Signals of selection and ancestry in independently feral Gallus gallus populations.

Recent work indicates that feralisation is not a simple reversal of domestication, and therefore raises questions about the predictability of evolution across replicated feral populations. In the present study we compare genes and traits of two independently established feral populations of chickens (Gallus gallus) that inhabit archipelagos within the Pacific and Atlantic regions to test for evolutionary parallelism and/or divergence. We find that feral populations from each region are genetically closer to one another than other domestic breeds, despite their geographical isolation and divergent colonisation histories. Next, we used genome scans to identify genomic regions selected during feralisation (selective sweeps) in two independently feral populations from Bermuda and Hawaii. Three selective sweep regions (each identified by multiple detection methods) were shared between feral populations, and this overlap is inconsistent with a null model in which selection targets are randomly distributed throughout the genome. In the case of the Bermudian population, many of the genes present within the selective sweeps were either not annotated or of unknown function. Of the nine genes that were identifiable, five were related to behaviour, with the remaining genes involved in bone metabolism, eye development and the immune system. Our findings suggest that a subset of feralisation loci (i.e. genomic targets of recent selection in feral populations) are shared across independently established populations, raising the possibility that feralisation involves some degree of parallelism or convergence and the potential for a shared feralisation 'syndrome'.

Complications after emergency laparotomy beyond the immediate postoperative period - a retrospective, observational cohort study of 1139 patients.

Mortality and morbidity occur commonly following emergency laparotomy, and incur a considerable clinical and financial healthcare burden. Limited data have been published describing the postoperative course and temporal pattern of complications after emergency laparotomy. We undertook a retrospective, observational, multicentre study of complications in 1139 patients after emergency laparotomy. A major complication occurred in 537/1139 (47%) of all patients within 30 days of surgery. Unadjusted 30-day mortality was 20.2% and 1-year mortality was 34%. One hundred and thirty-seven of 230 (60%) deaths occurred between 72 h and 30 days after surgery; all of these patients had complications, indicating that there is a prolonged period with a high frequency of complications and mortality after emergency laparotomy. We conclude that peri-operative, enhanced recovery care bundles for preventing complications should extend their focus on continuous complication detection and rescue beyond the first few postoperative days.

CREBBP and WDR 24 Identified as Candidate Genes for Quantitative Variation in Red-Brown Plumage Colouration in the Chicken.

Plumage colouration in birds is important for a plethora of reasons, ranging from camouflage, sexual signalling, and species recognition. The genes underlying colour variation have been vital in understanding how genes can affect a phenotype. Multiple genes have been identified that affect plumage variation, but research has principally focused on major-effect genes (such as those causing albinism, barring, and the like), rather than the smaller effect modifier loci that more subtly influence colour. By utilising a domestic × wild advanced intercross with a combination of classical QTL mapping of red colouration as a quantitative trait and a targeted genetical genomics approach, we have identified five separate candidate genes (CREBBP, WDR24, ARL8A, PHLDA3, LAD1) that putatively influence quantitative variation in red-brown colouration in chickens. By treating colour as a quantitative rather than qualitative trait, we have identified both QTL and genes of small effect. Such small effect loci are potentially far more prevalent in wild populations, and can therefore potentially be highly relevant to colour evolution.

Identification of a congenital dysthrombin, thrombin Quick.

A dysprothrombin designated prothrombin Quick, is isolated from the plasma of an individual with < 2% of normal functional prothrombin activity and 34% of the normal prothrombin level by immunologic assay. With Factor Xa or taipan snake venom as activators, a fragmentation pattern identical to that of normal prothrombin is observed on gel electrophoresis in dodecylsulfate. This evidence combined with the observed barium citrate adsorption of prothrombin Quick and the low activity suggests that the defect in prothrombin Quick is in the thrombin portion of the molecule. Thrombin Quick is isolated and comigrates with thrombin on dodecyl sulfate gel electrophoresis, either reduced or nonreduced. The activity of thrombin Quick on several biological substrates of thrombin is investigated. Relative to normal thrombin, thrombin Quick is 1/200 as active on fibrinogen and 1/20-1/50 as effective in activating Factors V and VIII and aggregating platelets. A complex with antithrombin III is detected by dodecyl sulfate gel electrophoresis. Further investigation with the active site titrant, dansylanginine-N-(3-ethyl-1,5-pentanediyl)amide showed that the thrombin Quick preparation has the same affinity for the titrant as thrombin, but apparently only 40% active sites per mole protein are titrable.

Expression and prognostic significance of platelet-derived growth factor and its receptors in epithelial ovarian neoplasms.

The expression of platelet-derived growth factor (PDGF), PDGF-alpha receptor (PDGFR alpha) and PDGF-beta receptor (PDGFR beta) was studied in normal ovaries and ovarian neoplasms by immunohistochemical analysis. PDGF was detected in tumor cells in 33 of 45 malignant tumor samples but in none of 20 benign tumors (P < 0.001) or 11 normal ovaries (P < 0.001). In borderline tumors, 4 of 7 tissues stained positive in tumor cells. PDGFR alpha was detected in tumor cells in 16 of 45 malignant tumors, while no epithelial staining was found in 16 benign tumors (P = 0.002) or in 10 normal ovaries (P = 0.023). In 1 of 7 borderline neoplasms, tumor cells expressed PDGFR alpha. Neither normal epithelium nor tumor cells stained positive with antibodies against PDGFR beta. Patients with ovarian cancer and PDGFR alpha-positive tumor cells demonstrated an overall shorter survival compared to those who had negatively stained tumors (P < 0.005). A similar correlation was found in patients having stage III ovarian cancer (P < 0.01), which further supports an independent role for PDGFR alpha as a prognostic factor. Thus, the concomitant expression of PDGF and PDGFR alpha in tumor cells is related to progression of malignant ovarian tumors, indicating a functional role of PDGF via autocrine growth stimulation.

The domesticated brain: genetics of brain mass and brain structure in an avian species.

As brain size usually increases with body size it has been assumed that the two are tightly constrained and evolutionary studies have therefore often been based on relative brain size (i.e. brain size proportional to body size) rather than absolute brain size. The process of domestication offers an excellent opportunity to disentangle the linkage between body and brain mass due to the extreme selection for increased body mass that has occurred. By breeding an intercross between domestic chicken and their wild progenitor, we address this relationship by simultaneously mapping the genes that control inter-population variation in brain mass and body mass. Loci controlling variation in brain mass and body mass have separate genetic architectures and are therefore not directly constrained. Genetic mapping of brain regions indicates that domestication has led to a larger body mass and to a lesser extent a larger absolute brain mass in chickens, mainly due to enlargement of the cerebellum. Domestication has traditionally been linked to brain mass regression, based on measurements of relative brain mass, which confounds the large body mass augmentation due to domestication. Our results refute this concept in the chicken.

Feralisation targets different genomic loci to domestication in the chicken.

Feralisation occurs when a domestic population recolonizes the wild, escaping its previous restricted environment, and has been considered as the reverse of domestication. We have previously shown that Kauai Island's feral chickens are a highly variable and admixed population. Here we map selective sweeps in feral Kauai chickens using whole-genome sequencing. The detected sweeps were mostly unique to feralisation and distinct to those selected for during domestication. To ascribe potential phenotypic functions to these genes we utilize a laboratory-controlled equivalent to the Kauai population-an advanced intercross between Red Junglefowl and domestic layer birds that has been used previously for both QTL and expression QTL studies. Certain sweep genes exhibit significant correlations with comb mass, maternal brooding behaviour and fecundity. Our analyses indicate that adaptations to feral and domestic environments involve different genomic regions and feral chickens show some evidence of adaptation at genes associated with sexual selection and reproduction.

A dose escalation study of ORG 10172 (low molecular weight heparinoid) in stroke.

An intravenous infusion of a low molecular weight heparinoid, with a reduced risk of hemorrhage, may be an alternative to heparin in the management of acute ischemic stroke. To evaluate this hypothesis, we studied the safety of the heparinoid, ORG 10172, in a dose-escalation study in 26 patients. The drug was administered as a loading bolus followed by a 7-day infusion in five rates with target anti-factor Xa levels from 0.2 to 1.0 U/ml. The drug was well tolerated; no major bleeding complications or thrombocytopenia occurred. There were no deaths or hemorrhagic transformation of cerebral infarctions. The results indicate that ORG 10172 at doses to achieve a level of 1.0 U/ml or less may be used safely in management of acute cerebral infarction.

Prothrombin Quick. A newly identified dysprothrombinemia.

The rarest of reported inherited plasmatic coagulopathies involve prothrombin. Only 10 families with significant reductions of this plasma protein (hypoprothrombinemia) have been observed. Even fewer, six families, have been found to have a functionally abnormal prothrombin (dysprothrombinemia) in their blood. An as yet undefined prothrombin abnormally has been recognized in eight other families. One of the first patients previously identified by Quick and his associates as having a defect in her plasma prothrombin has been shown to have about half the normal amount of prothrombin antigen but virtually no prothrombic function. We propose that this dysprothrombin be designated prothrombin Quick. An additional patient also first described by Quick was found to be truly hypoprothrombinemic--that is, to lack both functional and antigenic prothrombin. Briefly summarized are the other five families with dysprothrombinemia, nine with hypoprothrombinemia, and the eight in whom the defect has not been classified.

Modeling of therapeutic dialysis of cerebrospinal fluid by epidural cooling in spinal cord injuries.

Paraplegias of traumatic origin may be classified as primary or secondary. Secondary traumatic paraplegia (STP) is believed to result from an autodestructive process. Different authors have published results supporting or contradicting the therapeutic effects of durotomy alone or associated with exposed spinal cord and perfusion with a saline solution at normal or cold temperatures. It appears that although decompression and open dialysis might be beneficial, the surgical trauma over the injured region is detrimental. A method of local epidural spinal cord cooling has been developed and successfully used to treat STP. With this method, no surgical injury or damage is imposed on the dura, cerebrospinal fluid (CSF), or spinal cord. Furthermore, several of the beneficial effects attributed to hypothermia in the traumatized area are evident, including reduction of metabolic demands, edema, swelling, vasospasm, and blood pressure. Aware of the benefits that dialysis may have in STP, as well as of the encouraging results obtainable with local epidural spinal cord cooling, we hypothesized that this method of hypothermia may in some way trigger CSF dialysis. Based on this hypothesis, a model was developed approximating the behavior of the CSF in the situation where a cold source is applied to the dura. Using dimensionless analysis techniques, we predict that CSF under the cooled region of the dura undergoes convective motion, even in adverse situations where the spinal cord has swollen. Under steady-state conditions, the moving fluid forms several Bénard cells directly under the cold source. The size of these Bénard cells was estimated. The range of probe temperatures at which convective flow is generated was considered, as well as the relative benefits of hypothermia versus flow. Results of more rigorous analysis are discussed.

An intrinsic coagulation pathway inhibitor in a 3-year-old child.

An intrinsic coagulation pathway inhibitor with acitivty against factor XI was detected in the plasma of a 3-year-old child, following a respiratory infection. The inhibitor was present transiently, but it was clinically significant and was manifiested by extensive bruises. Abnormalities of clotting and the bruising tendency disappeared simultaneously without treatment before the inhibitor could be definitively characterized. It was possibly the consequence of an adenovirus infection, as an elevated adenovirus titer was demonstrated in the patient's blood. It is possible that inhibitors of this type are more common sequelae of viral infections than realized, and perhaps a systematic effort should be made to detect them and to define their etiology and mechanism of action.

Effects of protein kinase C inhibitors on thromboxane production by thrombin-stimulated platelets.

The purpose of these studies was to identify a possible role for protein kinase C in thromboxane production. The effects of four putative protein kinase C inhibitors were studied with platelet stimulation by thrombin (0.5-150 nM), Thrombin Quick I (1.5-500 nM) or a thrombin receptor (protease activated receptor-1) agonist peptide (TRAP) (5-120 microM). Thromboxane production was increased by the bisindolylmaleimide derivative, 2-[1-(3-dimethylaminopropyl)-1H-indol-3-yl]-3-(1H-indol-3-yl)-maleimi de (GF 109203X), unchanged by the inhibitors 12-(2-cyanoethyl)-6,7, 12,13-tetrahydro-13-methyl-5-oxo-5H-indolo (2,3-a) pyrrolo (3, 4-c)-carbazole (Gö 6976) and 5,21:12,17-dimetheno-18H-dibenzo[i, o]pyrrolo[3,4-l][1,8]diazacyclohexadecine-18,20(19H)-dione, 8-[(dimethylamino)methyl]-6,7,8,9,10,11-hexahydro-, monomethanesulfonate (379196), the latter of which is protein kinase C beta-selective, and decreased by 1-[6-[(3-acetyl-2,4, 6-trihydroxy-5-methylphenyl)methyl]-5,7-dihydroxy-2, 2-dimethyl-2H-1-benzopyran-8-yl]-3-phenyl-2-propen-1-one (rottlerin), an inhibitor selective for protein kinase C delta. These results indicate complex regulation of thromboxane synthesis in human platelets including a probable role for protein kinase C delta. The results taken together further suggest that GF 109203X may suppress negative feedback resulting from an unidentified kinase and that the classical protein kinase C isoforms alpha and beta do not have a significant role in regulating thromboxane production by platelets.

Effect of oral contraceptive use on platelet prothrombin converting (platelet factor 3) activity.

PIP: The attempt was made to determine if platelets obtained from women using oral contraceptives (OCs) have increased activity in support of Factor Xa-catalyzed prothrombin activation. 9 volumes of human blood were collected into 1 volume of 3.2% sodium citrate solution. Fibrinogen was prepared by the method of Straughn and Wagner. Prothrombin was assayed by the taipan sanke venom method, and antithrombin 3 was assayed as heparin cofactor activity by a modification of the method described by Henriksen and Owen. Subjects, including controls, were women between 20-30 years who were in good health and who had not ingested aspirin or other drugs within 10 days of the beginning of the experiment. Platelets from OC users, compared with controls, promote an increase in both rate and extent of prothrombin activation, with control and test groups falling into non-overlapping populations. The increased prothrombin activation correlates with an increase in antithrombin 3 consumption. Enhanced antithrombin 3 consumption explains the decreased serum antithrombin 3 levels among OC users. Considered with increased platelet lability and an increased tendency to aggregate, the increase in total available platelet factor 3 activity points to a specific role of the platelet in the increased risk of thromboembolic disease associated with OC use.^ieng

Ki-67 immunostaining and DNA flow cytometry as prognostic factors in epithelial ovarian cancers.

Samples from malignant epithelial ovarian tumors were examined for expression of Ki-67, DNA ploidy and S-phase fraction (SPF). In addition, normal ovary, benign and borderline tumors were immunostained with Ki-67 to examine its role in ovarian carcinogenesis and to estimate the prognostic significance. For both Ki-67, ploidy and SFP a significant correlation to survival in malignancy was observed. Furthermore, SPF above 10% showed significant correlation to decreased survival in both stage subgroups I-II and III-IV, while for aneuploidy a decrease in survival could be detected only in localized disease. Ki-67 expression was present in tumor cells in the main part of borderline and malignant tumors and even in 2 benign counterparts, which might indicate an active state in these commonly believed dormant neoplasms. In the malignant group the Ki-67 correlation to survival seemed to be independent of staining intensity, although the observed decay in survival seemed to be faster in the strongly stained group.

In Situ Hybridization Detection of TGF-ß mRNA.

Immunohistochemical techniques and molecular hybridization enable demonstration of specific proteins and DNA or RNA sequences, respectively. In situ hybridization is a variant of molecular hybridization that allows detection of specific DNA or RNA sequences in tissue sections or cell preparations, as well as in chromosome preparations and was first described by Pardue and Gall (1). Basically, a single-stranded probe of mRNA or DNA containing complementary sequences are hybridized to RNA or DNA in the sample. The probe is radioactively or nonisotopically labeled for localization and eventually quantification of the product.

Amylase activity and fast-migrating amylase isoenzymes in serum and cyst fluid from patients with ovarian neoplasms.

160 patients with various ovarian tumors were studied to establish whether total amylase activity and the occurrence of fast migrating amylase isoenzymes in serum could serve as indicators of ovarian cancer. It was found that patients with benign and malignant ovarian tumors could not be classified by means of total amylase activity. Electrophoretic separation of the amylases revealed fast-migrating forms in serum from 10 of 47 patients with malignant ovarian neoplasms; 8 of these 10, and altogether 19 of the 47 patients had a serous cystadenocarcinoma. Two of the 109 patients with benign ovarian tumors also showed the pattern with fast-migrating amylases; both of them had a serous cystadenoma. Four patients with borderline tumors showed normal amylase patterns. Tumor origin of these fast-migrating amylase forms in serum was substantiated by 1) amylase reactive cells detectable in tumor tissue, and 2) surgical removal of tumor followed by complete disappearance of the fast-migrating amylase forms in serum. Normal serum amylase patterns do not exclude the presence of a malignant ovarian tumor, but occurrence of these abnormal amylase forms in serum may indicate that an ovarian tumor is a cystadenocarcinoma.

Identification of the primary structural defect in the dysthrombin thrombin Quick I: substitution of cysteine for arginine-382.

A congenitally dysfunctional form of prothrombin, prothrombin Quick, was isolated from the plasma of an individual with less than 2% of normal prothrombin activity. Following activation of prothrombin Quick, two dysfunctional thrombins, thrombin Quick I and thrombin Quick II, were isolated. Functional characterization of thrombin Quick I indicated an increase in KM and a decrease in kcat, relative to thrombin, for release of fibrinopeptide A. Comparison of kcat/KM for thrombin Quick I to the value obtained for thrombin yielded a relative catalytic efficiency of 0.012 for thrombin Quick I [Henriksen, R. A., & Owen, W. G. (1987) J. Biol. Chem. 262, 4664-4669]. Lysyl endopeptidase digestor of reduced and S-carboxymethylated thrombin and thrombin Quick I has resulted in the identification of an altered peptide in this dysthrombin. Edman degradation of the isolated peptide has shown that the altered residue in this protein is Arg-382 which is replaced by Cys. This could result from a point mutation in the Arg codon, CGC, to yield TGC. Together, these results indicate that Arg-382 is a critical residue in determining the specificity of thrombin toward fibrinogen. Similar relative activities for thrombin Quick I in stimulating platelet aggregation, in the release of prostacyclin from human umbilical vein endothelium, and in the release of fibrinopeptide A suggest that these activities of thrombin share the same specificity determinants.