Effects of maternal diabetes on male offspring: high cell proliferation and increased activity of MMP-2 in the ventral prostate.

This study presents a comprehensive view of the histological and functional status of the prostate of adult rat offspring of mothers subjected to gestational diabetes induced by alloxan. The ventral prostate of male adult offspring of diabetic (DP) or normal (CP) mothers was evaluated for collagen fibres, cell death, fibroblasts, smooth muscle cells, cell proliferation, matrix metalloproteinases (MMPs), androgen receptors (AR), transforming growth factor ß1 (TGFß-1), catalase and total antioxidant activity. The prostates of DP animals were lower in weight than those of the CP group. The DP group also exhibited hyperglycaemia and hypotestosteronemia, higher cell proliferation and AR expression, a reduction in α-actin (possibly interfering with the reproductive function of the prostate), and enhanced activity of MMP-2, although the absolute content of MMP-2 was lower in this group. These findings were associated with increased TGFß-1 and decreased collagen distribution. The prostates of DP rats additionally exhibited reductions in catalase and total antioxidant activity. Thus, rats developing in a diabetic intrauterine environment have glycaemic and hormonal changes that impact on the structure and physiology of the prostate in adulthood. The increased AR expression possibly leads to elevated cell proliferation. Stromal remodelling was characterized by enhanced activity of MMP-2 and collagen degradation, even with increased TGFß-1 activation. These changes associated with increased oxidative stress might interfere with tissue architecture and glandular homeostasis.

High volume peritoneal dialysis vs daily hemodialysis: a randomized, controlled trial in patients with acute kidney injury.

There is no consensus in the literature on the best renal replacement therapy (RRT) in acute kidney injury (AKI), with both hemodialysis (HD) and peritoneal dialysis (PD) being used as AKI therapy. However, there are concerns about the inadequacy of PD as well as about the intermittency of HD complicated by hemodynamic instability. Recently, continuous replacement renal therapy (CRRT) have become the most commonly used dialysis method for AKI around the world. A prospective randomized controlled trial was performed to compare the effect of high volume peritoneal dialysis (HVPD) with daily hemodialysis (DHD) on AKI patient survival. A total of 120 patients with acute tubular necrosis (ATN) were assigned to HVPD or DHD in a tertiary-care university hospital. The primary end points were hospital survival rate and renal function recovery, with metabolic control as the secondary end point. Sixty patients were treated with HVPD and 60 with DHD. The HVPD and DHD groups were similar for age (64.2+/-19.8 and 62.5+/-21.2 years), gender (male: 72 and 66%), sepsis (42 and 47%), hemodynamic instability (61 and 63%), severity of AKI (Acute Tubular Necrosis-Index Specific Score (ATN-ISS): 0.68+/-0.2 and 0.66+/-0.2), Acute Physiology, Age, and Chronic Health Evaluation Score (APACHE II) (26.9+/-8.9 and 24.1+/-8.2), pre-dialysis BUN (116.4+/-33.6 and 112.6+/-36.8 mg per 100 ml), and creatinine (5.8+/-1.9 and 5.9+/-1.4 mg per 100 ml). Weekly delivered Kt/V was 3.6+/-0.6 in HVPD and 4.7+/-0.6 in DHD (P<0.01). Metabolic control, mortality rate (58 and 53%), and renal function recovery (28 and 26%) were similar in both groups, whereas HVPD was associated with a significantly shorter time to the recovery of renal function. In conclusion, HVPD and DHD can be considered as alternative forms of RRT in AKI.

Diabetes and pregnancy in Wistar rats: renal effects for mothers in the postpartum period.

In this study, diabetes mellitus (DM) was induced in Wistar rats during pregnancy and maintained in the postpartum period (PP) and we evaluated systolic blood pressure (SBP), glomerular filtration rate (GFR) and renal immunohistochemical and morphometric studies from different groups: G1 (non-pregnant control rats), G2 (non-pregnant diabetic rats), G3 (control mothers) and G4 (diabetic mothers). We found that there were no differences in relation to SBP, but there was a tendency for reduction in GFR from G4 compared with the other groups (G). There was increased total kidney weight/body weight ratio of G4 compared with other G. There were increase in glomerular tuft area in G3 and G4 compared with G1 and G2. G2 and G4 showed even higher percentage of cortical collagen. G3 showed increased glomerular proliferating cells compared with G1 and G2, while in G4 this number was smaller than G3. Cell proliferation was higher in the tubulointerstitial (TBI) compartment from G4. Glomerular and TBI α-smooth muscle actin expression was increased in G4 compared with other G. The glomerular p-p38 expression showed a pattern similar to proliferation cell nuclear antigen, with a reduction of p-p38 in G4 relative to other G. The immunoreactivity of p-JNK was higher in both the glomeruli and TBI compartment in G4 compared with G1, G2 and G3. The DM induced during pregnancy and maintained in the PP resulted in renal structural and functional changes to mothers. In addition, altered mitogen-activated protein kinase expression in association with these changes may play an important role in renal damage observed in the present investigation.

Low caloric and protein intake is associated with mortality in patients with acute kidney injury.

BACKGROUND: Acute renal injury (AKI) interferes greatly with nutritional status, affecting the metabolism of all macronutrients and increased mortality rates in hospitalized patients. Our objective was to evaluate the association of nutritional parameters (albumin, cholesterol, caloric and protein intake and nitrogen balance (NB)) with mortality in patients with AKI. METHODS: This is a prospective observational study that evaluated 595 consecutive patients over the age of 18 years with AKI, requiring enteral or parenteral feeding. At the time of the patient's enrollment, demographic and laboratorial data, caloric and protein supply and NB were recorded on the first day of referral to the nephrologist. All patients were followed throughout the hospital stay and mortality rate was also recorded. RESULTS: The medium age of patients with AKI was 64 (54-75) years, 64.5% male, 62% admitted to intensive care unit (ICU), 52% on dialysis and the majority (48%) were at stage 3 by AKIN. Length of stay and hospital mortality were 18 (10-31) days and 46%, respectively. Superior age, AKI severity, lower body weight and body mass index (BMI), higher need for dialysis, ICU admission and shorter hospital stay were associated with higher mortality. At logistic regression, caloric (OR: 0.946; CI:95%: 0.901-0.994; p:0.029) and protein intake (OR: 0.947; CI:95%: 0.988-0.992; p = 0.028) and serum albumin (OR: 0.545; CI:95%: 0.401-0741; p < 0.001) were associated with hospital mortality. Cholesterol (OR: 0.995; CI:95%: 0.991-1.000; p = 0.052) was not associated with increased mortality in the adjusted analysis. Analysis of the receiver operating characteristic (ROC) curve showed that calorie intake < 12 kcal/kg (AUC: 0.745; CI:95%: 0.684-0.765; p < 0.001) and protein intake < 0.5 g/kg (AUC: 0.726; CI:95%: 0.686-0.767; p < 0.001) were predictors of hospital mortality, as well as a negative NB < -6.47 g N/day (AUC: 0.745; CI:95%: 0.704-0.786; p < 0.001). CONCLUSIONS: In conclusion, low caloric and protein intake, negative NB and low albumin value are conditions associated with higher hospital mortality in patients with AKI.

Handgrip strength and weight predict long-term mortality in acute kidney injury patients.

INTRODUCTION: Surviving acute kidney (AKI) patients have a higher late mortality compared with those admitted without AKI. The negative impact of malnutrition on the early outcome of AKI patients has recently been confirmed by various studies. However, its impact after hospital discharge has not been studied. The objective of the study was to determine the role of anthropometric measurements and handgrip strength as predictors of mortality 180 days after discharge. METHODOLOGY: Eighty-two survivors AKI patients who were older than 18 y old and followed by AKI team were prospectively evaluated. Patient's characteristics were recorded, anthropometric measurements were taken, handgrip strength (HGS) was measured, subjective global assessment and bioimpedance were applied and blood samples were collected during hospitalization at first and last nephrologist evaluation and in after hospital discharge at 1 month, 3 and 6 months. Multivariable logistic regression was used to adjust confounding and selection bias. RESULTS: Age was 62.3 ± 14.7 years, prevalence of hospitalization in medical wards of 71.6%, index of severity of AKI (ATN-ISS) was 28% and late mortality rates was 25.6%. Risk factors associated with late mortality were the number of comorbidities (HR = 1.79, 95% CI = 1.45-2.46, p = 0.04), cancer (HR = 1.89, 95 CI% = 1.48-3.16, p = 0.01), sepsis (HR = 1.47, 95% CI = 1.18-2.38, p = 0.03), no recovery of renal function at hospital discharge (HR = 1.46, 95% CI = 1.02-2.16, p = 0.03), malnutrition at first evaluation (HR = 1.58, 95% CI = 1.14-2.94, p = 001), the HGS value at the moment of last evaluation by nephrologist (HR = 1.81, 95% CI = 1.17-2.31, p = 0.04) and gain weigh < 1 kg between the moment at first evaluation by nephrologist and one month after hospital discharge (HR = 1.95, 95 CI% = 1.29-3.3, p = 0.02). CONCLUSION: HGS and gain weight were identified as predictors of late mortality. Simple and ease methods can be applied in AKI patients during and after hospitalization to diagnose nutritionally patients who are at higher risk for poor prognosis and, consequently intervention measures can be performed to improve survival in long-term.

Increased expression of p38 mitogen-activated protein kinase is related to the acute renal lesions induced by gentamicin.

Mitogen-activated protein kinases (MAPK) may be involved in the pathogenesis of acute renal failure. This study investigated the expression of p-p38 MAPK and nuclear factor kappa B (NF-kappaB) in the renal cortex of rats treated with gentamicin. Twenty rats were injected with gentamicin, 40 mg/kg, i.m., twice a day for 9 days, 20 with gentamicin + pyrrolidine dithiocarbamate (PDTC, an NF-kappaB inhibitor), 14 with 0.15 M NaCl, i.m., twice a day for 9 days, and 14 with 0.15 M NaCl , i.m., twice a day for 9 days and PDTC, 50 mg kg(-1) day(-1), i.p., twice a day for 15 days. The animals were killed 5 and 30 days after the last of the injections and the kidneys were removed for histological, immunohistochemical and Western blot analysis and for nitrate determination. The results of the immunohistochemical study were evaluated by counting the p-p38 MAPK-positive cells per area of renal cortex measuring 0.05 mm2. Creatinine was measured by the Jaffé method in blood samples collected 5 and 30 days after the end of the treatments. Gentamicin-treated rats presented a transitory increase in plasma creatinine levels. In addition, animals killed 5 days after the end of gentamicin treatment presented acute tubular necrosis and increased nitrate levels in the renal cortex. Increased expression of p-p38 MAPK and NF-kappaB was also observed in the kidneys from these animals. The animals killed 30 days after gentamicin treatment showed residual areas of interstitial fibrosis in the renal cortex, although the expression of p-p38 MAPK in their kidneys did not differ from control. Treatment with PDTC reduced the functional and structural changes induced by gentamicin as well as the expression of p-p38 MAPK and NF-kappaB. The increased expression of p-p38 MAPK and NF-kappaB observed in these rats suggests that these signaling molecules may be involved in the pathogenesis of tubulointerstitial nephritis induced by gentamicin.

A novel dimeric fluoropyrimidine molecule behaves as a remote precursor of 5-fluoro-2'-deoxyuridine in human erythrocytes.

A new dimeric fluoropyrimidine molecule (5-fluoro-2'-deoxyuridilyl-(5'-->3')-5-fluoro-2'-deoxy-5'-uridylic acid, Compound 1) was chemically synthesized from two separately deblocked 5-fluoro-2'-deoxyuridine mononucleotide moieties. Other structurally related nucleotides, 5-fluoro-2'-deoxyuridine-5'-diphosphate (FdUDP), 5-fluoro-2'-deoxyuridine-5'-triphosphate (FdUTP) and 5-fluoro-2'-deoxyuridine-3',5'-bisphosphate were also synthesized. The structures of all synthesized molecules were verified by mass spectrometric analyses and were consistent with expected molecular mass values. The metabolic patterns of conversion of Compound 1 were investigated both in human erythrocyte lysates and in intact erythrocytes previously loaded with this molecule according to a highly conservative encapsulation procedure. In hemolysates, Compound 1 was transformed to 5-fluoro-2'-deoxyuridine (FUdR) and to 5-fluorouracil (FU) through the intermediate formation of 5-fluoro-2'-deoxyuridine-5'-monophosphate (FdUMP). In intact red cells, Compound 1 still generated FUdR (and to a lesser extent FU), that was then released outside. The conversion pathway involves a phosphodiesterase-catalysed hydrolysis of Compound 1 into two FdUMP molecules, followed by further dephosphorylation to FUdR and by partial conversion to FU. Unlike hemolysates, Compound 1-loaded intact erythrocytes featured transient formation of FdUDP and FdUTP, both metabolites representing storage compounds for the final and sustained production of FUdR and FU. Therefore, human erythrocytes can behave as bioreactors ensuring the time-controlled production and delivery of the two powerful antitumor drugs FUdR and FU from encapsulated Compound 1. This new molecule and other compounds as well (e.g. FdUDP and FdUTP) can be viewed as useful pre-prodrugs, exploitable for intraerythrocytic bioconversion reactions.

Renal artery clipping attenuates the progression of adriamycin nephropathy.

This study was designed to analyze the impact of diminished renal perfusion pressure due to renal clipping on the rat model of adriamycin nephropathy. Male Wistar rats, divided into four groups (n = 9 per group) were injected with saline as control (C), adriamycin 3 ml/kg (Ad), saline with the left renal artery clipped (Rv), and adriamycin plus clip (AdRv). After 24 weeks mean arterial pressure (MAP), inulin, and p-aminohippurate (PAH) clearances were performed to evaluate renal function. Morphologic analysis included histologic criteria of percentage of glomerulosclerosis and tubulointerstitial lesion index (TILI). The MAP (mm Hg) was similar between Rv (143+/-13) and AdRv (154+/-20), but higher (P < .05) than C (120 +/-8) and Ad (124+/-11). Inulin clearance (mL/min/ 100 g) in Ad (0.2+/-0.05) was smaller than in C (0.53+/-0.17) and Rv (0.4+/-0.01) (P < .05), and was at an intermediate level in AdRv (0.33+/-0.2). The level of PAH (mL/min/100 g) was normal at 1.76 in C, and diminished more in Ad (0.58) than in Rv (1.06) and AdRv (1.18) (P < .05). Both Ad and the AdRv nonclipped kidneys had the highest degree of glomerulosclerosis (33% and 25%) and TILI (7% and 8%), respectively, compared with C and Rv (both 0%), whereas the clipped kidneys displayed intermediate degrees (9% and 5%) (P < .05 v nonclipped). The data suggest that diminished perfusion pressure of the clipped kidney, by decreasing the intraglomerular pressure, protects the glomerulus from damage and attenuates the evolution of adriamycin nephropathy.

Association between hypervolemia and ventricular hypertrophy in hemodialysis patients.

BACKGROUND: Left ventricular hypertrophy (LVH) is a well-known predictor of cardiovascular mortality in patients who have end-stage renal disease and are maintained on hemodialysis (HD), and LVH is not always correlated with the severity of hypertension in these patients. The purpose of this study was to investigate the role of other factors contributing to LVH. METHODS: A total of 50 patients with HD were classified in three groups according to whether their LV mass index (LVMI) was higher than (n = 15), equal to (n = 20), or lower than (n = 15) that predicted by a formula based on their ambulatory blood pressure monitoring (ABPM). RESULTS: Subjects with higher LVMI than predicted had significantly greater inter-HD weight gain (3.4 +/- 0.8 v 2.7 +/- 0.8 and 2.6 +/- 05 kg, respectively, in the other two groups, P < .05), and subjects with lower LVMI than predicted had a tendency toward a more pronounced nocturnal dipping pattern of BP (P = .07 v the other two groups), although daytime and night-time average BP levels did not differ between groups. All other clinical and laboratory parameters were similar among the three groups except higher cardiac output and various indices of LVH, which were more pronounced in the group with higher LVMI by ABPM. This group had also the lowest survival rate over the 2 to 3 years of follow-up, with five deaths versus two in each of the other two groups. CONCLUSIONS: The data suggest that correct management of inter-HD weight gain by nutritional counseling and shorter inter-HD intervals may prevent LVH and improve survival independently of BP control.

Synthesis of ionones and carvone analogues: olfactory properties and preliminary toxicity assays.

Vilsmeier reagents react with alpha/beta-ionones and carvone to produce aldehydes 7-11 in a one-step procedure. The indene derivative 11, which came from the double iminoalkylation of carvone and ring closure with the elimination of dimethylamine, was practically odourless, while all the others had peculiar odours which were very different from the starting material. The cytotoxicity data of 9 and 10, which are the most promising potential perfume ingredients, are also reported.

Simultaneous HPLC determination of multiple components in a commercial cosmetic cream.

A high-performance liquid chromatographic method for the simultaneous determination of magnesium ascorbyl phosphate (I), imidazolidinylurea (II), a mixture of methyl-(III), ethyl-(IV), propyl-(V), butyl-(VI) parabens dissolved in phenoxyethanol, and ascorbyl palmitate (VII), was studied by using a cyano-propyl column and a methanol gradient at 220 and 240 nm. Calibration curves were found to be linear in the 0.05-5 mg ml(-1) range (compounds I, II, VII) and 0.9-160 mg ml(-1) (compounds III-VI). Linear regression analysis of the data demonstrates the efficacy of the method in terms of precision and accuracy. An extraction method is developed and validated in order to apply this chromatographic method to a commercial cosmetic cream. The precision of this method, calculated as the relative standard deviation (RSD) of the recoveries (1.57-2.21%) was excellent for all compounds I-VII.

Study of bioequivalence of magnesium and sodium valproates.

The in vivo bioavailability of magnesium valproate (500 and 1000 mg) enteric-coated tablets has been compared with that of sodium valproate (Depakine) (500 and 1000 mg) enteric-coated tablets. The two preparations were found to be bioequivalent; magnesium valproate appeared to be a drug without bioavailability problems and with reduced inter-subject variability, compared with that of sodium valproate. A reversed-phase HPLC method for the determination of valproates is described.

Characterization and quantitation of the active polynucleotide fraction (PDRN) from human placenta, a tissue repair stimulating agent.

The polydeoxyribonucleotide (PDRN) fraction is an extract which forms the active component in a new formulation of the drug Placentex (a tissue repair stimulating agent), obtained from human placenta through an original proprietory extraction method. From a comparison of the UV, NMR and IR spectra of this fraction (before and after nuclease treatment) with that of a similar standard (Sigma D1501), it was shown that the active substances in the PDRN fraction mainly consist of a mixture of DNA fragments. By gel electrophoresis, the molecular weights of the DNA fragments were shown to range from 50 to 2000 base pairs. Finally, an HPLC method is described, based on an anion-exchange material capable of determining the amount of PDRN in different batches of the extract, which varied from 80 to 90%.

Heterocyclizations under Vilsmeier conditions: an old method with interesting possibilities in drug research.

The Vilsmeier reaction is well known and widely used in substituting activated hydrogen with a formyl group into a great variety of molecules. This reaction is also a powerful synthetic tool used to obtain many heterocycle compounds, which may be difficult to reach or at times inaccessible. The more relevant data published in chemical literature in the last ten years and my personal experience are reported in the following with the aim of showing the valuable synthetic potential of this reaction.

Synthesis and HPLC assessment of N-nitrosoderivatives from six beta-adrenergic antagonists.

At first N-nitrosoderivatives from six beta-adrenergic antagonists of large consumption were prepared. Secondly the standard nitrosation reactions recommended by WHO were performed to get information about their possible in vivo production and the yields of N-nitrosoderivatives were evaluated by HPLC analyses. Pharmacological tests confirm the potential risk to man of the N-nitrosoderivatives studied.

HPLC analysis of theophylline: bioequivalence study of two sustained-release formulations at steady state.

Theophylline is at present one of the standard drugs used for asthma and obstructive respiratory diseases but still presents clinical problems due to its narrow therapeutic range. To provide additional studies in this important field, the in vivo bioavailability of Bronchoretard, a theophylline sustained-release preparation, not available at present in Italy, has been compared with that of Diffumal, formulation of large consumption. An isocratic reverse-phase HPLC method to assess the theophylline in plasma is described. The two formulations present good bioequivalence, though Bronchoretard seems to provide little therapeutic advantage.

[Synthesis and preliminary pharmacological screening of 2,4-disubstituted N,N-dialkyl-1,8-naphthyridine-3-carboxamides]. / Sintesi ed esame farmacologico preliminare di N,N-dialchil-1,8-naftiridin-3-carbossammidi 2,4-disostituite.

By treating at 100 degrees C 2-aminonicotinic acid with ethyl N,N-dialkylmalonamate (I) and phosphorus oxychloride N,N-dialkyl-4-chloro-1,2-dihydro-2-oxo-1,8-naphthyridine- 3-carboxamides (II) were obtained. The reaction of compounds (II) with an excess of refluxing phosphorus oxychloride afforded N,N-dialkyl-2,4-dichloro-1,8-naphthyridine-3-carboxamides (III), which in turn were treated at room temperature with excess primary amines to give a mixture of isomeric N,N-dialkyl-2-(alkylamino or cycloalkylamino)-4-chloro-1,8-naphthyridine-3-carboxamides (IV) and N,N-dialkyl-4-(alkylamino or cycloalkylamino)-2-chloro-1,8-naphthyridine-3-carboxamides (V). When this last reaction was performed at 160 degrees C, only N,N-dialkyl-2,4-bis(alkylamino or cycloalkylamino)-1,8-naphthyridine-3-carboxamides (VI) were obtained; under the same conditions (IV c) or (V c) reacted with methylamine to give isomeric 2,4-bis(alkylamino)derivatives (VII) or (VIII), respectively. Compounds (II b), (III b), (IV a,c,d), (V a,c,d) were submitted to a wide preliminary pharmacological screening. Some of them, depending on the structure, showed antihypertensive [(IV c)], anti-inflammatory [(IV c) greater than (III b)], or, in the behavioral test, anti-aggressive [(IV d) greater than (III b)] activity. Furthermore compound (III b) caused moderate inhibition of the 5-HT induced contraction of the guinea-pig ileum.

HPLC analysis of the nitrosation products of chlordiazepoxide.

Products originated from Chlordiazepoxide (I) hydrochloride/sodium nitrite interaction were analyzed by HPLC. The studied reactions were carried out in diluted hydrochloric acid solutions at pH values ranging between 0.5-5.0. Depending on the reaction pH values and molar ratios it was possible to find and assess variable amounts of the N-nitrosochlordiazepoxide (II), the dihydroquinazoline (III) and the lactam (IV). The highest degree of N-nitrosation was found at pH 3.5. At this pH value the yields of (II) were respectively 54.8% and 18.3% when the drug (I)/nitrite molar ratios were correspondingly 0.41 and 0.25. When the reaction was performed in concentrations which is possible to find in the gastric juice of patients taking (I) together with nitrite-rich foods the yield of (II) at pH 3.5 was 2.5%. Since in the meantime the genotoxicity of (II) was proved, "in vivo" formation of N-nitrosochlordiazepoxide (II) represents a potential risk not to be underestimated.

Synthesis in solution of oligodeoxynucleotides and some their 5'- and 3'-linked derivatives.

Antisense oligodeoxynucleotides (ODNs) and their derivatives are highly interesting tools to regulate gene expression and promising drugs for antiviral and anticancer therapy. In view of performing more extensive pharmacological trials requiring relatively great amounts of ODNs, we used a method of ODN synthesis derived from the phosphotriester approach which allow to prepare ODNs covalently linking cholesteryl residue (in 5' or 3') and phenazinium group (in 5'). HPLC purifications are discussed.

Synthesis and antirhinovirus activity of 3-(diethylamino)-5-phenylisoxazole derivatives.

Previous studies showed that some 2'-alkyloxyisoxazoles 2b-d obtained from 3-(diethylamino)-5-(2'hydroxy-4'-methoxyphenyl)isoxazole 2a were endowed with an interesting anti-group B rhinovirus activity (action on HRV-2 serotype). Other isoxazoles (WIN compounds) are well known to have anti-group A rhinovirus activity (action on HRV-14 serotype). To obtain an action similar to that of WIN compounds, starting from 2a, the 2'-acyl (3,4,5) and 2'alkyl (6,8) derivatives were synthesized. Also some Mannich bases (9,10) and bisisoxazoles (7,11,13,14) were studied. Though some of the tested compounds mainly exhibited anti-group B rhinovirus activity, their potency was less intense with respect to the above mentioned compounds 2b-d. The only N-methylpiperazinomethyl derivative 10 was slightly active against both tested serotypes.