Augmenter of liver regeneration: an important intracellular survival factor for hepatocytes

BACKGROUND/AIMS: Augmenter of liver regeneration (ALR), a protein synthesized and stored in hepatocytes, is associated with mitochondria, and possesses sulfhydryl oxidase and cytochrome c reductase activities. We sought to determine the effects of ALR depletion in hepatocytes by antisense oligonucleotide transfection. METHODS: Rat hepatocytes in primary culture were transfected with antisense oligonucleotide for ALR mRNA (ALR-AS) or scrambled oligonucleotide. Various analyses were performed at times up to 24h after transfection. RESULTS: Treatment with ALR-AS caused a decrease in ALR mRNA, cellular depletion of ALR protein primarily from mitochondria, and decreased viability. Flow cytometric analysis of ALR-AS-transfected hepatocytes stained with annexin-Vcy3 and 7-aminoactinomycin D revealed apoptosis as the predominant cause of death up to 6h; incubation beyond this time resulted in necrosis in addition to apoptosis. ALR-AS-transfection caused release of mitochondrial cytochrome c, activation of caspase-3, profound reduction in the ATP content, and cellular release of LDH. Inhibition of caspase-3 inhibited the early phase of ALR-AS-induced death but not the late phase that included ALR and LDH release. CONCLUSIONS: These results suggest that ALR is critically important for the survival of hepatocytes by its association with mitochondria and regulation of ATP synthesis.     

Insecticidal and larvicidal activities of medicinal plant extracts against mosquitoes

In recent years, use of environment friendly and biodegradable natural insecticides of plant origin have received renewed attention as agents for vector control because they are rich in bioactive chemicals, active against a limited number of species including specific target insects, and biodegradable. The present study was carried out to evaluate the adulticidal, repellent, and larvicidal activity of crude hexane, ethyl acetate, and methanol extracts of eight plants, viz. Aristolochia indica L., Cassia angustifolia Vahl, Diospyros melanoxylon Roxb., Dolichos biflorus L., Gymnema sylvestre (Retz) Schult, Justicia procumbens L., Mimosa pudica L., and Zingiber zerumbet L., were tested against adult and early fourth instar larvae of Culex gelidus Theobald and Culex quinquefasciatus Say (Diptera: Culicidae). The effective adult mortality was observed in methanol extract of A. indica, ethyl acetate extract of D. biflorus, and ethyl acetate and hexane extract of Z. zerumbet against C. gelidus and C. quinquefasciatus (LD₅₀ = 37.75, 78.56, 129.44, 86.13, 80.06, 112.42, 53.83, and 46.61; LD₉₀ = 166.83, 379.14, 521.50, 289.83, 328.18, 455.72, 181.15, and 354.50 ppm, respectively). Complete protections for 150 min were found in hexane and methanol extract of A. indica and Z. zerumbet at 1,000 ppm against mosquito bites. The highest larval mortality was found in the hexane extract of Z. zerumbet, ethyl acetate extract of D. biflorus, and methanol extracts of A. indica against C. gelidus (LC₅₀ = 26.48, 33.02, and 12.47 ppm; LC₉₀ = 127.73, 128.79, and 62.33 ppm) and against C. quinquefasciatus (LC₅₀ = 69.18, 34.76, and 25.60 ppm; LC₉₀ = 324.40, 172.78, and 105.52 ppm), respectively, after 24 h. The plant extracts are potential to be used as an ideal eco-friendly approach for the control of the Japanese encephalitis vector, C. gelidus, and lymphatic filariasis vector, C. quinquefasciatus.     

An approach to defining the upper safe limits of amino acid intake

The existing data on the safe upper limits of amino acid intake in humans is essentially observational; how much do individuals ingest and what side effects do they have? There are numerous studies in humans comparing the effects of high doses of amino acids given as protein bound vs. as free amino acids. These studies have shown that protein-bound amino acids have much less effect on plasma levels of the test amino acid, because protein intake stimulates protein synthesis as another sink for the increased amino acid intake. In practice, the highest amino acid intakes occur with free amino acid supplements that may be ingested by athletes who believe that the amino acids will benefit them in training and/or performance. Previously, in a piglet study, we were able to define the point at which maximal phenylalanine oxidation occurred, above which plasma phenylalanine concentration and body balance rose exponentially. We regard this value of maximal disposal (oxidation) of an amino acid as one metabolic marker of the upper limit of intake. Recently, others have demonstrated a similar maximal oxidation rate for leucine in rats. Based on these experimental data and the paucity of published human data in controlled experiments, we think that a systematic approach needs to be undertaken to define the maximal oxidation rate for all dietary indispensable amino acids and other amino acids that may be ingested in excess by humans. We believe that this will provide a rational basis to begin to define the upper limits of tolerance for dietary amino acids. However, some amino acids, such as threonine and methionine, will be more difficult to study, because they have more than 1 route of disposal or very complex metabolic regulation, in which case defining their upper limits will be more multifaceted.     

Individual amino acid requirements in humans: an update

PURPOSE OF REVIEW: To discuss recent amino acid requirement studies in adult humans and school-age children, primarily determined using the indicator amino acid oxidation technique. RECENT FINDINGS: Using the minimally invasive indicator amino acid oxidation model, requirements for most indispensable amino acids have been defined in adult humans. The estimates are comparable to the values obtained using the more elaborate 24-h indicator amino acid oxidation and balance model. The less-invasive indicator amino acid oxidation model has also been successfully applied to define requirements in healthy school-age children and children with disease. A recent adaptation of the indicator amino acid oxidation method to determine protein requirements in adult humans resulted in mean and safe values of 0.93 and 1.2 g protein/kg/day, respectively. These estimates are 40-50% higher than current recommendations and suggest an urgent need to reassess recommendations for protein intake in humans. SUMMARY: In summary, indicator amino acid oxidation is a robust technique, and has resulted in the definition of amino acid and protein requirements in adult humans and children. A wider application of the technique in other vulnerable populations across life stages and in other diseases is now possible.     

Gene therapy using genetically modified lymphocytes targeting VEGFR-2 inhibits the growth of vascularized syngenic tumors in mice

Immunotherapies based on adoptive cell transfer are highly effective in the treatment of metastatic melanoma, but the use of this approach in other cancer histologies has been hampered by the identification of appropriate target molecules. Immunologic approaches targeting tumor vasculature provide a means for the therapy of multiple solid tumor types. We developed a method to target tumor vasculature, using genetically redirected syngeneic or autologous T cells. Mouse and human T cells were engineered to express a chimeric antigen receptor (CAR) targeted against VEGFR-2, which is overexpressed in tumor vasculature and is responsible for VEGF-mediated tumor progression and metastasis. Mouse and human T cells expressing the relevant VEGFR-2 CARs mediated specific immune responses against VEGFR-2 protein as well as VEGFR-2-expressing cells in vitro. A single dose of VEGFR-2 CAR-engineered mouse T cells plus exogenous IL-2 significantly inhibited the growth of 5 different types of established, vascularized syngeneic tumors in 2 different strains of mice and prolonged the survival of mice. T cells transduced with VEGFR-2 CAR showed durable and increased tumor infiltration, correlating with their antitumor effect. This approach provides a potential method for the gene therapy of a variety of human cancers.     

Nasal manifestations of yaws

Yaws is an infectious, non-venereal disease of the tropical countries, which is caused by Treponema pertenue. Gangosa and goundou were seen commonly in cases of yaws in Africa, particularly in the early part of this century. After successful WHO mass treatment campaigns, these conditions are rarely seen now. A case of yaws with gangosa, goundou, and a nasopharyngeal carcinoma is presented here for its rarity.     

Childhood deafness in Malaysia.

One hundred and fifty-five children with childhood deafness were examined over a period of 4 years in order to assess the aetiology of hearing disorder. In 21 (13%) children, deafness was a sequel of meningitis. Perinatal pathology accounted for 34 (22%) cases. The aetiology of deafness was unknown in 44 (28.4%) children. The percentage of unknown causes can be reduced if the deafness is detected early. Hearing loss was diagnosed only in 30 (19%) children by the age of 2 years. The early detection of deafness can be achieved by screening the high risk infants and educating the general practitioners and health assistants.