Mental health & maltreatment risk of children with special educational needs during COVID-19.

BACKGROUND: Children with special educational needs (SEN) are more vulnerable during the COVID-19 pandemic with risk of poor mental wellbeing and child maltreatment. OBJECTIVE: To examine the impact of COVID-19 on the mental health of children with SEN and their maltreatment risk. PARTICIPANTS AND SETTING: 417 children with SEN studying at special schools and 25,427 children with typical development (TD) studying at mainstream schools completed an online survey in April 2020 in Hong Kong during school closures due to COVID-19. METHOD: Emotional/behavioural difficulties, quality of life and parental stress of children with SEN were compared with typically developed children using mixed effect model. Linear regression analyses were performed to explore factors associated with child emotional/behavioural difficulties and parental stress during the pandemic. Chi-square test was performed to detect the differences in maltreatment risk before and during COVID-19. RESULTS: Children with SEN had significantly poorer overall quality of life (68.05 vs 80.65, p < 0.01). 23.5% of children had at least one episode of severe physical assault and 1.9% experienced very severe physical assault during COVID-19. Rates of physical assault increased significantly (59.8% vs. 71.2% p < 0.001) while children with mental disorders had increased risk of severe physical assault comparing to those without mental disorders (RR = 1.58, ꭓ2 = 5.19 p = 0.023). CONCLUSION: Children with SEN had poorer mental health than typically developed children during the COVID-19 pandemic. Maltreatment risk for children with SEN is higher in comparison to pre-COVID-19 era. Surveillance of child maltreatment, continuity of medical and rehabilitation care to support children with SEN are essential during a disease pandemic.

The role of fruit bats in the transmission of pathogenic leptospires in Australia.

Although antileptospiral antibodies and leptospiral DNA have been detected in Australian fruit bats, the role of such bats as infectious hosts for the leptospires found in rodents and humans remains unconfirmed. A cohort-design, replicated survey was recently conducted in Far North Queensland, Australia, to determine if the abundance and leptospiral status of rodents were affected by association with colonies of fruit bats (Pteropus conspicillatus spp.) via rodent contact with potentially infectious fruit-bat urine. In each of four study areas, a 'colony site' that included a fruit-bat colony and the land within 1500 m of the colony was compared with a 'control site' that held no fruit-bat colonies and was >2000 m from the nearest edge of the colony site. Rodents were surveyed, for a total of 2400 trap-nights, over six sampling sessions between September 2007 and September 2008. A low abundance of rodents but a high carriage of leptospires in the rodents present were found to be associated with proximity to a fruit-bat colony. For example, means of 0·4 and 2·3 fawn-footed melomys (Melomys cervinipes) were collected/100 trap-nights at sites with and without fruit-bat colonies, respectively (P<0·001), but the corresponding prevalences of leptospiral carriage were 100% and 3·6% (P<0·001). Such trends were consistent across all of the sampling sessions but not across all of the sampling sites. Leptospires were not isolated from fruit bats by culture, and the role of such bats in the transmission of leptospires to rodents cannot be confirmed. The data collected do, however, indicate the existence of a potential pathway for transmission of leptospires from fruit bats to rodents, via rodent contact with infectious fruit-bat urine. Fruit bats may possibly be involved in the ecology of leptospires (including emergent serovars), as disseminators of pathogens to rodent populations. Stringent quantitative risk analysis of the present and similar data, to explore their implications in terms of disease prevalence and wildlife population dynamics, is recommended.

High-resolution melt-curve analysis of random amplified polymorphic DNA (RAPD-HRM) for the characterisation of pathogenic leptospires: intra-serovar divergence, inter-serovar convergence, and evidence of attenuation in Leptospira reference collections.

High-resolution melt-curve analysis of random amplified polymorphic DNA (RAPD-HRM) is a novel technology that has emerged as a possible method to characterise leptospires to serovar level. RAPD-HRM has recently been used to measure intra-serovar convergence between strains of the same serovar as well as inter-serovar divergence between strains of different serovars. The results indicate that intra-serovar heterogeneity and inter-serovar homogeneity may limit the application of RAPD-HRM in routine diagnostics. They also indicate that genetic attenuation of aged, high-passage-number isolates could undermine the use of RAPD-HRM or any other molecular technology. Such genetic attenuation may account for a general decrease seen in titres of rabbit hyperimmune antibodies over time. Before RAPD-HRM can be further advanced as a routine diagnostic tool, strains more representative of the wild-type serovars of a given region need to be identified. Further, RAPD-HRM analysis of reference strains indicates that the routine renewal of reference collections, with new isolates, may be needed to maintain the genetic integrity of the collections.

High-resolution melt-curve analysis of random-amplified-polymorphic-DNA markers, for the characterisation of pathogenic Leptospira.

A new test for pathogenic Leptospira isolates, based on RAPD-PCR and high-resolution melt (HRM) analysis (which measures the melting temperature of amplicons in real time, using a fluorescent DNA-binding dye), has recently been developed. A characteristic profile of the amplicons can be used to define serovars or detect genotypes. Ten serovars, of leptospires from the species Leptospira interrogans (serovars Australis, Robinsoni, Hardjo, Pomona, Zanoni, Copenhageni and Szwajizak), L. borgpetersenii (serovar Arborea), L. kirschneri (serovar Cynopteri) and L. weilii (serovar Celledoni), were typed against 13 previously published RAPD primers, using a real-time cycler (the Corbett Life Science RotorGene 6000) and the optimised reagents from a commercial kit (Quantace SensiMix). RAPD-HRM at specific temperatures generated defining amplicon melt profiles for each of the tested serovars. These profiles were evaluated as difference-curve graphs generated using the RotorGene software package, with a cut-off of at least 8 'U' (plus or minus). The results demonstrated that RAPD-HRM can be used to measure serovar diversity and establish identity, with a high degree of stability. The characterisation of Leptospira serotypes using a DNA-based methodology is now possible. As an objective and relatively inexpensive and rapid method of serovar identification, at least for cultured isolates, RAPD-HRM assays show convincing potential.

Cryopreservation of spermatozoa of black marlin, Makaira indica (Teleostei: Istiophoridae).

As a first step towards the development of a method for the cryopreservation of black marlin spermatozoa, this study investigated the effect of dimethylsulfoxide (DMSO) concentration and pellet size on post-thaw spermatozoal motility. Spermatozoa were recovered from the spermatic duct of testes retrieved post-mortem from four adult black marlin caught in the Coral Sea spawning grounds of Australia. Undiluted spermatozoa were stored on ice for 4 to 10 hours during transport to shore, then evaluated for motility after activation in seawater (1:10 v:v). Spermatozoa were prepared for cryopreservation in pellets by extension (1:3 v:v) in a defined fish Ringer's solution to give two final DMSO concentrations of 2.5% or 5.0%. Diluted spermatozoa were frozen directly on a dry ice block in pellet sizes of either 0.25 ml or 0.50 ml. Frozen pellets were thawed in a water bath at 40 degrees C for 60 seconds and assessed for post-thaw motility following activation in seawater. Spermatozoa recovered within 50 minutes of death and chilled on ice for 4 to 10 hours showed a mean (+/- SEM) motility immediately following activation of 91.6 +/- 7.9%. 50% of the spermatozoa remained motile for approximately 4 to 5 minutes. Following cryopreservation, mean motility declined significantly across all cryoprotectant and pellet size combinations (P < 0.001) but spermatozoa frozen in 2.5% DMSO showed higher motility than those frozen in 5.0% DMSO (P = 0.014). Pellet size had no effect on post-thaw motility (P = 0.179).

Flying foxes as carriers of pathogenic Leptospira species.

Recent serologic studies have identified flying foxes (Pteropus spp.) as carriers of leptospirosis; however, little is known about the role of flying foxes as carriers of pathogenic Leptospira spp. To determine if Australian Pteropus spp. are carriers of pathogenic Leptospira spp., TaqMan real-time polymerase chain reaction (PCR) was used to detect leptospiral DNA in kidney and urine specimens from four species of flying fox, including the spectacled flying fox (Pteropus conspicillatus), black flying fox (Pteropus alecto), grey-headed flying fox (Pteropus poliocephalus), and little red flying fox (Pteropus scapulatus). Of the 173 kidney samples tested, 19 (11%) were positive for leptospiral DNA. Positive individuals were detected in all four species; significant differences in prevalence were not detected between species, between species within the same geographic area, or between geographically separated samples from the same species. Of the 46 urine samples tested, 18 (39%) tested positive by PCR, confirming that flying foxes shed leptospires into the environment. The detection of leptospiral DNA in the kidneys and urine of flying foxes suggests that flying foxes are carriers of pathogenic Leptospira spp. No evidence collected in the present study, however, suggests that flying foxes pose a significant risk of leptospirosis to the wider community or that humans who are in regular, close contact with flying foxes are at risk for leptospirosis.

Diversity and Community Composition of Vertebrates in Desert River Habitats.

Animal species are seldom distributed evenly at either local or larger spatial scales, and instead tend to aggregate in sites that meet their resource requirements and maximise fitness. This tendency is likely to be especially marked in arid regions where species could be expected to concentrate at resource-rich oases. In this study, we first test the hypothesis that productive riparian sites in arid Australia support higher vertebrate diversity than other desert habitats, and then elucidate the habitats selected by different species. We addressed the first aim by examining the diversity and composition of vertebrate assemblages inhabiting the Field River and adjacent sand dunes in the Simpson Desert, western Queensland, over a period of two and a half years. The second aim was addressed by examining species composition in riparian and sand dune habitats in dry and wet years. Vertebrate species richness was estimated to be highest (54 species) in the riverine habitats and lowest on the surrounding dune habitats (45 species). The riverine habitats had different species pools compared to the dune habitats. Several species, including the agamid Gowidon longirostris and tree frog Litoria rubella, inhabited the riverine habitats exclusively, while others such as the skinks Ctenotus ariadnae and C. dux were captured only in the dune habitats. The results suggest that, on a local scale, diversity is higher along riparian corridors and that riparian woodland is important for tree-dependent species. Further, the distribution of some species, such as Mus musculus, may be governed by environmental variables (e.g. soil moisture) associated with riparian corridors that are not available in the surrounding desert environment. We conclude that inland river systems may be often of high conservation value, and that management should be initiated where possible to alleviate threats to their continued functioning.

Regulation of death promoter Bak expression by cell density and 17 beta-estradiol in MCF-7 cells.

In the current study we examined the regulation of Bak, a death promoter of an apoptotic pathway, in the human breast cancer cell line MCF-7. We observed a time-dependent increase in both Bak mRNA and protein levels which appeared to correlate well with the increase in cell density. We also found that treatment of cells with 17beta-estradiol resulted in inhibition of the time-dependent increases in Bak mRNA and protein. The effects of estradiol appeared to be via estrogen receptor as treatment of cells with progesterone did not effect Bak expression. Our study provides additional molecular evidence for (1) a link between apoptosis pathways and cell-cell and/or cell-cell matrix interactions and (2) a role for estradiol in the modulation of signals between apoptosis pathways and cell-cell and/or cell-cell matrix interactions.

Comparison of Immobilon-N membrane and other membranes for the detection of HIV-1 genome in high risk patients using PCR.

The polymerase chain reaction (PCR) was used for detection of the HIV-1 genome from the peripheral blood lymphocytes of high risk patients. The gag regions of HIV-1 (SK38-SK39) were chosen to amplify viral DNA and the amplified products were spotted onto membrane filters and hybridized with a 32P-labeled SK19 probe. Nitrocellulose, nylon and polyvinylidene difluoride (PVDF) membrane filters were used and compared in dot-blot hybridization. PVDF (Immobilon-N, trade name) filter membranes were demonstrated to be the best membranes on the basis of hybridization data and showed a stronger signal on autoradiograms than the other two types (nitrocellulose and nylon).

Lack of correlation between hepatic prostaglandin concentrations and DNA synthesis after the administration of phenobarbital and the peroxisome proliferator ciprofibrate in rats.

Peroxisome proliferators are a class of chemicals that induce and promote hepatic tumors in rodents. These compounds are not genotoxic, and the mechanism by which they induce and promote tumors is poorly understood. Phenobarbital (PB) also is a hepatic tumor promoter that produces a different natural history than peroxisome proliferators during the promotion of hepatocarcinogenesis. In addition, opposite effects on hepatic eicosanoid concentrations have been demonstrated previously. In this experiment, we examined whether higher hepatic eicosanoid concentrations correlated with the induction of DNA synthesis after the administration of PB or the peroxisome proliferator ciprofibrate (CIP). PB (0.05% in diet) or CIP (0.01% in diet) was fed to rats from 1-10 days. For the rats treated with CIP, the peroxisomal enzyme fatty acyl-CoA oxidase increased gradually from day 1 to day 10. PB treated rats had a higher cytochrome P450 2B1/2 activity over the entire course of feeding. Hepatic prostaglandins E2 and F2alpha concentrations were significantly reduced in the rats treated with CIP, while no significant differences were seen between the control and PB-treated rats. DNA synthesis was increased in both PB-treated and CIP-treated rats. These results show that higher eicosanoid concentrations do not correlate with the induction of hepatic DNA synthesis by CIP or PB.

Effect of the peroxisome proliferator ciprofibrate on hepatic cyclooxygenase and phospholipase A2 in rats.

Peroxisome proliferators, which include several hypolipidemic drugs, plasticizers and other chemicals, induce hepatic tumors in rodents. These chemicals alter the expression of enzymes involved in lipid metabolism, such as the cytochrome P450 4A family and peroxisomal beta-oxidation enzymes. Previous studies have shown that the peroxisome proliferator ciprofibrate reduces eicosanoid concentrations in rat livers and primary hepatocyte cultures, yet the mechanism is still unclear. In this study we examined cyclooxygenases 1 and 2 (COX-1 and COX-2) and cytosolic phospholipase A2 (cPLA2) to determine whether the rate-limiting enzymes in the eicosanoid synthetic pathway are altered by ciprofibrate. Rats were fed 0.01% ciprofibrate for 3, 6, or 10 days. Western analysis revealed that COX-2 protein was induced by ciprofibrate (up to 13-fold at day 10), but that calcium-dependent (Ca-D) cPLA2 protein was not different from controls. The enzyme activity of calcium-independent (Ca-I) cPLA2 in ciprofibrate-treated rats was increased 2-fold, whereas Ca-D cPLA2 and total COX activities were not affected. Using enzyme kinetics, we found that COX-1 (Ki = 143 microM) and Ca-I cPLA2 (Ki = 121 microM) were competitively inhibited by ciprofibrate, but the inhibition was not physiologically significant. COX-2 and Ca-D cPLA2 were not inhibited by ciprofibrate. These results show that ciprofibrate increases Ca-I cPLA2 enzyme activity and COX-2 protein expression.

Reduction of the concentrations of prostaglandins E2 and F2alpha, and thromboxane B2 in cultured rat hepatocytes treated with the peroxisome proliferator ciprofibrate.

Several hypolipidemic drugs, plasticizers and other chemicals induce peroxisome proliferation and hepatic tumors in rodents, but the mechanism by which they induce tumors is not fully understood. Their carcinogenic activity may be related to alterations in gene expression, such as induction of peroxisomal beta-oxidation enzymes or of the cytochrome P450 4A family. These enzymes metabolize lipids, including eicosanoids and their precursor fatty acids. Because eicosanoids likely play a role in the carcinogenic process, alterations in their concentration by xenobiotics may be important in their carcinogenic or promoting activities. In this study we used isolated hepatocytes to study if peroxisome proliferators alter the metabolism of prostaglandins (PG) and thromboxanes (Tx). Isolated rate hepatocytes were cultured for 4 days with 2 concentrations of ciprofibrate (CIP): 100 and 400 microM. Fatty acyl CoA oxidase activities of the 100 and 400 microM CIP treatment groups at the end of the experiment were increased 5.3 and 9.6 times, respectively. TxB2 and PGF2alpha concentrations in cultures treated with CIP were significantly lower than the control at days 3 and 4, whereas a lower concentration of PGE2 was seen at day 4 only. These studies show that PG and Tx concentrations in cultured hepatocytes are lowered by the peroxisome proliferator CIP.

Oxidative stability of conjugated linoleic acid isomers.

Conjugated linoleic acids (CLAs) have been shown to be a strong anticarcinogen in a number of animal models. Our previous study demonstrated that CLA as a whole was extremely unstable in air. The present study was undertaken further to examine the oxidative stability of individual CLA isomers using the combination of gas-liquid chromatography (GLC) and silver ion high-performance liquid chromatography (Ag-HPLC). It was found that CLA as a whole oxidized rapidly and more than 80% was degraded within 110 h in air at 50 degrees C. Four c,c-CLA isomers were most unstable followed by four c,t-CLA isomers. In contrast, four t,t-CLA isomers were relatively stable under the same experimental conditions. Both the oxygen consumption and the GLC analysis revealed that 200 ppm jasmine green tea catechins (GTCs) exhibited protection to CLA and were even stronger than 200 ppm butylated hydroxytoluene (BHT) when added to either CLA or canola oil containing 10% CLA. The present study emphasized that oxidative unstability of CLA should not be overlooked although CLA has many biological effects.

Feasibility and pharmacokinetics of carbamazepine oral loading doses.

The pharmacokinetics and adverse effects of an oral loading dose of carbamazepine administered in tablet or suspension form were studied. Patients on a hospital epilepsy unit who were to receive carbamazepine as a discharge medication were randomly assigned to receive either an oral 8-mg/kg loading dose of the tablet formulation or the same dose of the suspension on an empty stomach. Blood samples were drawn before and at intervals up to 12 hours after the loading dose. Adverse effects were evaluated subjectively and objectively. Total and free serum carbamazepine and carbamazepine-10, 11-epoxide (CBZE) concentrations were determined by high-performance liquid chromatography. Six adult patients were enrolled in and completed the study. All the patients achieved therapeutic total carbamazepine levels; the suspension group did so within two hours and the tablet group within five hours. Maximum serum carbamazepine concentrations ranged from 7.10 to 9.92 mg/L, area under the concentration-versus-time curve from 54.85 to 82.23 micrograms.hr/L, and terminal elimination half-life from 14.05 to 15.71 hours. Adverse effects were mild, few, and short-lived; none of the patients developed gastrointestinal toxicity. Adverse effects were not associated with total or free carbamazepine and CBZE concentrations or with total or free CBZE:carbamazepine ratios. An oral loading dose of carbamazepine 8 mg/kg achieved therapeutic levels within two hours when given as a suspension and within five hours when given as tablets and was well tolerated in all patients.

Genistein upregulates placental corticotropin-releasing hormone expression in lipopolysaccharide-sensitized mice.

Genistein is a phytoestrogen isolated from soya beans. Although soy products are staple food of Asian, the potential effect of genistein on reproduction has not been fully addressed. Lipopolysaccharide (LPS) is an endotoxin found in the cell membrane of gram-negative bacteria. It may cause inflammation and other immune responses. Previous study has shown that LPS may induce pre-mature birth in rodents. In the present study, effect of genistein on LPS-induced preterm birth was investigated. Pregnant ICR mice were gavaged with genistein at 40, 200 and 400 mg/kg body weight/day during E13 to E16. LPS was injected i.p. on E16.5 and the animals were sacrificed at E17. Compared to the control group, an increased incidence of early delivery was observed in the pooled mice under LPS treatment. A rising trend of incidence was also demonstrated dose-dependently with genistein co-treatment. Real-time RT-PCR indicated that the placental crh expression was highly induced by the co-administration of 400 mg/kg genistein and LPS. By contrast, neither genistein nor LPS alone could alter the expression. Increased plasma CRH concentration was also seen in the co-treatment groups. In addition, the mRNA expression of placental CRH-binding protein and plasma progesterone concentration were reduced in these groups. These results indicated that genistein might exacerbate the undesirable effect of LPS on pregnant mice by altering hormonal regulations.

Ultrastructure of the spermatozoon of Lepidogalaxias salamandroides and its phylogenetic significance.

The spermatozoon of Lepidogalaxias salamandroides possesses an acrosome (putative), one or two perforatoria (putative) but no nine-triplet centrioles. Two elongated mitochondria (12 micron long) are situated in parallel between the nucleus (20 micron long) and the axoneme (53 micron long). The above features are unique among other teleosts with internal fertilization. The presence of an "acrosome" in this primitive teleost supports the hypothesis that this structure has been secondarily lost in teleosts during evolution. The uncertainty of phylogenetic placement of this fish is reflected by its unique sperm ultrastructure.

Bcl-2 is not reduced in the death of MCF-7 cells at low genistein concentration.

Soy consumption has been associated with a lower incidence of breast cancer in Southeast Asia. Among the phytochemicals in soy, genistein has been suggested to be chemopreventive. Because genistein is an estrogen-receptor (ER) agonist, the chemopreventive mechanism has been attributed to its ability to compete with estrogen for receptor binding. In this study, we used an ER-positive cell line to investigate the effects of different genistein concentrations on the apoptotic response. The threshold concentration at which a significant number of cells underwent apoptosis was titrated to be 25 micromol/L. At or above this concentration, c-jun N-terminus kinase was activated and Bax and Bcl-2 expression were both elevated. The elevated Bcl-2 protein might neutralize the proapoptotic effect of Bax. Therefore, the mechanism of genistein-induced apoptosis at this concentration might rely largely on the stress pathway rather than the pathway mediated by the Bcl-2 family of proteins.