Downregulation of the expression of bone morphogenetic protein 7 in experimental pyelonephritis

Bone morphogenetic protein 7 (BMP 7) is a member of the transforming growth factor (TGF) beta superfamily and is involved in regeneration, repair, and development of specific tissues, for example kidney, gut, lens, and skeleton. BMP 7 has emerged as a renotrophic factor and experimental studies have shown its protective role against fibrotic processes. Tubulointerstitial changes are present in the pyelonephritic kidney which progresses to fibrosis. Renal fibrosis may lead to significant morbidity in the form of hypertension, proteinuria, and loss of renal function. The objective of this study was to investigate BMP 7 expression in experimental acute and chronic pyelonephritis models. Eighteen Wistar rats were injected with 0.1 mL solution containing E. coli ATCC 25922 10¹⁰ cfu mL^–1 into left renal medullae. Six rats were used as a sham group and were given 0.1 mL 0.9% NaCl. Pyelonephritic rats were sacrificed 24 h (group I, n=6), 1 week (group II, n=6), and 6 weeks (group III, n=6) after E. coli injection. Serum creatinine levels were analyzed. Renal tissues were studied histopathologically by use of hematoxylin and eosin and scored for diagnosis of pyelonephritis. BMP 7 expression was studied semiquantitatively by immunohistochemical staining. Acute (group I) and chronic (group II and group III) pyelonephritic histopathological changes were observed in experimental pyelonephritic groups. A gradual decrease in BMP 7 expression was observed in the tubulointerstitial and tubular area of the pyelonephritic kidneys, mildest in the acute pyelonephritic group and most severe in the chronic pyelonephritic 6th week group. A statistically significant difference was observed between tubulointerstitial BMP 7 expression by groups I and III (P=0.017) and by groups III and IV (P=0.000). Tubular BMP 7 expression was statistically significantly different between groups II and IV (P=0.009) and between groups III and IV (P=0.002). The data imply that BMP 7 has a major role in chronic pyelonephritis. Tubulointerstitial and tubular BMP 7 expression also had a significant negative correlation with fibrosis, tubular, atrophy, and vascular changes. Serum creatinine levels of the study group were all normal. We conclude that the decrease in renal BMP 7 expression in experimental chronic pyelonephritis is one of the factors responsible for fibrotic changes in persistent renal damage.     

Tricuspid valve perforation by permanent pacemaker lead--a case report

Tricuspid valve perforation with pacemaker lead is one of the extremely rare complications of transvenous pacemaker implantation. Approximately all reported cases have been diagnosed at autopsy. The authors present a case of tricuspid valve perforation caused by pacemaker lead that was diagnosed during cardiac surgery and treated successfully by removing the lead and suturing the tricuspid valve.     

Isolation, culture, and characterization of endothelial cells from mouse glomeruli

BACKGROUND: Cloned glomerular endothelial cells (GENC) have many potential uses and applications in immunologic and physiologic studies. Propagation of GENC has been difficult and available homogeneous GENC, particularly from mice, are limited. Herein we report isolation, cloning, propagation, and characterization of GENC from mice. METHODS: tsA58 immorto mice were used to isolate glomerular cells. Glomeruli were isolated by differential sieving, and decapsulated explants were cultured in permissive and optimal conditions for endothelial cells. The primary cells from glomerular outgrowths were expanded, taking advantage of the temperature-sensitive tsA58 gene, and then the cells were allowed to undergo spontaneous transformation. The cells were then sorted using anti-CD31 antibodies and their capacity to uptake acetylated-low-density lipoprotein (LDL). Individual subclones isolated by patch cloning were characterized using multiple markers. RESULTS: One of the homogeneous clones was morphologically endothelial-like, positive for CD31, CD106, CD62E, CD54, and acetylated-LDL uptake, formed tubes, and was negative for epithelial and mesangial cell markers. The functional properties of this GENC clone appeared to be intact, and signaling pathway was not altered. Two of the clones displayed the characteristics of either visceral epithelial or mesangial cells. CONCLUSION: The identified clones should have utility in multiple areas of investigation.     

Biochemical bone markers in nephrotic children

In this study we evaluated the effects of high-dose corticosteroid (CS) therapy and the character of the nephrotic syndrome (NS) itself on bones in patients with normal glomerular filtration rate. We measured serum osteocalcin (OC), alkaline phosphatase (ALP), intact parathyroid hormone (iPTH), 25-hydroxyvitamin D, calcium (Ca), phosphorus (P), and magnesium (Mg) levels, and urinary Ca and protein excretion in nephrotic children during the active phase before (group Ia) and after CS treatment (group Ib). The results were compared with age-matched control subjects. A significant increase in urinary Ca excretion was observed after CS treatment. Serum ALP, OC, and iPTH levels were within normal limits at the time of study entry. However, both serum OC and ALP levels showed a significant decrease after the completion of CS treatment (OC from 13.6±9.2 ng/ml to 6.7±5.2 ng/ml and ALP from 151.8±60.2 U/l to 116±43.8 U/l). 25-Hydroxyvitamin D levels increased to 17.2±8.9 g/l from 9.9±6.9 g/l after CS treatment. The effects of recurrent use of CSs were assessed by dividing nephrotic patients into two subgroups: infrequent relapsers (IFR) and frequent relapsers (FR). The cumulative dose of CS was 28,125 mg/m² for IFR and 105,000 mg/m² for FR. The changes in OC, ALP, and 25-hydroxyvitamin D levels after CS treatment were significantly different between IFR and FR. We conclude that high-dose CS treatment causes a decrease in bone formation, as shown by the changes in OC and ALP levels. 25-Hydroxyvitamin D levels remained lower than control subjects after CS therapy. The higher the cumulative dose of CS used the more marked the changes in biochemical bone markers. The contribution of FR to baseline 25-hydroxyvitamin D levels needs further study.     

Optimal Timing for Surgery After Adriamycin Treatment in Rats

Purpose To determine the optimal timing of surgery after adriamycin treatment, we investigated the time-related effect of adriamycin on wound healing over a long period.Methods We divided 119 female Sprague-Dawley rats into seven treatment groups. Group 1 was subjected to laparatomy only. All the other groups were given 8mg/kg adriamycin intravenously followed by laparotomy on the same day (group 2), 7 days later (group 3), 14 days later (group 4), 21 days later (group 5), 28 days later (group 6), or 35 days later (group 7). On postoperative day 7, the sutures were removed, abdominal bursting pressure was measured, and tissue samples were taken for histopathological evaluation and analysis of hydro-xyproline content.Results Bursting pressures were significantly lower in groups 3, 4, 5, and 6 than in group 1. The hydroxyproline content and histopathological evaluation supported these findings.Conclusions Our results showed that the optimal timing for surgery after adriamycin treatment is before the 7th day or after the 35th day. If surgery is performed between these days, there is a high risk of impaired wound healing.A preliminary study on this subject was accepted as a poster presentation at the Congress of the European Surgical Society of Oncology (ESSO) in 2002     

Efficiency of gamma probe and dual-phase Tc-99m sestamibi scintigraphy in surgery for patients with primary hyperparathyroidism

PURPOSE: The purpose of this study was to determine the value of the intraoperative gamma probe and the efficacy of dual-phase Tc-99m sestamibi imaging in patients with primary hyperparathyroidism. METHODS: Twenty-one patients with primary hyperparathyroidism were examined prospectively. Results of same-day dual-phase Tc-99m sestamibi scintigraphy and intraoperative gamma probe evaluations were compared with the intraoperative findings and histopathologic diagnoses. A 15-mm handheld gamma probe was used to measure gamma activity in the neck and upper mediastinum. Nuclear mapping by gamma probe showed a single quadrant of neck that emitted gamma radiation significantly greater than the other three quadrants, which correlated with the sestamibi scan. RESULTS: Dual-phase Tc-99m sestamibi scintigraphy determined and localized parathyroid lesions in 20 patients (sensitivity, 94%). Of the 20 parathyroid lesions removed, 15 were located in normal positions, whereas five were explored in ectopic sites (one within the thyroid, one in the anterior mediastinum, one in a retrotracheal position, one in the carotid sheath, and one in the retroesophageal region). Although the index of thyroid nodules varied from 15.8% to 22.9%, the index for parathyroid lesions was 77.3% to 112.8%. CONCLUSIONS: These results confirm that parathyroid lesions, especially at ectopic sites, can be treated successfully in shorter operative times with minimal complications with the help of the intraoperative gamma probe.     

The CC genotype of the C825T polymorphism of the G protein beta3 gene (GNB3) is associated with a high relapse rate in patients with chronic lymphocytic leukaemia

A C825T polymorphism has been described in the GNB3 gene which encodes the Gbeta3 subunit of heterotrimeric G proteins. The GNB3 825T allele is predictive of enhanced Gi protein activation. This study was performed to correlate genotypes of the C825T polymorphism with various clinical aspects of chronic lymphocytic leukaemia (B-CLL). The GNB3 genotype distribution in B-CLL patients was similar to that in other Caucasian populations, arguing against a role of the polymorphism in the susceptibility to develop B-CLL. No statistically significant differences were observed at diagnosis between patients with the CC genotype and homozygous or heterozygous T allele carriers with respect to age at disease onset, sex distribution, proportion of patients with CD38+ leukaemia or patients in Binet stage A, blood cell counts, degree of bone marrow infiltration or serum levels of lactate dehydrogenase, thymidine kinase or beta2-microglobulin. In a subgroup of 44 patients requiring chemotherapy, the median interval between diagnosis and first treatment and the response to treatment were similar in patients with CC or CT/TT genotypes. A statistically significant difference, however, was found in the proportion of patients relapsing and requiring second line chemotherapy (CC: 95%; CT/TT: 52%; p = 0.0043). The GNB3 genotype (p = 0.024) and age (p = 0.042) were identified as independent prognostic factors for a second therapy. Thus, the long-term success of the treatment appears to be correlated with the GNB3 genotype.