 EGC in 28 patients were as follows: man : female 27:1, average age 68.5 ± 13.1. Treatment outcomes were as follows: average of longer axis for lesion 26.5 ± 13.2 mm, procedure time 75.7 ± 44.1 minutes, ER rate 28/28(100.0%), ECR rate 19/28(67.8%), CR rate 7/28(25.0%). Adverse 上海皓元医药股份有限公司 events were as follows: perforation 1/28 (3.5%), delayed bleeding 2/28 (7.1%). there were no cases of severe bleeding during ESD procedure, severe stenosis, aspiration pneumonia, emergency surgery and disease-related death. Conclusion: ESD for total pathological diagnosis in excluded indication criteria has significance because ESD is safety and diagnosis of EGC has limitations. Key Word(s): 1. ESD included indication criteria Presenting Author: SHAHRIYAR GHAZANFAR Additional Authors: SAJIDA QURESHI, SAAD KHALID NIAZ Corresponding Author: SHAHRIYAR GHAZANFAR Affiliations: Dow University of Health Sciences, Dow University of Health Sciences Objective: To evaluate the success and complications of endoscopic balloon dilatation in patients with Achalasia Cardia, in a tertiary care setup.

19 to 0.94. Some of the highest CoAs were between fused females and their older speckled female offspring (which remained in their mother’s cluster). These older offspring

often had strong associations with their mother’s female associates and their older offspring. One speckled female lost her mother after 2000 and subsequently had strong associations (up to 0.74) with three other females in the Southern cluster. One http://www.selleckchem.com/products/ABT-263.html strong association between a fused mother and her mottled female offspring was observed and this pair had strong associations throughout the offspring’s development into adulthood (minimum CoA = 0.37, three times the population average). The majority of associations (59.0%–74.0%) were between different age classes in all years except 1997–1999, when it was 50.0%. Strong associations between speckled individuals were prevalent in all years except 1991–1993. CoA results indicated that reproductive status may have influenced strong female associations. In each pooled period, an average of about 30% of the strong female associations involved two reproductively active mottled and fused females. The majority (81%) of the strong associations were between reproductively active females in the same reproductive state (i.e., both had calves (majority), or both were pregnant, during

that time period). However there was no difference in average CoA of same reproductive state vs. different reproductive state (average CoA for both ~ 0.36). Out of all the possible combinations of mixed sex associations between

Selleckchem Caspase inhibitor individuals, 63.2%–68.0% were observed (CoA >0). This was higher than observed female-female associations, but lower than observed male-male associations. Strong CoAs ranged from 0.19 to 0.97, with the two highest (and several MCE lower strong CoAs) between females and their older speckled male offspring. The majority of associations were mixed age class (between 58.3%–72.9%) through 1999, but in 2000–2002 mixed age class associations comprised only 46.6%. Generally, strong mixed sex associations were between individuals of the same cluster. Only 21.3% of the observed mixed sex associations were between individuals from different clusters. One Northern male, Rivet, had strong associations with Central females in every pooled period, and one association with a Southern female in 2000–2002. No other Northern-Southern strong associations were observed. Although other Northern and Southern males had a few strong associations, the majority of cross cluster associations were between Southern or Northern females and Central males. There were speckled and young mottled males involved in cross cluster associations but the vast majority involved fused males. The percentage of males involved in mixed sex associations (85.7%–100.0%) was always larger than the percentage of females involved (73.1%–84.1%).

19 to 0.94. Some of the highest CoAs were between fused females and their older speckled female offspring (which remained in their mother’s cluster). These older offspring

often had strong associations with their mother’s female associates and their older offspring. One speckled female lost her mother after 2000 and subsequently had strong associations (up to 0.74) with three other females in the Southern cluster. One MG-132 strong association between a fused mother and her mottled female offspring was observed and this pair had strong associations throughout the offspring’s development into adulthood (minimum CoA = 0.37, three times the population average). The majority of associations (59.0%–74.0%) were between different age classes in all years except 1997–1999, when it was 50.0%. Strong associations between speckled individuals were prevalent in all years except 1991–1993. CoA results indicated that reproductive status may have influenced strong female associations. In each pooled period, an average of about 30% of the strong female associations involved two reproductively active mottled and fused females. The majority (81%) of the strong associations were between reproductively active females in the same reproductive state (i.e., both had calves (majority), or both were pregnant, during

that time period). However there was no difference in average CoA of same reproductive state vs. different reproductive state (average CoA for both ~ 0.36). Out of all the possible combinations of mixed sex associations between

selleck chemicals llc individuals, 63.2%–68.0% were observed (CoA >0). This was higher than observed female-female associations, but lower than observed male-male associations. Strong CoAs ranged from 0.19 to 0.97, with the two highest (and several 上海皓元 lower strong CoAs) between females and their older speckled male offspring. The majority of associations were mixed age class (between 58.3%–72.9%) through 1999, but in 2000–2002 mixed age class associations comprised only 46.6%. Generally, strong mixed sex associations were between individuals of the same cluster. Only 21.3% of the observed mixed sex associations were between individuals from different clusters. One Northern male, Rivet, had strong associations with Central females in every pooled period, and one association with a Southern female in 2000–2002. No other Northern-Southern strong associations were observed. Although other Northern and Southern males had a few strong associations, the majority of cross cluster associations were between Southern or Northern females and Central males. There were speckled and young mottled males involved in cross cluster associations but the vast majority involved fused males. The percentage of males involved in mixed sex associations (85.7%–100.0%) was always larger than the percentage of females involved (73.1%–84.1%).