Heterogeneity among septic shock patients in a set of immunoregulatory markers.

Immune activation is a regular feature of sepsis, but the incidence and nature of the ensuing inflammation-resolving and immunosuppressive component is less well understood. In this study, we compared immunoregulatory markers on blood leukocytes from patients with Gram-negative or Gram-positive sepsis or septic shock, and compared this to blood from patients with severe virosis or healthy controls. To this end, blood from 32 patients with sepsis, including ten cases with shock, and 12 patients with severe virosis were analysed by flow cytometry for the expression levels of monocyte HLA-DR, CD11c, CD14 and CD40, and for frequencies of CD163(+)-suppressive monocytes, HLA-DR(+) or CD40(+)-activated T cells and Tregs. Plasma cytokine levels were analysed as a functional measurement. Signs of immunosuppression dominated in the septic shock and Gram-positive sepsis groups, whereas monocyte activation was common in Gram-negative sepsis patients without shock. However, the main finding was the large inter-individual variation of immune activation and immunosuppression, with no correlation to prognosis among the shock patients. The pronounced inter-individual variation in the analysed monocyte and lymphocyte markers forms a strong argument that, when immunomodulatory treatment is considered in a sepsis patient, it should be personalised and guided by a detailed immune status assessment.

CT angiography followed by endovascular intervention for acute superior mesenteric artery occlusion does not increase risk of contrast-induced renal failure.

OBJECTIVES: Acute superior mesenteric artery (SMA) occlusion can be diagnosed in an early phase by computed tomography (CT) angiography, which is also a prerequisite for endovascular intervention. However, the issue of development of postoperative permanent renal failure due to contrast-induced nephropathy has not been evaluated. DESIGN: Retrospective MATERIALS: A total of 55 patients with acute SMA occlusion were retrieved from the in-hospital register during a 4-year period between 2005 and 2009. METHODS: Glomerular filtration rate was calculated as a simplified variant of Modification of Diet in Renal Disease Study Group (MDRD). RESULTS: Preoperative renal insufficiency was found in 52%; advanced state in one patient. Creatinine was lower (p = 0.018) at discharge (median: 71 micromol L(-1)), compared to admission (median: 76 micromol L(-1)), in the 32 survivors exposed to repeated iodinated contrast media (median: 54.7 g iodine). No patient died due to renal failure or needed dialysis after endovascular intervention. Endovascular intervention was associated with a higher survival rate (p = 0.001). CONCLUSION: Serious acute contrast-induced nephropathy was not found in patients diagnosed by CT angiography and treated by endovascular procedures for acute SMA occlusion. Elevated serum creatinine levels should not deter the clinician from ordering a CT angiography in patients with suspicion of acute SMA occlusion.

Leukocyte phenotype changes induced by specific immunotherapy in patients with birch allergy.

BACKGROUND: The underlying mechanisms of allergen-specific immunotherapy (SIT) are not fully understood. OBJECTIVES: The present study aimed to investigate how leukocyte phenotypes are affected by SIT. METHODS: Blood samples were taken from 10 patients with birch pollen--induced allergic rhinitis before, during, and immediately after SIT. Further samples were obtained after 1 year and 3 years. All samples were analyzed by flow cytometry and leukocyte differentiation. RESULTS: SIT caused a decrease in cell-bound immunoglobulin (Ig) E on granulocytes, along with a corresponding increase in the high-affinity IgG receptor. Accordingly, a lower level of allergen-specific IgE was found after 3 years. The treatment induced a decrease in neutrophil CD1 1b levels, a shift in monocyte subsets, and an increase in the number of activated T lymphocytes, manifested as an upregulation of CD69 and CD98, and an expansion of the CD4+CD25+ T-cell pool. CONCLUSION: The present study shows that the clinical effects of SIT are mirrored by systemic changes in cellular events and in antibodies, and offers new targets for immunomodulation.

Fast and simple screening for nutritional status in hospitalized, elderly people.

BACKGROUND: Undernutrition has been frequently reported among hospitalized elderly patients. The aim of this study was to evaluate the mini nutrition assessment (MNA) and a screening sheet for malnutrition (SSM) by full nutritional assessment (FNA) in elderly people, and to construct a shorter screening method by combining important questions from MNA and SSM. Having a screening tool as fast and simple as possible could increase its use in clinical routines. METHODS: FNA, MNA and SSM were carried out on 60 hospitalized patients (>65 years). Sensitivity and specificity for MNA and SSM were calculated in comparison with FNA. In order to construct a short and simple screening tool, questions from the two screening tools, which differed significantly between mal- and well-nourished patients, were used in a multivariate, stepwise linear regression. The regression model was simplified to be suitable in clinical routines. RESULTS: Malnourishment was diagnosed by FNA in 58.3% of the elderly patients, with no gender difference. Body mass index, unintended weight loss, recent surgery and loss of appetite were predictors of malnutrition in the regression model (R(2) = 60.1%). The sensitivity and specificity of the simplified regression model were 89 and 88%, respectively, which was more precise than MNA (77 and 36%) and SSM (89 and 60%). CONCLUSION: According to FNA, malnutrition is frequent in elderly hospitalized patients. Four questions are sufficient to conduct precise nutritional screening for malnutrition in elderly hospitalized patients. This new screening tool should be verified in other samples.

A costed profile audit of the surgical management of female stress incontinence.

BACKGROUND AND AIMS: To conduct a profile audit of three surgical treatments for urinary stress incontinence through the application of an episode costing process. METHODS: Four stage methodology: (1) construction or a profile of care for each of the surgical approaches (setting the standard); 2) calculation of a theoretical profile cost; (3) calculation of the observed costs from real patient episodes; (4) comparison of observed costs with profile costs, (comparing present practice with established standard). RESULTS: Profiles of care were constructed and compared with 39 actual in-patient episodes. Tension free vaginal tape (TVT Gynecare) is the cheapest modality of treatment in terms of both the expected profile cost and also observed (real patient) cost. Colposuspension is the most expensive form of treatment with real costs significantly greater than the expected profile. Clinical issues such as length of stay, duration of surgery, patient selection and complication rates were revealed through the exception reporting process. Length of stay is the main determinant of overall cost. CONCLUSION: It is possible to construct a costed and auditable standard of care for a surgical procedure. This standard can be compared with real patient costs calculated using the same methodology. Exception reporting based on differences between expected and real costs can be used to facilitate the audit of clinical practice. The technique is limited, however, by the need to collect accurate and detailed activity data.

Variations in the investigation and management of group B streptococcus in Scotland.

This article describes the practice of Scottish obstetricians in terms of their investigation and treatment of group B streptococcus (GBS). This was a postal questionnaire survey of all 125 consultant obstetricians in Scotland. We recorded indications for testing for GBS, categories of women to whom treatment is given empirically and following confirmed infection, type of antibiotic used, and timing and route of administration. No respondents screened all pregnant women but 97% screened some or all of those at highest risk. Three-quarters administered antibiotics empirically to women with intrapartum pyrexia. However, other high-risk groups were unlikely to receive treatment without confirmation of colonisation. Only one-third of respondents gave antibiotics to all women with confirmed GBS, and up to one-half withheld them from some colonised women in high-risk groups. Contrary to US guidelines, only 29% tested for GBS using low vaginal swabs and only 13% administered intrapartum antibiotics intravenously. There are wide variations in investigating and treating GBS throughout Scotland. It is likely that similar variations exist throughout the United Kingdom. UK guidelines are required to reduce variations and ensure appropriate and effective management.

Binding and detection of glycosaminoglycans immobilized on membranes treated with cationic detergents.

Immobilization of molecules on surfaces is used for preparative, quantitative, and qualitative studies. Glycosaminoglycans (GAGs) are strongly hydrophilic and negatively charged molecules that do not bind well to either polystyrene surfaces or hydrophobic blotting membranes. Hydrophobic membranes were derivatized with cationic detergents to become hydrophilic and positively charged. The ability of the polyvinylidene fluoride and nitrocellulose membranes to retain GAGs increased up to 12.8 microg per spot in the dot blot assay when the membrane was treated with a cationic detergent. Immobilized GAGs were stained with alcian blue, and the staining intensity was quantitated by scanning and densitometry. The derivatized membranes were used for solid-phase extraction of GAGs in blood plasma, urine, or cerebrospinal fluid. The detection sensitivity was equal for different types of GAGs but there was a slight negative interference from fibrinogen in blood plasma. The immobilized GAGs could also be released from the membrane using a nonionic detergent at high ionic strength. Recovery of different proteoglycan populations, separated by electrophoresis and detected by reversible staining with toluidine blue, was 70-100%.

Inflammatory bowel disease in Iceland, 1990-1994: a prospective, nationwide, epidemiological study.

BACKGROUND: Two separate nationwide, retrospective epidemiological studies of inflammatory bowel disease (IBD) in Iceland over a combined 40-year period, 1950-1989, have shown a continually rising incidence. The main objective of this study was to investigate the incidence of IBD prospectively for the 5-year period 1990-1994 to determine whether there still was an ongoing increase. METHODS: The retrieval of new cases of ulcerative colitis (UC) and Crohn's disease (CD) was based on a monthly review of all small and large intestinal tissue specimens with any type of inflammation submitted to all three departments of pathology in Iceland. All small intestinal X-ray records suggestive of CD were also reviewed. All possible new cases of IBD were then scrutinized by examination of the clinical records, using accepted criteria for confirmation or exclusion of IBD. RESULTS: A total of 215 cases of UC and 72 cases of CD were diagnosed, yielding a mean annual incidence for UC 16.5/100000 (95% confidence interval (CI) 14.4-18.9), and for CD 5.5/100000 (95% CI 4.3-7.0). The highest combined age related incidence for UC was 28.7/100000 in the group aged 30-39 years, and for CD 8.5/100000 in the groups aged 10-19 and 20-29 years. The most common extent in UC was proctosigmoid and the most frequent localization in CD was colonic only. CONCLUSIONS: This prospective study demonstrates a continuing and statistically significant increase in the incidence of both UC and CD in Iceland. The general changes identified for a few previously suggested environmental factors do not allow any firm conclusions as to their role in the observed increase in incidence. It is possible that there are some other causative factor(s).

Detection of epitopes in glycosaminoglycans immobilized on hydrophobic membranes.

Glycosaminoglycans (GAGs) are linear carbohydrate polymers expressed on all cell surfaces, and bind growth factors that recognize specific disaccharide sequences. Such sequences in the GAG chain are not genetically determined but may be assembled by the cell in response to environmental changes. GAGs are strongly hydrophilic and negatively charged molecules that do not bind well to either polystyrene surfaces or to hydrophobic blotting membranes. Cationic detergents were used to derivatize hydrophobic membranes to become hydrophilic and positively charged. Binding of GAGs to derivatized membranes was optimized regarding pH and ionic strength. Five different monoclonal antibodies (Mab) were used to detect sequence epitopes in immobilized GAGs. Parallel samples were stained with Alcian Blue and the staining intensities were quantitated by scanning and densitometry. By calculating the ratio between the antibody staining (epitope) and Alcian Blue staining (mass), the epitope density, i.e. the number of repetitive epitopes per mass, is obtained. The epitope density with each antibody was different with different GAGs. Some epitopes were common in GAG, i.e. highly repetitive epitopes. Some epitopes were rare and possibly expressed only once per GAG molecule, i.e. low degree of repetition. An epitope density profile was obtained when each sample was stained with all antibodies and their epitope densities calculated at the plateau level. The epitope profile is an indirect measure of the sequence variability in GAGs. Determination of epitope density and profile can be used to characterize a GAG population and to discriminate between different populations with similar chemical composition.

Prospective study of external cephalic version in Glasgow: patient selection, outcome and factors associated with outcome.

Data were collected prospectively on all 67 women who underwent an attempt at external cephalic version (ECV) over 1 year in the four Glasgow maternity hospitals. Ultrasonography was used in all women. However, tocolytics were used in only two (6%) nulliparous women despite published evidence of their efficacy. Only 25 (37%) women undergoing ECV had a free presenting part which is known to be associated with success. Seventeen (25%) women were less than 37 weeks pregnant despite spontaneous version being common at this stage. ECV was successful in only 26 (39%) women and only 18 (27%) had a vaginal cephalic delivery. These results compare unfavourably with published results of around two-thirds for both end-points. Although publication bias is likely, patient selection, under-usage of tocolytics and lack of experience may also be factors. Consideration should be given to a reduced number of operators who can maximise their throughput and expertise.

[Bronchoscopic diagnosis of lung malignancies.].

OBJECTIVE: To assess the diagnostic yield of fiberoptic bronchoscopy in patients with a variety of radiographic findings suspected of malignancy. MATERIAL AND METHODS: The study group was composed of all patients who underwent bronchoscopy in our hospital over a seven year period (1986-1993) where cytologic samples were obtained. Schematic drawings of endobronchial and radiographic findings done at the time of bronchoscopy were reviewed and the results of cytologic and histologic samples were compared for sensitivity and diagnostic accuracy. RESULTS: Bronchoscopy and cytologic sampling was performed on 189 patients. Tissue samples were obtained in 109 patients from visible endobronchial abnormalities or peripheral lesions using the transbronchial approach with fluoroscopic guidance. A total of 64 malignancies were diagnosed, 43 by bronchoscopy (67%). Among 58 patients who had primary malignancy of the lung, adenocarcinoma was the most frequent histologic type (50%) followed by squamous cell carcinoma (22%) and small cell carcinoma (17%). The diagnostic sensitivity of cytology was 30% whereas that of tissue biopsy was 70% among those patients where both tests were obtained. The two methods complemented each other to give a joint sensitivity of 76%. Among 13 patients with malignancy and normal endobronchial appearance, transbronchial biopsy was performed in 10 of which seven were diagnostic. Patients with primary malignancy who underwent surgery had a five year survival of 37% while the overall five year survival was 15.5%. CONCLUSIONS: In this study histology was much more sensitive than cytology as a sampling technique during bronchoscopy (p< 0.01). Adenocarcinoma has become the most frequent histologic type of primary lung malignancy in Iceland and overall survival among patients with primary tumors compares with recent international trends.

Variations in the management of singleton breech presentation throughout Scotland.

Breech presentation occurs in 3-5% of deliveries and can be managed by either a trial of vaginal breech delivery (TOVBD), external cephalic version (ECV) or Caesarean section. A postal questionnaire was completed by 82% of Scottish consultant obstetricians and revealed wide variations in practice. Eighteen percent never offered ECV. Among those who did consensus was lacking on some contraindications. One-quarter sometimes performed ECV before 37 weeks gestations despite the possibility of spontaneous version. Only 70% restricted ECV to one or more designated operators thereby maintaining levels of expertise. Variations were demonstrated in the use of tocolytics, and pre and post procedure investigations. Following failed ECV 28% considered a repeat attempt and 56% a TOVBD. TOVBD was not offered as first line management by one-fifth of respondents. Those who did varied in the pre-procedure investigations performed. Guidelines are required to ensure safe, consistent practice and avoid unnecessary Caesarean sections.

Differential expressions of mRNA for proteoglycans, collagens and transforming growth factor-beta in the human cervix during pregnancy and involution.

During pregnancy and involution, an extensive remodelling of the human cervical connective tissue occurs. This cervical ripening is one of the most pronounced physiological remodelling processes known in human connective tissue. To investigate how the remodelling is accomplished, the levels of mRNA for collagen I and III, versican and three small proteoglycans, biglycan, decorin and fibromodulin, were evaluated using Northern blots at different stages of cervical ripening. In the corresponding biopsies the concentration of collagen and of small and large proteoglycans were determined. The role of transforming growth factor-beta (TGF-beta) as a mediator of the remodelling process was also investigated. The concentration of collagen decreased and 1 week before partus, 50% of the nonpregnant level was attained. No further decrease was noted after partus. The mRNA for collagen I and III did, however, not decrease in the term pregnant cervix 1 week before partus. Only 20-30% decrease during the final ripening just before partus was recorded. Neither did the mRNA levels of the small proteoglycans change significantly during the ripening, despite an almost 50% decrease in the concentration of the small proteoglycans. The message for versican was, however, 5-fold increased at partus and then gradually returned to nonpregnant levels within 4 days after delivery. These changes corresponded to similar changes in the concentration of the large proteoglycan. Thus, the remodelling of the cervical connective tissue is achieved by two different mechanisms, on one hand an increased turnover of collagen and the small proteoglycans, on the other a changed transcription followed by an increased production of versican. During the involution 2- to 3-fold increases in the messages for collagen I and III, and the small proteoglycans, biglycan and decorin, corresponded to increases in the concentration of the small proteoglycans and non-extractable collagen. The message for TGF-beta was increased 2-fold immediately after delivery compared with the term pregnant state. Thus, TGF-beta may be of importance for the reconstruction of the cervix, which starts immediately after partus.

Quantitation of proteoglycans as glycosaminoglycans in biological fluids using an alcian blue dot blot analysis.

A method for quantitation of intact proteoglycans as GAGs in biological fluids (blood plasma, synovial fluid) or 4 M guanidine extracts of tissues has been published previously (S. Björnsson, Anal. Biochem. 210, 282-291, 1993). The method is based on the specific interaction between sulfated polymers and the tetravalent cationic dye Alcian blue at pH 1.5 in 0.4 M guanidine-HCl and in the presence of 0.25% Triton. The absorbance assay has a measuring range of 1-20 microgram of glycosaminoglycan (GAG) which is not sensitive enough to measure the low contents of proteoglycans in blood plasma, urine, or wound fluid. A dot blot assay is now described in which the Alcian blue-GAG complexes are collected on a polyvinylidene fluoride membrane, by filtration in a dot blot apparatus, and the stain is quantitated as reflectance by scanning and densitometry. The assay requires 10 microliter of sample and has a measuring range of 10-800 ng of GAG, corresponding to a concentration of 1-80 mg/liter, suitable for proteoglycans in biological fluids. The procedures for chemistry, scanning, densitometry, and curve fitting were each evaluated separately. The error contributed by chemistry accounted for a minor portion of the imprecision. The imprecision contributed by scanning was the most important source of within-run and between-run imprecision, and was caused by inequalities of the charge-coupled device along the scanning arm. Unexpectedly, curve fitting was also a major source of total imprecision in dot blot quantitation and differed with the type of equation used. The between-run imprecision calculated as CV (SD/mean . 100) was 13.0% at 8 mg/liter. The response of the assay was identical for six different commercial preparations of GAGs (chondroitin-4-sulfate, chondroitin-6-sulfate, dermatan sulfate, keratan sulfate, heparan sulfate, and heparin) despite differences in degree of sulfation known to exist. There was no positive or negative interference by blood plasma, apart from a slight negative interference on the quantitation of heparan sulfate. Analysis of 319 paired blood plasma and urine specimens from hospitalized patients showed a variation of plasma GAGs of 0.1-17.6 and urine-GAGs of 0.0-45.6 mg/liter. There was no correlation between plasma and urine GAG concentrations.

Inflammatory bowel disease in Iceland, 1980-89. A retrospective nationwide epidemiologic study.

BACKGROUND: A previous nationwide epidemiologic study of inflammatory bowel disease (IBD) in Iceland in 1950-79 showed a low but steadily rising incidence. The incidence of IBD in Iceland in 1980-89 was investigated and compared with reports from neighboring countries. METHODS: Cases were retrieved by a review of all small- and large-intestinal tissue specimens with any type of inflammation submitted to the only two departments of pathology in Iceland. All small-intestinal X-ray records suggestive of Crohn's disease (CD) in the three major hospitals were also screened. All hospital and outpatient records of cases suggestive of IBD were then reviewed using accepted criteria for confirmation or exclusion. RESULTS: The mean annual incidence of ulcerative colitis (UC) was 11.7/100,000, and that of CD 3.1/100,000. The highest age-specific incidence of UC was in the group 30-39 years old and for CD in the group 60-69 years old. The most frequent involvement at diagnosis of UC was proctitis only, in 54%, and in CD colon only, in 54.7% of the patients. CONCLUSIONS: This study shows a continuing increase in the incidence of both diseases. Compared with the period 1970-79, there has been an almost twofold increase in the mean annual incidence of UC and more than a threefold increase of CD, a statistically significant increase in both instances.

Decreased excretion of glycosaminoglycans in patients with primary glomerular diseases.

Urine glycosaminoglycans (GAG) concentrations were measured in 150 patients with primary glomerulonephritides: endocapillary glomerulonephritis, mesangial proliferative glomerulonephritis, IgA nephropathy, membranous glomerulonephritis and minimal change nephropathy, and in 63 healthy controls and 19 patients with diabetes nephropathy. The urine GAG to creatinine ratios (GCR) were significantly reduced (p < 0.01) in all the glomerulonephritides investigated (0.20 mg/mmol in endocapillary glomerulonephritis, 1.60 mg/mmol in mesangial proliferative glomerulonephritis, 1.74 mg/mmol in IgA nephropathy, 1.09 mg/mmol in membranous nephropathy, and 1.16 mg/mmol in minimal change nephropathy) compared to healthy controls (2.87 mg/mmol) but not compared to diabetes patients (1.17 mg/mmol). Also, the GCR in a group of 23 non-albuminuric glomerulonephritis patients (1.98 mg/mmol) was shown to be significantly decreased (p < 0.01) compared to healthy controls. Moreover, the GCR was significantly lower (p < 0.01) in endocapillary glomerulonephritis than in any of the other diseases studied. The GAG excretion per functioning glomerular area, calculated as fractional GAG excretion (FGE), was decreased in all the glomerulonephritides investigated compared to both healthy controls and diabetes nephropathy. The decreased GAG excretion in glomerulonephritides, obtained in the present study, might be a consequence of decreased synthesis or turnover of GAG in the functioning nephrons whereas the mechanisms for the reduced GAG excretion in diabetes nephropathy might be of a different nature. Urinary GAG excretion in this group of glomerular disorders and particularly in endocapillary glomerulonephritis, may lead to new approaches in non-invasive renal diagnostics and, particularly with regard to the differentiation of acute and chronic forms of glomerulonephritides.

Factors affecting women's knowledge of antenatal serum screening.

Antenatal screening for fetal anomaly is offered routinely in Glasgow. This study assessed pregnant women's knowledge of the test and implications of results. Questionnaires were completed by 574 women. Knowledge of the nature and uses of the test was superior to earlier studies Two-thirds knew that screening was undertaken for Down's syndrome, and 81% for spina bifida. The majority were aware of the sample used, ages invited and gestation at which it was undertaken. Knowledge of the likely results and implications was poorer. Three-quarters were unaware that 10% of results would suggest an increased risk. Half did not realise that positive results could occur without fetal abnormality, or negative results could be falsely reassuring. Socioeconomic deprivation was associated with poorer knowledge but not lower uptake. Written information was associated with superior knowledge and higher uptake. Leaflets should be provided prior to booking, at which time resultant queries can be answered.

Decreased excretion of urine glycosaminoglycans as marker in renal amyloidosis.

BACKGROUND: The diagnosis of renal amyloidosis is normally established by kidney biopsy. In order to advance the determination of the diagnosis and the initiation of the therapy, fast and cheap, non-invasive diagnostic techniques are required. METHODS: Urine excretion of glycosaminoglycans (GAG) was measured in 10 patients with AA amyloidosis and 5 patients with AL amyloidosis and compared to 25 controls with primary glomerular diseases and 22 healthy controls. The subjects with primary glomerular disease were matched with regard to their renal function and the degree of albuminuria. RESULTS: The median urine GAG to creatinine ratio and the median fractional GAG excretion were significantly decreased (P < 0.05) in both AA amyloidosis (0.21 mg/mmol and 0.053 respectively) and AL amyloidosis (0.33 mg/mmol and 0.077 respectively) compared to control patients with primary glomerular disease (1.73 mg/mmol and 0.336 respectively) and healthy controls (2.67 mg/mmol and 0.226 respectively). The urine GAG to creatinine ratio did not correlate to age, sex, serum creatinine, urine albumin, or to the plasma levels of acute phase proteins. CONCLUSIONS: The decreased GAG excretion in renal amyloidosis is probably caused both by diminished number of functioning nephrons, decreased GAG synthesis in functioning glomeruli, and the trapping of GAG by amyloid fibrils. Urinary GAG excretion may serve as an independent marker of renal amyloidosis. It may be used in diagnostic work-up of renal amyloidosis in patients with glomerular diseases and in screening of amyloidosis in patients with chronic inflammatory disorders, with or without signs of renal disease.