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PubMed1990

Simultaneous determination of ampicillin and sulbactam by liquid chromatography: post-column reaction with sodium hydroxide and sodium hypochlorite using an active hollow-fibre membrane reactor.

A high-performance liquid chromatographic method has been developed for the simultaneous determination of ampicillin (ABPC) and sulbactam (SBT) in serum and urine. The method involves separation of ABPC and SBT from the background components of serum and urine on a C18 column, post-column reaction with sodium hydroxide and sodium hypochlorite using an active hollow-fibre membrane reactor, and detection at 270 nm. At ABPC and SBT concentrations of 10 and 5 micrograms/ml in urine and serum samples, the precisions (relative standard deviations) were 0.9-2.5% (n = 8). The detection limits were 20 and 5 ng for ABPC and SBT, respectively, at a signal-to-noise ratio of 3.

Ampicillin↗
PubMedDate not supplied

Small bowel extrinsic denervation does not alter water and electrolyte absorption from the colon in the fasting or early postprandial state.

Small bowel transplantation (SBT) causes watery diarrhea. The decreases shown previously in absorption of water, electrolytes, and bile salts in the jejunum and ileum, although present, are not dramatic and seem not to be great enough to explain the diarrhea. Our aim was to determine changes in water and electrolyte absorption in the colon during fasting and postprandially in a canine preparation of jejunoileal extrinsic denervation, which serves as a model of jejunoileal autotransplantation. We hypothesized that colonic absorption of water and electrolytes decreases transiently in the colon after SBT. Six dogs had cannulas implanted in the colon to study absorption of an ileal-like basal electrolyte solution with or without 10 mmol/L glucose. Absorption during fasting and postprandially was measured before and 2 and 12 weeks after a validated preparation of jejunoileal extrinsic denervation. All dogs developed diarrhea after SBT. Net colonic absorptive fluxes of water and electrolytes in the colon did not change from baseline values at 2 or 12 weeks after extrinsic denervation, either during fasting or postprandially; glucose in the infusate did not alter absorptive fluxes during fasting or postprandially. Extrinsic denervation of the small intestine does not appear to alter colonic absorption of water or electrolytes during fasting or postprandially. These observations suggest that the neurally intact colon has a minimal role in the diarrhea after SBT.

Animals↗
PubMed2003

Establishment of a sequence-based typing system for BoLA-DRB3 exon 2.

A rapid, high-resolution sequence-based typing (SBT) system for BoLA-DRB3 exon 2 was developed. Amplification of the entire exon was achieved by a fully nested PCR with locus-specific primers and sequencing was performed directly on the PCR product. Heterozygous sequence data were obtained by automated sequence analysis of both alleles. Forward and reverse sequence data were assembled to improve identification of all heterozygous positions. Specific software (Haplofinder, Roslin Institute Software, Roslin, UK) was designed for allele assignment. Fifty-four females from a Holstein-Charolais resource herd cross, their 12 sires and five unrelated Holstein animals were used to establish the method. In parallel, these animals were typed by DRB3 polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) to confirm the results. Polymerase chain reaction-RFLP analysis defined 15 known types in the 71 animals, while SBT of the same animals showed 19 known alleles. Subsequently, 72 more animals from the same resource herd were typed by the established SBT method without PCR-RFLP typing. This SBT strategy and the Haplofinder software can be applied to the analysis of any polymorphic locus for which suitable locus-specific primers and allelic sequences are available.

Alleles↗
PubMed1999

Benefits of recruiting participants with friends and increasing social support for weight loss and maintenance.

To determine the benefits of social support for weight loss and maintenance, this study recruited participants (N = 166) either alone or with 3 friends or family members and then randomly assigned them to a standard behavioral treatment (SBT) or SBT with social support strategies. Participants recruited with friends had greater weight losses at the end of the 4-month treatment and at Month 10 follow-up. Both recruitment strategy and the social support manipulation affected treatment completion and weight-loss maintenance. In those recruited alone and given SBT, 76% completed treatment and 24% maintained their weight loss in full from Months 4 to 10. Among those recruited with friends and given SBT plus social support, 95% completed treatment and 66% maintained their weight loss in full.

Adult↗
PubMed1998

Diagnosis of pulmonary embolism by measurement of alveolar dead space.

OBJECTIVE: Pulmonary embolism (PE) gives rise to alveolar dead space, which can be measured with a single breath test for CO2 (SBT-CO2). The characteristics of the SBT-CO2 are different in PE and other common conditions giving rise to alveolar dead space, notably airways disease. An analysis of alveolar dead space focusing on the late part of the breath (fDlate) has been suggested as a method for diagnosis of PE. Our aim was to evaluate this technique by comparison with lung scintigraphy. METHODS: We randomly selected patients with clinical suspicion of PE. SBT-CO2 and lung scintigraphy were performed on the same day. The scintigraphies were reviewed and classified as high, intermediate and low probability of PE. RESULTS: Out of 223 patients able to be evaluated, there were 20 of the high, 29 of the intermediate and 174 of the low probability category. There were large differences between the means of fDlate in the high and the intermediate and in the high and the low categories. We obtained a sensitivity of 85% and a specificity of 93% for diagnosis of PE, based on high and low probability categories. If a patient with previous PE, but no scintigraphic evidence of current PE, is excluded the sensitivity increases to 90%. CONCLUSIONS: This study provides further support for the measurement of fDlate by the SBT-CO2 as a diagnostic test in patients with suspicion of PE. The test should be especially useful in small hospitals without access to pulmonary scintigraphy or pulmonary angiography.

Adult↗
PubMed2003

Functional dyspepsia, upper gastrointestinal symptoms, and transit in children.

OBJECTIVE: To assess the prevalence of abnormal gastric emptying and small bowel transit in children with functional dyspepsia at a tertiary care center, and the relationship between abnormal gastric and small bowel transit and symptoms in pediatric patients with functional gastrointestinal disorders. STUDY DESIGN: Patients were selected by a cross-sectional chart review based on the following inclusion criteria: (1) completion of scintigraphic study of the gastric emptying of solids at 2 hours (GE2), 4 hours (GE4), and small bowel transit at 6 hours (SBT) using a standardized egg meal labeled with 99mTechnetium sulfur colloid, and (2) gastrointestinal (GI) complaints without mucosal or organic disease. Logistic regression analysis was used to assess the association between the presence of upper GI symptoms, and each parameter of gastric and small bowel transit. RESULTS: Children with upper GI symptoms (n=96) were identified. Among 57 children with functional dyspepsia, 40% had slow SBT. Fast GE at 4 hours, and slow SBT were independently associated with bloating. Children with fast SBT were less likely to report abdominal pain. CONCLUSION: Incorporating assessments of gastric and small bowel transit may be useful in the evaluation of pediatric patients with upper GI symptoms and functional dyspepsia.

Abdominal Pain↗
PubMed2003

Expression of mucosal addressin cell adhesion molecule-1 (MAdCAM-1) during small-bowel graft rejection in rats.

BACKGROUND: Mucosal addressin cell adhesion molecule-1 (MAdCAM-1) is a critical endothelial adhesion molecule for lymphocyte trafficking to gut-associated lymphoid tissues (GALT) under physiological conditions. It is expressed on special postcapillary venules, that is the high endothelial venules (HEV) in GALT. In this study, we investigated changes in MAdCAM-1 expression during small-bowel graft rejection. METHODS: Orthotopic small-bowel transplantation (SBT) with portocaval drainage was performed from brown Norway (BN) rats to Lewis (LEW) rats. Isografted (LEW --> LEW) and untransplanted animals served as controls. Animals were killed on days 3, 4, 5, 6 and 7 after SBT. Cryostat sections (6 microm thick) were prepared from normal small-bowel tissues and small-bowel grafts, including Peyer patches (PPs). Indirect immunoperoxidase staining was performed using monoclonal antibodies (mAbs) against rat MAdCAM-1 (OST12). RESULTS: In the PPs of controls, MAdCAM-1 antibodies specifically stained the endothelial cells of HEV, which were predominantly located in the interfollicular areas. In the allografts on day 4 after SBT, the MAdCAM-1 expression was weaker on the HEV in the PPs than in the controls. In the lamina propria of controls, a faint expression of MAdCAM-1 on vessels was observed. On day 4 after allogeneic SBT, the MAdCAM-1 was more strongly expressed on the endothelial cells of the vessels at the base of the villi. As rejection developed, the MAdCAM-1 expression on the vessels progressed toward the villus tip. CONCLUSIONS: The change of MAdCAM-1 expression may be involved in the development of small-bowel graft rejection. The vessels at the base of villi, which is associated with lymphocyte recruitment, may become a site of intense immune reactivity in the early phase of small-bowel allograft rejection.

Animals↗
PubMed1997

External quality assessment of the serum bactericidal test: results of a methodology/interpretation questionnaire.

Two hundred microbiology laboratories in the UK took part in two separate experimental external quality assessment distributions related to the serum bactericidal test (SBT). In the first, Staphylococcus aureus NCTC 6571 (vancomycin MIC 1 mg/L), was tested against a human serum containing vancomycin 38 mg/L plus gentamicin 0.5 mg/L. In the second, Streptococcus oralis PAJ 112/4183 (penicillin MBC < or = 0.03 mg/L) and Streptococcus sanguis PAJ 107/4184 (penicillin MBC = 128 mg/L) were tested against human serum containing penicillin 15 mg/L. Respondents returned their laboratory results and a questionnaire on clinical interpretation and technical aspects. Most laboratories (194/199, 97.5%) recommend the use of the SBT in the management of infective endocarditis but only 48 (25.2%) often or always change therapy on the basis of the result. A wide range of interpretative criteria, definitions of bactericidal endpoints and methodologies are used. Performance in the first distribution was acceptable for 75% of laboratories but in the second only 34% could identify penicillin tolerance; 34 respondents reported an SBT result of < or = 2 for the tolerant strain, 81 laboratories reported one of > or = 16. Technical factors related to acceptable performance were: sonication of broth before counting the inoculum; knowing the inoculum size in cfu/mL; use of a 4-8 h broth culture to make the inoculum; incubation of recovery plates for > 36 h; use of a calibrated pipette to sample for surviving bacteria; use of measured volumes to add the inoculum. Use of uncalibrated pipettes or standard loops to recover survivors was related to poor performance. Microbiology departments in the UK should review the clinical need to perform the SBT in the light of their local circumstances and if they elect to continue to offer this test, revise their methodologies which could be producing misleading results when testing alpha-haemolytic streptococci.

Bacteriological Techniques↗
PubMedDate not supplied

Evidence that age-associated memory impairment is not a normal variant of aging.

The concept of age-associated memory impairment (AAMI) suggests that clinically recognized memory dysfunction can be a feature of normal aging. To determine whether AAMI represents a variant of normal aging, we longitudinally studied individuals meeting AAMI criteria for development of dementia. Two hundred two community-living individuals (mean age, 77 years) with or without mild memory impairment were assessed annually for an average of 3 years at the Washington University Alzheimer's Disease Research Center. At baseline, no individual was unequivocally demented, as defined by a Clinical Dementia Rating (CDR) score of 1 or greater. Modified National Institute of Mental Health criteria were used to identify individuals with AAMI who otherwise met a criterion for cognitive normality. The Short Blessed Test (SBT) was used as a measure of general cognitive function; conservative (SBT = 5) and permissive (SBT = 10) cutoff scores were used as indicators of cognitive normality. With the more permissive measure of cognitive normality, 59 (29%) of the 202 individuals met AAMI criteria. Progression to dementia by 3 years occurred in 42% of AAMI individuals versus 16% of the individuals who did not meet AAMI criteria. With the more restrictive SBT cutoff of 5, 22% of individuals met AAMI criteria; progression to dementia occurred in 31% of these individuals versus 9% of the individuals without AAMI. Survival times to dementia differed significantly between AAMI and non-AAMI groups defined by either cutoff score. Our findings indicate that individuals with AAMI have a three-fold greater risk for development of dementia than individuals who do not meet AAMI criteria. Hence, AAMI may represent a dementia prodrome rather than a benign variant of aging.

Aged↗
PubMed1997

Increased expression of tissue cytokines in graft-versus-host disease after small bowel transplantation in the rat.

BACKGROUND: Graft-versus-host disease (GVHD) occurs in the recipient after small bowel transplantation (SBT). Proinflammatory cytokines, such as tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma), and interleukin 6 (IL-6), may be important mediators of GVHD. Increased expression of these cytokines might precede the clinical manifestations of GVHD induced by SBT. METHODS: Heterotopic SBT was performed using Lewis donors into Lewis x Brown Norway F1 (LBN-F1) recipients. The isograft control was performed from LBN-F1 into LBN-F1. Animals were killed on the 5th and 11th postoperative day (POD). mRNA was isolated from recipient native small bowel, colon, spleen, liver, and mesenteric lymph nodes and from nonsurgical controls as baseline. Semiquantitative reverse transcriptase polymerase chain reaction was performed to amplify mRNA transcripts for TNF-alpha, IFN-gamma, and IL-6 using alpha32P-dATP incorporation. Clinical signs, histologic assessment, and cytokine expression were correlated. RESULTS: On POD 5, there were neither clinical signs nor histologic features of GVHD, but mRNA expression of TNF-alpha and IL-6 in small bowel, IL-6 in spleen, and IFN-gamma in mesenteric lymph nodes were significantly increased in allograft animals when compared with normal and isograft tissues. On POD 11, both the clinical signs and histologic features of GVHD were seen, and TNF-alpha and IL-6 in native small bowel, TNF-alpha in colon, IFN-gamma in spleen, and IL-6 in mesenteric lymph nodes were significantly increased in allograft animals when compared with that in normal and isograft tissues. CONCLUSIONS: In conclusion, TNF-alpha, IFN-gamma, and IL-6 expression precede clinical onset and histologic evidence of GVHD in specific tissues. Therefore, increased expression of these cytokines is correlated with the development of GVHD in this model of SBT.

Animals↗
PubMed2001

Changes in the composition of serum bile acid as a predictor of small bowel allograft rejection in rats.

Composition of bile acid was studied as a noninvasive rejection marker after small bowel transplantation (SBT). Orthotopic SBT were performed in rats, and they were divided into four groups: group 1, sham-operated rats; group 2, recipients with isografts; group 3, recipients with allografts treated with FK506; and group 4, recipients with untreated allografts. On postoperative days (POD) 5 and 7, the graft histology, intraluminal bacterial overgrowth, individual bile acids concentration of the recipient serum and bile were examined. On POD 5, early histological findings of acute rejection were observed in group 4, and the ratio of secondary bile acids of this group were significantly higher than the other groups. The bile acid changes were enhanced by bacterial overgrowth on POD 7. The ratio of secondary bile acids changed in relation to acute rejection after SBT, suggesting that they can be useful for early diagnosis of acute SBT allograft rejection.

Animals↗
PubMed2004

Early recurrence of ovarian serous borderline tumor as high-grade carcinoma: a report of two cases.

Ovarian serous borderline tumors (SBTs) are characteristically associated with an indolent course. Recurrences are often delayed and usually show morphologic features of SBT or low-grade serous carcinoma. Transformation to high-grade carcinoma has rarely been documented. We report 2 cases of ovarian SBTs that recurred early as high-grade carcinomas. The first was a 50-year-old woman treated surgically and with chemotherapy for a FIGO stage 1C SBT with microinvasion, who experienced a recurrence in an axillary lymph node at 27 months. The recurrent tumor consisted of well-differentiated papillary serous tumor that resembled the primary tumor and poorly differentiated serous carcinoma. The patient died of progressive disease 43 months after her initial presentation. The second case was a 61-year-old woman treated surgically and with chemotherapy for a stage 3C micropapillary SBT with noninvasive implants. Eighteen months later, an incisional hernia was found that contained high-grade sarcomatoid-type carcinoma with microscopic foci of better differentiated tumor that resembled the primary SBT. This patient is alive with disease 24 months after her initial presentation. Whereas the malignant potential of SBTs remains controversial, the cases described herein demonstrate that SBTs can behave unpredictably and may rarely transform into high-grade carcinoma.

Cystadenocarcinoma, Serous↗
PubMed2004

Pseudoneglect in solid-line versus character-line bisection tasks: a test for attention dominance theory.

OBJECTIVE: Normal subjects tend to bisect lines slightly to the left of the true midpoint, a phenomenon termed pseudoneglect. To test whether pseudoneglect relates to the right hemisphere's dominance for spatial attention or to the hemispheric difference in processing global-local stimulus properties, we administered conventional solid-line (SBT) and novel character-line (CBT) bisection tasks to normal subjects of different ages. METHODS: Normal subjects, consisting of 40 young and 40 older individuals, received 3 experimental tasks, a standard SBT and 2 types of CBT. Each subject completed 10 consecutive trials of each task presented in counterbalanced order between subjects. RESULTS: Across age groups, deviations on CBT were further to the right than those of SBT, and the leftward bias (pseudoneglect) was significant only in SBT. CONCLUSION: These results indicate that the bisection errors in normal subjects depend on the characteristics demanded by the specific task. Thus, our findings argue against the attention dominance theory and support a "task specificity" theory for pseudoneglect.

Adult↗
PubMed1995

An evaluation of a multicenter study on HLA-DPB1 typing using solid-phase Taq-cycle sequencing chemistry.

In HLA Class II genes, polymorphism is mainly located in the second exon. Most DNA based typing methods are confined to the identification of specific sequence motifs in the second exon. In contrast, Sequencing Based Typing (SBT) elucidates the entire exon 2 sequence for typing. Comparison of the obtained exon 2 sequence with an allele sequence library results in allele assignment. We tested the applicability of SBT using a protocol for amplification followed by solid phase Taq-cycle sequencing for HLA-DPB1 typing. A panel of 32 samples were typed by SBT at five test sites which are participating in the Sequencing Based Typing component of the 12th International Histocompatibility Workshop. The panel represents the existing polymorphism at all known polymorphic positions of exon 2, both in homozygous and heterozygous combinations. In this multicenter study we focused on the reliability of analyzing heterozygous sequences for HLA typing. A multi-sequence analysis approach, Polall, was developed to evaluate sequences obtained. The assignment of homozygosity and heterozygosity was validated by cluster analysis of chromatographic data of all sequences. Sequence characteristics were examined and considered for appropriate assignment. Differences in sequence characteristics that occurred between the test sites are considered in detail. The evaluation of data of 5 test sites reveals that Taq-cycle sequencing can reliably be performed for HLA-DPB1 SBT.

DNA-Directed DNA Polymerase↗
PubMed1997

HLA-A typing: comparison between serology, the amplification refractory mutation system with polymerase chain reaction and sequencing.

In this study we typed HLA-A polymorphisms by a new sequence-based typing (SBT) method, which involved one PCR reaction and four sequencing reactions covering exon 2 and exon 3. This method allowed complete identification of all known HLA-A alleles and revealed the presence of a new allele, named HLA-A*2608. We also introduced sequencing of exon 4 for some samples in order to discriminate the allelic pairs that are identical in exon 2 and 3, thus improving SBT resolution. Finally, we compared the results obtained by SBT with data obtained by serological typing and the amplification refractory mutation system (ARMS-PCR). Together, our results suggest that the SBT here described provides an optimal HLA-A typing technique that may be useful in selecting donor-recipient pairs in bone marrow transplantation between unrelated individuals.

Adult↗
PubMed1998

HLA-A towards a high-resolution DNA typing.

Sequencing-based typing (SBT) and sequence-specific oligonucleotide probing (PCR-SSOP) are DNA-based typing approaches to identify HLA-A alleles. In this study PCR-SSOP SBT have been evaluated and considered to reach a high-resolution typing. Based upon serological typing, 32 genomic samples were typed by SBT and PCR-SSOP Three main clusters of resolution could be defined. The advantage of the PCR-SSOP approach is the possibility to type numerous samples in a short time. SBT minimizes the number of ambiguous heterozygous combinations and often allows direct detection and identification of new alleles.

DNA↗
PubMed1998

A reliable and efficient high resolution typing method for HLA-C using sequence-based typing.

Serological typing of HLA-C has been poor and almost half of its alleles are serologically undetectable blanks in most populations. Therefore, DNA typing techniques have been used to identify and type for the HLA-C gene. Sequence-based typing (SBT) has proven a major typing strategy for highly polymorphic HLA genes. The technique enables direct identification of all sequence motifs without the need to continuously adjust primers. Here we describe a reliable solid-phase SBT strategy for HLA-C which can be used to distinguish all currently known HLA-C alleles without prior knowledge gained by low resolution typing. Exons 2 and 3 were amplified and sequenced and if necessary sequences of exons 1 and 5 were determined. A total of 257 individuals were typed for HLA-C using this protocol and 30 of the 42 known HLA-C alleles were detected. All heterozygous combinations found in this study were unambiguously discriminated. One hundred and forty-four individuals from the Dutch population were typed randomly. In this group Cw*0701 and *0702 were the most frequently detected alleles. Of the serological Cw blank alleles Cw*1203 was found to have the highest frequency (16%). From the total group 212 individuals were typed serologically and 106 were retyped with 97 selected antisera to further compare serological and molecular defined phenotypes. Discrepancies between serological typing and SBT are mainly attributable to the serologically Cw blank alleles Cw*12-18. The high resolution SBT protocol described will be a valuable tool for the identification of HLA-C alleles and the determination of the role of HLA-C in marrow and organ transplantation.

Alleles↗
PubMed2006

Impact of surveillance stool culture guided selection of antibiotics in the management of pediatric small bowel transplant recipients.

Surveillance stool cultures (SSC) have been used in immunocompromised populations to predict the organisms associated with invasive infections and aid in the selection of empiric antibiotic regimens. To evaluate the utility of this approach in pediatric small bowel transplant (SBT) recipients, we conducted a retrospective review of 33 patients who underwent SBT, 16 of whom had SSC done. In no case was the same organism isolated from SSC and subsequent blood, peritoneal fluid or wound cultures. In the first month post-transplantation, blood cultures were positive in 44% and 35% of patients that had and did not have SSC done, respectively (p = 0.73); peritoneal fluid cultures in 44% and 65% (p = 0.30); and wound cultures in 44% and 24% (p = 0.28). There were no significant differences among both groups in time to first infection, duration of ICU stay following SBT, graft survival or long-term patient survival. We conclude that SSC-guided antibiotic selection does not have a significant impact on the incidence of invasive infections in the first month following SBT or on specific indicators of patient outcome. This suggests that empiric antibiotic regimens should be selected based on clinical presentation and hospital flora and susceptibility patterns.

Anti-Bacterial Agents↗