bluuebunny/crossref_metadata_2025 huggingface dataset:
Intraoperative autologous blood recovery in prostate cancer surgery: in vivo validation using a tumour marker
Authors: Poli M. , Camargo A. , Villa L. , Moura R. , Colella R. , Deheinzelin D.
Published: October 2008

Background and Objective  Intraoperative autologous blood recovery during radical retro pubic prostatectomy has the potential of contamination with tumour cells. Its safety is proved by similar survival rates between allogeneic and autologous transfusion to oncology patients without standardization. Silencing of the gene encoding pi class of gluthatione‐S transferase is a specific and sensitive molecular marker for prostate cancer, because it is present in more than 90% of prostate tumours. Using such tumour marker, we aimed to demonstrate that viable tumour cells could be eliminated using leucodepletion filters followed by irradiation.

Materials and Methods  Fifty patients with pi class of gluthatione‐S transferase promoter hypermethylation in their primary prostate tumours were included in the analysis. Peripheral blood samples were collected during anaesthetic induction and recovered blood was collected throughout the surgery and then submitted to washing, leucoreduction and irradiation. Samples were analysed stepwise for the presence of promoter hypermethylation using real‐time methylation‐specific polymerase chain reaction.

Results  Positive hypermethylation was found in recovered blood (two samples), recovered and washed blood (three samples), and recovered washed and filtered blood (two samples). After filtration and irradiation of the recovered blood, this marker could not be detected in any of the cases analysed, suggesting the absence of viable tumour cells.

Conclusion  Even though the risk of disseminating tumour cells in prostate cancer surgery by intraoperative autologous blood recovery is not yet fully established, no tumour‐specific gene amplification was found after the association of blood filtration and irradiation, suggesting a significant reduction of such risk.
Blood transfusion requirements and independent predictors of increased transfusion requirements among adult patients on extracorporeal membrane oxygenation – a single centre experience
Authors: Ang A. L. , Teo D. , Lim C. H. , Leou K. K. , Tien S. L. , Koh M. B. C.
Published: October 2008

Background/Objectives  More adults undergo extracorporeal membrane oxygenation (ECMO) now. They have high transfusion requirements. This study described transfusion requirements of adults during ECMO in a single institution, and determined factors associated with high transfusion requirements.

Materials/Methods  Retrospective analysis was done on the amount of blood products received by adults during ECMO. Predictors of increased average daily transfusion requirements during ECMO and increased ECMO duration (which correlated positively with total transfusion requirements) were determined.

Results  Forty‐one patients (median age 50 years) underwent 42 ECMO sessions for respiratory failure (16·7%), cardiogenic shock (76·2%) or massive pulmonary embolism (7·1%). They received 569 red blood cells, 852 platelets, 126 fresh‐frozen plasma (FFP) and 220 cryoprecipitate in total during median ECMO duration of 5 (1–15) days. On multivariate analysis, average daily red blood cell transfusion increased with nadir haemoglobin (Hb) during ECMO (Hbnadir) of < 7·5 g/dl (P < 0·001). Average daily platelet transfusion increased with recent antiplatelet agents (P = 0·015) and maximum Hb decline of > 5·5 g/dl during ECMO (P = 0·011). Average daily platelet transfusion > 3 units was also associated with increased ECMO duration (P = 0·024). Average daily FFP transfusion was increased in patients with hypertension (P = 0·007) and Hbnadir < 7·5 g/dl (P = 0·050). Patients with sepsis (P = 0·009) or without surgery (P = 0·009) had increased ECMO duration, which correlated positively with total transfusion requirements during the entire ECMO session. ECMO improved mortality of patients with fulminant myocarditis, respiratory failure and massive pulmonary embolism.

Conclusion  Adult ECMO patients with lower Hbnadir require more daily red blood cell and FFP. Hypertension increases daily FFP requirements. Recent antiplatelet agents, larger Hb decline and longer ECMO duration increase daily platelet requirements. Patients with sepsis or on ECMO for medical reasons have longer ECMO duration, which is associated with total transfusion requirements. Some of these factors may be identified early to optimize blood product support.
Predictors for non‐compliance of notified hepatitis C virus‐positive blood donors with recommendation to seek medical counselling
Authors: Kerzman H. , Green M. S. , Shinar E.
Published: October 2008

Background  Notification of blood donors represents the commonest method of informing asymptomatic individuals of abnormal test results indicating exposure to hepatitis C virus (HCV) infection. Such notification is therefore important from both health and economic perspectives. This study aimed to identify predictors for non‐compliance of HCV‐positive blood donors with the National Blood Services recommendation to seek medical counselling.

Study Design and Methods  The current research is a cross‐sectional study. Telephone interviews were conducted with 201 blood donors identified as HCV positive following blood donation during 2001–2002 (40% response rate).

Results  About 25% of all the notified blood donors did not seek any counselling; 29% (44/150) of those who requested medical advice from their primary care physicians (general practitoner's) were not referred to specialists. Age, alcohol consumption and non‐practice of health‐promoting behaviour were independent predictors of non‐compliance with the blood services’ recommendation. In particular, smoking (odds ratio, 2·0; 95% confidence interval 1·0–4·2) and not undergoing professional teeth cleaning (odds ratio 2·8; 95% confidence interval 1·3–6·1) were found to be significant predictors of non‐compliance.

Conclusion  The study provides essential data regarding the extent and risk factors for non‐compliance of HCV‐positive blood donors with recommendation to seek medical advice. Our results can assist in identifying blood donors who would not seek counselling, based on demographic factors and past exposure to risk factors for HCV. Improvements in the notification process and additional training of general practitoners regarding the management of HCV disease are needed.
Blood group A1 and A2 revisited: an immunochemical analysis
Authors: Svensson L. , Rydberg L. , De Mattos L. C. , Henry S. M.
Published: October 2008

Background and Objective  The basis of blood group A1 and A2 phenotypes has been debated for many decades, and still the chemical basis is unresolved. The literature generally identifies the glycolipid chemical differences between blood group A1 and A2 phenotypes as being poor or no expression of A type 3 and A type 4 structures on A2 red cells, although this assertion is not unanimous.

Materials and Methods  Using purified glycolipids and specific monoclonal antibodies, we revisited the glycolipid basis of the A1 and A2 phenotypes. Purified glycolipids were extracted from four individual A1 and four individual A2 blood units. One blood unit from an A weak subgroup was also included. Monoclonal anti‐A reagents including those originally used to define the basis of A1 and A2 phenotypes were used in a thin layer chromatography – enzyme immunoassay to identify the presence of specific glycolipids.

Results  A type 3 glycolipid structures were found to be present in large amounts in all phenotypes. In contrast, the A type 4 glycolipid structure was virtually undetectable in the A2 phenotype, but was present in the A1 and A subgroup samples.

Conclusion  The major glycolipid difference between the A1 and A2 phenotypes is the dominance of A type 4 glycolipids in the A1 phenotype.
Re‐Introduction of Quínoa into Arid Chile: Cultivation of Two Lowland Races under Extremely Low Irrigation
Authors: Martínez E. A. , Veas E. , Jorquera C. , San Martín R. , Jara P.
Published: October 2008

Annual rainfall in Chile at 30°S decreased from 170 to 70 mm in the last century, forcing a search for new low‐rain adapted crops. Chenopodium quinoa Willd. was cultivated by pre‐Hispanic cultures, but it disappeared in this region since the Spanish conquest. Two quínoa landraces (Don Javi and Palmilla) were re‐introduced from lowlands of central Chile (34°S) evaluating seed saponin content and grain yields under low irrigation. Replicated assays were conducted in two sites with distinct microclimates after august (end of the rains in 2004 and 2005). Treatments included low (40–75 mm) and high (150–250 mm) irrigation and were distributed along the five cultivation months. Fertilization, with the humus of the worms, was carried out in the second season, as soils are poor in organic matter. Results showed significantly higher saponin content in the seeds of Don Javi landrace (1.2 %) with respect to Palmilla seeds (0.3 %). However, grain yields were not different between landraces under the same treatments. Yields were instead affected by microclimate, irrigation and fertilization. Although higher yields corresponded with higher irrigation, 2.6 tons ha−1 was obtained under high irrigation, but surprisingly, also under low irrigation in the more humid site. Yields of 2006 harvesting season (ca. 7 tons ha−1) were higher than that of the previous season (ca. 5.5 tons ha−1), mainly because of the addition of organic matter. We suggest that re‐introduction of Quinoa in arid Chile is feasible even under the prevailing conditions of low rainfall and deficient soils, but better yields will need some irrigation and addition of organic matter.
A case study of sewage discharge in the shallow coastal area of the Northern Adriatic Sea (Gulf of Trieste)
Authors: Mozetič Patricija , Malačič Vlado , Turk Valentina
Published: October 2008

A case study was carried out in 2000 in the shallow coastal area of the Northern Adriatic Sea (Gulf of Trieste) where untreated domestic sewage and industrial wastes are discharged at rate of 5500 m3·day−1. The sewage plume above the outfall was followed using faecal coliforms (FC) and overturning length scale (lT). The latter was rejected as a marker as the discharge conditions prohibit following the turbulence of sewage water. Intermittent sewage discharge is reflected in the minimal effect of eutrophication. Increase of phytoplankton biomass is thus only minor compared with the unpolluted area regardless of elevated concentrations of sewage‐derived nutrients (confirmed by correlation coefficients between FC and NH4+, TP, PO43−: 0.78, 0.71 and 0.67, respectively). Deteriorated trophic status, determined by the TRIX index, was observed only in the surface layer (average TRIX: 5.67). High FC content well above the regulation limit (up to 2.6 × 105 FC·100 ml−1) represents, therefore, the major negative impact of the improperly treated waste for the risk to human health.
Depth‐stratified community zonation patterns on Gulf of Alaska rocky shores
Authors: Konar Brenda , Iken Katrin , Edwards Matthew
Published: October 2008

Vertical zonation patterns have been considered ubiquitous in intertidal ecosystems but questions remain about their generality for individual taxonomic groups and over broad spatial scales, and whether they continue into adjacent shallow subtidal habitats. Taxon richness, invertebrate abundance, and macroalgal biomass were examined in the summer of 2003 along a vertical gradient in the rocky intertidal and shallow subtidal habitats around Kodiak Island, Kachemak Bay, and Prince William Sound, all within the Gulf of Alaska. Replicate samples of benthic organisms were taken in the high (∼ 7 m), mid (∼ 4 m) and low (∼ 0 m) intertidal (relative to MLLW), and at 1, 5, 10 and 15 m water depths at three sites in each region, and identified to the lowest possible taxonomic level. Our primary goals were to assess (1) how estimates of taxon richness, invertebrate abundance, and macroalgal biomass vary among intertidal heights and subtidal depths and (2) how general these patterns are when considered across the Gulf of Alaska. Our results show that when all invertebrates were considered together, most of the variation in taxon richness was accounted for by differences among depths (i.e. intertidal heights and subtidal depths) (∼ 51%), and among replicate samples within each depth (∼ 26%). Little to none of the variation was accounted for by differences among sites within each region (∼ 1%) or among regions themselves (∼ 0%). When considered across the Gulf of Alaska, total taxon richness and organism abundance were greatest in the low intertidal/shallow subtidal and decreased with increasing height/depth. When separated by phylum and examined together with macroalgae, variation in abundance and/or biomass among depths was significant and accounted for most of the variability. Differences among regions and sites within each region were not significant and accounted for little to none of the variance. Because the pattern of zonation varied among sites within each region, it reduced the generality of a single zonation pattern for the Gulf of Alaska. Likewise, when community composition was compared among depths, geographic regions and sites within each region using multivariate analyses, vertical zonation patterns were evident at a regional scale, but high variability in these patterns among sites within each region reduced the generality of these patterns.
Adaptations of araphid pennate diatoms to a planktonic existence
Authors: Kooistra Wiebe H. C. F. , Forlani Gandi , De Stefano Mario
Published: October 2008

Pennate diatoms are monophyletic. Their principal cell wall elements, called valves, are shaped like a ship’s hull. Within the pennates, the araphids are paraphyletic; they possess rimoportulae and pore fields located at the valve apices. The pore fields exude mucilage pads with which cells attach to one another to form chains. Many taxa use the pads also for attachment to substrata. Only a few genera are truly planktonic. The main question addressed in this study is whether the planktonic lifestyle is ancestral or derived. Phylogenies inferred from nuclear SSU rDNA gene sequences of diatoms indicated that the attached lifestyle is ancestral among the araphids, whereas a typically planktonic lifestyle seems to have developed at least three times and possibly four times independently. Acquisition of a planktonic lifestyle from benthic ancestry was accompanied by a reduction in the silicification of cell‐wall elements, but changes in morphological characters shared by all four clades were not detected. The reason why only three or four araphid pennate clades have adopted a planktonic lifestyle may be related to constraints associated to their sexual reproduction mode. Partner cells of opposite mating type align with one another and produce isogametes. These gametes lack flagella; they move to one another in an amoeboid fashion, which functions well on surfaces, but seems a liability in a turbulent water column. The planktonic lineages must have overcome this constraint, e.g. by sinking to the bottom, or aggregating, to perform sexual reproduction. Members of the four araphid pennate lineages are now common constituents of the plankton, suggesting that they are ecologically successful.
Genetic analysis of agronomic and fibre traits using four interspecific chromosome substitution lines in cotton
Authors: Saha S. , Jenkins J. N. , Wu J. , McCarty J. C. , Stelly D. M.
Published: October 2008

Backcrossed chromosome substitution lines (CS‐B) have been developed with a homologous pair of chromosomes or chromosome arms of Gossypium barbadense (3‐79) germplasm substituted for the homologous Gossypium hirsutum(TM‐1) chromosomes or chromosome segments. We report on agronomic and fibre trait performance of four backcrossed chromosome or chromosome arm substitution lines including chromosomes 01, 11sh (chromosome 11 short arm), 12 sh and 26 Lo (chromosome 26 long arm). Data for agronomic and fibre traits were collected from replicated field experiments at two different locations in 2 years, and analysed under an additive dominance genetic model. CS‐B 12sh had higher, while CS‐B 01 and CS‐B 26Lo had lower boll weight than TM‐1. The presence of significant negative additive effects for micronaire with CS‐B 01 and significant positive additive effects for elongation and fibre strength with CS‐B 11sh suggested the substituted chromosome arms of 3‐79 in these CS‐B lines were more likely carrying genes causing these effects. Results revealed that several CS‐B lines had significant homozygous and heterozygous dominance effects for different agronomic and fibre traits suggesting that specific CS‐B lines may be useful for improving agronomic and fibre traits in hybrid cottons. These CS‐B lines also provide novel genetic resources for improving upland cotton germplasm.
Lowering seed gossypol content in glanded cotton (Gossypium hirsutum L.) lines
Authors: Romano G. B. , Scheffler J. A.
Published: October 2008

Cottonseed is a rich source of high quality protein, but its value as an animal feed is limited by gossypol, a toxic polyphenolic compound contained in glands located throughout the plant. This compound helps protect the plant from pests. Totally glandless varieties have been developed, but not adopted as these plants are left vulnerable to pests. This study describes a breeding strategy to decrease the levels of gossypol in the seed while maintaining a high enough concentration of toxin in vegetative plant parts to offer protection from pests. Preliminary studies indicated that crosses between varieties with different gland densities and distributions produced a range of glanding patterns. By selecting within the resulting progeny, we have identified F7 generation progeny that have <0.30% total gossypol in the seed, while still possessing glands at critical locations on the vegetative plant parts. These new lines will be a valuable source of germplasm for developing low seed gossypol varieties. Seed from these varieties would provide a new source of inexpensive protein for animal feeding rations.
Pyramiding of genes conferring resistance to Tomato yellow leaf curl virus from different wild tomato species
Authors: Vidavski F. , Czosnek H. , Gazit S. , Levy D. , Lapidot M.
Published: October 2008

Tomato (Solanum lycopersicum) production in tropical and subtropical regions of the world is limited by the endemic presence of Tomato yellow leaf curl virus (TYLCV). Breeding programmes aimed at producing TYLCV‐resistant tomato cultivars have utilized resistance sources derived from wild tomato species. So far, all reported breeding programmes have introgressed TYLCV resistance from a single wild tomato source. Here, we tested the hypothesis that pyramiding resistances from different wild tomato species might improve the degree of resistance of the domesticated tomato to TYLCV. We have crossed TYLCV‐resistant lines that originated from different wild tomato progenitors, Solanum chilense, Solanum peruvianum, Solanum pimpinellifolium, and Solanum habrochaites. The various parental resistant lines and the F1 hybrids were inoculated in the greenhouse using viruliferous whiteflies. Control, non‐inoculated plants of the same lines and hybrids were exposed to non‐viruliferous whiteflies. Following inoculation, the plants were scored for disease symptom severity, and transplanted to the field. Resistance was assayed by comparing yield of inoculated plants to those of the control non‐inoculated plants of the same variety. Results showed that the F1 hybrids between the resistant lines and the susceptible line suffered major yield reduction because of infection, but all hybrids were more resistant than the susceptible parent. All F1 hybrids resulting from a cross between two resistant parents, showed a relatively high level of resistance, which in most cases was similar to that displayed by the more resistant parent. In some cases, the hybrids displayed better levels of resistance than both parents, but the differences were not statistically significant. The F1 hybrid between a line with resistance from S. habrochaites and a line with resistance from S. peruvianum (HAB and 72‐PER), exhibited the lowest yield loss and the mildest level of symptoms. Although the resistance level of this F1 hybrid was not statistically different from the level of resistance displayed by the 72‐PER parent itself, it was statistically better than the level of resistance displayed by the F1 hybrids between 72‐PER and any other resistant or susceptible line.
Common bean (Phaseolus vulgaris L.) lines devoid of major lectin proteins
Authors: Campion B. , Perrone D. , Galasso I. , Bollini R.
Published: October 2008

In common bean, the most abundant proteins of the lectin family (arcelin, phytohaemagglutinin and α‐amylase inhibitor –APA proteins) are major antinutritional factors of the seeds. Reduced protein digestibility and high toxicity at the intestinal level are the causes of their antinutritional effect. Bean lines producing seeds without the lectins mentioned above were developed to improve nutritional characteristics of bean seeds used for human consumption and, potentially, for animal feeding. The genetic trait was transferred from a wild bean accession into a number of bean cultivars. The 1 : 3 ‘lectin‐free/lectin‐containing seeds’ segregation ratio hypothesis was confirmed by chi‐square analysis in the F2 progenies. Four lectin‐free stable lines, P500, P501, 938 and 586/8 (white seeded), and 44 climbing breeding lines, have been developed. The ‘white seed coat’ trait, correlated with a reduced amount of tannins and polyphenols in bean seeds, has been introduced in our lectin‐free materials to further improve their nutritional characteristics. The field performance of P501, 938 and 586/8 and that of their lectin‐containing parents, was evaluated on 2007 in two locations. Dry seed yield, seed emergence, plant height, plant growth duration, average weight and germination capacity of the produced seeds were surveyed and statistically analysed. At the location of Battipaglia (Salerno), the dry seed yield value of the lectin‐free line 938 (3.74 t/ha) and that of one of its parent, the CIAT line BAT 881 (3.65 t/ha), were significantly higher than those of all other lines tested. Germination percentage of lectin‐free lines, recorded 3 days after sowing, was significantly higher than that of controls.
Seroprevalence of Canine Herpesvirus‐1 andBrucella canisin Finnish Breeding Kennels with and without Reproductive Problems
Authors: Dahlbom M , Johnsson M , Myllys V , Taponen J , Andersson M
Published: October 2008

Contents

We compared the serological status ofBrucella canisand canine herpesvirus‐1 (CHV‐1) in Finnish breeding kennels with and without reproductive problems. Dogs from kennels with reproductive problems had significantly higher CHV‐1 titres than dogs from kennels having no reproductive problems (p < 0.001). In dogs from kennels with reproductive problems 100% (32/32) had positive titres, whereas in dogs from kennels without reproductive problems 65% (22/34) had positive titres. The median titre for dogs from kennels with reproductive problems was 1 : 160 and for dogs from kennels without reproductive problems 1 : 80. The high prevalence of positive CHV‐1 titres in this study indicates that prevention of the disease is difficult and reinforces the need to minimize the reproductive problems caused by CHV‐1. All 388 dogs from 94 kennels had negativeB. canistitres.
Minimal External Masculinization in a SRY‐negative XX Male Podenco Dog
Authors: Buijtels JJCWM , De Gier J , Van Haeften T , Kooistra HS , Spee B , Veldhuis Kroeze EJB , Zijlstra C , Okkens AC
Published: October 2008

Contents

Normal mammalian sex differentiation takes place in three genetically controlled steps: chromosomal sex determination (XX or XY), gonadal differentiation and development of the phenotypic sex. Animals are considered to be sex reversed if chromosomal sex determination and gonadal development are not in agreement. In this report, sex reversal is described in a 1.5‐year‐old Podenco dog that was referred because of suspected recurrent growth of a previously removed os clitoridis in the vulva. With that exception the dog was phenotypically female, but had never been in oestrus and exhibited male behaviour. Abdominal ultrasonography showed a small tubular structure dorsal to the bladder, consistent with a uterus. An ovoid structure resembling a gonad was visible between the right kidney and inguinal canal. Plasma testosterone concentrations before and after GnRH administration indicated the presence of functional testicular tissue. Two testes, each with its epididymis and ductus deferens, and a complete bicornuate uterus were removed surgically. Cytogenetic analysis of peripheral blood lymphocytes showed a normal female karyotype (78, XX). These findings are consistent with the diagnosis of an XX male. PCR analysis of genomic DNA revealed that the SRY gene was absent. In summary, this report describes the first SRY‐negative XX male Podenco dog with an almost complete female phenotype despite high basal and stimulated plasma testosterone concentrations. It is hypothesized that the clinical observations in this dog may have been caused by reduced and delayed Müllerian‐inhibiting substance secretion and the absence of conversion of testosterone to dihydrotestosterone due to 5α‐reductase deficiency.
Immunohistochemical Localization of Oestrogen Receptors α and β, Progesterone Receptor and Aromatase in the Equine Placenta
Authors: Abd‐Elnaeim MMM , Derar IR , Wilsher S , Allen WR , Leiser R , Schuler G
Published: October 2008

Contents

The functions of placental oestrogens during equine pregnancy are still unclear. Yet, they may act predominantly as local regulators of growth and differentiation in the microplacentomes. Thus, expression patterns of oestrogen receptors (ERs) α and β were investigated in the microcotyledonary placenta from pregnant mares at 110, 121, 179, 199 and 309 days of gestation by immunohistochemistry. In microplacentomes, both the ER isoforms were detected in trophoblast (T) cells, chorionic villous stroma (FS), microcaruncular epithelium (ME) and microcaruncular stroma (MS). Proportions of positive cells were 38–91% (T), 11–41% (FS), 55–89% (ME), 17–51% (MS) for ERα and 66–76% (T), 21–37% (FS), 41–68% (ME) and 24–55% (MS) for ERβ. Between days 110 and 199, proportions of cells positive for progesterone receptor (PR) varied between 19% and 62% (T), 3% and 50% (CS), 15% and 46% (ME), and 4% and 33% (MS). At day 309, PR was virtually absent in T, CS and ME (percentages < 0.1), whereas in MS 14.3% of cells were still positive. The expression of ERs and PR in equine microplacentomes gives evidence for a role of placental steroids as regulators of placental growth, differentiation and function. The detection of ERα, ERβ and PR in foetal and maternal vascular tissue suggests that placental steroids are also involved in the control of placental angiogenesis and /or vascular functions. The co‐localization of ERs with aromatase in T suggests auto‐ or intracrine functions of oestrogens in this cell type.
Genetic Description of Factor XI Deficiency in Holstein Semen in Western Japan
Authors: Ghanem ME , Nishibori M
Published: October 2008

Contents

Factor XI deficiency was detected in Holstein cows and mummified foetuses in Japan; however, no report is available about the occurrence of Factor XI deficiency in Holstein semen in Japan. Five hundred cows in twelve dairy farms in Hiroshima Prefecture, Japan were under the study. Genomic DNA was extracted from the cows using a commercial DNA kits and screened to Factor XI mutation. Based on the information of the carrier cows found in the cattle population, four Holstein bulls were analysed for Factor XI mutation. DNA was extracted from bull’s semen using phenol chloroform method. Extracted genomic DNA of the bull’s semen was typed for Factor XI using specific polymerase chain reaction (PCR) primers. The resultant PCR was sequenced using big dye terminator sequencing method. The pedigree of the bulls was investigated. Furthermore, the inheritance of Factor XI mutation to next generation was estimated. Out of the 500 cows, five were heterozygous to Factor XI. Moreover, out of the four bulls, one was found to carry the mutation of Factor XI; it was also a complex vertebral malformation (CVM) carrier. In DNA sequencing, the insertion mutation of 76 bp of poly‐adenine that characterizes the Factor XI deficiency was detected in the carrier bull as well as the carrier cows. Pedigree analysis of the carrier bull revealed that his father and mother ID were 2247419A and 14189172A, respectively, that originated from USA Holstein. Out of six daughter cows born to the carrier bull, one cow (16.6%) inherited Factor XI mutation, while three of them (50.0%) inherited CVM mutation. Autosomal recessive genes that affect cow’s reproduction have a particular concern to dairy industry. To our knowledge this is the first report of Factor XI mutation in Holstein semen in Japan.
In vitro Fertilizing Capacity of Frozen‐thawed Bull Spermatozoa Selected by Single‐layer (Glycidoxypropyltrimethoxysilane) Silane‐coated Silica Colloidal Centrifugation
Authors: Thys M , Vandaele L , Morrell JM , Mestach J , Van Soom A , Hoogewijs M , Rodriguez‐Martinez H
Published: October 2008

Contents

Barriers to the use of density gradient centrifugation for preparing animal spermatozoa for artificial insemination (AI) include the scarcity of animal‐specific formulations and the daunting prospect of processing large volumes of ejaculate in small aliquots (1.5 ml extended semen). Recently, new colloid formulations have been tested in vitro in a modified procedure, centrifugation on a single layer of colloid. The present study investigated the fertilizing ability during in vitro fertilization (IVF) of frozen‐thawed bovine spermatozoa following centrifugation through a single layer of glycerolpropylsilane (GS)‐coated silica colloid with a species‐specific formulation (patent applied for; treatment, T). Controls (C) included centrifugation through gradients of either the same colloid (C1) or Percoll™ (C2). Sperm recovery surpassed 50% for both C1–C2 and T (n.s.). Mean values of various parameters of computerized analysis of sperm motility did not differ between T and C1 (n.s.), and only the proportions of path straightness and linearity were lower in T vs C2 (p < 0.05). In T, the mean (±SD) percentages of fertilization rate, blastocyst development rate and the total number of blastomeres were 58.1 ± 23.3%, 24.5 ± 14.3% and 94.6 ± 23.4%, respectively. The proportions did not differ significantly from controls (C1/C2). Therefore, centrifugation through a single layer of colloid offers an alternative method to density gradient centrifugation for selection of viable, potentially fertile frozen‐thawed bull spermatozoa. This single‐layer technique is gentle, versatile and convenient because it facilitates scaling‐up the process of sperm preparation to allow larger numbers of spermatozoa (for instance, whole ejaculates) to be processed for AI.
Cryotolerance of Bovine Blastocysts is Affected by Oocyte Maturation in Media Containing Palmitic or Stearic Acid
Authors: Shehab‐El‐Deen MA , Leroy JLMR , Maes D , Van Soom A
Published: October 2008

Contents

In this study, non‐esterified fatty acids (NEFAs) were added during in vitro maturation at concentrations measured previously in follicular fluid (FF) of high‐producing dairy cows in a negative energy status to evaluate their subsequent effect on the embryos cryotolerance. Oocytes were matured for 24 h in serum‐free media with or without (negative control) the addition of NEFAs dissolved in ethanol or ethanol alone (positive control). Matured oocytes were fertilized and cultured for 7 days in synthetic oviduct fluid medium supplemented with 5% FCS. Embryos that had at least reached the blastocyst stage were vitrified by open pulled straw (OPS) vitrification. Addition of palmitic (C16 : 0) or stearic acid (C18 : 0) during oocyte maturation had significant negative effects on embryo cryotolerance, whereas ethanol or oleic acid (C18 : 1) had no effect. These in vitro results suggest that high NEFA concentrations in FF during a period of negative energy balance in high‐yielding dairy cows can have carry‐over effects on embryo quality.
Ablation of Cx47 in transgenic mice leads to the loss of MUPP1, ZONAB and multiple connexins at oligodendrocyte–astrocyte gap junctions
Authors: Li Xinbo , Penes M. , Odermatt B. , Willecke K. , Nagy J. I.
Published: October 2008

Oligodendrocytes in CNS are linked to astrocytes by heterotypic gap junctions composed of Cx32 and Cx47 in oligodendrocytes and Cx30 and Cx43 in astrocytes. These gap junctions also harbour regulatory proteins, including ZO‐1 and ZONAB. Here, we investigated the localization of multi‐PDZ domain protein 1 (MUPP1) at these gap junctions and examined accessory proteins and connexins associated with oligodendrocytes in Cx47‐knockout mice. In every CNS region tested, punctate immunolabelling for MUPP1 was found on all oligodendrocyte somata in wild‐type mice. These MUPP1‐positive puncta were colocalized with punctate labelling for oligodendrocytic Cx32 or Cx47, and with astrocytic Cx30 or Cx43 at oligodendrocyte–astrocyte (O/A) gap junctions, but were not found at astrocyte–astrocyte gap junctions. In Cx47‐knockout mice, immunolabelling of MUPP1 and ZONAB was absent on oligodendrocytes, whereas some ZO‐1‐positive puncta remained. In Cx32‐knockout mice, MUPP1 and ZONAB persisted at O/A gap junctions. The absence of Cx47 in Cx47‐knockout mice was accompanied by a total loss of punctate labelling for Cx30, Cx32 and Cx43 on oligodendrocyte somata, and by a dramatic increase in immunolabelling for Cx32 along myelinated fibers. These results demonstrate MUPP1 at O/A gap junctions and Cx47‐dependent targeting of connexins to the plasma membranes of oligodendrocyte somata. Further, it appears that deficits in myelination reported in Cx47‐knockout mice may arise not only from a loss of Cx47 but also from the accompanied loss of gap junctions and their regulatory proteins at oligodendrocyte somata, and that loss of Cx47 may be partly compensated for by elevated levels of Cx32 along myelinated fibers.
Characterization of the cystine/glutamate transporter in the outer plexiform layer of the vertebrate retina
Authors: Hu Rebecca G. , Lim Julie , Donaldson Paul J. , Kalloniatis Michael
Published: October 2008

The cystine/glutamate transporter (Xc‐) is widely expressed in the central nervous system and is thought to play a role in glutamatergic neurotransmission by releasing low levels of glutamate. Although previous studies have localized the transporter throughout the retina, we now present results from localization and functional studies within the first synaptic layer, i.e. the outer plexiform layer. Using light and electron microscopy, we have localized the Xc‐ transporter to the ribbon complex of both rod and cone photoreceptors in the rat, cow, lamprey, chicken and monkey retina, suggesting that the pre‐synaptic expression of the Xc‐ transporter on the photoreceptor ribbon complex is phylogenetically preserved. The Xc‐ transporter does not appear to be located within the ribbon synapse of the inner plexiform layer. Developmentally, the evolution of distinct ribbon‐shaped Xc‐ labelling in the outer plexiform layer parallels the known morphological and electrophysiological maturation of photoreceptors. Using the cation channel probe agmatine, we tracked cation fluxes within bipolar cells and therefore indirectly determined the modulation of glutamate release from photoreceptors. Such cystine‐driven alteration in agmatine entry into bipolar cells can be modified by a specific metabotropic glutamate receptor 6 antagonist [(RS)‐α‐cyclopropyl‐4‐phosphonophenylglycine] and an Xc‐ transport inhibitor [(S)‐4‐carboxyphenylglycine]. The phylogenetic preservation of the transporter, its ultrastructural localization to the ribbon synapse and functional modulation of post‐receptoral neurons collectively support a role for the Xc‐ transporter in glutamate neurotransmission in the outer retina of vertebrates. We have therefore proposed a model of glutamate release in the photoreceptor synapse that incorporates the Xc‐ transporter, which complements the established vesicular‐mediated glutamate release.
Differential effects of lentiviral vector‐mediated overexpression of nerve growth factor and glial cell line‐derived neurotrophic factor on regenerating sensory and motor axons in the transected peripheral nerve
Authors: Tannemaat Martijn R. , Eggers Ruben , Hendriks William T. , De Ruiter Godard C. W. , Van Heerikhuize Joop J. , Pool Chris W. , Malessy Martijn J. A. , Boer Gerard J. , Verhaagen Joost
Published: October 2008

Even after reconstructive surgery, major functional impairments remain in the majority of patients with peripheral nerve injuries. The application of novel emerging therapeutic strategies, such as lentiviral (LV) vectors, may help to stimulate peripheral nerve regeneration at a molecular level. In the experiments described here, we examined the effect of LV vector‐mediated overexpression of nerve growth factor (NGF) and glial cell line‐derived neurotrophic factor (GDNF) on regeneration of the rat peripheral nerve in a transection/repair model in vivo. We showed that LV vectors can be used to locally elevate levels of NGF and GDNF in the injured rat peripheral nerve and this has profound and differential effects on regenerating sensory and motor neurons. For sensory neurons, increased levels of NGF and GDNF do not affect the number of regenerated neurons 1 cm distal to a lesion at 4 weeks post‐lesion but do cause changes in the expression of markers for different populations of nociceptive neurons. These changes are accompanied by significant alterations in the recovery of nociceptive function. For motoneurons, overexpression of GDNF causes trapping of regenerating axons, impairing both long‐distance axonal outgrowth and reinnervation of target muscles, whereas NGF has no effect on these parameters. These observations show the feasibility of combining surgical repair of the transected nerve with the application of viral vectors. Furthermore, they show a difference between the regenerative responses of motor and sensory neurons to locally increased levels of NGF and GDNF.
Alcohol self‐administration acutely stimulates the hypothalamic‐pituitary‐adrenal axis, but alcohol dependence leads to a dampened neuroendocrine state
Authors: Richardson Heather N. , Lee Soon Y. , O’Dell Laura E. , Koob George F. , Rivier Catherine L.
Published: October 2008

Clinical studies link disruption of the neuroendocrine stress system with alcoholism, but remaining unknown is whether functional differences in the hypothalamic‐pituitary‐adrenal (HPA) axis precede alcohol abuse and dependence or result from chronic exposure to this drug. Using an operant self‐administration animal model of alcohol dependence and serial blood sampling, we show that long‐term exposure to alcohol causes significant impairment of HPA function in adult male Wistar rats. Acute alcohol (voluntary self‐administration or experimenter‐administered) stimulated the release of corticosterone and its upstream regulator, adrenocorticotropic hormone, but chronic exposure sufficient to produce dependence led to a dampened neuroendocrine state. HPA responses to alcohol were most robust in ‘low‐responding’ non‐dependent animals (averaging < 0.2 mg/kg/session), intermediate in non‐dependent animals (averaging ∼0.4 mg/kg/session), and most blunted in dependent animals (averaging ∼1.0 mg/kg/session) following several weeks of daily 30‐min self‐administration sessions, suggesting that neuroendocrine tolerance can be initiated prior to dependence and relates to the amount of alcohol consumed. Decreased expression of corticotropin‐releasing factor (CRF) mRNA expression in the paraventricular nucleus of the hypothalamus and reduced sensitivity of the pituitary to CRF may contribute to, but do not completely explain, neuroendocrine tolerance. The present results, combined with previous studies, suggest that multiple adaptations to stress regulatory systems may be brought about by excessive drinking, including a compromised hormonal response and a sensitized brain stress response that together contribute to dependence.
The corpus callosum modulates spindle‐burst activity within homotopic regions of somatosensory cortex in newborn rats
Authors: Marcano‐Reik Amy Jo , Blumberg Mark S.
Published: October 2008

The corpus callosum, a major interhemispheric fiber tract, mediates communication between homotopic regions within the primary somatosensory cortex (S1). Recently, in 1‐ to 6‐day‐old rats, brief bursts of oscillatory activity – called spindle‐bursts (SBs) – were described in cortical somatosensory areas following sensory feedback from sleep‐related myoclonic twitches or specific peripheral stimulation. To determine whether interhemispheric communication via the corpus callosum modulates the expression of SBs during this early period of development, we investigated the spontaneous expression of SBs in unanesthetized 1‐ to 6‐day‐old rats as well as SBs evoked by plantar surface stimulation of the forepaw. We hypothesized that surgically disrupting transcallosal communication (i.e. with callosotomy) or unilateral pharmacological manipulation of S1 activity (e.g. by blocking muscarinic receptors) would alter S1 activity in one or both hemispheres. First, callosotomy doubled the rate of spontaneous, twitch‐related SBs in left and right S1s by reducing the interval between successive SBs. Second, unilateral infusion into the left S1 of the muscarinic receptor antagonist, scopolamine, inhibited SBs in response to right forepaw stimulation; importantly, SBs were now disinhibited in the right S1 to right forepaw stimulation, thus ‘unmasking’ an ipsilateral representation. Subsequent callosotomy reinstated contralateral SB responses in the left S1. Finally, tactile and proprioceptive stimulation produced dissociable neurophysiological S1 responses; specifically, SBs were produced in response to proprioceptive, but not tactile, stimulation. We conclude that the corpus callosum modulates functionally inhibitory interactions between homotopic regions in left and right S1s during the early developmental period when organized neurophysiological activity is first detected in the neocortex.
Insights on Escherichia coli biofilm formation and inhibition from whole‐transcriptome profiling
Authors: Wood Thomas K.
Published: October 2008

Biofilms transform independent cells into specialized cell communities. Here are presented some insights into biofilm formation ascertained with the best‐characterized strain, Escherichia coli. Investigations of biofilm formation and inhibition with this strain using whole‐transcriptome profiling coupled to phenotypic assays, in vivo DNA binding studies and isogenic mutants have led to discoveries related to the role of stress, to the role of intra‐ and interspecies cell signalling, to the impact of the environment on cell signalling, to biofilm inhibition by manipulating cell signalling, to the role of toxin/antitoxin genes in biofilm formation, and to the role of small RNAs on biofilm formation and dispersal. Hence, E. coli is an excellent resource for determining paradigms in biofilm formation and biofilm inhibition.
Autoinducers extracted from microbial mats reveal a surprising diversity of N‐acylhomoserine lactones (AHLs) and abundance changes that may relate to diel pH
Authors: Decho Alan W. , Visscher Pieter T. , Ferry John , Kawaguchi Tomohiro , He Lijian , Przekop Kristen M. , Norman R. Sean , Reid R. Pamela
Published: October 2008

Microbial mats are highly structured and diverse communities, and one of the earliest‐known life assemblages. Mat bacteria interact within an environment marked by strong geochemical gradients and fluctuations. We examined natural mat systems for the presence of autoinducers involved in quorum sensing, a form of cell–cell communication. Our results revealed that a diverse array of N‐acylhomoserine lactones (AHLs) including C4‐ to C14‐AHLs, were identified from mat extracts using mass spectrometry (MS), with further confirmation by MS/MS‐collision‐induced dissociation (CID), and additions of external standards. Microelectrode measurements showed that mats exhibited diel pH fluctuations, ranging from alkaline (pH 9.4) during daytime (net photosynthesis) to acidic (pH 6.8) during darkness (net respiration/fermentation). Under laboratory conditions, AHLs having shorter acyl‐chains were degraded within the time frame that daily alkaline pH (> 8.2) conditions exist in mats. Intensive sampling of mats after full day‐ or night‐time incubations revealed that accumulations of extractable shorter‐chain AHLs (e.g. C8‐ and C10‐AHLs) were significantly (P < 0.001) diminished during daytime. Our study offers evidence that stabilities of AHLs under natural conditions may be influenced by the proximal extracellular environment. We further propose that the ancient periodicity of photosynthesis/respiration in mats may potentially drive a mechanism for diel differences in activities of certain autoinducers, and hence bacterial activities mediated through quorum sensing.
Seasonal influence on heat‐resistant proteolytic capacity of Pseudomonas lundensis and Pseudomonas fragi, predominant milk spoilers isolated from Belgian raw milk samples
Authors: Marchand Sophie , Heylen Kim , Messens Winy , Coudijzer Katleen , De Vos Paul , Dewettinck Koen , Herman Lieve , De Block Jan , Heyndrickx Marc
Published: October 2008

Psychrotolerant bacteria and their heat‐resistant proteases play a major role in the spoilage of UHT‐processed dairy products. Summer and winter raw milk samples were screened for the presence of such bacteria. One hundred and three proteolytic psychrotolerant bacteria were isolated, characterized by API tests, rep‐PCR fingerprint analysis and evaluated for heat‐resistant protease production. Twenty‐nine strains (representing 79% of the complete collection) were further identified by 16S rRNA gene sequencing, rpoB gene sequencing and DNA–DNA hybridizations. A seasonal inter‐ and intra‐species influence on milk spoilage capacity (e.g. growth rate and/or protease production) was demonstrated. Moreover, this polyphasic approach led to the identification of Pseudomonas fragi and Pseudomonas lundensis (representing 53% of all isolates) as predominant producers of heat‐resistant proteases in raw milk. The role of Pseudomonas fluorescens, historically reported as important milk spoiler, could not unequivocally be established. The use of more reliable identification techniques and further revision of the taxonomy of P. fluorescens will probably result in a different perspective on its role in the milk spoilage issue.