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PubMed2006

Effect of storage in short--and long-term commercial semen extenders on the motility, plasma membrane and chromatin integrity of boar spermatozoa.

For artificial insemination (AI) in pigs, preservation of liquid boar semen at 16-20 degrees C is still common practice as sperm cryopreservation remains suboptimal in this species. To meet the different needs of the swine industry, several extenders have been developed to preserve semen in liquid form for short--and long-term storage. In the present study, three different commercial extenders devised for short-term (BTS+) or long-term preservation (MR-A and X-Cell), were used to test whether storage of semen from four mature, fertile boars at 17 degrees C for 96 h would affect sperm characteristics relevant for fertility, such as motility, membrane integrity and chromatin stability. Computer-assisted sperm analysis, and stainings with the acylated membrane dye SYBR-14/propidium iodide, and acridine orange in connection with flow cytometry were used to evaluate these variables. Percentages of total motile spermatozoa decreased slightly, but significantly, after 72-96 h. While membrane integrity values varied during the period of study, no significant changes in either membrane integrity or chromatin stability were, however, registered. This suggests a customary 96-day storage at 17 degrees C in these extenders was too short an interval to cause losses of integrity in nuclear DNA in the boar population studied.

Animals↗

PubMed1995

Extended workshifts and excessive fatigue.

Studies of overtime have pointed to fatigue as a potential factor producing, for example, a three-fold increase in accident rate after 16 h of work, increases in back injuries, hospital outbreaks of bacterial infection, or nuclear-power plant safety compromises. Fatigue has been measured more directly in studies of scheduled long workshifts, where performance decrements in both work-related tasks and laboratory-type behavioural tests have been observed, and significant loss of sleep and increases in subjective sleepiness have been reported. Analyses of accidents or injuries during scheduled extended workshifts, however, have produced equivocal results. Factors which could compound the fatiguing effects of extended workshifts, such as workload, noise, chemical exposure, or duties and responsibilities outside of the workplace, rarely have been studied systematically. It is concluded that extended workshift schedules should be instituted cautiously and evaluated carefully, with appropriate attention given to staffing levels, workload, job rotation, environmental exposures, emergency contingencies, rest breaks, commuting time, and social or domestic responsibilities.

Journal Article↗

PubMed2004

The effect of an extended hospital induction on perceived confidence and assessed clinical skills of newly qualified pre-registration house officers.

BACKGROUND: Recent studies raise concerns over the preparedness of newly qualified doctors for the role of the pre-registration house officer (PRHO). This study aimed to assess self-perception of preparedness, objective assessment of core clinical skills and the effect of an extended clinical induction programme prior to commencing full duties. METHODS: A group of 26 newly qualified doctors from 1 district general hospital underwent an extended 5-day, ward-based induction programme. The participants completed questionnaires on their own perceptions of their preparedness for PRHO duties and underwent an objective structured clinical examination (OSCE) of 4 core clinical skills prior to induction, on completion of induction and 1 month into working life. RESULTS: At the outset PRHOs had low perceptions of their own capabilities in all clinical scenarios and skills. Most perceptions improved after induction, although in 2 clinical areas they felt even less confident. One month into post there were significant improvements in all areas. Only 1 PRHO passed all 4 clinical skills assessments at the pre-induction assessment. Seven (26%) failed on 1 or more skills at the post-induction assessment. However, all participants were deemed competent in all skills at the 1-month assessment. CONCLUSION: Newly qualified doctors do not feel prepared for PRHO duties and objectively are not competent in basic clinical skills. An extended induction improves preparedness in some but not all clinical areas and improves performance of objectively assessed clinical skills.

Attitude of Health Personnel↗

PubMed1996

A 38 kDa precursor protein of aqualysin I (a thermophilic subtilisin-type protease) with a C-terminal extended sequence: its purification and in vitro processing.

The precursor of aqualysin I, an extracellular subtilisin-type protease produced by Thermus aquaticus, consists of four domains: an N-terminal signal peptide, an N-terminal pro-sequence, a protease domain, and a C-terminal extended sequence. In an Escherichia coli expression system for the aqualysin I gene, a 38 kDa precursor protein consisting of the protease domain and the C-terminal extended sequence is accumulated in the membrane fraction and processed to a 28 kDa mature enzyme upon heat treatment at 65 degrees C. The 38 kDa precursor protein is separated as a soluble form from denatured E. coli proteins after heat treatment. Accordingly, purification of the 38 kDa proaqualysin I was performed using chromatography. The purified precursor protein gave a single band on SDS-polyacrylamide gels. The precursor protein exhibited proteolytic activity comparable to that of the mature enzyme. The purified precursor protein was processed to the mature enzyme upon heat treatment. The processing was inhibited by diisopropyl fluorophosphate. The processing rate increased upon either the addition of mature aqualysin I or upon an increase in the concentration of the precursor, suggesting that the cleavage of the C-terminal extended sequence occurs through an intermolecular self-processing mechanism.

Amino Acid Sequence↗

PubMed2006

Structural basis for the extended substrate spectrum of CMY-10, a plasmid-encoded class C beta-lactamase.

The emergence and dissemination of extended-spectrum (ES) beta-lactamases induce therapeutic failure and a lack of eradication of clinical isolates even by third-generation beta-lactam antibiotics like ceftazidime. CMY-10 is a plasmid-encoded class C beta-lactamase with a wide spectrum of substrates. Unlike the well-studied class C ES beta-lactamase from Enterobacter cloacae GC1, the Omega-loop does not affect the active site conformation and the catalytic activity of CMY-10. Instead, a three-amino-acid deletion in the R2-loop appears to be responsible for the ES activity of CMY-10. According to the crystal structure solved at 1.55 A resolution, the deletion significantly widens the R2 active site, which accommodates the R2 side-chains of beta-lactam antibiotics. This observation led us to demonstrate the hydrolysing activity of CMY-10 towards imipenem with a long R2 substituent. The forced mutational analyses of P99 beta-lactamase reveal that the introduction of deletion mutations into the R2-loop is able to extend the substrate spectrum of class C non-ES beta-lactamases, which is compatible with the isolation of natural class C ES enzymes harbouring deletion mutations in the R2-loop. Consequently, the opening of the R2 active site by the deletion of some residues in the R2-loop can be considered as an operative molecular strategy of class C beta-lactamases to extend their substrate spectrum.

Amino Acid Sequence↗

PubMed1989

The absence of 21-OH A and C4 B genes is a constant characteristic of the "Sardinian" HLA extended haplotype A30 Cw5 B18 BfF1 DR3 DRw52 DQw2.

The genomic DNA of 25 unrelated individuals with HLA haplotype A30 Cw5 B18 BfF1 DR3 D w52 DQw2 was digested with the restriction enzyme Taq I and hybridized with cDNA probes for C4 and 21-OH genes. Eighteen individuals and a non-Sardinian reference cell (DUCAF) were homozygous for the entire extended haplotype, six individuals were heterozygous for the HLA-A locus and homozygous for the rest of the same haplotype. All of the 18 homozygous individuals, four of the six heterozygous subjects and the DUCAF cell revealed the absence of two fragments at 3.2 and 2.4 kb corresponding to the 21-OH A gene, and the absence of fragments corresponding to the C4 B "long" or "short" gene. The contemporary absence of the 21-OH A and C4 B genes seems to be a constant characteristic of this extended haplotype in the Sardinian population and suggests that this and other extended haplotypes bearing C4 A or C4 B null alleles could be ancestral haplotypes which never duplicated at these loci.

Alleles↗

PubMed1990

HLA extended haplotypes in childhood and adult onset HLA-DR4-associated arthropathies.

Alpha HLA-DR4 extended haplotypes have been established by family studies in children and adults with chronic arthritis, the children having polyarticular juvenile rheumatoid arthritis and the adults rheumatoid arthritis. These diseases are probably the same disorder presenting at different ages. The results demonstrate age-related differences in the frequency of HLA-DR4 extended haplotypes between the two groups of patients. The childhood onset was associated with the presence of the HLA-B44, SC30, DR4 extended haplotype, whereas the adult form was more strongly associated with the HLA-Bw62, SC33, DR4 haplotype. These results suggest that differences in the age at onset are influenced by the MHC in addition to the presence of the HLA DR4 gene itself.

Adult↗

PubMed1992

A high frequency of the A30, B18, DR3, DRw52, DQw2 extended haplotype in Sardinian celiac disease patients: further evidence that disease susceptibility is conferred by DQ A10501, B10201.

This study characterizes by serological and molecular methods the HLA class I and class II alleles in a group of celiac disease children, their parents and a control group of Sardinian descent. We found the DR3-DQw2 haplotype in all patients which was, in almost all cases (84%), associated with the HLA-A30, B18, DR3, DRw52, DQw2 extended haplotype named "Sardinian haplotype" because of its frequency (12-15%) in this Caucasian population. This is the first time that this DQw2-linked haplotype has been reported with such a high frequency in CD. However, no different distribution of "Sardinian haplotype" was found comparing CD patients with 91 haplotyped DQw2-positive controls. This finding indicates that the DQw2 antigen in Sardinians is almost always associated with the A30, B18, DR3, DRw52, DQw2 extended haplotype. The DQA1 and DQB1 second exon sequence analysis of the B18,DR3 and B8,DR3 haplotypes showed the DQA1*0501 and DQB1*0201 alleles which shared the already published sequences. DPB1 subtyping showed the DPB1*0301 allele more frequently (p less than 0.005) in CD patients but this difference was no longer significant when patients and controls, both heterozygous for the DR3-DQw2 haplotype, were compared. We suggest that the divergent HLA extended haplotypes and DP allele associated with CD, described in different Caucasian populations, can be explained by the particular DQw2 linkage disequilibrium in each population.

Adult↗

PubMed1996

C4 polymorphism and extended HLA haplotypes in Namibian San and Khoi and in South African Xhosa.

We studied C4A and C4B polymorphisms and HLA-B and -DR associations in the San, Khoi and Xhosa. C4A and C4B alleles were determined using conventional protein allotyping methods. The C4A*3, C4B*1 haplotype had a high frequency (30-55%) in all populations. The frequency of C4A*3, C4B*Q0 was 7-19%. The C4A*Q0, C4B*1 haplotype was frequent (15%) in the Khoi but very rare in the San (P < 0.001). C4A*12 A*91, C4B*Q0 was frequent in the Xhosa (15%) but rare in the San and Khoi (P < 0.001). Alleles C4A*5 and C4A*6, and the C4B*2 B*92 duplication were only found in the Xhosa. C4A alleles A*4, A*45, A*58, A*12, A*14, A*19 and the C4A*3 A*91 duplication were only found in the San/Khoi population group. In the San, fourteen extended haplotypes were found in a relatively high frequency (2-7%). In the Xhosa, one extended haplotype (B42, C4A*12 A*91, C4B*Q0, DR18) was found in a very high frequency (13%) and was characteristic for this group; five other extended haplotypes were found with a low frequency (< 3%).

Complement C4a↗

PubMed2006

Transperineal extended biopsy improves the clinically significant prostate cancer detection rate: a comparative study of 6 and 12 biopsy cores.

BACKGROUND: We evaluated the improvement in the rate of prostate cancer detection when using a 12-core transperineal biopsy protocol including transitional zone biopsy. METHODS: Between April 2003 and November 2004, 247 consecutive men underwent transperineal systemic 12-core biopsy of the prostate. Six cores were obtained at the peripheral zone, four at the transitional zone and two at the apex. We examined the cancer detection rate in each of the 12 cores, and also determined the improvement of cancer detection resulting from the extensive 12-core versus standard 6-core biopsy. RESULTS: Using the extensive 12-core biopsy, prostate cancer was detected in 98 cases (39.7%). Prostate-specific antigen (PSA), PSA density, the positive rate in digital rectal examination and transrectal ultrasound findings were significantly higher in the prostate cancer group than in the non-prostate cancer group, and prostate volume was larger in non-prostate cancer group. Every site showed almost the same positive rate, between 17.8 and 21.5%. There were 20 cases which were positive in the extended biopsy, but negative in the sextant. The detection improved significantly (20.4%). The improvement of cancer detection in extended biopsy was better in men with PSA levels of 10 ng/mL or less (28.9%), PSA density 0.3 or less (25.8%), negative digital rectal examination (23.3%), and negative transrectal ultrasound (21.6%). Of these twenty patients, no cases with insignificant tumor were detected in the six prostatectomy cases. In particular, three cases of the six were transitional-zone-only cancer. CONCLUSION: Transperineal extended 12-core biopsy including 4 transitional zone cores is a more useful procedure than transperineal 6-core biopsy. Routine transitional zone biopsy, that is different from transrectal biopsy, might be useful for detecting biologically significant cancer.

Aged↗

PubMed1988

Detection of N-terminally extended substance P but not of substance P in human cerebrospinal fluid: quantitation with HPLC-radioimmunoassay.

A reversed-phase HPLC system was used to concentrate and separate components of substance P-like immunoreactivity (SP-LI) from human CSF. When CSF was injected and fractions collected, no SP-LI could be detected by radioimmunoassay (RIA) at the retention time of SP or SP-sulfoxide. Instead, SP-LI was detected in later eluting fractions. This SP-LI reacted with two different antisera raised against the C-terminal part of SP, but not with an antiserum against the N-terminal part. A compound with similar properties was also found to be present in neutral extracts of rat dorsal spinal cord. When the late-eluting compound from human CSF was treated with trypsin and rechromatographed on HPLC, an immunoreactive component eluting at the position of SP could be detected with both the C- and N-terminally directed SP antisera. These results suggest that an N-terminally extended form of SP is present in human CSF. Trypsinization also gave two other compounds with affinity for the N- but not the C-terminally directed antisera. This may indicate that N-terminal fragments of SP extended at the N-terminus or SP molecules extended at both the N- and the C-terminus (i.e., preprotachykinins) also are present in human CSF. In 32 CSF samples from depressed patients, SP-LI was determined with a C-terminally directed antiserum with and without prior HPLC separation. SP itself could not be detected, but the late-eluting form of SP-LI could be quantitated in all samples by combined HPLC-RIA. In most samples, there was a relatively good agreement between the SP-LI levels measured with and without HPLC.

Antibody Specificity↗

PubMed2004

The arachidonic acid 5-lipoxygenase inhibitor nordihydroguaiaretic acid inhibits tumor necrosis factor alpha activation of microglia and extends survival of G93A-SOD1 transgenic mice.

Familial forms of amyotrophic lateral sclerosis (ALS) can be caused by mutations in copper, zinc-superoxide dismutase (SOD1). Mice expressing SOD1 mutants demonstrate a robust neuroinflammatory reaction characterized, in part, by up-regulation of tumor necrosis factor alpha (TNFalpha) and its primary receptor TNF-RI. In an effort to identify small molecule inhibitors of neuroinflammation useful in treatment of ALS, a microglial culture system was established to identify TNFalpha antagonists. Walker EOC-20 microglia cells were stimulated with recombinant TNFalpha, with or without inhibitors, and the cell response was indexed by NO2- output. Three hundred and fifty-five rationally selected compounds were included in this bioassay. The arachidonic acid 5-lipoxygenase (5LOX) and tyrosine kinase inhibitor nordihydroguaiaretic acid (NDGA), a natural dicatechol, was one of the most potent non-cytotoxic antagonists tested (IC50 8 +/- 3 microm). Investigation of the G93A-SOD1 mouse model for ALS revealed increased message and protein levels of 5LOX at 120 days of age. Oral NDGA (2500 p.p.m.) significantly extended lifespan and slowed motor dysfunction in this mouse, when administration was begun relatively late in life (90 days). NDGA extended median total lifespan of G93A-SOD1 mice by 10%, and life expectancy following start of treatment was extended by 32%. Disease-associated gliosis and cleaved microtubule-associated tau protein, an indicator of axon damage, were likewise reduced by NDGA. Thus, TNFalpha antagonists and especially 5LOX inhibitors might offer new opportunities for treatment of ALS.

Administration, Oral↗

PubMed2004

Investigation of extended-spectrum beta-lactamases produced by clinical isolates of Klebsiella pneumoniae and Escherichia coli in Korea.

AIMS: Isolates obtained from various regions in Korea in 2002 were identified and their susceptibility to extended-spectrum cephalosporins, monobactams and/or cephamycins was studied along with any production of extended-spectrum beta-lactamases (ESBLs). METHODS AND RESULTS: Bacteria identified by the conventional techniques and Vitek GNI card were Klebsiella pneumoniae and Escherichia coli. Using disk diffusion and double-disk synergy tests, we found that 39.2% of strains produced ESBLs. About 52% of isolates transferred resistance to ceftazidime by conjugation. Banding patterns of PCR amplification with the designed primers showed that 837- and 259-bp fragments specific to bla(TEM) genes were amplified in 63.3% of strains. 929- and 231-bp fragments (bla(SHV)), 847- and 520-bp fragments (bla(CMY)), 597- and 858-bp fragments (bla(CTX-M)) were amplified in 61.5, 17.3 and 7.7% of strains respectively. About 51.9% of strains contained more than two types of beta-lactamase genes. Especially, one strain contained bla(TEM), bla(CMY) and bla(CTX-M) genes. SIGNIFICANCE: Resistance mechanisms to beta-lactams, comprising mostly ESBL production, lead to the resistance against even recently developed beta-lactams in enterobacteria, which is now a serious threat to antibiotic therapy. The high prevalence of bla(CMY) genes and multidrug-resistant genes may also make therapeutic failure and lack of eradiation of these strains by extended-spectrum cephalosporins or cephamycins.

Anti-Bacterial Agents↗

PubMed2006

Design and synthesis of compounds that extend yeast replicative lifespan.

This past decade has seen the identification of numerous conserved genes that extend lifespan in diverse species, yet the number of compounds that extend lifespan is relatively small. A class of compounds called STACs, which were identified as activators of Sir2/SIRT1 NAD+-dependent deacetylases, extend the lifespans of multiple species in a Sir2-dependent manner and can delay the onset of age-related diseases such as cancer, diabetes and neurodegeneration in model organisms. Plant-derived STACs such as fisetin and resveratrol have several liabilities, including poor stability and relatively low potency as SIRT1 activators. To develop improved STACs, stilbene derivatives with modifications at the 4' position of the B ring were synthesized using a Horner-Emmons-based synthetic route or by hydrolyzing deoxyrhapontin. Here, we describe synthetic STACs with lower toxicity toward human cells, and higher potency with respect to SIRT1 activation and lifespan extension in Saccharomyces cerevisiae. These studies show that it is possible to improve upon naturally occurring STACs based on a number of criteria including lifespan extension.

Cell Line↗

PubMed2006

The effect of extended-release metoprolol succinate on C-reactive protein levels in persons with hypertension.

The objective of this study was to determine whether 3 months of treatment with extended-release metoprolol succinate would reduce C-reactive protein (CRP) levels. Seventy-five patients aged 30-65 years with uncontrolled hypertension were treated with extended-release metoprolol at 25-50 mg, titrated up to 100-200 mg daily. CRP was evaluated at baseline and at 1 and 3 months. In the 61 hypertensive patients who completed the study, CRP decreased from 6.2+/-7.5 mg/L at baseline to 5.4+/-7.0 mg/L (p=0.03) at 1 month and showed no further change at 3 months (5.6+/-6.5 mg/L; p=0.13). The 13 patients who received 200 mg of extended-release metoprolol had a 32% decline in CRP from 7.0+/-9.0 mg/L to 4.8+/-6.6 mg/L (-2.2 mg/L) (p=0.005) over the 3-month period, whereas lower doses did not reduce CRP (p>0.05). Age, race, sex, and change in blood pressure were not related to the reduction in CRP in multivariate analysis. If CRP evolves into a confirmed modifiable risk factor, a beta blocker such as metoprolol may be a useful addition to pharmacotherapy options.

Adrenergic beta-Antagonists↗

PubMed2004

Treatment of full-thickness skin defect with concomitant grafting of 6-fold extended mesh auto-skin and allogeneic cultured dermal substitute.

The aim of this clinical study was to evaluate an allogeneic cultured dermal substitute (CDS) as a biological dressing for highly extended mesh auto-skin grafting. The subjects were five patients with extensive deep burn wounds. Allogeneic CDS was prepared by seeding fibroblasts on a spongy matrix of hyaluronic acid and atelo-collagen. Six-fold extended auto-skin graft was applied to the debrided wound, on which allogeneic CDS was placed. A conventional ointment-gauze dressing was used to protect the CDS. The CDS was applied repeatedly at intervals of 5-7 days. In all cases, the wounds were closed by successful take of mesh auto-skin graft and prompt epithelization from the grafted skin. The skin on the grafted area had a cicatrix appearance, but was soft and thin, maintaining good quality. The application of 6-fold extended auto-skin graft in conjunction with allogeneic CDS is an effective method for treatment of extensive severe burn wounds.

Adult↗

PubMed2006

Extended duration of thromboprophylaxis in acutely ill medical patients: optimizing therapy?

Summary. Patients who are hospitalized for an acute medical illness are at risk of venous thromboembolism (VTE). Current evidence-based guidelines recommend prophylaxis with unfractionated heparin or low-molecular-weight heparin in acutely ill medical patients who are admitted to hospital with congestive heart failure, severe respiratory disease, or who are bedridden with an additional VTE risk factor. The need for thromboprophylaxis is therefore clear in this patient population; however, the optimal duration of prophylaxis in these patients is less clear. In patients undergoing orthopedic or cancer surgery, extended-duration prophylaxis has been shown to be superior to placebo. To date, however, no large-scale clinical trials have assessed the benefits of extended-duration prophylaxis in acutely ill medical patients. This review therefore focuses on the VTE risk profile of acutely ill medical patients, examines the currently available literature for evidence of a potential benefit of extended-duration prophylaxis in these patients, and provides a rationale for the testing of such a hypothesis in a randomized clinical trial.

Acute Disease↗

PubMedDate not supplied

Extended and total simultaneous aortic replacement: latest technical modifications and improved results with thirty-four patients.

Aortic disease frequently requires extended and multiple resections. Occasionally, resection of the entire aorta may be indicated. At our Institution, from 1982 to 1994, 34 patients were operated upon for extended and total simultaneous aortic replacement. In seven patients, the aorta was replaced from valve to bifurcation; in 27, the aortic valve was included. Operations were performed with circulatory arrest under profound hypothermia. As the first step, the aortic valve and ascending aorta are replaced and the coronary arteries are reconnected, following which the aortic arch is reconstructed. Meanwhile, a second surgical team proceeds to open the thoracoabdominal aorta and tie up the intercostal orifices. If circulatory arrest is likely to exceed 60 minutes, the aortic graft is clamped and upper body perfusion (1000 cc/min) is begun. Finally, the thoracoabdominal aorta is fully replaced. Cardiopulmonary bypass (CPB) with rewarming is resumed only after the operation has been completed. Thirty-four patients survived operation; five died within 1 month for an overall mortality of 14.7%. No mortality occurred in the most recent nine operations. No permanent spinal neurological deficits occurred. Total simultaneous aortic replacement for treatment of extended aortic disease may be reasonable using our approach.

Aorta, Abdominal↗