Arginase deficiency is characterized by hyperargininemia and infrequent episodes of hyperammonemia. Human patients suffer from neurological impairment with spasticity, loss of ambulation, seizures, and severe mental and growth retardation. In a murine model, onset of the phenotypic abnormality is heralded by weight loss beginning around day 15 with death occurring typically by postnatal day 17 with hyperargininemia and markedly elevated ammonia. The goal of this study was to address the development of a gene therapy approach for arginase deficiency beginning in the neonatal period. Lifespan extension, body weight, circulating amino acids and ammonia levels were examined as outcome parameters after gene therapy with an adeno-associated viral vector expressing arginase was administered to mice on the second day of life (DOL). One-hundred percent of untreated arginase-deficient mice died by DOL 24, whereas 89% of the adeno-associated virus (AAV)-treated arginase deficient mice have survived for >8 months. While animals at 8 months demonstrate elevated glutamine levels, ammonia is less than three times that of controls and arginine levels are normal. These studies are the first to demonstrate that AAV-based therapy for arginase deficiency is effective and supports the development of gene therapy for this and the other urea cycle disorders.

These results suggest that peripheral CB1R blockade improves obesity-induced insulin resistance by suppressing adipose tissue inflammation via the NLRP3 inflammasome.

Histone deacetylase 2 (HDAC2) is aberrantly regulated and plays a pivotal role in the development of hepatocellular carcinoma (HCC) through regulation of cell-cycle components at the transcriptional level, but the underlying mechanism leading to oncogenic HDAC2 remains unknown. In this study, we show that expression of CK2a (casein kinase II a subunit) was up-regulated in a large cohort of human HCC patients, and that high expression of CK2a was significantly associated with poor prognosis of HCC patients in terms of five-year overall survival. It was also found that CK2a over-expression positively correlated with HDAC2 over-expression in a subset of HCCs. We observed that treatment with epidermal growth factor (EGF) elicited an increase in CK2a expression and Akt phosphorylation, causing induction of HDAC2 expression in liver cancer cells. It was also observed that ectopic expression of dominant-negative CK2a blocked EGF-induced HDAC2 expression, and that ectopic CK2a expression attenuated the suppressive effect of Akt knockdown on HDAC2 expression in liver cancer cells. Targeted disruption of CK2a influenced the cell cycle, causing a significant increase in the number of liver cancer cells remaining in G 2 /M phase, and suppressed growth via repression of Cdc25c and cyclin B in liver cancer cells. Taken together, our findings suggest the oncogenic potential of CK2a in liver tumorigenesis. Furthermore, a regulatory mechanism for HDAC2 expression is proposed whereby EGF induces transcriptional activation of HDAC2 by CK2a/Akt activation in liver cancer cells. Therefore, this makes CK2a a promising target in cancer therapy.

Corresponding author's email: reffros@labiomed.org: Exhaled breath condensates (EBC) can provide information regarding inflammatory mediators in the fluid lining the airways. RationaleAlthough this noninvasive procedure avoids the risks and costs associated with bronchoalveolar lavage, EBC samples are variably diluted from ~5,000 to 15,000 times by water vapor and they can be contaminated with saliva.These factors were addressed with a pilot study of 10 subjects (5 healthy nonsmokers, 2 healthy smokers, 3 asthmatics) from Methods: whom EBC, saliva, and blood were collected in duplicate on two separate days. EBC was obtained using methods developed for field use, with salivary contamination minimized by insertion of upwardly inclined respiratory tubing between the mouthpiece and condenser. EBC and salivary concentrations of the biomarkers adenosine, adenosine monophosphate (AMP), phenylalanine, and tyrosine plus the dilution marker urea were measured using liquid chromatography-tandem mass spectrometry (LC-MS/MS), and salivary contamination was assessed by comparing salivary and EBC amylase concentrations with a sensitive, fluorometric assay. Airway lining fluid (ALF) concentrations were calculated by multiplying EBC concentrations by the ratios between plasma and EBC concentrations of urea.Salivary contamination of most EBC samples was minimal (1±17% of the respiratory droplet volumes) but exceeded 10% in 5 Results: samples from 3 subjects. EBC biomarkers and urea were readily detected, and although intra-individual variability was relatively high (average coefficients of variation = 0.44 to 0.65), it was reduced by averaging duplicate collections and using plasma:EBC urea ratios to calculate ALF concentrations: Adenosine: 0.41±0.29 μM SD, AMP= 0.43±0.35 μM, phenylalanine= 9.5±5.7 μM and tyrosine = 15.1±10.3 μM. As previously observed, ALF adenosine concentrations in asthmatics exceeded those in normals (p<0.05). Mean concentrations of urea were similar in saliva and plasma, suggesting that salivary samples may be used instead of plasma values, thereby avoiding discomfort of blood collection. Unexpectedly, the calculated concentrations of adenosine, AMP, tyrosine and phenylalanine in the ALF were similar to those measured in the saliva, suggesting that similar mechanisms regulate these parameters.We conclude that EBC can be successfully collected and analyzed using conventional portable equipment, modified to Conclusions: minimize salivary contamination. However, sensitive measures of biomarkers, as well as salivary and dilutional indicators remain essential to accurately determine airway biomarker concentrations.

Abstract. Human Cripto-1, a membrane-bound protein, plays an important role during early embryogenesis and has oncogenic properties, including cell transformation and enhancement of invasion. Cripto-1 is up-regulated in various malignant tissues and premalignant lesions. However, Cripto-1 expression in intraductal papillary mucinous neoplasms (IPMNs) has yet to be reported. This study aimed to investigate Cripto-1 expression in IPMNs and evaluate the expression patterns according to the histological grade or phenotypic subclassification. Cripto-1 expression was evaluated by immunohistochemistry using 37 IPMN tissue samples and real-time RT-PCR analysis of seven frozen samples. Cripto-1 was up-regulated in 59.5% of IPMNs. Cripto-1 was positively stained in 3 of 4 (75%) adenomas, 12 of 19 (63.2%) borderline neoplasms, 5 of 11 (45.5%) non-invasive carcinomas and 2 of 3 (66.7%) invasive carcinomas. There was no correlation between Cripto-1 overexpression and the histological grade (P>0.05). Cripto-1 expression was significantly increased in pancreatobiliary-(4/5, 80%) and gastric-type (13/19, 68.4.2%) IPMNs compared with those of the intestinal type (2/10, 20%; P<0.01). Cripto-1 mRNA expression was higher in gastricand pancreatobiliary-type IPMNs than in intestinal ones, supporting the immunohistochemical results. It is concluded that Cripto-1 overexpression is involved in the tumorigenesis of gastric-and pancreatobiliary-type IPMNs.