The association between haemorrhage and markers of endothelial insufficiency and inflammation in patients with hypoproliferative thrombocytopenia: a cohort study.

In daily haematological practice, predicting bleeding in thrombocytopenic patients is difficult, and clinicians adhere to transfusion triggers to guide patients through the aplastic phase of chemotherapy. Platelet count is not the only determinant of bleeding and additional mechanisms for impending haemostasis are likely. Beside clot formation, platelets are essential for the maintenance of integrity of vascular beds. We therefore prospectively investigated associations between biomarkers for endothelial damage (urine albumin excretion) and inflammation (C-reactive protein) and bleeding (WHO grading) in 88 patients with 116 on-protocol episodes. We found an increase in grade 2 bleeding with a higher urine albumin/creatinine ratio one day after the measurement [odds ratio (OR) 1·24 for every doubling of the ratio, 95% CI 1·05-1·46, P-value 0·01] and a 29% increase in the odds of grade 2 bleeding for every doubling of serum C-reactive protein (CRP) (95% CI 1·04-1·60, P-value 0·02) after correction for morning platelet count. The 24 h post-transfusion corrected count increment (CCI24 ) showed a significant association with these biomarkers: increasing urine albumin/creatinine ratio and CRP were associated with lower CCI24. We report two inexpensive and easy-to-apply biomarkers that could be useful in designing a prediction model for bleeding risk in thrombocytopenic patients.

1H NMR-based metabonomic revealed protective effect of Moutan Cortex charcoal on blood-heat and hemorrhage rats.

Moutan Cortex charcoal (MCC), the processed root bark of Paeonia suff ;ruticosa Andrews (Paeoniaceae), is a kind of traditional Chinese medicine (TCM) and has been used for treating blood-heat and hemorrhage(BHH)syndrome in China for thousands of years. In order to explore potential metabolic mechanism, 1H NMR-based metabonomics technique was applied to evaluate the effect of MCC on metabolic changes in plasma and urine of BHH rat models. Serum and urine samples were obtained from male SD rats with normal group, model group and MCC group for study. Based on 1H NMR spectra obtained from plasma and urine samples, principal component analysis (PCA) and orthogonal partial least-squares discriminant analysis (OPLS-DA) models were capable of distinguishing the three group. And the 13 pharmacodynamic biomarkers of MCC were identified in the plasma and urine. The results showed that BHH induced great metabolic disorders in plasma and urine metabolisms. However, MCC could reverse the imbalanced metabolites by alanine, aspartate and glutamate metabolism and citrate cycle (TCA cycle) pathway, and its effect was also confirmed by the general signs and pharmacodynamics assessments. The results indicated that NMR-based metabolomic profiling method is sensitive and specific enough to evaluate the MCC efficacy and mechanism of action on BHH syndromes.

Laparoscopic kidney biopsy in dogs: Comparison of cup forceps and core needle biopsy.

OBJECTIVE: To investigate the feasibility of laparoscopic kidney biopsy with cup biopsy forceps in dogs (CupBF), and to compare to the use of a core biopsy needle (CoreBN). STUDY DESIGN: Experimental; randomized, controlled design. ANIMALS: Eight healthy, adult Beagle dogs. METHODS: Dogs were randomized to undergo laparoscopic biopsy of the right kidney using either 5 mm CupBF or a 16 gauge CoreBN. Intraoperative hemorrhage of the biopsy site was monitored. Biopsy quality was evaluated for tissue fragmentation and crushing, presence of renal cortex with or without medulla, and number of glomeruli. Postoperative packed cell volume, urinalysis, and ultrasonographic appearance of the biopsy site were evaluated. RESULTS: Biopsy specimens were obtained by both techniques and reliable hemostasis was achieved with direct compression in all dogs. The histologic score for CupBF biopsies was not significantly different from CoreBN biopsies. One CoreBN biopsy contained both renal cortex and medullar, while all CupBF biopsies contained cortex only. The mean (SD) number of glomeruli was significantly higher in CupBF biopsies [60 (9.1)] than CoreBN biopsies [26 (4.3)]. There was no gross hematuria, perirenal hematoma, or hydronephrosis in any dog postoperative. CONCLUSION: Laparoscopic kidney biopsy in dogs using 5 mm cup biopsy forceps is feasible with minimal risk and more glomeruli obtained compared to laparoscopic kidney biopsy using 16 gauge core biopsy needles.

[THE QUANTIFICATION OF SULPHATED GLYCOSAMINOGLYCANS IN RAT URINE IN EXPERIMENTAL HEMORRHAGIC CYSTITIS].

The paper presents the study of the excretion of sulfated glycosaminoglycans (GAG) in the urine of rats in experimental hemorrhagic cystitis induced by cyclophosphamide and treated with glycosaminoglycan replacement therapy. Rats were given intraperitoneal injections of cyclophosphamide at a dose of 100 mg per 1 kg body weight and subsequently treated with intragastric administration of the combined preparation of glycosaminoglycans containing glucosamine hydrochloride and chondroitin sulfate at a dose of 10 and 100 mg per 1 kg of body weight. Within 24 or 72 hours after cystitis induction there was a statistically significant increase in urinary GAG excretion. The study also found a decrease (from 1.34 to 1.22 mg/dL) in urinary GAG within 0 to 72 hours following induction of acute cystitis without glycosaminoglycan therapy. In the subchronic model of inflammation in the bladder, upon repeated administration of low doses of cyclophosphamide (50 mg/kg), decrease in urinary GAG within 0 to 72 hours (1,32±0,13 mg/dL) as well as increased excretion after 96 hours at a concentration of 2,29±0,13 mg/L after initiation cystitis were found.

Efficacy and Safety of Dabigatran Etexilate vs. Warfarin in Asian RE-LY Patients According to Baseline Renal Function or CHADS2 Score.

BACKGROUND: In Asian patients in RE-LY, dabigatran etexilate (DE) was as effective as warfarin, with a significantly lower bleeding risk. We evaluated the relationship between baseline renal function or CHADS2 score and efficacy or safety outcomes in these patients. METHODS AND RESULTS: Asian patients (n=2,782) were categorized according to baseline renal function or CHADS2 score, and efficacy and safety outcomes were analyzed for DE (110 mg and 150 mg b.i.d.) vs. warfarin. There was an increase in the rates of stroke/systemic embolism and major bleeding with worsening renal function and CHADS2 score. For stroke/systemic embolism (primary efficacy endpoint), there was no treatment interaction for dabigatran at either 110 or 150 mg b.i.d. compared with warfarin related to patients' baseline renal function (Pinteraction=0.56 for DE 110 mg and 0.62 for DE 150 mg vs. warfarin) or CHADS2 score (Pinteraction=0.68 for DE 110 mg and 0.31 for DE 150 mg vs. warfarin). For major bleeding, there was no treatment interaction by creatinine clearance category observed for either dose (Pinteraction=0.60 and 0.62 for DE 110 mg and DE 150 mg, respectively). Baseline CHADS2 score had no significant effect on bleeding event rates with DE vs. warfarin. CONCLUSIONS: Bleeding and stroke rates in Asian patients varied according to renal function and CHADS2 score, but the relative benefits of DE over warfarin were preserved when analyzed by subcategories.

Isolation of an X-factor-dependent but porphyrin-positive Escherichia coli from urine of a patient with hemorrhagic cystitis.

An Escherichia coli isolate was recovered from a 92-year-old female patient with urinary tract infection. Gram-stained preparation of the urine sediment manifested some gram-negative rod-shaped cells, and the urine specimen culture yielded nonhemolytic colonies on sheep blood agar plate. However, no visible colonies appeared on modified Drigalski agar plate. The isolate was finally identified as an X-factor-dependent E. coli. The interesting finding was that the isolate revealed a positive reaction for porphyrin test despite the requirement of hemin. This finding suggested that some pyrrol-ring-containing porphyrin compounds or fluorescent porphyrins had been produced as chemical intermediates in the synthetic pathway from δ-amino-levulinic acid (ALA), although the isolate should be devoid of synthesizing hems from ALA. This was the first clinical isolation of such a strain, indicating that the E. coli isolate should possess incomplete synthetic pathways of hems from ALA.

Blood, and not urine, BK viral load predicts renal outcome in children with hemorrhagic cystitis following hematopoietic stem cell transplantation.

BK virus is a significant cause of hemorrhagic cystitis after hematopoietic stem cell transplantation (HSCT). However, its role in nephropathy post-HSCT is less studied. We retrospectively evaluated clinical outcomes in pediatric HSCT patients with hemorrhagic cystitis. Although most of these patients had very high urine BK viral loads (viruria), patients with higher BK plasma loads (viremia) had significant renal dysfunction, a worse clinical course, and decreased survival. Patients with a peak plasma BK viral load of >10,000 copies/mL (high viremia) were more likely to need dialysis and aggressive treatment for hemorrhagic cystitis compared to patients with ≤ 10,000 copies/mL (low viremia). Conversely, most patients with low viremia had only transient elevations in creatinine, and less severe hemorrhagic cystitis that resolved with supportive therapy. Overall survival (OS) at 1 year post-HSCT was 89% in the low viremia group and 48% in the high viremia group. We conclude that the degree of BK viremia, and not viruria, may predict renal, urologic, and overall outcome in the post-HSCT population.

Elevated urinary catecholamines and adrenal haemorrhage mimicking phaeochromocytoma.

A 51-year-old woman was admitted with left-sided flank pain initially thought to be renal colic. However, a CT urogram was normal. During the course of the admission the pain persisted and she developed severe sustained hypertension. A repeat CT scan of the abdomen revealed a 5×3 cm left adrenal abnormality consistent with haemorrhage, not seen on the original scan. Further assessment revealed elevated urine catecholamines and a short synacthen test showed a suboptimal cortisol response. The diagnosis was initially considered as a phaeochromocytoma, she received phenoxybenzamine with good resolution of hypertension and was referred for surgical opinion. However, serial urinary catecholamine concentrations returned to within the normal range and the diagnosis was revised to adrenal infarction and haemorrhage due to antiphospholipid syndrome. This case illustrates the importance of recognising adrenal infarction as a potential cause of 'pseudophaeochromocytoma'.

The incidence of hemorrhagic cystitis and BK-viruria in allogeneic hematopoietic stem cell recipients according to intensity of the conditioning regimen.

The influence of BK-viruria, donor background, and conditioning on the development of hemorrhagic cystitis was examined in 90 allogeneic hematopoetic stem cell transplant patients, of whom 15 developed hemorrhagic cystitis. Thirty-two patients had related and 58 had unrelated donors, while 44 received full, and 46 received reduced intensity conditioning (RIC). BK-viruria was more common in patients with hemorrhagic cystitis than in those without (p<0.01), and hemorrhagic cystitis was less common in patients with related donors than in those with unrelated donors (p=0.02). Finally, hemorrhagic cystitis and BK-viruria were less common in patients receiving RIC, rather than full conditioning (p<0.01 and p<0.01, respectively).

Occult bleeding in three different antithrombotic regimes after myocardial infarction. A WARIS-II subgroup analysis.

INTRODUCTION: Warfarin, aspirin, and the combination of these, have all proven to be efficacious in preventing future events after myocardial infarction. The accompanying bleeding tendency is a concern. The aim of the present study was to compare the occurrence of occult bleeding and iron deficiency during these treatment modalities. METHODS: The 267 patients who had survived a myocardial infarction were randomly assigned in the Warfarin Aspirin Reinfarction Study to treatment with aspirin 160 mg/day, or warfarin (INR 2.8-4.2), or aspirin 75 mg/day plus warfarin (INR 2.0-2.5). The patients were screened for the occurrence of occult bleeding in faeces and urine after 3 months. Haemoglobin and iron metabolism parameters were measured at baseline, after 3 months, and at the end of the 4 years follow-up. RESULTS: The number of occult bleeding in faeces was 19 (7.1%) and in urine 29 (10.9%). There were no intergroup differences (p=0.45 and 0.39, respectively). In the occult bleeders, a second test showed 3 (1.1%) positive samples in faeces and 9 (3.4%) in urine. Further investigation revealed 2 cases of malignant disease. Haemoglobin and iron status variables were all within normal limits after 3 months and after 4 years in all treatment groups. CONCLUSIONS: Long-term treatment with aspirin, warfarin, or both, in the present doses and levels of anticoagulation did not lead to anemia or iron deficiency. The occurrence of occult bleeding in faeces and urine was a temporary phenomenon in most patients. Only macroscopic bleedings during these treatment modalities were of clinical importance, and screening for occult bleeding was of limit value.

BKV infection and hemorrhagic cystitis after allogeneic bone marrow transplant.

Hemorrhagic cystitis (HC) is a well-known complication after allogeneic bone marrow transplant (BMT) and can be related to adenovirus or human polyomavirus BK (BKV) infections. In this study a group of 20 patients after allogeneic BMT has been examined. BMT urine samples were analysed for the presence of Adenovirus and BKV DNAby means of polymerase chain reaction (PCR). 5/20 BMT patients developed HC after BMT. The presence of BKV DNA in urine samples was evident in 3/15 patients without HC and in 5/5 patients with HC. In 2/5 HC-patients the BKV DNA was not found after therapy with Cidofovir and Ribavirin. The search for adenovirus DNA in all samples was negative. The analysis of BKV non-coding control region (NCCR) isolated from urine samples revealed a structure very similar to the archetype in all samples. The RFLP (Restriction Fragment Length Polymorphism assay) showed the presence of BKV subtypes I and IV, with the prevalence of subtype I (4/5). This study supports the hypothesis that HC is mainly related to BKV rather than to adenovirus infection in BMT patients. Moreover, since BKV subtype I was predominant, it is reasonable to hypothesize that a specific BKV subtype could be associated with the development of HC.

[Evaluation of a new method for diagnosing the origin of urinary bleeding by the morphological characteristics of urinary red blood cells].

In this study, we attempted to develop a new method for diagnosing the origin of urinary bleeding by the morphological characteristics of urinary red blood cells (RBC). Seventy-five samples were divided into five types by individual features using phase-contrast microscopy. It was revealed that the ratios of type III, namely acanthocytes, and IV, namely donut-shaped RBC, were significantly higher in patients with glomerular bleeding than those with non-glomerular bleeding. Acanthocytes seemed to be specific to glomerular bleeding, but some urinary samples from patients with glomerular bleeding did not show acanthocytes. Therefore, we suggest that the detection of a combination of acanthocytes and donut-shaped RBC in a urine sample is useful for the diagnosis of glomerular bleeding.

Urine cytology in renal glomerular disease and value of G1 cell in the diagnosis of glomerular bleeding.

The objectives of the present study were to evaluate the cytology of urine sediments in patients with glomerular diseases, as well as the value of G1 dysmorphic erythrocytes (G1DE) or G1 cells in the detection of renal glomerular hematuria. Freshly voided urine samples from 174 patients with glomerular diseases were processed according to the method used for semiquantitative cytologic urinalysis. G1DEs (distorted erythrocytes with doughnut-like shape, target configuration with or without membranous protrusions or blebs), non-G1DEs (distorted erythrocytes without the above-mentioned morphologic changes), normal erythrocytes (NEs), and renal tubular cells (RTCs) were evaluated. Erythrocytic casts (ECs) were counted and graded as abundant (>1 per high-power field) or rare (1 per 5 high-power fields). G1DE/total erythrocyte ratios were calculated by counting 200 erythrocytes including G1DEs, non-G1DEs, and NEs. Only abundant NEs were found in 13 cases; abundant G1DEs, non-G1DEs, NEs, and no ECs in 95 cases; abundant NEs, non-G1DEs, and ECs and no G1DEs in 31 cases; and abundant NEs, G1DEs and non-G1DEs, and rare ECs in 35 cases. In 130 cases in which G1DEs were present, the G1DE/total erythrocyte ratios varied from 10% to 100%. This parameter was greater or equal to 80%, 50%, 20%, and 10% in 58 (44.6%), 29 (22.3%), 28 (21.5%), and 15 (11.5%) patients, respectively. In all cases, the number of RTCs was within normal limits or slightly increased, and a variable number of non-G1DEs were present in 161 cases. Thus, abundant ECs and/or G1DEs with a G1DE/total erythrocyte ratio of 10-100% proved to be specific urinary markers for renal glomerular diseases.

[Evaluation of dysmorphic red cells in the urinary sediment].

The usefulness of the morphologic examination of urinary red cells was first described by Birch and Fairley, who reported that the dysmorphic red cells(morphologically variable) were the marker for glomerular bleeding and the isomorphic red cells(morphologically uniform) for non-glomerular bleeding. They also noted that healthy individuals had dysmorphic cells, indicating a glomerular source. The relation between red blood cell morphology and the origin of hematuria has been confirmed subsequently by numerous clinical studies. Although the reports varied on the ratio(10-100%) of dysmorphic cells, their findings have been supported by many investigators. In 1991 Köhler noted that acanthocytes(AC) were the most characteristic red cell type for glomerular bleeding and that acanthocyturia > or = 5% was a good predictive marker for glomerular bleeding. Moreover the red cell was easily recognized. On the other hand, in 1993 Fairley and Birch reported that in glomerulonephritis, erythrocytes varied markedly in size, shape, and hemoglobin content, and that the pattern of morphology indicates the source of bleeding, not the morphology of individual cells. Both observations were confirmed in our in vitro experiments. To clarify the causative mechanism of acanthocyturia, the normal washed erythrocytes were successively exposed to two kinds of solutions simulating conditions in the distal tubules(105 mOsm, pH 5.5, NaCl 37 mmol/l, KH2PO4 1.0 mmol/l, Urea 0.9 g/l) and the collecting tubules(a mixture of NaCl and KH2PO4 solutions with the osmolality between 299 and 1192 mOsm). Various degrees of hemolytic process appeared in the first solution. However, AC appeared only in the second solutions with 9 mmol/l of KH2PO4 and 390 mOsm or more osmolality or with 725 mOsm and 4.1 mmol/l or more of KH2PO4. This result supported the nephron passage theory in the formation of AC in glomerular diseases.

Monitoring furosemide in racehorses participating in an EIPH program.

Analytical procedures were developed to monitor furosemide concentrations in post-race serum and urine samples obtained from horses participating in an exercise-induced pulmonary haemorrhage (EIPH) program. High performance liquid chromatography with ultraviolet light detection proved a reliable, sensitive method for measuring urinary furosemide concentrations up to 12 h after administration of either 150 or 250 mg of the drug to race horses. However, this method was unreliable for determination of serum furosemide concentration. High performance liquid chromatography with fluorescence detection proved a reliable, sensitive method for measuring serum furosemide concentration in horses administered 250 mg of the diuretic, permitting detection of approximately 5-10 ng/ml 6 h after treatment. This method was applied to field conditions where furosemide was administered to horses (between 150 and 250 mg intravenously) 4 h prior to the race. Analytical results assisted in establishing a threshold concentration of 85 ng/ml for serum furosemide. It was found that serum furosemide concentrations are a valid measure of compliance with furosemide administration in the EIPH program.