Identification of 16 novel mutations in the argininosuccinate synthetase gene and genotype-phenotype correlation in 38 classical citrullinemia patients

Classical citrullinemia (CTLN1), a rare autosomal recessive disorder, is caused by mutations of the argininosuccinate synthetase (ASS) gene, localized on chromosome 9q34.1. ASS functions as a rate-limiting enzyme in the urea cycle. Previously, we identified 32 mutations in the ASS gene of CTLN1 patients mainly in Japan and the United States, and to date 34 different mutations have been described in 50 families worldwide. In the present study, we report ASS mutations detected in 35 additional CTLN1 families from 11 countries. By analyzing the entire coding sequence and the intron-exon boundaries of the ASS gene using RT-PCR and/or genomic DNA-PCR, we have identified 16 novel mutations (two different 1-bp deletions, a 67-bp insertion, and 13 missense) and have detected 12 known mutations. Altogether, 50 different mutations (seven deletion, three splice site, one duplication, two nonsense, and 37 missense) in 85 CTLN1 families were identified. On the basis of primary sequence comparisons with the crystal structure of E. coli ASS protein, it may be concluded that any of the 37 missense mutations found at 30 different positions led to structural and functional impairments of the human ASS protein. It has been found that three mutations are particularly frequent: IVS6-2A>G in 23 families (Japan: 20 and Korea: three), G390R in 18 families (Turkey: six, U.S.: five, Spain: three, Israel: one, Austria: one, Canada: one, and Bolivia: one), and R304W in 10 families (Japan: nine and Turkey: one). Most mutations of the ASS gene are "private" and are distributed throughout the gene, except for exons 5 and 12-14. It seems that the clinical course of the patients with truncated mutations or the G390R mutation is early-onset/severe. The phenotype of the patients with certain missense mutations (G362V or W179R) is more late-onset/mild. Eight patients with R86H, A118T, R265H, or K310R mutations were adult/late-onset and four of them showed severe symptoms during pregnancy or postpartum. However, it is still difficult to prove the genotype-phenotype correlation, because many patients were compound heterozygotes (with two different mutations), lived in different environments at the time of diagnosis, and/or had several treatment regimes or various knowledge of the disease.     

Immunomodulatory effects of mesenchymal stem cells on leukocytes with emphasis on neutrophils

Mesenchymal stem cells (MSCs) are a population of multipotent cells with the ability of expansion and plastic-adherence in vitro. MSCs can differentiate into chondrocytes, osteocytes and adipocytes; they lack co-stimulatory molecules and have small amount of MHC-I that makes no immunogenicity. These characteristics are empowering MSCs’ huge in vivo applications. In addition, MSCs possess the ability of regulating the immune responses in many diseases. Many studies have shown that MSCs have immunosuppressive as well as immunoenhancing properties such as inhibition of T-lymphocytes proliferation and cytokines production which lead to the balance of Th1 and Th2. Some other immunomodulatory features of MSCs are increasing suppressive capacity of T_reg, reducing activity of B-lymphocytes and immunoglobulins secretion, inhibition of dendritic cells maturation and antigen presenting capacity, and inhibition of NK-cells activity. MSCs also exert inhibitory effects on neutrophil apoptosis and reduce reactive oxygen species production. The purpose of this paper is to focus on the MSCs? effects on immune cells, especially neutrophils.     

Effect of heating and enzymatic hydrolysis on casein cow milk sensitivity in Moroccan population

The objectives of the present work were first to evaluate the sensitivity to cow raw milk of the population of Fez, and then to study the effect of heating and pepsin hydrolysis on the allergenicity of casein. A cross-sectional study was carried out in Fez Hospitals, in which 1000 patients were recruited to establish a sera bank used to evaluate specific IgE to cow milk and to casein. Then, we evaluated the reaction of human IgE to heated and pepsin-hydrolysed casein. The results showed that 11.5% of the population studied self-reported reactions to foods. From them, 3.6% reported allergy to milk. Evaluation of specific IgE to cow raw milk showed that 11.9% of patients presented higher specific IgE levels. The treatments of casein indicated that both heating and pepsin hydrolysis totally decreased its binding on the human IgE.     

Effects of two dietary exogenous multi-enzyme supplementation,Natuzyme<Superscript>®</Superscript> and beta-mannanase (Hemicell<Superscript>®</Superscript>), on growth and blood parameters of Caspian salmon (<Emphasis Type="Italic">Salmo trutta caspius</Emphasis>)

Recent increases in feed ingredient costs have motivated the fisheries industry to identify technologies that will improve feed utilisation and reduce the cost per pound of gain. The effects of two supplemental exogenous enzymes (Natuzyme® and Hemicell®) on the growth performance in Caspian salmon (Salmo trutta caspius) were examined over an 8-week feeding trial. After the experimental period, the survival rate ranged from 91.33?±?1.15 % in controls to 96.67?±?1.15 % in the group that received 0.5 g Natuzyme® kg^?1?+?0.5 g Hemicell® kg^?1 (NH) in their diet and there was a statistical difference between experimental and control groups (p?<?0.05). Growth rate was significantly higher in the NH group (1.01?±?0.01) than the other groups (Sig.?=?0.00). The best feed conversion rate (0.64?±?0.01) was in the NH group and it was significantly lower than the control group, the 0.5 g Natuzyme® kg^?1 group, and the 0.25 g Hemicell® kg^?1 group (Sig.?=?0.03). The best final body weight (80.68?±?5.27) was observed in the NH group. Also, WBC count (7,716.67?±?348.80 N/mm³) was significantly higher in the NH group compared to the control (6,916.67?±?194.10 N/mm³; p?<?0.05). No difference was observed in haematocrit%, haemoglobin, red blood cell, mean corpuscular volume, mean corpuscular haemoglobin, and mean corpuscular haemoglobin concentration (p?>?0.05). The results suggested that enzyme supplementation caused significant improvement on growth performance and feed utilisation in Caspian salmon.     

Gastroprotective effect of mummy on induced gastric ulcer in rats

Mummy was used in traditional medicine as a remedy for inflammation, articular injuries, rheumatism, bone fractures, wounds and back pains. In previous research, the healing effects of mummy on a rabbit’s tibial fracture were studied, and the results supported its efficacy. In consideration of claims and suggestions by traditional medicine about the effectiveness of mummy on wound healing, the present study was aimed at investigating the protective effect of mummy on gastric ulcer induced by ethanol. Young male albino rats (NMRI strain, n?=?21, 190–230 g body weight) were randomly divided into three experimental groups: treatment group, positive control group and negative control group, each containing seven rats. All animals had been exposed to starvation for 24 h and then received oral ethanol (1 ml/200 g body weight). The treatment group received 100 mg/kg/day mummy; the positive control group received 200 mg/kg/day cimethidine, and the negative control group received 5 ml/kg/day distilled water for 2 weeks by gavages. One hour after the last treatment, all animals were killed, and their stomachs were dissected. The number and length of ulcers were determined using stereomicroscope; then graticule and the ulcer index and curative ratio were calculated. Data was analysed using one-way ANOVA and Tukey’s post hoc test. The administration of mummy significantly decreased the gastric ulcer score and ulcer index in the treatment group, compared with the positive control and negative control groups. The curative ratio in the treatment group was significantly higher than the negative control and the positive control groups (89.26 versus 87.79 %, respectively). The results showed excellent effects of mummy on wound healing after the ethanol-induced gastric ulcer in rats.     

The negative predictive value of p16INK4a to assess the outcome of cervical intraepithelial neoplasia 1 in the uterine cervix.

The immunohistochemical expression of p16 in formalin-fixed and paraffin-embedded histological sections was evaluated in a retrospective study comprising a low-grade group of 100 cases of cervical intraepithelial neoplasia (CIN) 1, a high-grade group of 50 cases of CIN 2 to 3, and a benign group of 50 cases of normal tissue or benign lesions in the uterine cervix. The cases were consecutive within each group and had a minimum follow-up period of 5 years. Positive reaction for p16 was detected in all cases in the high-grade group and in only 3 cases in the benign group. In the low-grade group, a total of 9 cases had to be excluded. The remaining 91 cases in the low-grade group showed positive reaction for p16 in 65 cases (71%), including 23 cases that progressed to a high-grade lesion, 36 cases that revealed normal cytological and/or histological picture during the follow-up period, and 6 cases that persisted as CIN 1. A total of 26 cases (29%) in the low-grade group showed negative reaction for p16. All but one of these p16 negative cases in the low-grade group had a benign or normal outcome. This case showed a high-grade lesion in the follow-up period and was probably a high-grade lesion from the beginning and so underestimated as CIN 1. These results reveal that the negative predictive value of p16 to predict the outcome of the cases of CIN 1 is as high as 96%, which strongly suggest an important role of p16 in the assessment of this type of lesion.     

Protective effect of kombucha tea against acetaminophen-induced hepatotoxicity in mice: a biochemical and histopathological study

Acetaminophen overdose causes severe hepatotoxicity leading to liver failure in experimental animals and humans. This study was undertaken to evaluate the protective effect of kombucha tea (KT) against acetaminophen-induced hepatotoxicity. Forty male Balb/c mice were divided into four equal groups: (1) the control group, (2) KT-treated group, (3) acetaminophen-treated group, and (4) KT/acetaminophen-treated group. All mice in group 4 were given KT orally for 7?days before a single hepatotoxic dose of acetaminophen (1,000?mg/kg orally). Activities of liver marker enzymes in serum; aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH), and alkaline phosphatase (ALP); and total protein (TP), albumin (ALB), and direct and total bilirubin levels were determined. Acetaminophen challenge caused significant increases in the levels of bilirubin and liver enzymes (AST, ALT, ALP, and LDH), while TP and ALB levels were reduced significantly. Histopathologic assessments showed that severe glycogen storage in hepatocytes, hepatocellular degeneration and necrosis, mononuclear cell infiltration in portal area, dilation of central veins, and capillarization also reduced in KT/acetaminophen group compared to acetaminophen-treated mice. In conclusion, these findings suggest that KT has protective effect on acetaminophen-induced hepatotoxicity.     

A liver transplant center experience with liver dialysis in the management of patients with fulminant hepatic failure: a preliminary report

Among extracorporeal liver support devices, liver dialysis is cleared by the U.S. Food and Drug Administration to be used for the management of fulminant hepatic failure (FHF). The outcomes of patients following liver dialysis need to be clearly evaluated. Among the 25 patients with FHF admitted to the Liver ICU between May 2000 and November 2002, 12 underwent liver dialysis, including 6 men and 6 women, of mean age 32 years. The causes of FHF were identified as acetaminophen (n = 10), herbal medications (n = 1) and autoimmune disease (n = 1). At presentation, the mean total bilirubin was 9.35 mg/dL (range, 0 to 1.3), mean ALT 3015 U/L (range, 0 to 48), mean AST 3457 (range, 0 to 42), mean ammonia 98 micromol/L (range, 10 to 60) and mean INR 1.88. A control group including 13 patients (2 men and 11 women), of mean age 27.8 years mean total bilirubin 5.66, mean ALT 3494, mean AST 3528, mean ammonia 113 and mean INR 3, were not treated with liver dialysis, due to the lack of machine availability or physician's choice. The causes of FHF were acute hepatitis B (n = 1), acetaminophen (n = 10) or unknown (n = 2). There was no statistically significant difference in the baseline characteristics of the two groups (P >.05). Among the liver dialysis group, 1 patient died, 2 underwent OLTx, and 9 were discharged home. Among the control group; 4 patients died, 2 underwent OLTx, and 7 were discharged home. Preliminary results seem to support survival benefit among patients who underwent liver dialysis compared to non-liver dialysis; however, further randomized control trials are warranted to verify this observation.     

Reactive and reparative fibrillar collagen remodelling in the hypertrophied rat left ventricle: two experimental models of myocardial fibrosis

STUDY OBJECTIVE--The aim was to compare the temporal sequence and structural relationship between perivascular and interstitial fibrosis and microscopic scarring seen in the left ventricle in response to either a transient or sustained stimulus to fibrosis. DESIGN--In 72 male Wistar rats (250-350 g) the transient stimulus model was based on the administration of isoprenaline (500 micrograms.kg-1) while the sustained stimulus model was produced by abdominal aortic banding with right renal artery constriction. Serial sections of myocardium were examined and compared at 4 and 12 weeks in each model and to corresponding controls. EXPERIMENTAL MATERIAL--The collagen specific stain, Sirius Red F3BA, was used to determine collagen volume fraction and the fibrillar nature of the fibrous tissue response seen by light microscopy. MEASUREMENTS AND MAIN RESULTS--Following isoprenaline a stable reparative fibrosis of the endomyocardium and increase in collagen volume fraction was seen without an interstitial or perivascular fibrosis of the non-involved myocardium. In unilateral renal ischemia, on the other hand, a progressive perivascular fibrosis was evident throughout the myocardium and from which fibrillar collagen extended into the extracellular space between muscle bundles creating an interstitial fibrosis; microscopic scarring of the endomyocardium became evident at 12 weeks. CONCLUSIONS--The reactive perivascular fibrosis of intramyocardial coronary arteries seen in renovascular hypertension is a progressive process that leads to an interstitial fibrosis and eventual microscopic scarring. In contrast, the endomyocardial scarring that follows isoprenaline induced myocyte necrosis is stable and intramural vessels in remote regions are not involved.