Abscisic acid and stress induced by salt: Effect on the phenylpropanoid, L-ascorbic acid and abscisic acid metabolism of strawberry fruits.

Strawberry is one of the most popular fruits because of its sensorial and functional properties. However, strawberry crop is sensitive to salt stress conditions. Despite plants have plasticity, high salt concentrations induce molecular, biochemical, and physiological responses in plants. There is evidence that the abscisic acid (ABA) hormone acts as a signaling molecule under stress conditions; however, the molecular mechanisms involved in the synthesis and homeostasis of ABA and in the induction of phytochemical antioxidant compounds under stress conditions remain unclear. In this study, the effect of stress induced by NaCl (salt stress - SS), with or without the simultaneous application of ABA, on the ABA, phenylpropanoids and L-ascorbic acid (AsA) metabolisms were evaluated. The physiological aspects (Na, Cl and proline concentration, photosynthetic variables) were also studied. The results showed that salt stress increases the Na and Cl content in the leaves, affects photosynthetic variables and triggers the production of proline, pelargonidin-3-O-glucoside, total phenolic compounds and AsA content, alongside the upregulation of several genes from the phenylpropanoid and flavonoid pathways. These effects were accompanied by the induction of compounds and transcripts related to ABA biosynthesis, conjugation and catabolism. Otherwise, the exogenous application of ABA in salt stressed plants promotes a shift in gene expression and metabolism to mitigate the stress. Therefore, salt stress affects the metabolism of ABA, phenylpropanoids and AsA in strawberry increasing phytochemical composition which is strongly associated with an ABA-dependent mechanism.

Synthesis and characterization of cyclohexane-1R,2R-diamine-based Pt(iv) dicarboxylato anticancer prodrugs: their selective activity against human colon cancer cell lines.

Three pairs of asymmetric dicarboxylato derivatives based on the cisplatin and oxaliplatin-like skeletons have been synthesized de novo or re-synthesized. The axial ligands consist of one medium-chain fatty acid (MCFA), namely clofibrate (i.e. 2-(p-chlorophenoxy)-2-methylpropionic acid, CA), heptanoate (HA) or octanoate (OA), respectively, and an inactive acetato ligand that imparts acceptable water solubility to such conjugates. Stability tests provided evidence for the partial formation of two hydrolyzed products, corresponding to two monoaqua diastereomers derived from the substitution of an equatorial chlorido ligand with a water molecule. The complexes have been tested on three different colon cancer cell lines having different histological history, and also on the cisplatin-sensitive A2780 ovarian cancer cell line for comparison. This allowed the evaluation not only of the increase in activity on passing from Pt(ii) to Pt(iv) derivatives, but also the selectivity towards colon cancer cells brought about by the cyclohexane-1R,2R-diamine carrier ligand.

[Possible use of the growing liver biological set for hepatic recovery after toxic damage (an experimental study)]. / Vozmozhnosti biologicheskoi kombinatsii, poluchennoi iz rastushchei pecheni, dlia ee vosstanovleniia pri toksicheskom povrezhdenii (éksperimental'noe issledovanie).

The lack of acceptable pharmacological approaches for restoration of the injured liver is associated with complex of mechanisms involved in hepatic regeneration and with difficulty of the target selection. The aim of this research was to study the hepatoprotective function of the extract from both the growing and regenerating liver containing a natural set of factors crucial for the hepatic restoration. Extracts from both regenerating liver of rats after 70% hepatic resection and the growing liver of neonatal pigs were obtained using own original technique. The set of resultant extracts was named as the hepatic regeneration set (HRS). HRS fractionation was carried out using the Toyopearl HW-50S sorbent. The efficiency of HRS and its fractions was estimated using a model of the mouse liver thioacetamide injury and monitoring hepatic enzyme activity in blood serum. The activities of AST and ALT in intact animals were 50 U/l and 80 U/l, respectively; after thioacetamide administration they increased to 2059±212 U/l and 4280±440 E/l, respectively (p<0.05). Treatment of injured animals with HRS from the rat regenerating liver resulted in a significant decrease of transaminase activities to 924±148 U/l (AST; p<0.05) and 1633±308 U/l (ALT; p<0.05). A similar effect was observed after treatment with HRS from the neonatal pig liver: the AST decreased to 937±138 U/l (p<0.05), while ALT activity decreased to 1710±237 U/l (p<0.05). HRs fractionation resulted in identification two active fractions characterized by much higher (8-29) hepatotropic effect that that of the whole extract. These fractions contained peptide/protein components with the range of molecular mass of 3-60 kDa (fraction 1) and 3-25 kDa (fraction 2a). Fraction 1 also contained some polynucleotides in fraction 1. Subsequent studies of these fractions exceeding the hepatotropic effect of original HRS is clearly needed to identify their individual components by immunochromatography methods, ELISA, MRM mass spectrometry and quantitative PCR.

How to obtain Pt(iv) complexes suitable for conjugation to nanovectors from the oxidation of [PtCl(terpyridine)].

Oxidation of [Pt(II)Cl(terpy)]+ (terpy = 2,2':6',2''-terpyridine) has been attempted with several oxidizing agents and under different experimental conditions in order to obtain a Pt(iv) complex suitable for the conjugation to nanovectors to be used in drug delivery targeting for anticancer therapy. The best compromise in terms of yield and purity of the final complex was obtained by microwave-assisted reaction at 70 °C in 50% aqueous H2O2 for 2 h. Under these conditions the quantitative formation of [Pt(IV)Cl(OH)2(terpy)]+ was observed. The subsequent synthetic steps were, (i) functionalization of [Pt(IV)Cl(OH)2(terpy)]+ in the axial position with succinic anhydride to obtain [Pt(IV)Cl(OH)(succinato)(terpy)]+ and (ii) reaction of the latter with nonporous silica nanoparticles (SNPs) with an external shell containing primary amino groups to obtain a nanovector able to transport the Pt(iv) antitumor prodrug in the form of a conjugate Pt-SNP. Finally, the antiproliferative activity and cell accumulation of [Pt(II)Cl(terpy)]+, [Pt(IV)Cl(OH)2(terpy)]+, and the Pt-SNP conjugate were measured on three cancer cell lines. Despite highly effective accumulation of Pt-SNP in cells, a modest increase in activity was observed with respect to the molecular species. Further experiments showed that the Pt-SNP conjugate can release [Pt(II)Cl(terpy)]+ upon reduction, but this metabolite may undergo hydrolysis, and the resulting aquo complex could coordinate once again the free amino groups of the SNPs. In the resulting tetraamine form, the Pt(ii) complex conjugated to the SNPs cannot completely perform its antiproliferative activity.

Cisplatin and valproate released from the bifunctional [Pt(IV)Cl2(NH3)2(valproato)2] antitumor prodrug or from liposome formulations: who does what?

The cisplatin-sensitive human ovarian cancer cells A2780 have been challenged with cationic liposomes containing the single drug cisplatin or valproate or their combination with an approximate 1 : 2 molar ratio, i.e. the same ratio present in preformed (OC-6-33)-diamminedichloridobis(valproato)platinum(iv), that releases such metabolites by intracellular Pt(iv) → Pt(ii) reduction. The results of this comparison confirm that valproate barely penetrates cells unless it is transported by liposomes or it is coordinated to a lipophilic Pt(iv) assembly. The two drugs have a synergistic action, cisplatin being the more potent antiproliferative agent. Even if the preformed (OC-6-33)-diamminedichloridobis(valproato)platinum(iv) releases cisplatin and valproate in the same amount as the liposome formulation, the Pt(iv) derivative is more active. This important feature, common to all Pt(iv) complexes having very lipophilic carboxylates, is attributable to their propensity to remain in cells and to continuously bind DNA, unlike cisplatin that is partially removed from cells by efficient efflux pathways.

Functionalized nonporous silica nanoparticles as carriers for Pt(iv) anticancer prodrugs.

Nonporous silica nanoparticles (SNPs) with an external shell containing primary amino groups were proposed as potential delivery systems for Pt(iv) antitumor prodrugs. Spherical SNPs containing two different external arms, i.e. 3-aminopropyl and N-(6-aminohexyl)aminomethylene, of around 125 nm hydrodynamic diameter were loaded with two different cisplatin-based Pt(iv) complexes, namely (OC-6-44)-diamminedichloridoethoxidosuccinatoplatinum(iv) and (OC-6-44)-diamminedichloridoacetylamidosuccinatoplatinum(iv), through the formation of amide bonds between the pendant amino groups on SNPs and the free carboxylic group of the complexes. In the presence of the N-(6-aminohexyl)aminomethylene arm, the Pt(iv)-SNP conjugates showed a negligible (unwanted) Pt release by hydrolysis, whereas in the presence of ascorbic acid the reduction of Pt(iv) → Pt(ii) caused the substantial release of the active metabolite cisplatin. Conjugate Pt(iv)-SNP exhibited better antiproliferative activity on the Pt-sensitive A2780 human ovarian cancer cell line than the parent cisplatin and their free Pt(iv) precursors, due to their more efficient cellular uptake, likely by endocytosis.

The suicidality continuum in a large sample of obsessive-compulsive disorder (OCD) patients.

BACKGROUND: Obsessive-compulsive disorder (OCD) has a chronic course leading to huge impact in the patient's functioning. Suicidal thoughts and attempts are much more frequent in OCD subjects than once thought before. AIM: To empirically investigate whether the suicidal phenomena could be analyzed as a suicidality severity continuum and its association with obsessive-compulsive (OC) symptom dimensions and quality of life (QoL), in a large OCD sample. METHODS: Cross-sectional study with 548 patients diagnosed with OCD according to the DSM-IV criteria, interviewed in the Brazilian OCD Consortium (C-TOC) sites. Patients were evaluated by OCD experts using standardized instruments including: Yale-Brown Obsessive-Compulsive Scale (YBOCS); Dimensional Yale-Brown Obsessive-Compulsive Scale (DYBOCS); Beck Depression and Anxiety Inventories; Structured Clinical Interview for DSM-IV (SCID); and the SF-36 QoL Health Survey. RESULTS: There were extremely high correlations between all the suicidal phenomena. OCD patients with suicidality had significantly lower QoL, higher severity in the "sexual/religious", "aggression" and "symmetry/ordering" OC symptom dimensions, higher BDI and BA scores and a higher frequency of suicide attempts in a family member. In the regression analysis, the factors that most impacted suicidality were the sexual dimension severity, the SF-36 QoL Mental Health domain, the severity of depressive symptoms and a relative with an attempted suicide history. CONCLUSIONS: Suicidality could be analyzed as a severity continuum and patients should be carefully monitored since they present with suicidal ideation. Lower QoL scores, higher scores on the sexual dimension and a family history of suicide attempts should be considered as risk factors for suicidality among OCD patients.

Safety and efficacy of plasmid DNA expressing two isoforms of hepatocyte growth factor in patients with critical limb ischemia.

VM202, a plasmid DNA that expresses two isoforms of hepatocyte growth factor, may elicit angiogenic effects that could benefit patients with critical limb ischemia (CLI). In a phase 2, double-blind trial in 52 CLI patients, we examined the safety and potential efficacy of intramuscular injections of low-dose (n=21) or high-dose (n=20) VM202 or placebo (n=11) in the affected limb (days 0, 14, 28 and 42). Adverse events and serious adverse events were similar among the groups; no malignancy or proliferative retinopathy was seen. In exploratory efficacy analyses, we found no differences in ankle or toe-brachial index, VAS, VascuQuol or amputation rate among the groups. Complete ulcer healing was significantly better in high-dose (8/13 ulcers; P<0.01) versus placebo (1/9) patients. Clinically meaningful reductions (>50%) in ulcer area occurred in high-dose (9/13 ulcers) and low-dose (19/27) groups versus placebo (1/9; P<0.05 and P<0.005, respectively). At 12 months, significant differences were seen in TcPO2 between the high-dose and placebo groups (47.5 ± 17.8 versus 36.6 ± 24.0 mm Hg, respectively; P<0.05) and in the change from baseline among the groups (P<0.05). These data suggest that VM202 is safe and may provide therapeutic bioactivity in CLI patients.

Therapy side-effects and predictive factors for preterm delivery in patients undergoing tocolysis with atosiban or ritodrine for threatened preterm labour.

The objective of this study was to compare the safety and efficacy of atosiban and ritodrine in the treatment of threatened preterm labour (TPL) and to analyse the predictive factors of preterm delivery. We retrospectively sampled data on 380 women hospitalised for TPL (24-35 weeks' gestation), in our clinic between 2004 and 2007. All were subjected to tocolysis with ritodrine and/or atosiban. Data were analysed using R (version 2.12.1), considering p < 0.05 as significant. We had 69 women treated with atosiban, 242 treated with ritodrine and 69 treated with ritodrine changed for atosiban, if adverse effects occurred. In the multivariate logistic regression, the use of atosiban vs ritodrine does not play any role in delaying delivery after 48 h or 7 days, whereas the cervical change at the digital examination, high contractions pre/post-therapy ratio, pPROM, cervical length and fibronectin result as predictive factors for both delivery before 48 h or 7 days. Maternal adverse drug effects were significantly more frequent in patients treated with ritodrine, and one single case of pulmonary oedema was observed. We found fewer side-effects in the atosiban than in the ritodrine group and no difference in efficacy. Moreover, the most predictive factors for preterm delivery were fibronectin test, pPROM, digital vaginal examination and uterine contraction persistence. We believe that predictive capacity of these tests could give the opportunity for targeting therapy and limiting drug side-effects and cost.

[The classification of obstructive jaundice severity].

It was analyzed the results of examination and surgical treatment of 338 patients with obstructive jaundice. The patients were operated in 2004-2012 years. 148 patients had benign jaundice and 190 patients had malignancy genesis of jaundice. It was determined laboratory data (bilirubin total, protein total, prothrombin index) as well as complications of obstructive jaundice (cholangitis, renal failure, hepatic encephalopathy, gastrointestinal bleeding, sepsis) indicating to the severity of obstructive jaundice and it was taken into consideration the malignant origin of jaundice. The selected attributes were assigned marks from 1 to 3. Each complication as well as factor of malignancy was evaluated two times higher than mark of bilirubin. The grade of jaundice severity (A, B, C) was determined by combination of the marks that identified the attributes. It was analyzed the influence of prothrombin index and blood serum protein on the results of severity grade formation of obstructive jaundice with the postoperative results (lethality, complications). It was done for simplification of classification. The absence of significant differences in assessing of the severity obstructive jaundice class with and without attributes of a "total protein" and "prothrombin index". To simplify the classification and to get more reliable information about the difference of postoperative results between the grades (A, B, C) are possible by exception of total protein attributes and prothrombin index.

[Rate of biliary ducts' decompression by the tumorous obstructive jaundice].

Results of biliary decompression were analyzed in 185 patients with malignant obstructive jaundice. Among them 85 patients underwent nasobiliary drainage, 37 - percutaneous transhepatic bile drainage and 63 - cholecystectomy. Dynamics in biochemical indices of blood serum, cholangiomanometry and jaundice response to decompression according to T. Shimizu et K.Yoshida's formula in our modification were evaluated for estimation of bile flow rate. It is ascertained that application of nasobiliary drainage leads to low-rate decompression, percutaneous transhepatic bile drainage and cholecystectomy - to high-rate decompression. Low-rate decompression enables to decrease significantly the frequency of "fast decompression syndrome" and complications, particularly augmenting of liver dysfunction, multiple organ failure and lethality in post-decompression period related to them.

[Postoperative biliary complications after liver resections].

Postoperative biliary complications (BC) after liver resections carried out from 1990 to 2009 were analyzed in 216 patients. Among them - 139 women and 77 men aged 15-79 years (mean age 49,8±0,9 years). Complications in early postoperative period were observed in 110 patients (50,9%); among them BC - in 27 patients (12,2%), bilomas - in 6, external biliation - in 14, abscesses - in 7. Lethal outcomes were not observed. Average duration of hospital stay amounted 41,3±12 days. Relationship between frequency of BC and age, malignancy of neoplasm and extent of liver resection was estimated. All patients were cured using low invasive techniques. Recurrent operations were not carried out.

[Intraoperative injuries of bile ducts].

125 patients with injuries of bile ducts were observed from 1988 to 2008 years. Minor injuries (ducts in the bed of the gall bladder) were registered in 64 patients. Magistral bile ducts were injured in 61 cases. Analysis revealed 3 key points of the successful treatment of of such patients: type of the injury (cut, dissection, side wound), localization of the defect (intra- and extrahepatic ducts) and participation of a highly qualified surgeon.

[Intrahepatic cholestasis of pregnancy: diagnosis and obstetric management]. / Colestasi intraepatica della gravidanza: diagnosi e management ostetrico.

AIM: The aim of this study was to suggest a possible obstetric management of patients with intrahepatic cholestasis of pregnancy (ICP). METHODS: We performed a retrospective analysis enrolling 44 women with ICP deliveries at our Obstetrics and Gynaecology clinic of the University of Udine between January 2005 and July 2008. Data on patient age, body mass index, weight, gestational age, parity, symptoms, comorbidity, value of liver test, APGAR score and fetal weight were prospectively recorded in a computed database and clinical folder. RESULTS: The intensification of maternal and fetal surveillance in patients with ICP (liver function tests, ultrasound and cardiotocography), can significantly reduce perinatal mortality (no case of stillbirth in ours study) but, inevitably, increases the cesarean delivery rate (65.1%), the induction of labor rate (38.4%) and the preterm delivery rate (58.13 %, median gestational age 35 sg+/-1.46). CONCLUSION: Obstetric management of ICP consist of weighing the risk of premature delivery against the risk of sudden death in utero. To date, no ideal method of fetal surveillance has been determined for ICP; the intrauterine deaths are thought to occur suddenly and fetal cardiac monitoring cannot forecast an acute event. Nevertheless, we think that a management strategy, inclusive of induction of labor at 37 weeks, can reduces the risk of fetal death. In most severe cases, no responsive to ursodeoxycholic acid and S-adenosylmethionine treatment, delivery has been initiated even before the 37 weeks, as soon as lung maturity has been established.

[The role of biliary decompression and correction of dysbiosis in the treatment of obstructive jaundice].

The article providing in-depth analysis of pathogenesis of obstructive jaundice shows that this disease is manifest not only as changes at the hepatic level (cholestasis, cholehemia, cholangitis, cholangio- and lymphovenous shunts, hepatic encephalopathy) but also as marked dysbiotic disturbances due to anacholia and toxic metabolites that cause bacterial translocation and endotoxemia complicating liver insufficiency. Based on the literary data and original observations, a new scheme for the treatment of obstructive jaundice is proposed including simultaneous correction of both components of hepatoenteric turnover, also, it permits to improve the outcome of the postoperative period.

[Relationship of tumor necrosis factor alpha expression with activation of sphingomyelinase and lipid peroxidation after removal of cholestatic factor].

The goal of this work was to study the expression of tumor necrosis factor alpha (TNFalpha), sphingomyelin cycle activation, and lipid peroxidation (LPO) processes after the removal of a cholestatic factor in the liver subjected to different durations of cholestasis. Restored bile flow after a 9-day hepatic cholestasis normalized sphingomyelinase (SMase) activity and levels of TNFalpha and LPO products. The removal of a cholestatic factor after a 12-day cholestasis did not normalize the studied parameters: SMase activity and the levels of TNFalpha and LPO products remained much higher compared to control. A significant positive correlation between TNFalpha expression, SMase activity, and LPO rate has been revealed. The obtained data indicate that hepatocyte apoptosis after bile outflow restoration in late cholestasis can be due to the activation of the sphingomyelin cycle, LPO, and TNFalpha expression. The synergistic interaction can sharply increase the proapoptotic capacity of each of these factors since TNFalpha activates SMase and LPO, SMase activity depends on the LPO rate, while ceramide, an SMase-produced secondary messenger of apoptosis, can induce oxidative stress.

[Chronic pancreatitis: resective and decompressive procedures].

PURPOSE: Definition of resective and decompressive operations in chronic pancreatitis. MATERIALS AND METHODS: A retrospective analyses of surgical management of 51 chronic pancreatitis patients is carried out. 24 patients underwent longitudinal pancreaticojejunostomy (PA), 27--pancreatic head ( PH) resective procedures: Beger--5, modified Frey (PH intraparenchimatose resection-- IR, supposing removal of fibrous masses from the ventral and greater part of the dorsal PH)--22. According to the character of the PH lesion the patients were divided in 3 groups: head dominant (13), total (11) and cystic pancreatitis (20). RESULTS: One early postoperative death developed after reoperation for adhesive intestinal ileus in patient with PA. No mortality was noted after IR. Remote results in terms of 4.8+/-0,7 years were studied in 32 (71%) patients underwent PA and IR. IR lead to good and satisfactory results. After PA poor results came to 5 (29.5%). CONCLUSIONS: Combination of PH IR and PA is indicated in head dominant and total chronic pancreatitis patients. In cases of pseudocysts and dilation of the main pancreatic duct (>6-7 cm) decompressive variant of procedure cysto-pancreato-jejunostomy is preferable.

[Causes, diagnosis and surgical treatment of strictures of lobar and segmental hepatic ducts].

Causes of strictures of lobar and segmental ducts after their injuries during open and laparoscopic cholecystectomy in 53 patients were analyzed. For correction of bile outflow precision non-wireframe (n=22) and wireframe (n=20) anastomoses were used. In 10 patients a combined anastomosis was established. In 1 patient the external drainage of hepatic ducts was performed. After surgery 3 patients died. 1-15 year long-term results were studied in 48 (96%) patients. Good results were achieved in 29 (60.5%), satisfactory -- in 13 (27%) patients. Recurrences of the strictures were diagnosed in 6 patients. It is concluded that dynamic control is necessary in patients operated on for bile ducts injuries. Underestimation of remittent cholangitis leading to biliary cirrhosis worsens prognosis of the disease.

[Changes in sphingomyelinase activity, tumor necrosis factor alpha level, and lipid peroxidation rate in the course of development of cholestatic liver injury].

Changes in sphingomyelinase activity, tumor necrosis factor alpha expression, and lipid peroxidation rate in the course of development of cholestatic liver injury have been studied. The same type phase shifts in the parameters analyzed were observed, which included a marked decrease at the early stages of cholestasis (days 3-6) and a pronounced increase at the later stages (days 12-16), i.e., under the conditions of developed pathology. There is a significant positive linear correlation between tumor necrosis factor alpha expression, sphingomyelinase activity, and lipid peroxidation rate during cholestatic injury. The changes detected may reflect balance between the effects of the two major bile components--bilirubin, which is accumulated in the liver at the early stages of cholestasis, and bile acids, whose influence dominates at the later stages of pathologic process. Our results indicate that tumor necrosis factor alpha overexpression, the sphingomyelin cycle activation, and lipid peroxidation intensification may cause apoptosis of hepatocytes at the late stages of cholestasis.