The aim of this study was to determine whether specific organisms in the tissue-associated microbiota are associated with inflammatory pouch complications.Inflammatory complications following ileal pouch-anal anastomosis (IPAA) for ulcerative colitis (UC) are common and thought to arise through mechanisms similar to Patients having previously undergone IPAA were recruited from Mount Sinai Hospital. Clinical and demographic information were collected and a pouchoscopy with biopsy of both the pouch and afferent limb was performed. Patients were classified based on post-surgical phenotype into four outcome groups: familial adenomatous polyposis controls (FAP), no pouchitis, pouchitis, and Crohn’s disease-like (CDL). Pyrosequencing of the 16S rRNA V1-V3 hypervariable region, and quantitative PCR for bacteria of interest, were used to identify organisms present in the afferent limb and pouch. Associations with outcomes were evaluated using exact and non-parametric tests of significance.Bacteroidetes were detected significantly less frequently (P<0.0001) in the inflammatory outcome groups (pouchitis and CDL) compared to both FAP and no pouchitis. Conversely, Proteobacteria were detected more frequently in the inflammatory groups (P=0.01). At the genus level, organisms associated with outcome were detected less frequently among the inflammatory groups compared to those without inflammation. Several of these organisms, including Bacteroides (P<0.0001), Parabacteroides (P≤2.2x10-3), Blautia (P≤3.0x10-3) and Sutterella (P≤2.5x10-3), were associated with outcome in both the pouch and afferent limb. These associations remained significant even following adjustment for antibiotic use, smoking, country of birth and gender. Individuals with quiescent disease receiving antibiotic therapy displayed similar reductions in these organisms as those with active pouch inflammation.Analysis at the phylum level indicated that Specific genera are associated with inflammation of the ileal pouch, with a reduction of typically ubiquitous organisms characterizing the inflammatory phenotypes. Escherichia coli (AIEC) and Mycobacterium avium subspecies paratuberculosis as potential contributors to pathogenesis .,20.19,20DNeasy Blood and Tissue Kit  with an additional bead beating step was used to extract microbial DNA. Barcoded primers specific for the V1-V3 hypervariable region of the 16S rRNA molecule were used for 454 pyrosequencing using the GS FLX Titanium assay  [mothur (Supplementary Methods).Biopsies were thawed and processed in two batches at the Department of Animal Science, University of Manitoba . The QIAGEN he, USA) . SequencTaxonomic groups which were detected in less than 5% of the samples were excluded from further analysis. Analyses were performed separately on pouch and afferent limb samples in order to avoid falsely inflating significance. Genus level results were analyzed using presence-absence and frequency data with exact and non-parametric methodologies (Supplementary Methods). Confirmation of results and adjustment for confounding variables was performed using exact logistic regression. FDR corrected p-values below 0.05 were considered significant. All statistical analyses were conducted using STATA version 11.1  and R version 2.13.1 . Additional analyses were carried out using the Linear Discriminant Analysis (LDA) Effect Size (LEfSe)  tool  [2 transformed and analyzed using the Kruskal-Wallis test. Additionally, primers specific for the Bacteroides genus were used to confirm pyrosequencing results by evaluating the correlation between dichotomized (Matthew’s) and frequency (Spearman) pyrosequencing and qPCR data.Organisms previously implicated in bowel inflammation including er pairs . RelativMethods) . Results274 patients were recruited to this study. To ensure a balanced study design, 78 were selected for analysis based on phenotype, with 71 classified into the four primary outcome groups. Seven patients were excluded from the initial analysis having been diagnosed with chronic pouchitis or CDL which was effectively treated with antibiotics or other medications (quiescent). In total, there were 18 patients in the FAP group, 19 no pouchitis, 15 pouchitis, and 19 CDL. Among those classified as CDL, two had only perianal fistulae and an inflammation score of zero in both the pouch and afferent limb (no evidence of inflammation at the time of endoscopy). One additional patient included in this group had evidence of only mild inflammation in both the pouch and afferent limb , with additional features of the CDL phenotype. Inflammatory activity, as measured by our objective inflammatory score, was low among individuals in the FAP and no pouchitis groups, and higher in both the pouchitis and CDL groups in the pouch (P<0.01). In the afferent limb, only individuals in the CDL group displayed evidence of inflammation (P<0.01) .P=0.007) among individuals who were in the FAP group. The majority of patients (54%) had taken antibiotics,  at some point following their pouch procedure . Antibiotic use at the time of pouchoscopy, or during the month immediately prior is documented in Interestingly, C-reactive protein (CRP) levels were similarly elevated among each of the UC groups, compared to individuals with FAP (P=0.002) . There wThe pyrosequencing experiment provided 557,215 (mean of 7143 sequences per sample) raw sequences with a median read length of 453 basepairs range 250-531). However, following quality trimming and chimera checking, 345,466 (mean of 4429 sequences per sample) high quality reads remained. To ensure that our stringent inclusion criteria did not adversely affect our ability to detect organisms in our samples, Good’s coverage was calculated for each sample, and was on average greater than 92% for all outcome groups. A single sample from the pouchitis group had coverage which was below 80%. Diversity, measured using the Shannon  and the 0-531. HoFirmicutes, Proteobacteria, Bacteroidetes, and Actinobacteria were the most diverse phyla present with the majority of genera detected belonging to one of these groups  . Similar trends were observed between the pouchitis and FAP group, although not reaching statistical significance.The e groups . Firmicurganisms . Additio=0.0001) . ResultsIn total, 287 genera were detected. Of these 83 were present in more than 5% of samples. There was no statistical difference in genera positivity or abundance between sampling locations (pouch vs. afferent limb). However, to avoid bias from including multiple samples from the same individual, analysis was conducted separately on the different biopsy sites.P≤0.05, with five  remaining significant after FDR correction for multiple testing (Pcorr≤0.05)  remaining significant after correction  . In the rrection . These gMoryella in the pouch and Anaerococcus in the afferent limb did not remain significant after multiple testing correction (Bacteroides (P=2.5x10-5) and Parabacteroides (P=2.5x10-4) less commonly detected among inflamed samples . The results were similar to those in the previous analysis, although rrection . When we samples .Bacteroides, Blautia, and Sutterella were positively associated with outcome, as were, Parabacteroides and Moryella in only the pouch samples (Pcorr<0.05)  . In eachLactobacillus in both the pouch and afferent limb . Also associated with antibiotic use in the afferent limb were Dorea (P=4.0x10-4) and Ralstonia (P=1.4x10-4), with Ralstonia detected more commonly among individuals who were on antibiotics, and Dorea less common among individuals on these medications. Several additional genera, including those which were associated with outcome were shown to be marginally effected by antibiotic use at a nominal significance threshold (P<0.05), however did not remain significant after correction for multiple testing. Genera abundance data showed similar results, with Lactobacillus and Haemophilus comprising an increased proportion of the microbiome among individuals on antibiotics in both the pouch and afferent limb, and Ralstonia, detected at higher frequency in the afferent limb. Dorea was detected at a lower frequency in the afferent limb among individuals on antibiotics. No other medication use appeared to significantly influence microbial composition, although few patients were available for this analysis.Across all groups, use of one or more antibiotics at the time of the pouchoscopy or during the preceding month was positively associated with the presence of P<0.05) with the exception of Moryella in the pouch and Parabacteroides in the afferent limb (53.5% of our population had taken antibiotics at some point in the past (pre- or post-colectomy) . To verient limb . All preBacteroides, Sutterella and Blautia, remained significantly associated with outcome (P<0.01). Moryella, and Parabacteroides were associated with outcome only in the pouch and Dorea and Anaerococcus in the afferent limb  which was detected at higher frequency among inflammatory groups, and Proteobacteria detected more often among individuals with CDL . Furthermore, detection rates among the outcome groups were similar with Bacteroides detected less frequently in the pouchitis and CDL groups than in either FAP or no pouchitis groups (P<0.01). Additional organisms associated with outcome included the Roseburia spp. / Erectale.  group (pouch) and Clostridium cluster IV (afferent limb). These organisms were present in increased numbers in the FAP group compared to both pouchitis and CDL groups . L groups .P<0.01) were observed between the quiescent and FAP groups in several genera in both the pouch (Bacteroides and Blautia) and the afferent limb  in both the dichotomized and frequency analyses , it is present at low abundance (mean 0.8%). Fusobacteria, on the other hand, were detected in only 28% of samples, yet among those individuals in whom these organisms were detected they were dominant members of the bacterial community. This contrasts reports from others [At the phylum level, significantly decreased levels of ical IBD ,28,29,30m others , and canClostridia group XIVa, Blautia, Moryella and Dorea were convincingly associated with outcome, detected more frequently and in greater abundance among individuals without inflammatory complications. Interestingly, a recent study of healthy subjects showed that Blautia was one of the only genera detected in samples from all individuals, suggesting that it is a common component of a healthy gut microbiota [Clostridia-related organisms detected [Bacteroides and Parabacteroides were also strongly associated with outcome. These closely related genera have both been previously implicated in pouch inflammation [Bacteroides have also been previously observed in some studies in inflamed UC and CD tissue compared to healthy controls, while other studies have shown increases in this organism to be associated with disease [Bacteroides species, specifically B. fragilis, have been shown to modulate immune function, stimulating immune tolerance through Toll-like receptor (TLR) 2 [Sutterella was the only genus from the Proteobacteria phylum associated with outcome, and in contrast with previous studies suggesting a potential pathogenic role for this organism in IBD or no association at all [Proteobacteria and the class Bacilli among the inflamed groups, however this association was less dramatic. Further study investigating the relevance of these groups of organisms is necessary to determine the significance of this finding.Phylum level comparisons, while interesting, are of limited use in predicting biological function. We subsequently demonstrated that specific genera are associated with pouch outcome among our patient cohort. Among the crobiota . Furtherdetected ,33. Bothammation ,28. Decr disease ,35,36,37 (TLR) 2 . Suttern at all , was detFprausnitzii.  or AIEC and inflammatory outcome in our cohort. However, our results do support previous observations that additional members of the Clostridium group [Roseburia spp./E. rectale may be associated with pouch inflammation.We specifically evaluated whether organisms previously associated with bowel inflammation, including CD, were also involved in pouch inflammation to determine whether similar biological processes might be of importance in both disorders. Contrary to previous studies, we did not observe an association between um group , and RoThe use of subjects who had an IPAA for FAP, allowed us to evaluate changes occurring in the pouch microbiome which were relevant specifically to individuals with an inherited predisposition to intestinal inflammation. Individuals with FAP rarely develop pouch inflammation, despite the fact that their digestive tract anatomy is identical to those with UC. Additionally, subclinical inflammation may be present in individuals with pre-colectomy UC, without a clinical diagnosis of pouchitis, as our finding of elevated levels of CRP among this group may suggest. This subclinical inflammation has important clinical and experimental implications, and is reflected in our observations illustrating substantial changes between the microbiome of individuals with inflammatory pouch complications compared to those with FAP. More moderate, yet similar alterations occur in the microbiome of individuals with UC when comparing those with and without inflammation. This likely reflects underlying differences in host genetic predisposition in UC patients resulting in specific alterations in the pouch microbiome that characterize the inflammatory phenotypes.While there were no statistical differences in medication use between outcome groups at the time of or immediately prior to pouchoscopy, it is conceivable that medications, particularly antibiotics, may have impacted the tissue associated microbiota. Yet the described genera were detected significantly less frequently among individuals in the inflammatory outcome groups even when individuals who had taken antibiotics in the preceding month or six months were excluded from the analysis. This would suggest that the effect of inflammation on the microbiome supersedes the impact of antibiotic use. Alternatively, if shifts in the microbiome of the pouch precede and possibly induce pouch inflammation, then the use of antibiotics may not result in long term beneficial effects on bacterial composition or lead to a reversion to a “healthy” pouch microbiome as would be suspected by the typically antibiotic responsive nature of pouchitis. This may in turn be responsible for the often recurrent nature of pouchitis following treatment. It is also possible that past antibiotic use (beyond one month prior to procedure) leads to permanent changes to the microbiome which are detectable long after cessation of medications. Studies evaluating the long-term effect of antibiotics on intestinal microbial composition have shown mixed results with some studies showing a relatively quick reversion to a profile similar to the pre-medication state , while oInterestingly, we did not observe differences in the microbial composition of the pouch compared to the afferent limb among patients regardless of the inflammatory status of either site. The anatomical alterations resulting from the IPAA procedure, namely the surgically induced continuity between the pouch and afferent limb, likely promotes similar bacterial community structure between these two locations. Previous studies have documented the occurrence of variability in the microbiome between inflamed and non-inflamed neighbouring sites, although none were consistent across many individuals . This suOf particular interest, is the observation that the seven individuals who were taking medication for chronically inflamed pouch phenotypes, and who had successfully achieved resolution of their symptoms, did not demonstrate a change in their microbial composition to resemble that of individuals with healthy pouches. While it appears that the samples from individuals with quiescent disease were most similar to those from individuals with pouchitis, it is conceivable that with a larger sample size differences between these groups may have been observed. Additionally, changes at lower taxonomic levels (ie. species), which were undetectable in this experiment could also have been associated with different phenotypes. It is interesting to speculate, however, that perhaps the reason for patients’ continual requirement for medical therapy is their inability to be colonized by the ‘beneficial organisms’ described by our group.Clostridia XIVa group  produce butyrate, a known anti-inflammatory molecule [Bacteroides produce vitamin K, which is associated with decreased levels of serum inflammatory markers  [The majority of associations described in this report suggest that a loss of organism diversity is associated with inflammation. It is possible that individuals with pouch inflammation may undergo a loss of specific anti-inflammatory organisms that have important roles in modulating the immune response. Many of the organisms detected less frequently in the inflammatory groups have been previously shown to have anti-inflammatory potential: members of the molecule ,45, and D40 etc) . AlternaD40 etc) ,47,48. SD40 etc) ,49. In tOur experiments have demonstrated that a reduction in specific organisms is associated with inflammatory outcomes. These changes are characteristic of both pouchitis and the Crohn’s disease-like phenotype, and include several organisms which have been previously associated with non-surgical IBD. These observations demonstrate that the role of these microbes in bowel inflammation may be common and independent of surgical history or anatomic location of disease. Our previous findings showing that IBD-associated genetic polymorphisms are also important in pouch pathology , in conjSequence accession numbers and availabilitySequences generated in this study are publically available (dbGaP accession number: phs000659.v1.p1).Figure S1Distribution of inflammatory activity scores through each of the phenotypic outcome groups.Mean values for each group are indicated with a red line.(BMP)Click here for additional data file.Figure S2Phylum level comparisons between four outcome groups for the pouch and afferent limb.FAP=familial adenomatous polyposis, CDL=Crohn’s disease-like.(BMP)Click here for additional data file.Figure S3Pcorr<0.05) following removal of individuals without evidence of inflammation at study pouchoscopy from the CDL group.Proportion of patients positive for genera which were significantly associated with outcome (* represent pairwise comparisons which were significant (Pcorr <0.05) after correction. A) Pouch samples. B) Afferent limb samples. I.S. = Incertae sedis.(BMP)Click here for additional data file.Figure S4Pcorr<0.05) in A) pouch and B) afferent limb.Mean and standard error of the frequency of genera significantly associated with outcome  Bacteroides, ii) other significant organisms.(BMP)Click here for additional data file.Figure S5Pcorr<0.05) in the preliminary analysis, among the cohort of patients on no antibiotic therapy (n=57).Proportion of patients positive for genera which were significantly associated with outcome (* represent pairwise comparisons which were significant (Pcorr <0.05) after correction. A) Pouch samples. B) Afferent limb samples. I.S. = Incertae sedis.(BMP)Click here for additional data file.Figure S6A) Cladograms demonstrating the results obtained through LDA Effect Size (LEfSe) analysis.Highlighted results are those which were increased in the corresponding group. B) Differential abundance of organisms detected at significantly different frequencies via LEfSe.(PDF)Click here for additional data file.Figure S72 transformed relative abundance of organisms of interest from real-time quantitative PCR in the A) pouch and B) afferent limb.logSignificant results are marked with an astrix (Pcorr<0.05). FAP=familial adenomatous polyposis, CDL=Crohn’s disease-like.(BMP)Click here for additional data file.Figure S8Proportion of individuals in each outcome group positive for genera which were previously associated with pouch inflammatory outcomes.FAP=familial adenomatous polyposis, Quiescent=seven individuals requiring long-term medical therapy to maintain remission. Lines represent associations between the quiescent group and others which reached nominal significance (P<0.05). A) Pouch samples. B) Afferent limb samples.(BMP)Click here for additional data file.Table S1(PDF)Click here for additional data file.Table S2(PDF)Click here for additional data file.Table S3(PDF)Click here for additional data file.Table S4(PDF)Click here for additional data file.Table S5(PDF)Click here for additional data file.


Increasing alcohol consumption among older individuals in the UK is a public health concern. Although brief interventions (BIs) can effectively reduce heavy drinking in older age groups generally, social and contextual factors may influence implementation and effectiveness. Identities offer a useful theoretical concept to explain why the potential for public health messages to reduce rates of heavy drinking in this sector has not been realised, and can inform preventive approaches. A qualitative study explored older people’s reasoning about drinking in later life and how this interacted with health concerns, to inform future, targeted prevention in this group. A diverse sample of older adults in North East England (ages 51-90) participated in interviews  and three focus groups (participants n=27). Data were analysed using grounded theory and discursive psychology methods. Older adults portrayed drinking less alcohol as an appropriate response if one experienced impaired health. However continued heavy drinking could be presented as normal behaviour for someone experiencing relative wellbeing in later life, or if ill health was construed as unrelated to alcohol consumption. When talking about alcohol use older people oriented strongly towards opposed identities of normal or problematic drinker, defined by propriety rather than health considerations. These identities were flexible in older people’s talk since they could be applied to either moderate or heavy drinking. Older people displayed scepticism about health advice on alcohol when avoiding stigmatised identity as a drinker. Findings indicate that older UK adults do not recognise unhealthy drinking as distinct from dependent drinking, and are highly sensitive to stigma when discussing alcohol consumption. Preventive strategies should encourage older people’s identification of heavy drinking as neither healthy nor synonymous with dependence. BIs for heavy drinkers in later life should be tailored to address lay reasoning that is resistant to recognition of health risks.


In 2000 the NHS plan sought to double the number of patients entering cancer trials within 3 years. This was achieved through the creation of the National Cancer Research Network in 2001. By 2004, approximately 10.9% of all cancer patients were entered into a trial. However, recruitment of Black, Minority and Ethnic (BME) patients into clinical trials has been reported to be under-represented compared to the split of ethnicity both within the reported incidence of disease and the reported total population. Mason found a two-fold under representation of South Asians within breast cancer trials . The under-representation of any patient group within a clinical trial, specifically an ethnic one, can bias trial results, and subsequent extrapolation into the general population.In the USA, a significant under-representation of Hispanics, Blacks and Asians was found when comparing ethnic proportions recruited within oncology trials with existing cancer cases  and the USA National Institute of Health Revitalisation Act (1993).Preliminary results of the review have revealed a paucity of published evidence from the UK, with the majority of the articles meeting the inclusion criteria originating from the USA. Final results will be presented.


UV-Vis shows significant bathochromic shift in benzoyl upon coordination as well as IR well illustrates the peak shift of C=O group and formation of a O–Cr(III) bond. Likewise, 1H NMR studies clarify that Cr(III) metal ion replaces the 5OH proton hence; 5-hydroxy-4-keto site is employed by morin in chelation to form six-membered stable ring system out of three available chelating sites. In addition, TG-DTA denotes the presence of coordinated and crystalline water molecules. The melting point of the complex was found to be 389°C by DSC. In addition, Cr(III)-morin complex was found to be a more potent antioxidant than morin as evaluated by DPPH• and FRAP methods.The complex formation between Cr(III) and morin was carried out in methanol and confirmed by analytical characterization using UV-Vis, IR,  Flavonoids have recently attracted great interest as potential therapeutic agents against a variety of diseases like those involving radical damage. These polyphenolic compounds, ubiquitous in higher plants, are commonly major dietary constituents. The biological and medicinal properties of flavonoids have been reviewed extensively, with wealth of data on their activity as reducing agents, hydrogen-donating antioxidants, and singlet oxygen quenchers –5. They in vitro and in vivo. It is also reported that morin could modulate the activities of the metabolic enzymes, including cytochrome P450 Cl·2H2O: C, 43.20; H, 4.11%, respectively. It was soluble in H2O, MeOH, EtOH, DMSO, DMF, acetone, and n-hexane and completely insoluble in DCM and chloroform.In a 50 mL two-necked round-bottomed flask equipped with electromagnetic stirrer and a thermometer, solid morin (0.6044 g or 0.2 mmol) was dissolved in MeOH and stirred until it was completely dissolved within 15 minutes and the solution became clear yellow colored and then we quickly added solid CrClμL of each solution was mixed with 2 mL of the solution of DPPH• free radical having concentration of 0.1 mM. The decline in absorbance of DPPH• solution made by a sample compound was noted with the gape of every 5 minutes in the order of 0, 5, 10, 15, 20, 25, and 30 minutes till reaching the steady state. Finally, we calculated the percent scavenging (SC%) for each sample by using formula (The antioxidant activity for morin and the synthesized complex of Cr(III)-morin was studied spectrophotometrically by DPPH• method. DPPH• itself is a stable commercially available free radical. Its solution in methanol is violet upon reduction by an antioxidant it changes to corresponding yellow color. The absorbance of DPPH• is noted at 515 nm, which decreases by addition of an antioxidant sample. For the measurement of relative antioxidant activity of both of the compounds, they were prepared of appropriate concentration in methanol . Their cFerric reducing power of morin and the Cr(III)-morin complex was also studied by using the method of Oyaizu . In thisMorin is one of the important members of flavonoid family which exhibits two distinct peaks in the electronic spectrum, where peak I appears at 359 nm for cinnamoyl part and simultaneously band II emerges at 263 nm for benzoyl part  by analy31. In addition, structurally similar quercetin was also found to preferentially bind Fe(III) at 5-hydroxy-4-keto site studied by UV-Vis and cyclic voltammetry 3 complex which is reduced to Fe2+ and changed into yellow color (cuvettes) as shown in Absorbance of samples increased with increase in their concentrations. The higher reducing capacity originates from the higher amounts of polyphenolics (reductones) present and therefrom the antioxidant potential of the investigated compounds can be measured . The redt 700 nm . Thus coAt the same time, the study proves that the complexation between morin and metal ion takes place, which may also be very useful to remove many toxic metal ions from body at specific conditions through metal chelation therapy and it can also be useful to supply the essential metal ions to body if it is deficient to any essential metal ion.1H NMR studies. From DPPH radical scavenging as well as ferric reducing power studies, it has been proved that complex of Cr (III)-morin is a more powerful antioxidant than the morin molecule alone. In addition, the study shows very nice impact for designing anticancer drugs, combined with their potential cytotoxic and antioxidant activities, which can be targeted selectively against cancer cells and increase their therapeutic index and additional advantages over other anticancer drugs.It has been concluded from the study that Cr(III) forms complex from 4C=O and 5-OH groups, which is proved from various techniques such as UV-Vis, IR, and


Brain stem arteriovenous malformations (AVMs) are rare and their clinical management is controversial. A location in highly eloquent areas and a greater risk of radionecrosis are both serious issues for radiosurgery of this entity. We report a case of a pontine AVM treated successfully with gamma knife therapy. At 3 years angiographic follow-up, imaging demonstrated complete thrombosis and there were no new neurological deficits, and at 7 years clinical follow-up, the patient continued to be neurologically stable. Although all treatments carry risk of neurological compromise, gamma knife therapy may offer the best treatment option for brain stem AVMs as seen in the case presented herein. This case illustrates a rare case of holo-pontine AVM tolerating gamma radiation with complete angiographical response and minimal neurological sequalae. Arteriovenous malformations (AVM) are congenital vascular malformations with direct arterial to venous connections without an intervening capillary network Doppman . The abrThe main diagnostic tools for these pathologic entities are magnetic resonance imaging (MRI), CT angiogram, and angiography , where single treatment of a pontine AVM with overlapping isodoses led to complete obliteration without any new neurological deficit.Any treatment of brain stem AVMs offers considerable risk for neurological compromise. Radiosurgery in highly selected cases may offer a treatment option with reasonable risks.


Background: The gold standard of the management of rectal cancer in the middle and lower parts is low anterior resection with coloanal anastomosis. About 50% of the patients undergoing this procedure might experience some complications because of the low capacity of the neorectum. The aim of this study was to evaluate ileal J-pouch interposition as a neorectum between the anal canal and the remaining colon in comparison to coloanal anastomosis and transverse coloplasty.Methods: Twelve dogs, weighing 23-27 kg, were divided into three groups. After laparotomy, the volume of the primary rectum was measured so that it could be compared with that of the neorectum at the end of the study. After rectal resection in Group A, the colon was directly anastomosed to the anus. In Group B, a 5-cm longitudinal incision was made 2 cm proximal to the anastomosis and was sutured transversely (coloplasty). In Group C, a 5-cm ileal J-pouch was interposed between the colon and anus. After 8 weeks, the neorectum was evaluated for volume, radiology, and pathology.Results: All the samples were alive until the end of the study. The healing of the anastomotic lines was acceptable  in all. The mean volume expansion was 20.9% in Group A, 21.7% in Group B, and 118.2% in Group C, with the latter being significantly higher than that of the other groups (P=0.03). Colon J-pouch and coloplasty after proctectomy in some situations have not been performable. This study evaluated the performance of ileal J-pouch interposition.Conclusion: This study showed that ileal J-pouch interposition might produce an acceptable reservoir function and that it seems feasible and safe in selected cases. The neorectum is constructed with the remaining left colon. However, almost all of the patients with straight coloanal anastomosis have tolerated some complications of defecation after low anterior resection such as urgency, frequency, soiling, anastomotic line stenosis because of the vascular tension of the sigmoid colon, and incontinence due to the low capacity of the neorectum.Accordingly, in this animal pilot study, we tried to construct the neorectum with the interposition of the ileal J-pouch between the anal canal and the remaining colon. We thereafter evaluated the efficacy of the ileal J-pooch as a neorectum in comparison to straight coloanal anastomosis and transverse coloplasty. MaterialsThe present study was performed in the Animal House of Shiraz University of Medical Sciences (South of Iran) in August, September, and October 2011 and was approved by the Ethics Committee of Shiraz University of Medical Sciences for the use of laboratory animals.   In this study, 12 German shepherd dogs, weighing 23-27 kg, were selected and divided into three equal groups. Surgical Procedure1 In all the cases, the rectum was resected 2 centimeters above the dentate line. The proximal margin was resected at the level of the sacral promontory. Total mesorectal excision was done.1,6 After the resection of the rectum, in Group A the colon was directly anastomosed to the anus. In Group B, however, a 5-cm longitudinal incision was made 2 cm proximal to the anastomosis and was transversely sutured (coloplasty).3 In Group C, 10 centimeters of the terminal ileum with the main branch of the ileocolic vessel was separated.7 After the creation of a J-pouch by the terminal ileum, the pouch was applied in the pelvis and anastomosis was done with Prolyn 3/0. All the dogs were anesthetized with 17 mg/kg of thiopental intravenously. After intubation, anesthesia was continued through a mixture of halothane and oxygen up to the end of the operation. Hydration of the samples was maintained by 1000 cc of D/S during the operation. Additionally, half of a pen-and-strep vial (3 million units of penicillin plus 3 g of streptomycin) and a 1 g of Keflin were injected preoperatively. Then via a midline laparotomy incision, a clamp was placed 10 cm proximal to the anus. An orotracheal tube (OT tube) was entered into the rectum, and 10 ml of air was injected into its cuff. In order to prevent leakage, a purse-string suture was placed around the anus with nylon 1. Moreover, the volume of the primary rectum was measured to be compared with that of the neorectum at the end of the study. To measure the volume of the rectum, N/S solution was injected into the rectum in the form of free fluid through a bottle, which was placed 100 cm above the anus level. After fullness of the rectum, the volume of the primary rectum was measured and recorded. In fact, the volume of the primary rectum was equal to the volume of the injected N/S plus the 10 ml of air injected into the OT tube. Afterwards, the rectum was emptied, the OT tube was also removed, and the intended operations were performed in each group.Postoperative ManagementAfter irrigation and hemostasis, the abdominal wall was closed in layers and tetracycline was sprayed on the wound. The animals were kept NPO and were given intravenous fluid (D/S) for 3 days with half of a pen-and-strep vial intramuscularly per day for 10 days postoperatively. After 3 days, a soft food diet without bones was started for them for 7 days. Then, they were given normal food up to the end of the study. The samples were kept in the same condition for 8 weeks. Afterwards, the volume of the neorectum was measured and recorded. Furthermore, the neorectum was removed for pathological and radiographic evaluations.Statistical AnalysisA non-parametric Mann-Whitney U test with SPSS (version 18) was used for data analysis. Significance level was considered as 0.05. All the dogs were alive until the end of the study with a good condition. According to the pathological reports, the healing of the proximal anastomosis was acceptable in all the three groups under study. Inflammation and ulceration was detected in the samples taken from the pouch . Repair After biopsy, all the samples were filled by contrast and evaluated by an expert radiologist. In comparison, the volume increase in the pouch group  was markThe dogs’ weights in the three groups under study were not markedly different. The primary volume of the rectum, volume after 8 weeks (end of the study), and volume increase for each dog were measured. The volume increase in each group was also calculated .Considering Group A (the control group), the percentage of the increase in the volume of the rectum (the volume of the primary rectum in comparison to the volume of the neorectum at the end of the study) was as follows:A1: 150cc            180cc (20% ↑)A2: 150cc             200cc (33% ↑)A3:140cc              150cc (7.1% ↑)A4: 170cc             210cc (23.5% ↑)Moreover, the mean volume increase in Group A was measured as 20.9%.The percentage of the volume increase in the place of the rectum in Group B (the coloplasty group) was as follows:B1: 160cc           180cc (12.5% ↑)B2: 130cc          150cc (15% ↑)B3: 140cc          180cc (28.5% ↑)B4: 130cc         170cc (31% ↑)In addition, the mean volume increase in Group B was equal to 21.7%.Finally, the percentage of the volume increase in the place of the rectum in Group C (J-pouch) was as follows:C1:170cc           350cc (106% ↑)C2:155cc           380cc (145% ↑)C3:150cc           300cc (100% ↑)C4: 90cc            200cc (122% ↑)Also, the mean volume increase in Group C was 118.2%.3 This situation occurs in the following conditions:Although colon J-pouch is the best method of operation after removing the rectum, J-pouch coloanal anastomosis was not possible in 26.2% of low rectal cancer patients who had undergone low ant resection plus total mesorectal excision.3Narrow pelvic, Bulky sphincter, Diverticulitis, Insufficient colon length, Pregnancy, Complex surgery, Distant metastasisNowadays, the low ant resection operation, accompanied by total mesorectal excision (TME) is considered the standard treatment for rectal cancers. Moreover, in order to decrease the complications of the direct anastomosis of rectum to anus, various operational methods such as side-to-end coloanal anastomosis, coloplasty, colonic J-pouch, and even ileocecal pouch interposition have been used.Considering the comparison of the primary volume to the volume after the operation, the findings of the present study revealed no significant difference between the volume increase in Group A (control) and Group B (coloplasty) . Therefore, one can conclude that coloplasty operation has no superiority over coloanal anastomosis, which is a simple operation, and, consequently, it causes no more significant increase in the place of the rectum.In Group C (J-pouch), however, a highly significant volume increase was observed compared to the control group . The volume increase in Group C (J-pouch) was also significantly different from that of Group B (the coloplasty group) (P=0.030). Comparison of the pathology slides showed healing at the place of the anastomosis in all the three groups. However, the amount of inflammation in Group C (in the place of the pouch) was more in comparison to the place of the neorectum in the other two groups. Furthermore, coloplasty can be used as an appropriate treatment option since it is not accompanied by early dysfunction, which occurs after straight coloanal anastomosis, and long-term problems as well as the problems related to pouch evacuation, which occur after performing the colon J-pouch.7,8Moreover, in comparison to colon J-pouch, the chance of clinical or radiological leakage is higher in coloplasty. Therefore, the blood flow is lower at the place of the proximal anastomosis and, particularly, the anterior area in the place distal to the performance of coloplasty.8 conducted a study and used the cecum reservoir as the neorectum by maintaining the neurovascular part of the cecum and ileum. They came to the conclusion that this method of operation was safe and practical and that it provided acceptable physiological results up to 6 months after the operation.  In a study, the chance of leakage and stricture in the CP operation was shown as 7% and 14%, respectively.9 In the first year after CP and colon J-pouch operations, stool fragmentation may occur, which causes the patients to defecate in 15-minute intervals. Of course, the patients may take this situation for the increase in the number of defecations by mistake.10 Mantyh11 conducted a study and revealed that the functional results were similar in both the CP group and the colon J-group.In 1996 on Flüe et al.7 In comparison to straight coloanal anastomosis or CP, colon J-pouch has less chance of leakage. This is due to the better blood flow in the direction of the proximal anastomosis, which is shown through the laser Doppler technique.12 Colon J-pouch can increase the volume of the rectum, especially when the pouch is long; nonetheless, the increase in the length of the pouch can decrease the motility.1 In fact, colon J-pouch must be designed in a way that the pouch is not more than 5-6 cm because the increase in its length can lead to the problems related to evacuation.6Nowadays, after removing the rectum, colon J-pouch operation is known as the best way for connecting the colon to the anus.13When the length of the colon J-pouch is less than 6 cm, the findings of anorectal manometry, tolerable volume and compliance, and maximum rectal volume of the initial sensation are similar in both the CP group and the J-pouch group. However, in case the length of the colon J-pouch is more than 6 cm, these parameters are better in the colon J-pouch group compared to the CP group.In spite of the fact that both CP and J-pouch have advantages as well as disadvantages and colon J-pouch is considered as the best method of operation, colon J-pouch cannot be performed on all cases. Therefore, another method of operation must be utilized in such cases. The results of the present study revealed that ileal J-pouch can be done in the previous location of the rectum and that it provides an appropriate volume. More studies are, however, needed to be conducted on the issue.Colon J-pouch reconstruction after rectal resection is not a suitable procedure in several cases. Therefore, this study evaluated the possibility of the creation of an ileal J-pouch interposition in an animal model and evaluated the volume of the neorectum. The present study is an animal study with a small sample size. If larger studies demonstrate that ileal J pouch interposition can safely create an acceptable reservoir function, this technique can be performed as a new procedure in selected cases.


Objectives. (1) To determine the frequency of maternal near miss, maternal near miss incidence ratio (MNMR), maternal near miss to mortality ratio and mortality index. (2) To compare the nature of near miss events with that of maternal mortality. (3) To see the trend of near miss events. Design. Audit. Setting. Kasturba Hospital, Manipal University, Manipal, India. Population. Near miss cases & maternal deaths. Methods. Cases were defined based on WHO criteria 2009. Main Outcome Measures. Severe acute maternal morbidity and maternal deaths. Results. There were 7390 deliveries and 131 “near miss” cases during the study period. The Maternal near miss incidence ratio was 17.8/1000 live births, maternal near miss to mortality ratio was 5.6 : 1, and mortality index was 14.9%. A total of 126 cases were referred, while 5 cases were booked at our hospital. Hemorrhage was the leading cause (44.2%), followed by hypertensive disorders (23.6%) and sepsis (16.3%). Maternal mortality ratio (MMR) was 313/100000 live births. Conclusion. Hemorrhage and hypertensive disorders are the leading causes of near miss events. New-onset viral infections have emerged as the leading cause of maternal mortality. As near miss analysis indicates the quality of health care, it is worth presenting in national indices. Our target in reducing maternal mortality by 75% has not been met with [g system .SAMM has been studied extensively in the recent past as a complement for maternal mortality and also to evaluate the quality of obstetric care in that particular institution. This concept is superior over maternal death in drawing attention to surviving women's reproductive health and lives and is equally applicable in developing countries as well as developed countries. In many developed countries, maternal mortality has fallen to single digits whereas near miss cases are more and hence useful in evaluation of the present system. Moreover, they have the advantage of not being as rare as maternal deaths for providing adequate information, as well as still being rare enough not to overload clinicians and data collection personnel within the facility . Till recently there were no set criteria for identification of these cases for routine implementation, and wider application of this concept was limited . But in Maternal near miss case is defined as “a woman who nearly died but survived a complication that occurred during pregnancy, childbirth, or within 42 days of termination of pregnancy” . In our study, we aimed to determine the frequency of maternal near miss, MNM incidence ratio (MNRM), maternal near miss to mortality ratio, and mortality index. Our second objective was to analyze the nature of near miss events and compare the causes of near miss cases with that of maternal mortality. We also saw the trend of near miss events and maternal deaths in two years.An audit of maternal near miss cases from January 2011 to December 2012 was undertaken. Ours is a tertiary care institution with six primary health centers attached to it. It is a referral hospital for both public and private hospitals in Udupi and three other surrounding districts in Karnataka. In addition to providing twenty-four-hour emergency obstetric services, the hospital also provides antenatal care and delivery services for both low and high risk pregnant women. Hospital has 24-hour facility for blood component therapy. High dependency unit (HDU) in labor room complex and intensive care ICU with 24-hour facility for multidisciplinary specialty also function well. Potentially life threatening conditions were diagnosed, and those cases which met WHO 2009 criteria for near miss were selected. WHO criteria included a set of clinical, laboratory, and management-based criteria. Maternal mortality during the same period was also analyzed. Patient characteristics including age, parity, gestational age at admission, booked  , while low index suggests better quality of health care. (MI = MD/(MNM + MD) × 100.The following near miss indices were calculated. (1) MNM incidence ratio refers to the number of maternal near miss cases per 1,000 live births (LB). MNM IR = MNM/LB. (2) Maternal near miss: mortality ratio: Proportion between maternal near miss cases and maternal deaths. Higher ratio indicates better care. MNM: 1MD. (3) Mortality index: Number of maternal deaths divided by the number of women with life threatening conditions, expressed as a percentage.  During the period of audit there were a total of 7390 deliveries and 7330 live births, 131 near miss cases, and 23 maternal deaths.  Primiparas were slightly more in the near miss group. Majority of the patients (57.2%) were in third trimester at a near miss event, whereas, in the maternal death group, the number of postnatal patients with puerperal sepsis following section was high (2012). A huge burden of maternal near miss cases 96.2% and 86.96% of maternal deaths were referred.A total of 755 potentially life threatening conditions were identified of which 131 were near miss cases. Maternal near miss incidence ratio is 17.8/1000 live births. Maternal near miss to mortality ratio is 5.6 : 1. The mortality index is 14.9%. A total of 62.6% of the cases required ICU admission. Among the causes of near miss events, hemorrhage was the leading cause with 44.2%, and hypertension was 23.6%. Third among the group was sepsis, and the last was cardiac disease. Sepsis was the leading cause of maternal mortality at our setup, followed by hemorrhage, cardiac disease, and hypertension. We had 5 cases of sepsis in the year 2011, and 7 in 2012. In 2011 we had 3 cases of H1N1 infection with ARDS, one patient with viral pneumonia and other with pyelonephritis. In 2012 there were five cases after cesarean delivery with sepsis, one H1N1 and hepatitis E infection. This accounted for the high mortality due to sepsis.Obstetric deaths represent the quality of maternal care. But for the present scenario it may not reflect the global situation with regard to obstetric care. Hence new “near miss” criteria take over maternal mortality ratio. Although near miss criteria were in vogue for some years, lack of uniformity was the hindrance. WHO criteria, 2009  are uniqA study by Jayarathnam et al.  represenSAMM study from Brazil was in ICU setting only, while our study included ICU, high dependency unit, and labor room and hence represent all cases of near miss. Our study results were comparable to the studies of other developing countries, but these studies did not consider the WHO criteria .The maternal near miss incidence ratio (MNMR) was 17.8/1000 live births in our hospital. Studies done in the developing countries show the same trend and vary from anywhere between 15–40/1000 live births , 17, 18.The maternal mortality ratio at our setup was 313/100000 live births. The Brazilian study showed a similar mortality rate of 260/100000 live births . In otheThe near miss to mortality ratio was 5.6 : 1, which means for every five to six life threatening conditions there was one maternal death. Higher ratios indicate better care. Syrian study showed a ratio of 60 : 1 and study done in Nepal showed a ratio of 7.2 : 1 , 12. Thi Ours is a tertiary referral center covering three other districts in and around Udupi, with most of the cases being referred in an already moribund state. The delays in referrals are a major cause of morbidity and mortality. Establishment of a tertiary care in each district is essential. Delayed diagnosis, inappropriate transfer, and inadequate utilization of resources might have been the cause for maternal morbidities and mortalities in our study. Along with increased awareness of one's own health, health education may go a long way in improving the quality of obstetric care. In Limitation of the study is that as ICU facilities were available few potentially life threatening conditions before going on to near miss might have been selected. It is a single audit; data collection spanning over a few years would give a true picture of the improvement in obstetric care and also the long-term effects of SAMM.  Hemorrhage and hypertensive disorders are the leading causes of near miss events. New-onset viral infections have emerged as the leading cause of maternal mortality. The trend of near miss events and maternal deaths is the same in the two years. As near miss analysis indicates quality of health care, it is worth presenting in national indices.


Video encoding for multimedia services over communication networks has significantly advanced in recent years with the development of the highly efficient and flexible H.264/AVC video coding standard and its SVC extension. The emerging H.265/HEVC video coding standard as well as 3D video coding further advance video coding for multimedia communications. This paper first gives an overview of these new video coding standards and then examines their implications for multimedia communications by studying the traffic characteristics of long videos encoded with the new coding standards. We review video coding advances from MPEG-2 and MPEG-4 Part 2 to H.264/AVC and its SVC and MVC extensions as well as H.265/HEVC. For single-layer  video, we compare H.265/HEVC and H.264/AVC in terms of video traffic and statistical multiplexing characteristics. Our study is the first to examine the H.265/HEVC traffic variability for long videos. We also illustrate the video traffic characteristics and statistical multiplexing of scalable video encoded with the SVC extension of H.264/AVC as well as 3D video encoded with the MVC extension of H.264/AVC. Network traffic forecasts, such as the Cisco Visual Networking Index , predictThe encoders that are used to compress video before network transport have significantly advanced in recent years. These video encoding advances have important implications for the network transport of encoded video. The purpose of this paper is to first give an overview of the recent developments in video coding. Then, we examine the traffic characteristics of long videos encoded with the recently developed video coding standards so as to illustrate their main implications for the transport of encoded video in communication networks.This paper covers three main areas of video coding advances: (i) efficient encoding of conventional two-dimensional 2D) video into a nonscalable video bitstream, that is, a video bitstream that is not explicitly designed to be scaled  during network transport, (ii) scalable video coding, that is, video coding that is explicitly designed to permit for scaling  so as to make the video traffic adaptive (elastic) during network transport, and (iii) efficient nonscalable encoding of three-dimensional 3D) video. For conventional 2D video, we start from the video coding standards MPEG-2 and MPEG-4 Part 2 and outline the advances in video coding that have led to the H.264/MPEG-4 Advanced Video Coding (H.264/AVC) standard   − 1 frame periods compared to the classical B frame prediction structure.If the dyadic hierarchical B frame structure of H.264/AVC and H.265/HEVC is employed, it imposes additional constraints on the timing of the frame transmissions compared to the classical B frame prediction employed in the preceding MPEG-2 and MPEG-4 Part 2 standards. Generally, a given frame can only be encoded MPEG-2 and MPEG-4 provide scalable video coding into a base layer giving a basic version of the video and one or several enhancement layers that improve the video quality. The quality layering can be done in the dimensions of temporal resolution (video frame frequency), spatial resolution , or SNR video quality. The layering in the SNR quality dimension employs coarse quantization (with high QP) for the base layer, and successively finer quantization  for the enhancement layers that successively improve the SNR video quality. These layered scalability modes permit scaling of the encoded video stream at the granularity of complete enhancement layers; for example, a network node can drop an enhancement layer if there is congestion downstream. MPEG-4 has a form of sublayer SNR quality scalability referred to as fine grained scalability (FGS). With FGS there is one enhancement layer that can be scaled at the granularity of individual Bytes of video encoding information. With both MPEG-2 and MPEG-4, the flexibility of scaling the encoded video stream comes at the expense of a relatively high encoding overhead that significantly reduces the RD efficiency of the encoding and results in very limited adoption of these scalability modes in practice.Similar to the preceding MPEG standards, the Scalable Video Coding (SVC) extension of H.264/AVC  providesH.264 SVC has a novel Medium Grain Scalability (MGS) that splits a given SNR quality enhancement layer of a given video frame into up to 16 MGS layers that facilitate highly flexible and RD efficient stream adaption during network transport.E = 25. When encoding this enhancement layer, the 16 coefficients resulting from the discrete cosine transform of a 4 × 4 block are split into MGS layers according to a weight vector (a similar splitting strategy is employed for larger blocks). For instance, for the weight vector W = , the 16 coefficients are split into six MGS layers as follows. The lowest frequency coefficient is assigned to the first MGS layer m = 1, the next two higher frequency coefficients are assigned to the second MGS layer m = 2, and so on, until the four highest frequency coefficients are assigned to the sixth MGS layer m = 6, the highest MGS layer in this example. For network transport, the base layer and each MGS layer of a given frame are encapsulated into a so-called network adaptation layer unit (NALU).As for all SNR quality scalable encodings, the base layer of an MGS encoding provides a coarse quantization with a relatively high QP; for example, B = 35 or 40. MGS encodings have typically one enhancement layer providing a fine quantization with a relatively small QP; for example, 8 in 1–B7 and B9–B15. It is therefore recommended to drop MGS layers first from the B frames without any dependent frames, that is, the odd-indexed B frames in the highest layer in When scaling down an MGS video stream at a network node, dropping MGS layers uniformly across the frame sequence results in low RD efficiency of the downscaled stream. This is due to the dependencies in the B frame hierarchy. Specifically, dropping an MGS layer from a B frame that other B frames depend on, for example, frame BAlternatively, MGS encodings can be conducted so that each NALU is assigned a priority ID between 0 indicating lowest priority and 63 indicating highest priority for RD efficiency. These priority IDs can be assigned by the video encoder based on RD optimization. For downscaling an MGS stream with priority IDs, a network node first drops MGS layers (NALUs) with priority ID 0, then priority ID 1, and so on.Sony HD video for the priority ID stream scaling. We compare the RD curves of the MGS streams with cascaded QPs (C) with the RD curve from single-layer encoding, whereby all encodings have a GoP structure with one I frame and β = 15 B frames with the prediction structure illustrated in E = 25, in In E = 25) gives somewhat lower RD efficiency and substantially higher traffic variability at the encoder output than the corresponding B = 35, E = 25 encoding. That is, the increased adaptation flexibility of the B = 40, E = 25 encoding comes at the expense of reduced RD efficiency and very high traffic variability reaching CoV values above 3.5 at the encoder output.Similar to the single-layer encoding, we observe from the comparison of MGS streams encoded without and with cascaded QPs in Figures E = 25 MGS, C encoding, the CoV values of the MGS encoding are close or slightly below the single-layer CoV values.We observe from Cmin plots in The 3D video employs views from two slightly shifted perspectives, commonly referred to as the left view and the right view, of a given scene. Displaying these two slightly different views gives viewers the perception of depth, that is, a three-dimensional (3D) video experience. Since two views are involved, 3D video is also sometimes referred to as stereoscopic video. The concept of employing multiple views from different perspectives can be extended to more than two views and is generally referred to as multiview video.3D video streaming requires the transport of the two sequences of video frames resulting from the two slightly different viewing perspectives over the network to the viewer. Since the two views capture the same scene, their video frame content is highly correlated. That is, there is a high level of redundant information in the two views that can be removed through encoding (compression). The Multiview Video Coding (MVC) standard builds on the inter-coding techniques that are applied across a temporal sequence of frames in single-layer video coding to extract the redundancy between the two views of 3D video. More specifically, MVC typically first encodes the left view and then predictively encodes the right view with respect to the left view.One approach to streaming the MVC encoded 3D video is to transmit the encoded left and right views as a frame sequence with twice the frame rate of the original video, that is, left view of first video frame (from first capture instant), right view of first video frame, left view of second video frame, right view of second video frame, and so on. Since the right view is encoded with respect to the left view, it is typically significantly smaller (in Bytes) and the sequence of alternating left and right views result in high traffic variability, as illustrated in the next section.multiview frame for transmission. The sequence of multiview frames has then the same frame rate as the original video.Another MVC streaming approach is to aggregate the left and right views from a given video frame (capture instant) into one An alternative encoding approach for 3D video is to sequence the left and right views to form a video stream with doubled frame frequency and feed this stream into a single-view video encoder, such as H.264 SVC or H.265/HEVC. This approach essentially translates the interview redundancies into redundancies among subsequent frames. Similar to MVC encoding, the two encoded views for a given capture instant can be transmitted sequentially or aggregated.Yet another encoding alternative is to downsample (subsample) the left and right views to fit into one frame of the original video resolution. For instance, the 1920 × 1080 pixels of left and right views are horizontally subsampled to 960 × 1080 pixels so that they fit side-by-side into one 1920 × 1080 frame. This side by side approach permits the use of conventional 2D video coding and transmission systems but requires interpolation at the receiver to obtain the left and right views at the original 960 × 1080 pixel resolution.β = 7 B frames between successive I and P frames and 16 frames  per GoP. We observe from Alice video compared to the IMAX video and (ii) that the QP cascading improves the RD efficiency by up to about 0.5 dB for the Alice video and almost 1 dB for the IMAX video. These different RD efficiency levels and RD efficiency increases with QP cascading are due to the different content of the two videos. The IMAX video is richer in texture and motion and thus “more difficult” to compress and higher RD efficiency gains can be achieved with improved coding strategies.In IMAX video gives higher frame size variability and that (ii) QP cascading increases the frame size variability, mirroring the above observations for single-view (2D) video. We also observe from We observe from Recent studies  indicateWe have given an overview of modern video coding standards for multimedia networking. We have outlined the advances in the main video coding standards for nonscalable (single-layer) video and scalable video. For single-layer video, we gave an overview of H.264/AVC and H.265/HEVC. We compared their rate-distortion (RD) and rate variability-distortion (VD) characteristics before and after smoothing as well as link bitrate requirements. This comparison included the first study of the traffic variability (before and after smoothing) and statistical multiplexing characteristics of H.265/HEVC encoding for long videos as well as an original study of the effects of cascading of quantization parameters (QPs) for the different levels of the hierarchical dyadic B frame prediction structure of H.264/AVC. We found that the advances in the video coding standards have led to increased RD efficiency, that is, higher video quality for a prescribed mean video bitrate, but also substantially increased traffic variability at the encoder output (which is effectively mitigated through smoothing). We also found that elementary smoothing with moderately sized buffers and statistical multiplexing during network transport translate the RD improvements of H.265/HEVC into commensurate reductions of the required network link bitrate .For scalable video coding, we gave a brief overview of H.264 SVC Medium Grain Scalability (MGS) and the scaling of an encoded H.264 SVC MGS video bitstream in a network node. We compared the RD and VD characteristics (before and after smoothing) as well as the link bitrate requirements of the scaled H.264 MGS stream with corresponding single-layer encodings. We illustrated that H.264 SVC MGS streams can be flexibly scaled in the network in the lower region of the quality adaptation range while maintaining RD efficiency and link bitrate requirements very close to the unscalable single-layer encodings. For 3D video, we outlined the encoding and streaming of the two views and examined the RD and VD characteristics of the streams.The H.264/AVC video coding standard formed the basis for the development of the highly efficient scalable video coding (SVC) extension as well as extensions for stereo and multiview video coding suitable for 3D video . Similar


Total Mg concentrations in serum and cardiac ventricular tissues were measured by atomic absorption spectroscopy. Intracellular free Mg2+ concentration ([Mg2+]i) of ventricular myocytes was measured with the fluorescent indicator furaptra. Mg2+ transport rates, rates of Mg2+ influx and Mg2+ efflux, were estimated from the rates of change in [Mg2+]i during Mg loading/depletion and recovery procedures. In Mg-deficient rats, the serum total Mg concentration (0.29±0.026 mM) was significantly lower than in control rats (0.86±0.072 mM) after 4–6 weeks of Mg deficiency. However, neither total Mg concentration in ventricular tissues nor [Mg2+]i of ventricular myocytes was significantly different between Mg-deficient rats and control rats. The rates of Mg2+ influx and efflux were not significantly different in both groups. In addition, quantitative RT-PCR revealed that Mg deficiency did not substantially change mRNA expression levels of known Mg2+ channels/transporters  in heart and kidney tissues. These results suggest that [Mg2+]i as well as the total Mg content of cardiac myocytes, was well maintained even under chronic hypomagnesemia without persistent modulation in function and expression of major Mg2+ channels/transporters in the heart.To study possible modulation of Mg In cardiac myocytes, Mg2+ regulates Ca2+ and K+ channels 2+ release from sarcoplasmic reticulum 2+ sensitivity of intracellular buffer sites 2+ homeostasis might cause cardiovascular diseases, such as arrhythmias and heart failure.Intracellular Mg2+ concentration ([Mg2+]i) is in the range of 0.8–1.0 mM + influx . We have reported that Mg2+ efflux through Na+/Mg2+ exchange is activated by a slight increase in [Mg2+]iRegarding the physiological condition of rat ventricular myocytes, the intracellular free Mg2+ channels/transporters have been identified in eukaryotes. Among them, the melastatin subfamily 6 and 7 of the transient receptor potential cation channels  2+ channels/transporters implicated in Mg2+ homeostasis of mammalian cells. It has been reported that the function and expression of such Mg2+ channels/transporters are modified by extracellular and intracellular levels of Mg2+. In mammalian epithelial cells (HC11), low [Mg2+]i and high [Mg2+]i accelerated, respectively, Mg2+ influx and efflux. When the cells were incubated in Mg2+-deprived medium, TRPM6 mRNA and protein levels were upregulated 2+, whereas TRPM7 expression remained unaltered In the last decade, several Mg2+ transport functions and related gene expressions in cardiac myocytes. We unexpectedly found that neither Mg2+ transport rates nor mRNA expressions of major Mg2+ channels/transporters were significantly altered in rats fed a Mg-deficient diet for 4–6 weeks, in spite of severe hypomagnesemia. Portions of this work have been reported in abstract form In this study, we fed rats a Mg-deficient diet, and examined changes in MgAll experimental procedures involving animals were approved in advance by the institutional Animal Care and Use Committee of Tokyo Medical University (Permit Number: S-23013), and were performed in accordance with the “Guidelines for Proper Conduct of Animal Experiments” approved by the Science Council of Japan.Male Wistar rats  were fed either a control diet . After chest opening, a blood sample (3–5 ml) was collected from the left ventricular cavity by puncture, and the heart was quickly excised. Blood samples and heart ventricles were immediately processed for atomic absorption spectroscopy (AAS) to analyze mineral concentrations. For isolation of ventricular myocytes, the aorta of the excised heart was cannulated for Langendorff perfusion and subsequent enzymatic dispersion of single cells 3) and 20% trichloroacetic acid (TCA) to deproteinize. After centrifugal separation, the supernatant was diluted with 0.4N HNO3 and 8% TCA to determine concentrations of Mg, Ca, Na and K using a Spectra 880 atomic absorption spectrometer . Mg and Ca contents of ventricular tissues were determined by AAS after wet-digestion of ventricles with trace element-grade HNO3 and hydrogen peroxide  and appropriate dilution with 0.1N hydrochloric acid.Total mineral concentrations in serum and tissues were measured by AAS. Serum was treated with 1N nitric acid . After the measurement of background fluorescence and indicator loading by incubation with 5 µM furaptra AM  in normal Tyrode’s solution for 15 min at room temperature, the AM ester was washed out with Ca2+-free Tyrode’s solution that contained 0.1 mM K2EGTA in place of 1.0 mM CaCl2 of normal Tyrode’s solution , because in vivo parameters for calibration of furaptra fluorescence (in terms of [Mg2+]i) have been determined at 25°C 2+]i according to the equation:D is the dissociation constant, and Rmin and Rmax are R values at zero [Mg2+] and saturating [Mg2+], respectively. We used the parameter values previously estimated in rat ventricular myocytes at 25°C: KD = 5.30 mM, Rmin = 0.969, and Rmax = 0.223 All fluorescence measurements were carried out at 25°C, except during Mg2+]i caused by net flux (sum of influx and efflux) of Mg2+ across the cell membrane, experimental protocols were designed to minimize Mg2+ efflux for Mg2+ influx measurements (or to minimize Mg2+ influx for Mg2+ efflux measurements). We used the following two protocols to evaluate the rates of Mg2+ influx.As we measured the changes in [Mg2+ by incubation in a Mg-loading solution that contained 24 mM Mg2+ .The myocytes were loaded with Mg mM Mg2+  at ∼25°C2+ by incubation in the Mg-depleting solution  to 0.2–0.5 mM. In pilot experiments the rate of decrease in [Mg2+] i was significantly higher at 35°C than at 25°C, thus we chose 35°C (rather than 25°C) to minimize possible cell damage caused by prolonged exposure to the high-K solution.The myocytes were depleted of Mgsolution  for 20 m2+]i level was maintained , [Mg2+]i started to rise and reached a plateau in ∼2 h near the initial basal level ,The bathing solution was then switched to the Mg-free NMDG solution  for 30–4ined see . When thal level . We foll2+]i recovery is likely caused by the influx of Mg2+, the first derivative of the recovery function (2) is thought to reflect the rate of Mg2+ influx,constant . Because2+]i (t)/dt at time 0, −A/τ, as an index of the initial rate of Mg2+ influx. Note that initial [Mg2+]i of Mg2+ recovery was 0.2–0.5 mM, at which the Na+-dependent Mg2+ efflux activity was thought to be negligible 2+ in Mg-loading solution  induced a rapid decrease of [Mg2+]i2+]i was estimated by linear regression of data points spanning 120 s after the addition of extracellular Na+, and it was considered to reflect the rate of Mg2+ efflux, as previously reported 2+]i and 1 mM [Mg2+]o), Mg2+ influx is thought to be negligible, because the Mg2+ influx rate measured in the absence of extracellular Na+ was much lower even under reversed Mg2+ gradient (1–1.5 mM [Mg2+]i and 93 mM [Mg2+]o) After the myocytes were loaded with Mgsolution  to reachTotal RNA was extracted from segments of cardiac ventricles and kidneys using the SV total RNA Isolation System , following the manufacturer’s protocols. The heart and the kidney were quickly excised from anesthetized rats, and stored in RNA-later® solution  until used. High Capacity RNA to cDNA kit  was used for reverse transcriptase reactions. The expression levels of TRPM6, TRPM7, MagT1, SLC41A1 and ACDP2 were determined by quantitative real-time PCR in each sample using ABI 7500 Real-Time PCR system (Life Technologies). The expression level of the house-keeping gene GAPDH was used as an internal control. Primers and probes used for target genes were TaqMan Gene Expression Assays  purchased from Applied Biosystems, Life Technologies.t-test (or 2-way ANOVA) with the significance level set at p<0.05.Linear and nonlinear least-squares fittings were performed with the program Origin . Statistical values are expressed as the mean±SEM. Differences between groups were analyzed by Student’s two-tailed When rats were fed with the Mg-deficient diet for a prolonged period, their growth was retarded. Average body weight of the Mg-deficient diet group was significantly lower than that of the control diet group after feeding for 5 weeks or longer . Also, fThe serum Mg concentration quickly decreased after starting the Mg-deficient diet, and was significantly lower than the control rats after 2 weeks of feeding . ThereafIn addition to Mg, we measured the serum concentrations of Ca, Na and K of rats fed the control diet and those receiving the Mg-deficient diet for 4–6 weeks . In Mg-d2+ concentration . We therefore measured [Mg2+]i of ventricular myocytes isolated from control rats and Mg-deficient rats, using the fluorescent indicator furaptra . For myocytes incubated in Ca2+-free Tyrode’s solution that contained 1 mM Mg2+, basal levels of [Mg2+]i were within the previously reported range 2+]i values were not significantly different between the two groups (2+]o). However, because the perfusion solutions used for cell isolation and fluorescence measurements contained 1 mM Mg2+, one could argue that [Mg2+]i of myocytes isolated from Mg-deficient rats might rise to the normal level during isolation and fluorescence measurement procedures. To rule out this possibility, we isolated the myocytes using solutions that contained only 0.2 mM Mg2+, and fluorescence measurements were carried out at 0.2 mM [Mg2+]o (Mg2+ concentration of Ca2+-free Tyrode’s solution was reduced to 0.2 mM). [Mg2+]i values thus obtained were still within the normal range, and were not significantly different between the two groups (2+]o). It should be also noted that [Mg2+]i of myocytes isolated from Mg-deficient rats under the low Mg2+ condition (0.2 mM [Mg2+]o) was not significantly different from [Mg2+]i of myocytes isolated from control rats under the normal Mg2+ condition (1 mM [Mg2+]o). Thus, it is likely that [Mg2+]i of ventricular myocytes is sustained within the normal range even under the prolonged Mg deficiency for 4–6 weeks.Although the total Mg contents of ventricular tissues were not altered under serum Mg deficiency, redistribution of intracellular Mg could lead to changes in free Mg2+ influx via the cell membrane is facilitated and/or Mg2+ efflux is suppressed under serum Mg deficiency. We therefore examined the activities of Mg2+ transport across the cell membrane and the expression levels of known Mg2+ channels/transporters in the following sections.Overall, the results suggest that Mg deficiency significantly alters neither total concentration nor free concentration of intracellular Mg in cardiac ventricles, in spite of the marked reduction of serum Mg by 66% . This ap2+  and 0.57±0.038 mM (n = 10) in the cells isolated from, respectively, the Mg-deficient rats and the control rats. The decrements of [Mg2+]i were not significantly different between the two groups.During the 20-min exposure to the Mg-depleting solution , [Mg2+]i2+]i was lowered by incubation of the myocytes in Mg2+-free solutions  and fitted curves (solid lines) obtained from two myocytes : one isolated from a Mg-deficient rat (open circles and a red line) and the other isolated from a control rat (filled circles and a black line). Note that the time courses of the [Mg2+]i recovery were nearly superposable in these myocytes. The average values of the influx rate were 0.22±0.05 µM/s and 0.27±0.04 µM/s for the myocytes isolated from the Mg-deficient rats and the control rats, respectively , but the addition of extracellular Na+ induces a rapid decrease in [Mg2+]i . The functional characteristics of the Na+-dependent Mg2+ efflux are consistent with the Na+/Mg2+ exchange which extrudes Mg2+ in exchange for Na+ influx It has been demonstrated that Mg+-dependent Mg2+ efflux activity, a major efflux pathway in ventricular myocytes. It should be noted that the Mg2+ efflux is negligible at the basal [Mg2+]i (∼0.9 mM), but it is activated by higher [Mg2+]i at half-maximal activation at 1.5 mM 2+]i dependence, comparisons of the rates of Mg2+ efflux should be made at comparable [Mg2+]i levels. 2+]i recordings in two myocytes isolated from control rats (A) and Mg-deficient rats (B). After Mg loading of these myocytes for 3 h  and those from Mg-deficient rats , and the initial rates of decrease in [Mg2+]i upon Na+ addition were also not significantly different between these two groups (2+ efflux).We evaluated the Na2+ channels/transporters, TRPM6, TRPM7, MagT1, SLC41A1 and ACDP2, in cardiac ventricle and kidney tissues excised from control rats and Mg-deficient rats  measurements of [Mg2+]i, 2) functional assays of Mg2+ influx and efflux rates, and 3) quantification of mRNA expression of known Mg2+ channels/transporters. We found that none of these measurements showed a significant alteration in rats fed a Mg-deficient diet for 4–6 weeks, although the serum levels of Mg fell to 1/3 in comparison with control rats fed a Mg-containing diet. These results suggest that [Mg2+]i as well as total Mg content of cardiac myocytes is tightly regulated, under altered driving force for Mg2+ across the cell membrane, without persistent modulation of functions and expressions of major Mg2+ channels/transporters.It has been reported that in mice fed a Mg-deficient diet chronic Mg deficiency does not significantly alter the Mg content of the heart tissue, in spite of severe hypomagnesemia 2+ transport functions were influenced during cell isolation and manipulation procedures, it is unlikely that exposure of cells to high [Mg2+]o (during Mg-loading) or low [Mg2+]o (during Mg-depletion) overrides changes in cell membrane expression of channel/transporter proteins  previously induced by Mg deficiency in vivo based on the following findings: 1) Even at the very beginning of the Mg-loading, time courses of the rise in [Mg2+]i  were similar between the cells isolated from the Mg-deficient rats and those isolated from the control rats  in the initial 20 min of Mg-depletion were not significantly different between the two groups (see Results).Although we cannot exclude the possibility that Mgrol rats . 2) The 2+ channels/transporters are transiently upregulated by the onset of hypomagnesemia, and are normalized in chronic hypomagnesemia. On the other hand, prolongation of Mg-deficiency for a very long period (>6 weeks) also could significantly affect cellular Mg2+ handling. It is therefore important in future studies to follow the time courses of changes in function and expression of Mg2+ channels/transporters, as well as intracellular magnesium concentrations , during Mg deficiency.Goytain and Quamme 2+]i is maintained by the balance between Mg2+ influx and Mg2+ efflux. As a Mg2+ efflux pathway, we have studied active Mg2+ transport in exchange with Na+ influx  in rat ventricular myocytes + nearly completely abolishes Mg2+ efflux activity +/Mg2+ exchange is thought to be the major pathway for Mg2+ efflux in cardiac myocytes. The Mg2+ transport rate by the Na+/Mg2+ exchange is critically dependent upon [Mg2+]i; the transport rate is almost null at resting [Mg2+]i, but a slight increase in [Mg2+]i markedly activates the transport with half-maximal activation of ∼1.5 mM 2+ influx is thought to be passive via Mg2+ permeable channels . It has been shown that conductance of TRPM7 channels increases with the reduction of [Mg2+]i2+]i dependence of Mg2+ influx is also shown in 2+-depleted cells, [Mg2+]i recovery is initially fast, but it slows as [Mg2+]i approaches the basal level, reaching a plateau near the basal level  and other accompanying changes, rather than by changes in [Mg2+]i.In studies with Mg-deficient animals, it is often assumed that a decrease in [Mg


Adult prairie voles’ drinking patterns were monitored in a lickometer apparatus that recorded each lick a subject exhibited during continuous access to water and 10% ethanol during periods of isolation, pair housing of high and low drinkers, and subsequent isolation. Analysis of fluid consumption confirmed previous results that high drinkers typically decrease ethanol intake when paired with low drinkers, but that a subset of voles do not decrease. Analysis of bout structure revealed differences in the number of ethanol drinking bouts in the subpopulations of high drinkers when paired with low drinkers. Lickometer drinking patterns analyzed by visual and by cross-correlation analyses demonstrated that pair housing did not increase the rate of subjects drinking in bouts occurring at the same time. The length of the avpr1a microsatellite did not predict susceptibility to peer influence or any other drinking behaviors. In summary, subpopulations of high drinkers were identified, by fluid intake and number of drinking bouts, which did or did not lower their ethanol intake when paired with a low drinking peer, and these subpopulations should be explored for testing the efficacy of treatments to decrease ethanol use in groups that are likely to be responsive to different types of therapy.Peer influences are critical in the decrease of alcohol (ethanol) abuse and maintenance of abstinence. We previously developed an animal model of inhibitory peer influences on ethanol drinking using prairie voles and here sought to understand whether this influential behavior was due to specific changes in drinking patterns and to variation in a microsatellite sequence in the regulatory region of the vasopressin receptor 1a gene ( Excessive alcohol (ethanol) use in the United States contributes to over 80,000 deaths per year (apps.nccd.cdc.gov/DACH_ARDI). Therefore, it is extremely important to understand all factors contributing to excessive ethanol drinking, as well as those that contribute to decreases in drinking. Peer influences can lead to increases in ethanol drinking in some cases, and to decreases in others. Both types of influence can be crucial on the path to either alcohol abuse  or abstiMicrotus ochrogaster) exhibit increased preference not only for their sexual partner, but also to their same-sex cage mates  influence of voles drinking low doses of ethanol on voles drinking high doses of ethanol was specific to ethanol, and was not observed with other palatable fluids . The decavpr1a) contains a microsatellite region upstream of the transcription start site, which is polymorphic . Adult male and female voles (n = 95) were used in these studies, ranging from 58 to 95 days of age at the start of the experiment.Prairie voles were bred in our colony at the Portland Veterans Affairs Medical Center Veterinary Medical Unit. All procedures were approved by the Institutional Animal Care and Use Committee and adhered to the guidelines put forth in the National Institutes of Health Guide for the Care and Use of Laboratory Animals. Voles were weaned around 21 days of age and housed in same-sex sibling pairs, with females and males housed in different rooms, until beginning the experiment. Voles were housed under 14L:10D lighting conditions and had continuous Figure 1. The plastic cage bottom that surrounded the wire rack was 16.8 cm × 27.6 cm and 5.4 cm high  and had bedding, food, and a nestlet available, so the subjects were not required to be on the wire rack when they were not drinking. The plastic cage top was 17 cm high and in addition to the holes for the drinking spouts, there were holes in the lid and openings along the bottom for air circulation . The cages used for pair housing were identical except that they were twice as wide, with separate lids for each half, and a wire mesh down the center that divided the cage into two equal compartments but allowed the subjects visual, olfactory, vocal, and some tactile contact, similar to what has been described by us previously  and water in the other, and they had continuous access to these solutions throughout the rest of the experiment. Fluid volumes were recorded every 24 h, and the position of the bottles relative to one another was counterbalanced across pairs and switched every 2 days. Fluids were replaced every 2 days. After recording fluid volumes each day, and changing fluids every second day, the lickometer recording began and continued for 22 h.After 4 days of access to ethanol in isolation, subjects were categorized as consistent high, medium, or low drinkers, dependent on the amount of ethanol they consumed (g/kg/day) and the preference ratio for ethanol over water, using identical criteria to a previous study . Specifi2 inhalation, and tail tissue samples were taken for genetic analysis.Following the final isolation period, voles were euthanized by COEthanol intake and preference were calculated for each day based on fluid volumes consumed. Average measures for each housing period were compared by two-way repeated measures ANOVA with high and low drinkers as a between-subjects variable. Further analyses were done by splitting high drinkers into a group of animals that decreased their drinking level category during the 4 days of pair housing with a low drinker and a group of animals that did not, and comparing ethanol intake on each day of isolation and pair housing. Drinking data from days two and six are not presented in order to correspond with the lickometer data (see below), but data from each of these days were very consistent with the respective surrounding days. Bonferroni post-tests were used to determine specific group differences. As in a previous study , there wTo validate the lickometer, water and ethanol volume consumed were each compared with the number of licks registered for each subject, and analyzed using a Pearson’s correlation.The lickometer data were analyzed as described previously  by custoThe lickometer data were then processed using custom-designed software  which first rescaled the data from 10 ms to 1 s resolution. Cumulative lick plots for each pair on the last day of isolation and pairing were examined, since the subjects would have had the most time to establish stable drinking patterns under each housing condition. The number of bouts occurring in temporal proximity (≤15 min apart) was determined using a standardized visual assessment. The number of close bouts, and the number of close bouts normalized to the lowest number of bouts exhibited by one member of the pair, were compared using two-way repeated measures ANOVA with change in drinking as a between-subjects factor and housing as the repeated measure.The data processed through u2615 were then analyzed for each pair by a cross-correlation analysis (R for Mac OS). The correlations were compared between the last day of isolation and the last day of pair housing. The presence or absence of a significant correlation for each day was noted, as well as the lag time and degree of correlation  for each significant correlation. The lag time range was limited to ±10 min, in order to analyze only behaviors that occurred close together in time. This metric indicated which subject followed the other in drinking, and was examined in conjunction with individual pair data indicating which subject changed intake.DNA was extracted from each subject’s tail tissue sample using a DNeasy Blood and Tissue Extraction Kit . The V1aR microsatellite sequence was amplified using a variation of a previously published PCR technique . We usedThe samples were each read by a 3130xl Genetic Analyzer , by the Oregon Clinical and Translational Research Institute Core Laboratory at Oregon Health & Science University (OHSU). The microsatellite length was determined for each allele for each subject with approximately 3 base pair resolution.t-tests were performed to compare between animals that had short or long average microsatellite length. A number of dependent variables were tested  and a Bonferroni correction for multiple comparisons was applied to yield the corrected threshold for significance α = 0.005.Microsatellite allele lengths were not normally distributed due to a highly leptokurtotic sample, which could not be normalized by any transformation. Thus, correlations could not be conducted using the collected data. Instead, a median split was applied to the data and High drinkers were paired with low drinkers, leading to a total of 32 pairs that completed the experiment, while medium drinkers did not continue past the initial isolation phase. Of these, 14 pairs were female and 18 pairs were male.F = 45.71; p <0.0001) and exhibited higher intakes  = 40.85; p <0.0001), in accordance with their categorization. There was a significant effect of housing conditions on preference  = 4.91; p = 0.009) but not intake  = 0.82; p = 0.441). As seen in our previous study  = 6.94; p = 0.0014) and intake  = 4.48; p = 0.013). Planned Bonferroni post-tests revealed that high drinkers decreased their ethanol preference from baseline (isolation 1) to paired housing and isolation 2 , as well as the intake level , while low drinkers did not significantly change .As expected, high drinkers had a significantly higher preference for ethanol than low drinkers . There was a main effect of group on ethanol preference  = 51.65, p < 0.0001) and intake  = 34.47, p < 0.0001), a main effect of day on preference  = 10.26, p < 0.0001) and intake  = 7.66, p < 0.0001), and an interaction between the group and housing on preference  = 6.40, p < 0.0001) and intake  = 9.86, p < 0.0001). Post hoc tests revealed that the low drinkers had significantly lower ethanol preference and intake than both groups of high drinkers on each day of isolation (p < 0.001), while during pair housing, both the low drinkers and the high group that changed had significantly lower ethanol intake than the high drinkers that did not change (p < 0.001). On day 4 of isolation, the high-change group had a significantly lower preference for ethanol than the high-no change group (p < 0.05), while still remaining significantly higher than the low drinkers, as described above.Also as in the previous study , the behFigure 4). Analysis of the bouts of ethanol consumption revealed one notable difference between high drinkers that did not change ethanol intake when paired with low drinkers, and high drinkers that did change, out of six different parameters assessed . Since the software could not analyze data from subjects with one or fewer drinking bouts, 10 low drinkers and one high drinker were not included in the analysis of other features besides the number of bouts. There was a main effect of group on the number of bouts  = 10.69; p = 0.0001), interbout interval  = 5.71; p = 0.006), and lick rate  = 5.30; p = 0.009). There was also a main effect of day on the number of bouts  = 12.82; p < 0.0001), interbout interval  = 5.33; p = 0.0001), and lick rate  = 11.17; p < 0.0001). Most notably, there was an interaction effect of group and day on the number of bouts  = 3.06; p = 0.001).Volumes of ethanol and water consumed each day correlated very well with the number of licks recorded for each subject , while the high-change group was never significantly different from the low group. The high-change group did have significantly fewer drinking bouts than the high-no change group on days 5 (p < 0.001) and 7 (p < 0.05) during pair housing .Figure 6) revealed that while there were occurrences of ethanol drinking bouts close together in time for pairs of animals, the frequency of close bouts was not significantly different between isolation  and pair housing , or between pairs that did not change drinking levels compared to those who did, and there was no interaction between the two factors when either the number  or proportion  of close bouts was assessed.Visual analysis of the cumulative lick graphs  or housed together . Additionally, there was no consistent difference in the presence or absence of correlations between pairs that exhibited changes in drinking behavior and those that did not (Table 1). Contrary to our hypothesis, there was no consistent directionality of the lag time of cross-correlations in pairs that changed their drinking level: in pairs where high drinkers changed to low drinkers, there was not a greater presence of a negative lag time that would indicate the low drinker “leading” the high drinker .Cross-correlation analyses revealed that over two-thirds of the pairs exhibited a significant correlation between ethanol drinking patterns regardless of whether they were physically isolated . However, there was a statistical trend for a positive correlation between the proportion of close bouts by visual assessment  and the strength of cross-correlations .There was no significant correlation between the number of close bouts by visual assessment and the strength of cross-correlations : initial ethanol preference, initial ethanol intake, change in ethanol preference or intake from isolation to pair housing, pair housing to subsequent isolation, or overall change from the beginning to the end of the experiment. There was a difference within high drinkers, where subjects with long alleles had a greater decrease in ethanol preference from the beginning to the end of the experiment than those with short alleles , but this difference did not remain significant when adjusted for multiple comparisons.There was no significant difference in drinking behavior between subjects with short or long average microsatellite lengths on any measure of behavior  microsatellite fragment observed here was different than what has previously been reported by others. The length of the vasopressin receptor 1a (avpr1a microsatellite to modulate social influence on ethanol drinking. A recent study in human adolescents demonstrated no role of a different repeat region, a variable number tandem repeat in the dopamine D4 receptor gene, in the effects of friends’ drinking levels on subjects’ drinking ( In addition to differences in allele length in the samples, the present experiment did not find effects of the microsatellite length on any measure of prairie voles’ ethanol drinking behavior, or on the propensity to change ethanol intake when paired with a peer. There was an indication of an effect of the longer microsatellite length corresponding to a greater change in ethanol preference following the effect of a peer influence, but this effect did not remain significant following adjustment for multiple comparisons. Thus, this trend should only be considered suggestive of the potential for the drinking .The effects of the V1aR microsatellite length reported by others appear to be very specific to particular tests and environments. For example, microsatellite length was correlated with the receptor expression level in various brain regions, and several of these regions were then correlated with measures of partner preference in the laboratory test , but notavpr1, rather than its length, could be a better predictor gene expression and, ultimately, behavior.One possible reason for effects that may be difficult to detect has previously been proposed by others : while tThe present study shows that while high drinkers decrease their ethanol intake when paired with low drinkers, it is not due to matching patterns of drinking, and the behavioral changes cannot be predicted by the length of the microsatellite polymorphism in the vasopressin receptor 1a. Other behaviors and specific genetic polymorphisms that may affect peer-influenced ethanol drinking may be studied in the future. This study demonstrates new methods for examining data from fluid consumption studies where social influences can be assessed using visual and cross-correlational analyses. Most importantly, this study shows that subpopulations of high drinkers that decrease their ethanol intake can be identified based on changes in intake levels and bout number when paired with a low drinker. This provides a model system in which the efficacy of potential therapies can be tested using groups which are likely to respond to different types of treatments. It will be important to examine whether subpopulations of human alcohol drinkers can be identified with similar means, and to explore whether they are similarly responsive to social or other types of treatments to decrease alcohol drinking; then further testing of this animal model of alcohol drinking can be used to elucidate specific mechanisms of action and responses to treatments that can inform treatment of humans with alcohol use disorders.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.