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.

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.

Staphylococcal bacteremia in a "Germ-free" unit.

The rate of carriage and infections due to strains of Staphylococcus aureus were evaluated in adults with acute leukemia in isolators characterized by laminar air flow and barrier isolation. Patients were randomly given antimicrobial prophylaxis with oral nonabsorbed antibiotics and a nasal antibiotic ointment. In four years S aureus was isolated from the nostrils or other sites in 36 patients. Persistent isolation was noted in 24 patients. Suppression of gut flora was associated with a higher carriage rate of S aureus. Five episodes of bacteremia due to S aureus occurred at the nadir of leukopenia induced by chemotherapy. Death occurred within five days in the three patients whose peripheral white blood cell count did not rise. Patient isolation and suppression of gut flora helped reduce infections due to Pseudomonas sp and fungi, but S aureus emerged as a life-threatening pathogen.

Murine myeloid leukemia: colony formation in vitro.

Normal and myeloid leukemic spleen cells from RF mice were cultured in vitro in plasma clots. In situ histochemical staining and karyotypic analysis of the colonies formed in the clot revealed that the colonies produced by leukemic and normal progenitors were indistinguishable morphologically and cytochemically. Colonies of leukemic origin were identified by in situ karyotypic analysis, a method not previously utilized in studies of hematopoietic proliferation in semi-solid matrices.

Natural killer cell function and malignant cell phenotype in hairy cell leukaemia.

We have followed for 33 months the changes that occurred in natural killer (NK) cell numbers and activity in a patient (A) with hairy cell leukaemia (HCL), using a single cell assay and a microcytotoxicity assay. The composition of the peripheral blood mononuclear cell population and malignant cell phenotype were also analysed. During this period he received treatment with interferon and his grossly enlarged spleen was removed. Four further patients were also studied, two were splenectomized and all had received treatment with interferon. In four of the five patients studied there was an apparent link between low NK activity and presence of a tumour-infiltrated spleen, and in the fifth patient, who was aleukemic and had no splenomegaly, NK function was related to disease activity. There was no correlation between NK activity and the number of target binding (TB) cells in these five patients. IFN had little direct effect on overall NK activity, but the proportion of killing cells among TB cells was increased. Three patients showed binding of several cells to a single target. Further analysis revealed that in the patients most of the TB cells were not CD56-positive NK cells, in contrast to TB cells from normal subjects. In patient A a large proportion (84%) of TB cells were identified as malignant cells and in patient E 15% of TB cells were malignant cells. The phenotype of the malignant cells was: CD19+, HLA-DR+ and CD25(Tac)+, except for patient A. In this patient the hairy cells were positive for the NK marker CD56 as well as the monocyte marker CD14. Furthermore, a change occurred in phenotype as only later samples carried CD25. It is concluded that the level of NK function correlates closely with disease activity in HCL and that competitive target cell binding by malignant cells may be one cause of depressed NK-cell function in hairy cell leukaemia.

Immunohistochemical characterization of endometrial lymphoid cell populations in women infected with human immunodeficiency virus.

OBJECTIVE: To determine whether lymphocytic infiltration of the endometrium accompanies human immunodeficiency virus (HIV) infection. METHODS: Endometrial samples from 12 HIV-infected women and from rigorously matched controls were examined. The following markers were used: common leukocyte antigen (CD45), T lymphocytes (CD3), monocytes-macrophages (CD68), and CD4 and CD8 lymphocytes. Cell counts were performed without knowledge of HIV status. Factors considered in relation to these markers were menstrual symptoms, pelvic pain, peripheral blood CD4+ count, and time since seroconversion. RESULTS: Histology showed conventional features of chronic endometritis in only one case. In the remainder, the endometrium of HIV-infected women, compared with controls, showed an increase in CD45 cells (P < .02) and an increase in CD3 staining cells (P < .05). This appeared to be restricted to those with menstrual symptoms, and this group also had lower peripheral blood CD4 counts. There was no difference in cells of the monocyte-macrophage series (CD68). In contrast to control samples, CD4 lymphocytes were infrequent or absent in the endometrium of HIV-infected women, regardless of peripheral blood CD4 count or presence of menstrual symptoms; however, this was not universal, as one sample showed an area of dense CD4 cell infiltration. The ratio of CD4 to CD8 was reduced in HIV-seropositive samples compared with controls (P < .02). CONCLUSION: We hypothesize that chronic endometritis of a nonclassical form may be common in advancing HIV disease, possibly directed against HIV-infected cells or self-determined antigens. This could be associated with morbidity and may represent a reservoir of infection. Endometrial depletion of CD4 cells is a common, but not universal, feature and may be independent of immune compromise.

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).

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 the use of folic acid supplements in pregnant women in Glasgow.

BACKGROUND: Use of folic acid supplements preconception, and during the first trimester, is associated with a reduced incidence of first and subsequent neural tube defects. The Department of Health guidelines recommend the use of folic acid supplements by all women planning a pregnancy. AIM: To ascertain the proportion using folic acid supplements and the factors affecting their use. METHOD: Questionnaires were distributed postpartum to the 515 women who delivered normal babies in three maternity units in Glasgow over a four-week period. RESULTS: Forms were completed by 487 (95%) women. Only 57% took supplements at some point during their pregnancy, and only 21% took them before conception. Failure to take supplements was significantly associated with unplanned pregnancy, younger age, and previous pregnancies. Lack of awareness of the potential benefits associated with folic acid use was the commonest reason cited by women for not taking supplements. CONCLUSIONS: Increased health education through health care professionals and mass media campaigns can improve awareness and thereby increase the use of supplements in planned pregnancies. However, 42% of women in our study had unplanned pregnancies. Intake of folic acid supplements in this group can only be increased by improvements in dietary intake within the population as a whole, and by fortification of commonly ingested foods.

Labetalol therapy in pregnancy induced hypertension: the effects on fetoplacental circulation and fetal outcome.

We prospectively studied the effects of oral labetalol therapy in patients with moderate to severe pregnancy induced hypertension (PIH). The outcome variables were blood pressure control, effect on umbilical artery flow velocity waveforms (UAFVW) and fetal outcome. Forty-two patients were recruited, all had moderate to severe PIH. The mean systolic blood pressure (SBP) and diastolic blood pressure (DBP) on entry were 154 +/- 7 mmHg and 104 +/- 5 mmHg, respectively. All had significant proteinuria. After 1 week on labetalol therapy, 85% of patients had their blood pressure controlled. The reduction in both SBP and DBP was statistically significant. There were no significant changes in UAFVW, Resistance Index (RI), uric acid or platelets. The mean fetal age on entry was 246 +/- 10 days while gestation at delivery was 258 +/- 17 days. The mean birth weight was 2712 +/- 609 g. No perinatal mortality occurred in this study. Labetalol is an effective drug in controlling blood pressure and does not adversely affect the UAFVW. No neonatal problems were attributed directly to the drug. Fetal outcome was satisfactory despite the 12 fetuses that were growth-retarded. Labetalol allows safe prolongation of pregnancies complicated by PIH.

The effect of acute and chronic antihypertensive therapy on maternal and fetoplacental Doppler velocimetry.

Maternal and feto-placental Doppler flow velocity waveforms were studied during acute and chronic antihypertensive therapy in patients with pregnancy-induced hypertension. Eight primigravidae were acutely treated with oral nicardipine. Diastolic blood pressure fell at 30, 45 and 60 min after nicardipine. The uteroplacental systolic/diastolic ratio rose significantly at 30 min, but this change was no longer apparent at 60 min. Umbilical artery and maternal brachial artery systolic/diastolic ratios were unchanged. Fifteen patients with mild pre-eclampsia were chronically treated with oral pindolol. Diastolic blood pressure fell significantly within 24 h. The uteroplacental systolic/diastolic ratio rose 3 days after pindolol. Brachial artery or umbilical artery systolic/diastolic ratios were unchanged. A control group of 15 patients with untreated mild pre-eclampsia showed a significant rise in uteroplacental and umbilical artery systolic/diastolic ratios within 7 days of starting recordings. In patients with pregnancy-induced hypertension, acute and chronic blood pressure reduction was associated with no change in umbilical artery or maternal brachial artery Doppler systolic/diastolic ratios and a transient rise in the uteroplacental systolic/diastolic ratio.

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.

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 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.

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.

Inflammatory bowel disease in Iceland during a 30-year period, 1950-1979.

Thirty-three patients with Crohn's disease (CD) and 318 with ulcerative colitis (UC) were identified among the 250,000 inhabitants of Iceland over a 30-year period. The present incidence of CD is 0.9/10(5)/year and of UC 7.4/10(5)/year.