Frequent COL4 mutations in familial microhematuria accompanied by later-onset Alport nephropathy due to focal segmental glomerulosclerosis.

Familial microscopic hematuria (FMH) is associated with a genetically heterogeneous group of conditions including the collagen-IV nephropathies, the heritable C3/CFHR5 nephropathy and the glomerulopathy with fibronectin deposits. The clinical course varies widely, ranging from isolated benign familial hematuria to end-stage renal disease (ESRD) later in life. We investigated 24 families using next generation sequencing (NGS) for 5 genes: COL4A3, COL4A4, COL4A5, CFHR5 and FN1. In 17 families (71%), we found 15 pathogenic mutations in COL4A3/A4/A5, 9 of them novel. In 5 families patients inherited classical AS with hemizygous X-linked COL4A5 mutations. Even more patients developed later-onset Alport-related nephropathy having inherited heterozygous COL4A3/A4 mutations that cause thin basement membranes. Amongst 62 heterozygous or hemizygous patients, 8 (13%) reached ESRD, while 25% of patients with heterozygous COL4A3/A4 mutations, aged >50-years, reached ESRD. In conclusion, COL4A mutations comprise a frequent cause of FMH. Heterozygous COL4A3/A4 mutations predispose to renal function impairment, supporting that thin basement membrane nephropathy is not always benign. The molecular diagnosis is essential for differentiating the X-linked from the autosomal recessive and dominant inheritance. Finally, NGS technology is established as the gold standard for the diagnosis of FMH and associated collagen-IV glomerulopathies, frequently averting the need for invasive renal biopsies.

Renal Effects of SGLT-2 Inhibitors and Other Anti-diabetic Drugs: Clinical Relevance and Potential Risks.

Type 2 diabetes mellitus (T2DM) is a metabolic disease affecting an increasing percentage of general population worldwide. Patients with T2DM are frequently characterized by impaired renal function, primarily as a result of diabetic kidney injury, but also by other contributing factors, such as hypertension, atherosclerosis, and medications. Sodium-glucose cotransporter (SGLT)-2 inhibitors have emerged as a new, promising class of antidiabetic agents with actions that seem to extend beyond their hypoglycemic effect.

Relationship between adiposity, adipokines, inflammatory markers and lipid profile in hemodialysis patients.

OBJECTIVES: Our aim is to study the correlations of leptin and adiponectin with inflammation markers, body composition and lipid profile in end stage renal disease (ESRD) patients. PATIENTS AND METHODS: Phase angle values and fat mass as calculated using BIA, Malnutrition-Inflammation Score (MIS), leptin, adiponectin, IL-6, IL-8 triglycerides, cholesterol and other common serum markers' concentrations were analyzed using simple and multiple linear regression models in 47 hemodialysis patients. RESULTS: In contrast to leptin, adiponectin is inversely correlated to BMI and fat mass in hemodialysis patients. Triglycerides were the only parameter that retained its statistical correlation significance with adiponectin in the multiple regression model. CONCLUSIONS: Fat mass is of important consideration when calculating adipokines levels and their possible correlations with other variables. The inverse correlation of adiponectin with triglycerides levels should be further delineated due to the important role of vascular diseases in total mortality and morbidity of ESRD patients.

Refractory and massive lymphocele in a transplant patient with encapsulating peritoneal sclerosis treated with a single infusion of bleomycin: a case report.

A 26-year-old Caucasian man developed a large lymphocele after living-related kidney transplantation necessitating repeated drainage of large volumes every other day owing to ureteral compression. An open laparotomy for internal drainage was unsuccessful because of severe encapsulating peritoneal sclerosis. Prolonged external drainage with a catheter was inefficient. Repeated fine-needle aspirations of large volumes were needed to maintain ureteral patency over a period of 4 months. Finally, a single instillation of bleomycin immediately and effectively treated the lympocele with no relapse over the following 5 years. The presence of encapsulating peritoneal sclerosis seemed to be an obstacle to surgical treatment.

Early treatment with glucocorticoids or cyclophosphamide retains the slit diaphragm proteins nephrin and podocin in experimental lupus nephritis.

Renal podocytes and their slit diaphragms ensure the integrity of renal basement membrane and prevent urinary protein loss. We have previously reported that decreases of the podocyte slit diaphragm proteins nephrin and podocin represent early events in the podocytopathy of lupus nephritis (LN). We asked whether immunosuppressive agents such as glucocorticoids and cyclophosphamide may have direct effects on podocytes. We assessed in New Zealand Black/New Zealand White (NZB/W) F1 LN mice glomerular nephrin and podocin expression and localization by the use of Western blot and immunofluorescence; mRNA levels were measured by real-time polymerase chain reaction (PCR) and renal histology by light and electron microscopy. Early treatment with glucocorticoids and cyclophosphamide halted the histologic alterations associated with LN, preserving podocyte foot processes. Nephrin and podocin protein expression significantly increased in both glucocorticoid and cyclophosphamide groups as early as after three months of therapy. Real-time PCR revealed similar enhancement in nephrin and podocin mRNA levels after three to six months of treatment. This study documents that early treatment in experimental LN with glucocorticoids or cyclophosphamide preserves slit diaphragm proteins in podocytes and halts histological changes of the glomeruli, thus raising the possibility of a direct protective effect of these drugs on podocytes.

X-linked Alport syndrome in Hellenic families: phenotypic heterogeneity and mutations near interruptions of the collagen domain in COL4A5.

The X-linked Alport syndrome (ATS) is caused by mutations in COL4A5 and exhibits a widely variable expression. Usually ATS is heralded with continuous microhematuria which rapidly progresses to proteinuria, hypertension and chronic or end-stage renal disease (ESRD) by adolescence, frequently accompanied by sensorineural deafness and ocular complications. Milder forms of ATS also exist. We studied 42 patients (19M, 23F) of nine Hellenic families suspected clinically of X-linked ATS who presented with marked phenotypic heterogeneity. We identified mutations in COL4A5 in six families. Two males with nonsense mutation E228X reached ESRD by ages 14 and 18. Frameshift mutation 2946delT followed the same course with early onset renal involvement and deafness. However, two males with the milder missense mutation G624D, reached ESRD after 39 years and one patient showed thin basement membrane nephropathy (TBMN). Another 5/8 affected males with missense mutation P628L also developed ESRD between 30 and 57 years, while three exhibit only mild chronic renal failure (CRF). The data support previous findings that certain mutations are associated with milder phenotypes and confirm that mutation G624D may be expressed as TBMN with familial hematuria. Similar conclusions apply for missense mutation P628L. Interestingly, mutations G624D and P628L are near the 12th natural interruption of COL4A5 triple helical domain, which may explain the milder phenotype.

Podocyte main slit diaphragm proteins, nephrin and podocin, are affected at early stages of lupus nephritis and correlate with disease histology.

Renal podocytes and their slit diaphragms ensure the integrity of the renal basement membrane that forms the barrier to urinary protein loss. A putative disruption of the slit diaphragm and its main protein components, nephrin and podocin, may be implicated in the pathogenesis of lupus nephritis (LN). We studied the glomerular protein expression of nephrin and podocin in NZB/W LN mice by Western blot and immunofluorescence; mRNA levels were measured by real-time PCR. Human kidney biopsies of class II (n = 5), IV (n = 4), V (n = 7) LN were evaluated for nephrin expression by immunohistochemistry. Glomerular protein expression of nephrin and podocin were significantly reduced in NZB/W LN, starting from the earlier stages (mild mesangial LN) and becoming pronounced at advanced histological forms (focal and diffuse proliferative LN). Nephrin and podocin mRNA levels were substantially decreased in diffuse proliferative disease. Decreased expression of both proteins correlated with electron microscopy findings of distorted slit diaphragms. In patients with LN, nephrin was decreased particularly in diffuse proliferative LN. The main slit diaphragm proteins, nephrin and podocin, are affected from the earlier stages of LN and their expression correlates with disease histology. Our findings suggest a novel role of podocytes and their structures in immune-mediated nephritis.

The epidemiology of primary systemic vasculitides involving small vessels in Crete (southern Greece): a comparison of older versus younger adult patients.

BACKGROUND: The frequency of primary systemic small vessel vasculitides (PSV) varies among different geographic regions and age categories. We studied PSV in patients from middle-eastern Crete (Greece), and compared clinical characteristics in younger (<65 years) versus older (> or = 65 years) adult patients. METHODS: The records of 67 patients (33 younger, 34 older adults) diagnosed with PSV during 1995-2003 who were referred to a mixed secondary/tertiary care University Hospital in Crete were reviewed. Data on clinical manifestations, diagnosis, therapy, and adverse outcomes (end stage renal disease, death) during a median follow-up of 6 (range 0-12) years were recorded. Multivariate regression analysis was applied to identify independent predictors for adverse outcomes. RESULTS: The overall annual incidence of PSV was 19.5/million (95% confidence interval [CI] 15.7-23.4), 48.9/million (95% CI 33.8-63.9) in older and 12.4/million (95% CI 7.7-17) in younger adults. Microscopic polyangiitis was more prevalent in older patients (65%) and Wegener's Granulomatosis in younger patients (52%). Thirty-one percent of older patients developed end-stage renal disease as compared to 11% of younger patients (p=0.103). Mortality rates were 60% in older patients and 19% in younger patients (p=0.001). In multivariate regression analysis age (Beta=0.33 per 1-year, p=0.005), serum creatinine (Beta=0.29 per 1-mg/dL, p=0.011), and lung involvement (Beta=0.36, p=0.002) at the time of diagnosis were independent predictors for end stage renal disease and/or death. CONCLUSION: This study documents increased frequency and significant mortality of PSV among older people in Crete, with MPA being the most prevalent type. Age, serum creatinine, and lung involvement are important predictors for adverse outcome in these patients.

Cardiac manifestations of inborn errors of metabolism.

AIM: The aim of the study was to investigate the frequency and type of cardiac manifestations in a defined group of patients with inborn errors of metabolism. This paper also explores the key role of cardiac manifestations in the diagnosis of inborn errors of metabolism in daily practice. METHODS: Out of the 287 patients with the potential for inborn errors of metabolism who had been referred to the University Hospital of Heraklion (202 children and adolescents and 85 adults), 41 were found to have a variety of cardiac manifestations, including cardiomyopathy, cardiomegaly, atrioventricular conduction disorders and coronary artery disease. RESULTS: In 15 out of the 41 patients a diagnosis of inborn errors of metabolism was established, while the total number of patients with inborn errors of metabolism was 60 out of the 287. In 6 out of the 15 patients the major symptoms were from the cardiovascular system and 7 of them were adults with symptoms initiating in childhood. CONCLUSION: The cardiac findings consist of a neglected area in the diagnosis of the inborn errors of metabolism. Neurologists, pediatricians and internists should cooperate with cardiologists in managing people with unexplained cardiac symptoms and signs and be aware that several inborn errors of metabolism are associated with cardiac abnormalities and mild neurologic findings.

Impedance cardiography derived cardiac output in hemodialysis patients: a study of reproducibility and comparison with echocardiography.

BACKGROUND: Hemodialysis patients experience a variety of hemodynamic abnormalities that contribute to cardiovascular disease mortality which is the leading cause of death in these patients. Impedance cardiography has been utilized in order to monitor cardiac hemodynamics with lower cost and inconvenience, but it has not been appropriately validated in the hemodialysis population. AIM: We repeatedly used impedance cardiography to assess short- (48 hours) and long-term (15 days) reproducibility of cardiac output measurements and we compared baseline impedance cardiography measurements with echocardiographic measurements. PATIENTS AND METHODS: We studied 109 stable hemodialysis patients, aged 59.70 +/- 11.97 years being on hemodialysis for 67.59 +/- 40.15 months, on a non-dialysis day. Cardiac output was obtained with the BioZ impedance cardiography system (Cardiodynamics, San Diego, Ca, USA). Baseline echocardiography was performed using a Hewlett-Packard Sonos 2500 (Andover, Mass., USA). RESULTS: The values of impedance cardiography derived cardiac output were 5.28 +/- 0.79, 5.27 +/- 0.75 and 5.25 +/- 0.74 l/min at baseline (107 patients), 48 hours (107 patients) and 15 days (98 patients) respectively, showing high reproducibility. Bland and Altman analysis estimated that bias at 48 hours and at 15 days were: -0.013 (95% confidence intervals = -0.045 to 0.019) and 0.028, (95% confidence intervals = -0.044 to 0.101), respectively. In addition baseline impedance cardiography derived cardiac output was significantly correlated with the echocardiographic derived cardiac output (r = 0.9, p < 0.0001). CONCLUSION: Impedance cardiography is a simple non invasive technique for cardiac output estimation in hemodialysis patients which has high reproducibility when performed under controlled conditions, and is closely correlated with echocardiographic measurements of cardiac output.

Alteration of proximal aorta biophysical properties in patients with end stage renal disease.

OBJECTIVE: To present a novel, non-invasive echocardiographic application to assess the structural and functional properties of the complex composition of the proximal aorta in patients with end stage renal disease (ESRD). METHODS: 71 haemodialysis patients (mean (SD) age 61.3 (9.3) years, dialysis duration 79.2 (51.6) months) and 62 age matched controls were studied. From the suprasternal view, the distance between ascending and descending aorta was measured with two dimensional ultrasound. The aortic flow wave transit time was measured with pulsed wave Doppler. M mode echocardiography, with simultaneous blood pressure estimates, was used to assess the diameters of the aortic annulus and of the ascending aorta. Pulse pressure, pulse wave velocity (PWV), pressure strain elastic modulus, characteristic impedance, and beta index were calculated. RESULTS: Patients had increased pulse pressure (68.0 (7.2) v 51.4 (5.0) mm Hg, p < 0.001), PWV (6.1 (1.1) v 3.9 (0.6) m/s, p < 0.001), characteristic impedance (174 (58) v 111 (31) m/s.cm2, p < 0.001), pressure strain elastic modulus (872 (254) v 541 (140) mm Hg, p < 0.001), and beta index (8.9 (3.4) v 5.5 (1.4), p < 0.001) compared with controls. In patients PWV was correlated with age and time on haemodialysis (r = 0.44, p < 0.001, and r = 0.51, p < 0.001, respectively). CONCLUSION: A novel application of duplex ultrasound of the proximal aorta showed that patients with ESRD have impaired proximal aortic function compared with controls. The data indicate that these non-invasive measurements can be used to describe status and change in aortic biophysical properties and may be used as a marker for cardiovascular disease risk.

Evaluation of ciprofloxacin in the treatment of Pseudomonas aeruginosa infections.

The efficacy and safety of ciprofloxacin in the treatment of Pseudomonas aeruginosa infections was evaluated in 72 patients suffering from upper urinary tract infection (19 patients), deep soft tissue infection (16), chronic osteomyelitis (12), abscess (7), chronic otitis media (6), otitis externa (3) and bronchopneumonia (9). Forty-eight patients received an oral dose of 500 mg or 750 mg b.i.d. and five patients an i.v. dose of 200 mg b.i.d., while 19 patients were given both oral and parenteral doses. The duration of therapy ranged from seven days to more than four months. The MICs of ciprofloxacin for the Pseudomonas aeruginosa strains isolated were in the range less than 0.06-2 mg/l; 36% of the strains were resistant to all other available antibiotics. At follow-up after a minimum of six months the clinical success rate was 75% and the infecting organism was permanently eradicated in 49% of the patients. In nine patients the organism developed resistance, particularly when the initial MIC was higher than 0.5 mg/l. No significant adverse reactions were observed. Ciprofloxacin is the first antipseudomonal antimicrobial agent which can be administered orally and therefore fulfills a need in chemotherapy.

Experience with ciprofloxacin in the treatment of various infections caused mainly by Pseudomonas aeruginosa.

Ciprofloxacin, a new quinolone carboxylic acid derivative with an enhanced spectrum of activity including P. aeruginosa, was given to 42 patients, 26 males and 16 females, ranging in age from 12 to 75 years. They were suffering from upper urinary tract infection (15), abscesses (hepatic 1, intra-abdominal 5, retroperitoneal 1, soft tissue, deep soft tissue infection (9), chronic otitis media in exacerbation (3), chronic osteomyelitis in exacerbation (3), bronchopneumonia (3) and otitis externa (1). Pathogens included P. aeruginosa (29), E. cloacae (7), P. mirabilis, E. coli (3) and S. marcescens (1), with MICs for ciprofloxacin ranging from 0.003-2 micrograms/ml. Over half of the isolates were multiresistant, also to amikacin, with almost all the Pseudomonas strains resistant to carbenicillin and the ureidopenicillins. In 21 patients ciprofloxacin was given orally at a dose of 500 mg or 750 mg 12-hourly, in 5 patients i.v. at a dose of 200 mg 12-hourly; while in 16 patients treatment was started i.v. and was continued by the oral route. Seventeen patients were given ciprofloxacin for 7-14 days, 5 for 15-22 days, 4 for 23-28 days, 8 for 29-42 days, and 8 for greater than 42 days. Treatment response was considered clinically as cure in 30 (71.5%) patients, improvement in 8 (19%) and failed in 4 (9.5%). Pathogens were eradicated during treatment in 34 (81%), persisted in 8 (19%) and recurred in 11 (26.2%) patients. Development of resistance was observed in one patient only. Adverse reactions in 12 patients were minimal and self-limited. It was concluded that ciprofloxacin is a very promising new antimicrobial which merits further clinical trials in systemic infections.