Serum glomerular permeability activity in patients with podocin mutations (NPHS2) and steroid-resistant nephrotic syndrome

A plasma factor displaying permeability activity in vitro and possibly determining proteinuria has been hypothesized in idiopathic focal segmental glomerulosclerosis (FSGS). In vitro permeability activity (P(alb)) was determined in sera of five patients with autosomal recessive steroid-resistant nephrotic syndrome (NPHS2), an inherited condition indistinguishable from idiopathic FSGS on clinical grounds, but in which proteinuria is determined by homozygous mutations of podocin, a key component of the glomerular podocyte. All patients had presented intractable proteinuria with nephrotic syndrome; four developed renal failure and received a renal allograft. For comparison, sera from 31 children with nephrotic syndrome were tested. Pretransplant P(alb) was high in all cases (mean 0.81 +/- 0.06), equivalent to levels observed in idiopathic FSGS. Overall, P(alb) did not correlate with proteinuria. The posttransplant outcome was complicated in two patients by recurrence of proteinuria after 10 and 300 d, respectively, that responded to plasmapheresis plus cyclophosphamide. P(alb) levels were high at the time of the recurrence episodes and steadily decreased after plasmapheresis, to reach normal levels in the absence of proteinuria after the seventh cycle. In an attempt to explain high P(alb) in these patients, putative inhibitors of the permeability activity were studied. Coincubation of serum with homologous nephrotic urine reduced P(alb) to 0, whereas normal urine did not determine any change, which suggests loss of inhibitory substances in nephrotic urine. The urinary levels of the serum P(alb) inhibitors apo J and apo E were negligible in all cases, thus suggesting that other urinary inhibitors were responsible for the neutralizing effect. These data indicate that P(alb) is high in NPHS2, probably resulting from loss of inhibitors in urine. Lack of correlation of P(alb) with proteinuria suggests a selective loss of inhibitors. As in idiopathic FSGS, proteinuria may also recur after renal transplantation in NPHS2 patients, and post-transplant proteinuria is associated with high P(alb). The relationship between elevated P(alb) and proteinuria in NPHS2 remains to be determined.     

Renal and cardiovascular protection in type 2 diabetes mellitus: angiotensin II receptor blockers

Aggressive treatment of hypertension is effective in reducing both microvascular and macrovascular complications in type 2 diabetes, and target BP less than 130/85 or 130/80 mmHg are now recommended. Inhibition of renin angiotensin aldosterone system (RAAS) plays an essential role in the treatment of hypertension and diabetes-related complications. Studies focusing on renal end-points suggest that angiotensin-converting enzyme inhibitors (ACE-I) are more effective than other traditional agents in reducing the onset of clinical proteinuria in both type 1 and type 2 diabetic patients with incipient nephropathy, mainly in normotensive ones (secondary prevention). However, several small trials in type 2 diabetic patients with overt nephropathy (tertiary prevention) failed to demonstrate a specific renoprotective role for ACE-I, at variance with type 1 diabetes. Three recent large trials address the question of whether angiotensin II receptor blockers (ARB) prevent the development of clinical proteinuria or delay the progression of nephropathy in type 2 diabetes. The IRMA study showed that irbesartan is more effective than conventional therapy in preventing the development of clinical proteinuria and in favoring the regression to normoalbuminuria for comparable BP control in patients with incipient nephropathy. The IDNT and RENAAL trials showed that ARB are more effective than traditional antihypertensive therapies in reducing progression toward end-stage renal failure (ESRF) in type 2 diabetic patients with overt nephropathy independently of changes in BP. Moreover, a reduction in hospitalizations for heart failure was demonstrated for ARB-treated patients compared with placebo. Furthermore, the LIFE study showed that losartan is more effective than conventional therapy in reducing cardiovascular morbidity and mortality in a cohort of diabetic patients with hypertension and left ventricular hypertrophy. In conclusion, ARB seem to be effective in both preventing renal damage and reducing progression toward ESRF in type 2 diabetic patients. Thus, the guidelines for the prevention and treatment of diabetic nephropathy are now changed. In type 1 diabetes ACE-I are the first-choice drug; in type 2 diabetes, ARB are considered first-choice drugs in secondary prevention as well as ACE-I and have been now elected the unique first-choice drug in tertiary prevention of ESRF. Finally, ARB should be considered as the first-choice drug in cardiovascular prevention too, as well as ACE-I.     

Tumor angiogenesis in lymph node-negative rectal cancer: Correlation with clinicopathological parameters and prognosis

BACKGROUND: Intratumoral microvessel density (MVD) could be used as a prognostic factor in colorectal cancer. We retrospectively analyzed the value of microvessel count in predicting the clinical outcome of stage I and II (Dukes A and B) rectal cancer patients. METHODS: Eighty-four patients who had undergone curative resection of lymph node-negative rectal cancer were included. Tumor type and differentiation, the depth of local invasion, venous invasion, the character of the invasive margin, and the degree of lymphocytic infiltration were evaluated for each tumor specimen. Immunohistochemical staining for the CD31 endothelial antigen was performed to highlight the microvessels. RESULTS: The median value of MVD was 45 microvessels. Low MVD (microvessels < or = 45) was observed in 41 patients (48.8%), and high MVD (>45) was found in 43 (51.2%). The presence of conspicuous lymphocytic infiltration was significantly associated with increased vessel density. With uni- and multivariate survival analysis MVD did not show any prognostic significance. The character of the invasive margin was the only parameter with independent prognostic value. CONCLUSIONS: MVD does not seem to provide any additional prognostic information when compared with standard histopathological parameters in lymph node-negative rectal cancer. It is likely that the strong association between MVD and the presence of conspicuous lymphocytic infiltration may interfere with its predictive value.     

Toward refinement of the colony-forming unit-granulocyte/macrophage clonogenic assay: inclusion of a metabolic system.

This work represents a first attempt to refine the colony-forming unit-granulocyte/macrophage (CFU-GM) clonogenic assay by incorporating liver microsomes and co-factors as a metabolic system into the in vitro test system in response to an ECVAM recommendation. From the comparison of results obtained with the CFU-GM clonogenic assay currently used and with the new experimental protocol, different toxicity on granulocyte/macrophage precursors was demonstrated, when drugs with a known metabolism in vivo were tested.     

Intracerebroventricular injection of prostaglandin E(1) changes concentrations of biogenic amines in the posterior hypothalamus of the rat

Since the posterior hypothalamus (PH) plays a key role in the control of body temperature, the aim of this study was to evaluate the changes in adrenaline, noradrenaline and dopamine levels in the PH during the hyperthermia induced by prostaglandin E(1) (PGE(1)). The concentration of adrenaline, noradrenaline and dopamine in the PH, the firing rate of the sympathetic nerves innervating interscapular brown adipose tissue (IBAT), IBAT and colonic temperatures (T(IBAT) and T(C)) were monitored in 12 urethane-anaesthetized male Sprague-Dawley rats before and after an intracerebroventricular injection of 500 ng PGE(1) dissolved in 2 microl of 0.9% NaCl saline solution or only saline. The catecholamines were collected using a microdialysis probe and quantified by HPLC. The results showed that PGE(1) caused a significant increment in the concentration of adrenaline from 15. 83+/-2.69 to 34.95+/-3.9 ng ml(-1) and of dopamine from 35.15+/-4.48 to 55.68+/-6.21 ng ml(-1). A significant decrease in the level of noradrenaline from 18.75+/-2.05 to 8.56+/-2.26 ng ml(-1) was registered. The firing rate of sympathetic nerves to IBAT was increased from 100+/-0% to 204.83+/-15.22% by PGE(1). T(IBAT) and T(C) rose respectively from 36.91+/-0.15 degrees C to 38.88+/-0.29 degrees C, and from 36.7+/-0.15 degrees C to 38.13+/-0.36 degrees C after the injection of PGE(1). The changes in adrenaline and noradrenaline occurred during the first 20 min as did the changes in temperature and firing rate, while the change in dopamine was delayed until 21-60 min after the PGE(1) injection. No significant change of analyzed variables was found in the control rats. These findings suggest that these biogenic amines of the PH are involved in the control of the sympathetic and thermogenic changes induced by PGE(1).     

Subcellular localization of β-catenin and APC proteins in colorectal preneoplastic and neoplastic lesions

Adenomatous polyposis coli (APC) is a tumor suppressor gene whose main function is the destabilization of β-catenin, a key effector of the Wnt signaling pathway. This gene is defective in familial adenomatous polyposis (FAP), a dominantly inherited disease, but inactivation of APC has been reported also in most sporadic colorectal tumors and it is considered an early event in colorectal tumorigenesis. The aim of the present study was to evaluate the intracellular ultrastructural distribution of β-catenin and APC proteins in epithelial cells of normal colorectal mucosa, aberrant crypt foci (ACF, an early premalignant lesion) and cancer. We used the immunogold electron microscopic method to identify both proteins. Normal colonic epithelial cells showed a strong membranous expression of β-catenin and lacked cytoplasmic and nuclear expression. Normal cells showed APC localization pattern characterized by diffuse nuclear expression and along the plasma membrane. In ACF and in carcinoma an absent or reduced membranous expression of β-catenin was associated with an increased nuclear and cytoplasmatic expression. In aberrant crypt foci and carcinoma, APC was evident inside the nucleus and at the level of cell-cell junctions, but it was decreased in the cytoplasm. This method allowed the accurate localization of proteins of the Wnt signaling pathway in the early steps of colorectal carcinogenesis. The similar pattern of subcellular distribution of APC and β-catenin in dysplastic ACF and colorectal cancer suggests that ACF are precursor lesions of sporadic and FAP-associated colorectal carcinoma.     

Loss of pRb2/p130 expression is associated with unfavorable clinical outcome in lung cancer.

Altered expression of cell cycle regulators represents a frequent event in both small cell and non-small cell lung cancer (NSCLC). Despite several studies that reported involvement of tumor suppressor genes, such as p53 and pRb, in the development and progression of lung cancer, contrasting opinions exist about the prognostic role of this protein in this neoplasm. We developed an immunohistochemical assay suitable for the detection of pRb2/p130, the last discovered member of the retinoblastoma gene family, on formalin-fixed and paraffin-embedded sections. We evaluated the immunohistochemical expression of pRb2/p130 in 135 lung cancer specimens, and performed Western blot analysis in a subset of 30 corresponding tumor lysates. A high correlation between immunohistochemical data and Western blot results (P = 0.0004) was found. We statistically analyzed the relationship between overall survival (OS) time and pRb2/p130 expression according to the different histological types in 105 patients. We did not find any correlation between pRb2/p130 expression and OS in small cell lung cancers, whereas in NSCLCs a direct relationship between pRb2 and OS was found in both adenocarcinoma (P = 0.0002) and squamous cell carcinoma (P = 0.0002) histotypes. According to univariate analysis, pRb2/p130 was a prognostic factor of which the lost or reduced expression correlated with a shorter OS (P < 0.0000). At multivariate analysis, pRb2/p130 expression was an independent predictor of OS (P = 0.0001) when considered together with histotype. This study demonstrates for the first time the potential independent prognostic value of pRb2/p130 expression on formalin-fixed, paraffin-embedded sections from lung cancer patients. pRb2/p130 immunoreactivity can be used to predict OS in patients with NSCLC and, therefore, may represent a new prognostic marker.     

Internal jugular vein patency after lateral neck dissection: a prospective study

In spite of anatomical preservation of the internal jugular vein (IJV), an occlusion rate of the vessel of up to 30% has been documented after selective or modified radical neck dissections. The aim of the present prospective study was to evaluate the patency of the IJV following selective lateral neck dissection (LND) in 34 patients affected by squamous cell carcinoma of the upper aerodigestive tract who underwent surgery concomitantly on the primary site and the neck. Eighteen patients received unilateral and 16 bilateral LND, for a total of 50 IJVs. Postoperative radiotherapy on the neck was delivered in four patients with histologic evidence of micro-extracapsular spread; the impact of this variable on IJV patency was assessed by the Fisher test. A preoperative baseline study of vein patency and flow by ultrasonography (US) was obtained. Postoperative controls were scheduled at 1 week, 1 month and 3 months following surgery. No patient developed either wound infection or a pharyngocutaneous fistula, and no signs or symptoms of IJV occlusion were observed during the postoperative course. At the first US control, 25 IJVs (50%) did not present any alteration in patency, and 23 (46%) and 2 (4%) had a reduced or absent flow, respectively. At the second and third controls, 33 (66%) and 45 (90%) of the IJVs presented with normal flow, respectively. At the end of the study, none of the patients showed evidence of occlusion. Postoperative radiotherapy did not have a statistically significant impact on IJV patency ( P=0.09). In conclusion, long-term IJV occlusion after LND has to be considered an exceedingly rare event with negligible morbidity. However, alterations of IJV flow frequently occur in the immediate postoperative course.