Epidemiology and seasonality of human parainfluenza serotypes 1-3 in Australian children.

BACKGROUND: Parainfluenza viruses are significant contributors to childhood respiratory illness worldwide, although detailed epidemiological studies are lacking. Few recent Australian studies have investigated serotype-specific PIV epidemiology, and there is a paucity of southern hemisphere PIV reports. We report age-stratified PIV hospitalisation rates and a mathematical model of PIV seasonality and dynamics in Western Australia (WA). METHODS: We used linked perinatal, hospital admission and laboratory diagnostic data of 469 589 children born in WA between 1996 and 2012. Age-specific rates of viral testing and PIV detection in hospitalised children were determined using person time-at-risk analysis. PIV seasonality was modelled using a compartmental SEIRS model and complex demodulation methods. RESULTS: From 2000 to 2012, 9% (n = 43 627) of hospitalised children underwent PIV testing, of which 5% (n = 2218) were positive for PIV-1, 2 or 3. The highest incidence was in children aged 1-5 months (PIV-1:62.6 per 100 000 child-years, PIV-2:26.3/100 000, PIV-3:256/100 000), and hospitalisation rates were three times higher for Aboriginal children compared with non-Aboriginal children overall (IRR: 2.93). PIV-1 peaked in the autumn of even-numbered years, and PIV-3 annually in the spring, whereas PIV-2 had inconsistent peak timing. Fitting models to the higher incidence serotypes estimated reproduction numbers of 1.24 (PIV-1) and 1.72 (PIV-3). CONCLUSION: PIV-1 and 3 are significant contributors towards infant respiratory hospitalisations. Interventions should prioritise children in the first 6 months of life, with respect to the observed autumn PIV-1 and spring PIV-3 activity peaks. Continued surveillance of all serotypes and investigation into PIV-1 and 3 interventions should be prioritised.

Unmasking latent inhibition in humans.

Presentations of a to-be-conditioned stimulus (CS) on its own impairs subsequent learning when that CS is paired with an unconditioned stimulus (US). Evidence for this latent inhibition (LI) effect in humans is said to require a "masking task" that diverts attention from the CS during preexposure. We present three experiments that demonstrate LI in humans without masking. Subjects performed a computerised task, making speeded responses to an imperative cue (the US) presented within a continuous stream of stimuli. During preexposure, a to-be-CS was presented 20 times among other stimuli, but excluding the US. Instructions ensured subjects actively monitored all stimuli at this time. This was immediately followed by the training phase, which included the US, the preexposed CS, and a novel CS. Both CSs were reliably followed by the US, but these associations were incidental to the instructed task. Nonetheless, some subjects learned the CS-US associations, responding faster when the US followed a CS than when it was unsignalled. All three experiments also found evidence for LI, in that subjects learned the novel CS-US association sooner than the preexposed CS-US association. We conclude that humans can show LI even when actively attending to the CS during preexposure.

Ibudilast reduces oxaliplatin-induced tactile allodynia and cognitive impairments in rats.

Chemotherapy can cause serious neurotoxic side effects, such as painful peripheral neuropathies and disabling cognitive impairments. Four experiments examined whether Ibudilast, a clinically approved neuroimmune therapy, would reduce tactile allodynia and memory impairments caused by oxaliplatin in laboratory rats. Rats received an intraperitoneal injection of oxaliplatin (6mg/kg i.p.) or vehicle and were assessed for tactile allodynia 3 or 5days after injection, memory impairments in the novel object and novel location recognition tests 10-12days after injection, and fear conditioning 14days after injection. Ibudilast (7.5mg/kg) or vehicle was administered prior to oxaliplatin (Experiments 1 and 3) or prior to behavioural testing (Experiments 2 and 4). Ibudilast treatment prior to oxaliplatin prevented the development of tactile allodynia and memory impairments. Ibudilast treatment prior to behavioural testing reduced oxaliplatin-induced tactile allodynia, memory impairments, and impaired renewal of fear conditioning. These results suggest that Ibudilast could be an effective treatment against oxaliplatin-induced neuropathies and cognitive impairments.

WWOX, the FRA16D gene, behaves as a suppressor of tumor growth.

We recently reported the cloning of WWOX, a gene that maps to the common fragile site FRA16D region in chromosome 16q23.3-24.1. It was observed that the genomic area spanned by WWOX is affected by chromosomal translocations and homozygous deletions. Furthermore, the high incidence of allelic loss in breast, ovarian, prostate, and other cancers affecting this region suggests that WWOX is a candidate tumor suppressor gene. Expression of WWOX is highly variable in breast cancer cell lines, with some cases showing low or undetectable levels of expression. In this report, we demonstrate that ectopic WWOX expression strongly inhibits anchorage-independent growth in soft agar of breast cancer cell lines MDA-MB-435 and T47D. Additionally, we observed that WWOX induces a dramatic inhibition of tumorigenicity of MDA-MB-435 breast cancer cells when tested in vivo. We also detected the common occurrence of aberrant WWOX transcripts with deletions of exons 5-8 or 6-8 in various carcinoma cell lines, multiple myeloma cell lines, and primary breast tumors. These aberrant mRNA forms were not detected in normal tissues. Interestingly, we further observed that proteins encoded by such aberrant transcripts display an abnormal nuclear localization in contrast to the wild-type WWOX protein that localizes to the Golgi system. Our data indicate that WWOX behaves as a potent suppressor of tumor growth and suggest that abnormalities affecting this gene at the genomic and transcriptional level may be of relevance in carcinogenesis.

Heterozygosity for the mouse Apex gene results in phenotypes associated with oxidative stress.

Apurinic/apyrimidinic endonuclease is a key enzyme in the process of base excision repair, required for the repair of spontaneous base damage that arises as a result of oxidative damage to DNA. In mice, this endonuclease is coded by the Apex gene, disruption of which is incompatible with embryonic life. Here we confirm the embryonic lethality of Apex-null mice and report the phenotypic characterization of mice that are heterozygous mutants for the Apex gene (Apex+/-). We show that Apex heterozygous mutant cells and animals are abnormally sensitive to increased oxidative stress. Additionally, such animals manifest elevated levels of oxidative stress markers in serum, and we show that dietary supplementation with antioxidants restores these to normal levels. Apex+/- embryos and pups manifest reduced survival that can also be partially rescued by dietary supplementation with antioxidants. These results are consistent with a proposed role for this enzyme in protection against the deleterious effects of oxidative stress and raise the possibility that humans with heterozygous mutations in the homologous HAP1 gene may be at increased risk for the phenotypic consequences of oxidative stress in cells.

Cytotoxic effects of 10% citric acid and EDTA-T used as root canal irrigants: an in vitro analysis.

EDTA-T and 10% citric acid used as root canal irrigants lead to more visible dentinal tubules than 5% sodium hypochlorite associated with 3% hydrogen peroxide. However, these cleansing agents must be compatible with apical periodontal tissue. We analyzed the cytotoxicity of 10% citric acid and EDTA-T in cultured fibroblasts using Trypan blue. The solutions were diluted to 1%, 0.1%, and 0.01% and applied to NIH 3T3 cell cultures. Cells grown on fresh DMEM served as a control. After 0, 6, 12, and 24 h (short-term assay, viability) and 1, 3, 5, and 7 days (long-term assay, survival), the cells were counted using a hemocytometer. In short-term tests, cell viability ranged from 85% to 99% for all experimental groups with no statistical differences when compared with control cultures, except for the group treated with 1% EDTA-T, which caused a progressive decrease in cell viability. In long-term tests, all cultures increased in number from day 1 to the end of the experimental period, showing no inhibition of cell proliferation, except for the cultures treated with 1% EDTA-T, which totally prevented cell growth. All dilutions of 10% citric acid were more biocompatible than EDTA-T. Cultures treated with citric acid had a higher percentage of viable cells in the short-term assays, and the cells retained their self-renewal capacity.

Effects of estrogen on global gene expression: identification of novel targets of estrogen action.

The important role played by the sex hormone estrogen in disease and physiological processes has been well documented. However, the mechanisms by which this hormone elicits many of its normal as well as pathological effects are unclear. To identify both known and unknown genes that are regulated by or associated with estrogen action, we performed serial analysis of gene expression on estrogen-responsive breast cancer cells after exposure to this hormone. We examined approximately 190,000 mRNA transcripts and monitored the expression behavior of 12,550 genes. Expression levels for the vast majority of those transcripts were observed to remain constant upon 17beta estradiol (E2) treatment. Only approximately 0.4% of the genes showed an increase in expression of > or =3-fold by 3 h post-E2 treatment. We cloned five novel genes (E2IG1-5), which were observed up-regulated by the hormonal treatment. Of these the most highly induced transcript, E2IG1, appears to be a novel member of the family of small heat shock proteins. The E2IG4 gene is a new member of the large family of leucine-rich repeat-containing proteins. On the basis of architectural and domain homology, this gene appears to be a good candidate for secretion in the extracellular environment and, therefore, may play a role in breast tissue remodeling and/or epithelium-stroma interactions. Several interesting genes with a potential role in the regulation of cell cycle progression were also identified to increase in expression, including Pescadillo and chaperonin CCT2. Two putative paracrine/autocrine factors of potential importance in the regulation of the growth of breast cancer cells were identified to be highly up-regulated by E2: stanniocalcin 2, a calcium/phosphate homeostatic hormone; and inhibin-beta B, a TGF-beta-like factor. Interestingly, we also determined that E2IG1 and stanniocalcin 2 were exclusively overexpressed in estrogen-receptor-positive breast cancer lines, and thus they have the potential to serve as breast cancer biomarkers. This data provides a comprehensive view of the changes induced by E2 on the transcriptional program of human E2-responsive cells, and it also identifies novel and previously unsuspected gene targets whose expression is affected by this hormone.

WWOX, a novel WW domain-containing protein mapping to human chromosome 16q23.3-24.1, a region frequently affected in breast cancer.

Studies were conducted with the final goal of identifying genes of interest mapping to the chromosome region 16q23.3-24.1, an area commonly affected by allelic losses in breast cancer. To this end we generated a detailed physical map of the genomic region spanning between sequence-tagged site markers D16S518 and D16S516. To identify candidate genes, we used shotgun genomic sequencing as well as isolation and analysis of transcripts mapping to the area of interest. We identified and cloned a novel gene, the genomic structure of which spans the whole region of interest. We named this gene WWOX because it contains two WW domains coupled to a region with high homology to the short-chain dehydrogenase/reductase family of enzymes. The ORF of WWOX is 1245 bp long, encoding a 414-amino acid protein. This gene is composed of nine exons. We performed a mutation screening of WWOX exons in a panel of breast cancer lines, most of which are hemizygous for the 16q genomic region indicated. We found no evidence of mutations, thus indicating that WWOX is probably not a tumor suppressor gene. However, we observed that one case of homozygous deletion as well as two previously described translocation breakpoints map to intronic regions of this gene. We speculate that WWOX may span the yet uncharacterized common fragile site FRA16D region. In expression studies we found overexpression of WWOX in breast cancer cell lines when compared with normal breast cells and tissues. The highest normal expression of WWOX was observed in hormonally regulated tissues such as testis, ovary, and prostate. This expression pattern and the presence of a short-chain dehydrogenase/reductase domain and specific amino acid features suggest a role for WWOX in steroid metabolism. Interestingly, the presence of WW domains in the structure of WWOX indicate the likelihood that this protein physically interacts with other proteins. The unique features of WWOX and its possible association with cancer processes make it an interesting target for further investigation.

A new classification system predicting keratomalacia after trauma in vitamin A deficiency: sodium citrate does not prevent disease progression.

PURPOSE: The purpose of this study was to develop a classification system to predict keratomalacia after trauma in vitamin A-deficient eyes and to determine whether citrate impedes polymorphonuclear leukocyte infiltration into the cornea, thus preventing keratomalacia. METHODS: Preliminary classification studies showed that a 7.0-mm corneal epithelial scrape, before clinical findings of corneal xerosis, did not induce keratomalacia. Primary studies were conducted concurrently on the same animals to develop the classification system and test the effect of citrate in vitamin A deficiency. A 7.0-mm corneal epithelial scrape was performed on vitamin A-deficient eyes in various stages of corneal xerosis and treated as follows. Experiment 1: group 1, 10% citrate drops; group 2, phosphate buffer solution (PBS) drops; experiment II: group 3, drops and subconjunctival injection of 10% citrate; group 4, drops and subconjunctival injection of PBS. RESULTS: Corneal abrasion in eyes with 2+ corneal xerosis yielded keratomalacia in 50% of cases; the remainder healed with xerotic epithelium. Eighty-three percent of eyes with > 2+ xerosis developed keratomalacia after corneal abrasion, whereas only 7.1% of eyes with < 2+ xerosis advanced to this stage. In experiment I, 27% of citrate-treated eyes and 38% of PBS-treated eyes developed keratomalacia (not significant). In experiment II, two of six citrate-treated eyes perforated and one eye developed keratomalacia. One of six control PBS eyes perforated and four developed keratomalacia. CONCLUSION: We correlated the degree of corneal xerosis with the occurrence of keratomalacia after corneal trauma. This led to the development of a classification scale that is of research and clinical significance. Additionally, citrate did not significantly reduce keratomalacia or perforation in the vitamin. A-deficient eye.

Case-control study of brain tumor deaths among employees at a chemical manufacturing plant.

Recent studies have reported an increased risk of death from primary intracranial neoplasms among employees in the petrochemical industry. One of them, a NIOSH/OSHA county-based case-control study, suggested a twofold risk for having ever been employed at the Texas Division of Dow Chemical U.S.A. Using the case-control approach, we compared 24 brain tumor cases among employees at this large, diverse chemical plant in Texas with two sets of age-, race-, and sex-matched controls to determine if a risk could be associated with job assignment in a particular process area, with presumptive exposure to a major process chemical, or with a number of other occupationally related and unrelated variables. Results implicated no specific area or chemical. The only significant findings were an elevated odds for being hired prior to 1950 and a negative correlation with employment greater than 10 years. Further, preliminary calculations of expected deaths from primary intracranial neoplasms among the employee cohort suggested there was no excessive risk.