Apoptosis,cytokine and chemokine induction by non-structural 1 (NS1) proteins encoded by different influenza subtypes

Background

Influenza pandemic remains a serious threat to human health. Viruses of avian origin, H5N1, H7N7 and H9N2, have repeatedly crossed the species barrier to infect humans. Recently, a novel strain originated from swine has evolved to a pandemic. This study aims at improving our understanding on the pathogenic mechanism of influenza viruses, in particular the role of non-structural (NS1) protein in inducing pro-inflammatory and apoptotic responses.

Methods

Human lung epithelial cells (NCI-H292) was used as an in-vitro model to study cytokine/chemokine production and apoptosis induced by transfection of NS1 mRNA encoded by seven infleunza subtypes (seasonal and pandemic H1, H2, H3, H5, H7, and H9), respectively.

Results

The results showed that CXCL-10/IP10 was most prominently induced (> 1000 folds) and IL-6 was slightly induced (< 10 folds) by all subtypes. A subtype-dependent pattern was observed for CCL-2/MCP-1, CCL3/MIP-1α, CCL-5/RANTES and CXCL-9/MIG; where induction by H5N1 was much higher than all other subtypes examined. All subtypes induced a similar temporal profile of apoptosis following transfection. The level of apoptosis induced by H5N1 was remarkably higher than all others. The cytokine/chemokine and apoptosis inducing ability of the 2009 pandemic H1N1 was similar to previous seasonal strains.

Conclusions

In conclusion, the NS1 protein encoded by H5N1 carries a remarkably different property as compared to other avian and human subtypes, and is one of the keys to its high pathogenicity. NCI-H292 cells system proves to be a good in-vitro model to delineate the property of NS1 proteins.

     

STING Millennium Suite: integrated software for extensive analyses of 3d structures of proteins and their complexes

Background  

The integration of many aspects of protein/DNA structure analysis is an important requirement for software products in general area of structural bioinformatics. In fact, there are too few software packages on the internet which can be described as successful in this respect. We might say that what is still missing is publicly available, web based software for interactive analysis of the sequence/structure/function of proteins and their complexes with DNA and ligands. Some of existing software packages do have certain level of integration and do offer analysis of several structure related parameters, however not to the extent generally demanded by a user.     

Polymer optoelectronic structures for retinal prosthesis

This commentary highlights the effectiveness of optoelectronic properties of polymer semiconductors based on recent results emerging from our laboratory, where these materials are explored as artificial receptors for interfacing with the visual systems. Organic semiconductors based polymer layers in contact with physiological media exhibit interesting photophysical features, which mimic certain natural photoreceptors, including those in the retina. The availability of such optoelectronic materials opens up a gateway to utilize these structures as neuronal interfaces for stimulating retinal ganglion cells. In a recently reported work entitled “A polymer optoelectronic interface provides visual cues to a blind retina,” we utilized a specific configuration of a polymer semiconductor device structure to elicit neuronal activity in a blind retina upon photoexcitation. The elicited neuronal signals were found to have several features that followed the optoelectronic response of the polymer film. More importantly, the polymer-induced retinal response resembled the natural response of the retina to photoexcitation. These observations open up a promising material alternative for artificial retina applications.     

Genome-scale metabolic reconstruction and <Emphasis Type="Italic">in silico</Emphasis> analysis of methylotrophic yeast <Emphasis Type="Italic">Pichia pastoris</Emphasis> for strain improvement

Background  

Pichia pastoris has been recognized as an effective host for recombinant protein production. A number of studies have been reported for improving this expression system. However, its physiology and cellular metabolism still remained largely uncharacterized. Thus, it is highly desirable to establish a systems biotechnological framework, in which a comprehensive in silico model of P. pastoris can be employed together with high throughput experimental data analysis, for better understanding of the methylotrophic yeast's metabolism.     

Thermophysiological responses of human volunteers to whole body RF exposure at 220 MHz

Since 1994, our research has demonstrated how thermophysiological responses are mobilized in human volunteers exposed to three radio frequencies, 100, 450, and 2450 MHz. A significant gap in this frequency range is now filled by the present study, conducted at 220 MHz. Thermoregulatory responses of heat loss and heat production were measured in six adult volunteers (five males, one female, aged 24-63 years) during 45 min whole body dorsal exposures to 220 MHz radio frequency (RF) energy. Three power densities (PD = 9, 12, and 15 mW/cm(2) [1 mW/cm(2) = 10 W/m(2)], whole body average normalized specific absorption rate [SAR] = 0.045 [W/kg]/[mW/cm(2)] = 0.0045 [W/kg]/[W/m(2)]) were tested at each of three ambient temperatures (T(a) = 24, 28, and 31 degrees C) plus T(a) controls (no RF). Measured responses included esophageal (T(esoph)) and seven skin temperatures (T(sk)), metabolic rate (M), local sweat rate, and local skin blood flow (SkBF). Derived measures included heart rate (HR), respiration rate, and total evaporative water loss (EWL). Finite difference-time domain (FDTD) modeling of a seated 70 kg human exposed to 220 MHz predicted six localized "hot spots" at which local temperatures were also measured. No changes in M occurred under any test condition, while T(esoph) showed small changes (< or =0.35 degrees C) but never exceeded 37.3 degrees C. As with similar exposures at 100 MHz, local T(sk) changed little and modest increases in SkBF were recorded. At 220 MHz, vigorous sweating occurred at PD = 12 and 15 mW/cm(2), with sweating levels higher than those observed for equivalent PD at 100 MHz. Predicted "hot spots" were confirmed by local temperature measurements. The FDTD model showed the local SAR in deep neural tissues that harbor temperature-sensitive neurons (e.g., brainstem, spinal cord) to be greater at 220 than at 100 MHz. Human exposure at both 220 and 100 MHz results in far less skin heating than occurs during exposure at 450 MHz. However, the exposed subjects thermoregulate efficiently because of increased heat loss responses, particularly sweating. It is clear that these responses are controlled by neural signals from thermosensors deep in the brainstem and spinal cord, rather than those in the skin.     

Oncogenic comparison of human papillomavirus type 58 E7 variants

Human papillomavirus 58 (HPV58) ranks the second or third in East Asian cervical cancers. Current studies on HPV58 are scarce and focus on the prototype. Previously, we identified the three most common circulating HPV58 E7 strains contained amino acid alterations: G41R/G63D (51%), T20I/G63S (22%) and T74A/D76E (14%) respectively. Among them, the T20I/G63S variant (V1) had a stronger epidemiological association with cervical cancer. We therefore suggested that V1 possessed stronger oncogenicity than the other two variants. Here, we performed phenotypic assays to characterize and compare their oncogenicities with HPV58 E7 prototype. Our results showed that overexpression of V1 conferred a higher colony‐forming ability to primary murine epithelial cells than prototype (P < 0.05) and other variants, implicating its higher immortalising potential. Further experiments showed that both V1 and prototype enhanced the anchorage‐independent growth of NIH/3T3 cells (P < 0.001), implicating their stronger transforming power than the two other variants. Moreover, they possessed an increased ability to degrade pRb (P < 0.001), which is a major effector pathway of E7‐driven oncogenesis. Our work represents the first study to compare the oncogenicities of HPV58 E7 prototype and variants. These findings deepened our understanding of HPV58 and might inform clinical screening and follow‐up strategy.     

Thermophysiological consequences of whole body resonant RF exposure (100 MHz) in human volunteers

Thermophysiological responses of heat production and heat loss were measured in seven adult volunteers (six males and one female, aged 31-74 years) during 45 min dorsal exposures of the whole body to 100 MHz continuous wave (CW) radio frequency (RF) energy. Three power densities (PD) (average PD = 4, 6, and 8 mW/cm(2); whole body specific absorption rate [SAR] = 0.068 [W/kg]/[mW/cm(2)]) were tested in each of three ambient temperatures (T(a) = 24, 28, and 31 degrees C), as well as in T(a) controls (no RF). A standardized protocol (30 min baseline, 45 min RF or sham exposure, 10 min baseline) was used. Measured responses included esophageal and seven skin temperatures, metabolic heat production, local sweat rate, and local skin blood flow. No changes in metabolic heat production occurred under any test condition. Unlike published results of similar exposures at 450 and 2450 MHz, local skin temperatures, even those on the back that were irradiated directly, changed little or not at all during 100 MHz exposures. The sole exception was the temperature of the ankle skin, which increased by 3-4 degrees C in some subjects at PD = 8 mW/cm(2). During the 45 min RF exposure, esophageal temperature showed modest changes (range = -0.15 to 0.13 degrees C) and never exceeded 37.2 degrees C. Thermoregulation was principally controlled by appropriate increases in evaporative heat loss (sweating) and, to a lesser extent, by changes in skin blood flow. Because of the deep penetration of RF energy at this frequency, effectively bypassing the skin, these changes must have been stimulated by thermal receptors deep in the body rather than those located in the skin.     

The occurrence of two species of <Emphasis Type="Italic">Macroramphosus</Emphasis> (Gasterosteiformes: Macroramphosidae) in Japan: morphological and ecological observations on larvae,juveniles, and adults

Earlier opinions that Macroramphosus is monotypic are refuted, with two species apparently occurring in Japan (tentatively identified as M. gracilis and M. scolopax). In postsettlement young and adults, the former is characterized by a dark slender body (vs. red-orange and deep) and short second dorsal fin spine with a smooth posterior margin (vs. long spine with a serrated margin). Food habits also differ between the two species, which are either plankton or benthos feeders. Two types of Macroramphosus larvae and juveniles occurring at the surface were recognized, one having a straight ventral body profile of the body (identified here as M. gracilis) and the other having a notch in the anal region. The dark body of postsettlement M. gracilis is considered to be a retention of the character suited to the neustonic distribution of the larval and juvenile stages, the species remaining to ca. 40mm in standard length (SL) in that habitat (vs. to ca. 12mm SL in M. scolopax).