Di-n-butyl phthalate diminishes testicular steroidogenesis by blocking the hypothalamic-pituitary-testicular axis: relationship with germ cell apoptosis in Japanese quail.

Although di-n-butyl phthalate (DBP) induces germ cell apoptosis, the underlying mechanism is not yet clear in quail. In this study, prepubertal quails were given a single dose of 500mg kg-1 DBP by gavage and were then killed 3, 6 and 24h after treatment. There was a significant reduction in intratesticular testosterone (ITT) concentrations and testicular steroidogenic enzyme mRNA expression and a significant increase in germ cell apoptosis in DBP-treated compared with control quails at all time points. Maximum apoptosis was detected 6h after treatment and the maximum reduction in testosterone concentrations was at 3h. To investigate whether DBP suppressed testicular steroidogenesis by affecting the hypothalamic-pituitary-testicular axis, we analysed pituitary LH subunit ß (Lhb) mRNA expression and serum LH concentrations. At all time points, pituitary Lhb expression and serum LH concentrations were significantly decreased following DBP treatment. The present observations suggest the possibility that DBP blocked LH secretion from the hypothalamus and/or pituitary, thereby decreasing LH stimulation of Leydig cells and reducing ITT concentrations. DBP-induced decreases in ITT concentrations may cause changes to the physical structure of Sertoli cells, which, in turn, may induce germ cell apoptosis.

Changes of Land Use and Land Cover with the Diversity of Fishes, Aquatic Plants, and Bird's Species at Wetland Ecosystem.

Bangladesh is rich in wetland biodiversity with aquatic plants, fishes, and birds. Mohanganj Upazila is known as the capital of lower Bangladesh. The present study focuses on the changes of land use and land cover (LULC) with a diversity of species that are being least concerned (LC), vulnerable (VU), and endangered (EN). Over the last two decades, the wetland species of Mohanganj were gradually declined. Our results showed that 19 fish, 4 aquatic plants, and 7 bird species were LC in 2015. Among the fish and aquatic plant species, 6 fish species (Wallago attu, Ompok pabda, Channa punctate, Chitala chitala, Salmostoma phulo, and Corica soborna) and 2 aquatic plant species (Nymphaea nouchali and Nymphaea lotus) were VU during the dry and rainy season of 2017 and 2019, respectively. In the dry season of 2019, 4 fish species (W. attu, O. pabda, C. punctate, and Ch. chitala), 2 aquatic plant species (N. nouchali and N. lotus), and 7 bird species (Anas platyrhynchos, Ardeola grayii, Gyps bengalensis, Alcedo atthis, Phalacrocorax fuscicollis, Porphyrio porphyria, and Larus ridibundus) were EN. Among the species, W. attu, N. nouchaii, G. bengalensis, P. porphyria, and L. ridibundus were extremely endangered categories. Changes in LULC, the establishment of settlements for the increasing population, indiscriminate use of pesticides, environmental pollutions, and climate change are the potential reasons for declining trends of wetland biodiversity. Stern actions on land use policy, expansion of organic agriculture, bioremediation of industrial effluents, and adoption of sustainable environmental policies should be taken by the Government of Bangladesh for immediate conservation of wetland biodiversity.

Minimizing the Energy Hole Problem in Wireless Sensor Networks: A Wedge Merging Approach.

The Energy hole problem, a common phenomenon in wireless sensor networks, significantly decreases the lifetime of any deployed network. Some of the popular techniques to minimize such problems are using mobile sinks instead of static sinks, extending the transmission range dynamically, and deploying redundant sensor nodes near the base station/sink. The major drawback to these techniques are that energy holes may still be created at some point due to their static nature of deployment, despite having the overall residual energy very high. In this research work, we adopt a new approach by dividing the whole network into equiangular wedges and merging a wedge with its neighboring wedge dynamically whenever individual residual energy of all member nodes of a wedge fall below a threshold value. We also propose an efficient Head Node (HN) selection scheme to reduce the transmission energy needed for forwarding data packets among Head Nodes. Simulation results show that WEMER, our proposed WEdge MERging based scheme, provides significantly higher lifetime and better energy efficiency compared to state-of-the-art Power-Efficient Gathering in Sensor Information Systems (PEGASIS) and contemporary Concentric Clustering Scheme (CCS), and Multilayer Cluster Designing Algorithm (MCDA).

Role of anlotinib-induced CCL2 decrease in anti-angiogenesis and response prediction for nonsmall cell lung cancer therapy.

BACKGROUND: Anlotinib has been demonstrated in clinical trials to be effective in prolonging the progression-free survival (PFS) and overall survival (OS) of refractory advanced nonsmall cell lung cancer (NSCLC) patients. However, the underlying molecular mechanisms and predictive biomarkers of anlotinib are still unclear. METHODS: A retrospective analysis of anlotinib administered to 294 NSCLC patients was performed to screen for underlying biomarkers of anlotinib-responsive patients. Transcriptome and functional assays were performed to understand the antitumour molecular mechanisms of anlotinib. Changes in serum CCL2 levels were analysed to examine the correlation of the anlotinib response between responders and nonresponders. RESULTS: Anlotinib therapy was beneficial for prolonging OS in NSCLC patients harbouring positive driver gene mutations, especially patients harbouring the epithelial growth factor receptor (EGFR)T790M mutation. Moreover, anlotinib inhibited angiogenesis in an NCI-H1975-derived xenograft model via inhibiting CCL2. Finally, anlotinib-induced serum CCL2 level decreases were associated with the benefits of PFS and OS in refractory advanced NSCLC patients. CONCLUSIONS: Our study reports a novel anti-angiogenesis mechanism of anlotinib via inhibiting CCL2 in an NCI-H1975-derived xenograft model and suggests that changes in serum CCL2 levels may be used to monitor and predict clinical outcomes in anlotinib-administered refractory advanced NSCLC patients using third-line therapy or beyond.

Minimizing the Adverse Effects of Asymmetric Links: A Novel Cooperative Asynchronous MAC Protocol for Wireless Sensor Networks.

As Wireless Sensor Networks (WSNs) grow in popularity, researchers are now focusing more on some challenging issues that significantly degrade overall performance, such as energy hole mitigation, link asymmetry minimization, etc. Link asymmetry is a problem that arises when the coverage distance between two adjacent nodes varies. It creates an obstacle to overcome when designing an efficient Medium Access Control (MAC) protocol for WSNs with low duty-cycling. This phenomenon poses an especially difficult challenge for receiver-initiated asynchronous MAC protocols, which are popular due to their relatively higher energy efficiency. Exploiting the benefits of cooperative communication has emerged as one of the viable solutions to overcome this limitation. Cooperative communication in WSNs has received a lot of attention in recent years. Many researchers have worked to create a MAC layer supporting cooperative communication. However, the association of cooperative communication with an asymmetric link is not studied in the literature. In this research work, COASYM-MAC, a cooperative asynchronous MAC protocol for WSNs, is proposed based on a receiver-initiated MAC protocol that uses the fact that nodes have alternate paths between them to reduce link asymmetry. A key feature of the proposed protocol is that the optimal helper node is selected automatically in case of link asymmetry. Simulations exhibited that COASYM-MAC performs significantly better than a state-of-the-art MAC protocol for WSNs that handles asymmetric links, ASYM-MAC.

SAFE-MAC: Speed Aware Fairness Enabled MAC Protocol for Vehicular Ad-hoc Networks.

Highly dynamic geographical topology, two-direction mobility, and varying traffic density can lead to fairness issues in Vehicular Ad-hoc Networks (VANETs). The Medium Access Control (MAC) protocol plays a vital role in sharing the common wireless channel efficiently between vehicles in a VANET system. However, ensuring fairness between vehicles can be a challenge in designing MAC protocols for VANET systems. The existing protocol, IEEE 802.11 DCF, ensures that the packet transmission rate for a particular vehicle is directly proportional to the amount of time a vehicle spends within a service area, but it does not guarantee that faster vehicles will be able to send the minimum number of packets. Other existing MAC protocols based on IEEE 802.11 are able to provide a minimum amount of data transmission regardless of velocity, but are unable to provide an amount of data transmission that is more proportionate to the time a vehicle spends in the service area. To address the above limitations, we propose a Speed Aware Fairness Enabled MAC (SAFE-MAC) protocol that calculates the residence time of a vehicle in a service area by using mobility metrics such as position, direction, and speed to synthesize the transmission probability of each individual vehicle with respect to its residence time. This is achieved by dynamically altering the values of parameters such as minimum contention window, maximum backoff stage, and retransmission limit in the MAC protocol. We then develop an analytical model to compare the performance of our proposed protocol with contemporary MAC protocols. Numerical analysis results show that our proposed protocol significantly improves fairness among the speed-varying vehicles in VANET.

Different screening tests and milk somatic cell count for the prevalence of subclinical bovine mastitis in Bangladesh.

Identification of cows with subclinical mastitis (SCM) is an important tool for sustainable dairying and implementing effective mastitis control strategies. A total of 892 quarters milk samples from 228 lactating cows were screened by California mastitis test (CMT), White side test (WST), Surf field mastitis test (SFMT), and somatic cell count (SCC) to study the prevalence of bovine SCM in some selected areas of Bangladesh. Out of 228 cows, 148 (64.9%), 138 (60.5%), 132 (57.9%), and 164 (71.9%) were found positive for SCM by CMT, WST, SFMT, and SCC, respectively. The prevalence of bovine SCM was diagnosed 45.7, 40.2, 36.6, and 29.6% in Chittagong, Sirajgonj, Mymensingh, and Gazipur districts, respectively, based on a combination of all tests. The overall quarter-wise prevalence of SCM was 45.7, 43.5, 41.2, and 55.0% for CMT, WST, SFMT, and SCC. Single quarters and left front quarters were more prone to SCM (P < 0.05). Friesian crossbred cows (56.4%), BCS 2.0-2.5 (55.4%), and parity 4-6 (52.4%), the late lactation stage (5-8 months; 64.7%) and high yielding cows (16-20 L/day; 65.3%) were more susceptible to SCM (P < 0.05). The sensitivity of the CMT, WST, SFMT, and SCC was 65.8, 57.9, 51.0, and 82.5%; specificity 76.2, 72.4, 69.5, and 89.4%; percentage accuracy 70.0, 64.8, 59.9, and 85.2%; positive predictive value 75.2, 69.8, 64.9, and 92.7%, respectively. The categories of CMT reactions were strongly correlated with SCC (P < 0.05). Kappa value of SCC was higher than that of other tests (SCC>CMT>WST>SFMT). Thus, CMT was concluded to be the most accurate (r = 0.782) field diagnostic test after laboratory test like SCC (r = 0.924). However, the use of any single test may not be reliable in diagnosing SCM, while the result of CMT supported by SCC might be used effectively to pinpoint diagnosis of SCM in dairy animals than alone.

Unveiling distinct genetic features in multidrug-resistant Escherichia coli isolated from mammary tissue and gut of mastitis induced mice.

Escherichia coli is one of the major pathogens causing mastitis in lactating mammals. We hypothesized that E. coli from the gut and mammary glands may have similar genomic characteristics in the causation of mastitis. To test this hypothesis, we used whole genome sequencing to analyze two multidrug resistant E. coli strains isolated from mammary tissue (G2M6U) and fecal sample (G6M1F) of experimentally induced mastitis mice. Both strains showed resistance to multiple (>7) antibiotics such as oxacillin, aztreonam, nalidixic acid, streptomycin, gentamicin, cefoxitin, ampicillin, tetracycline, azithromycin and nitrofurantoin. The genome of E. coli G2M6U had 59 antimicrobial resistance genes (ARGs) and 159 virulence factor genes (VFGs), while the E. coli G6M1F genome possessed 77 ARGs and 178 VFGs. Both strains were found to be genetically related to many E. coli strains causing mastitis and enteric diseases originating from different hosts and regions. The G6M1F had several unique ARGs (e.g., QnrS1, sul2, tetA, tetR, emrK, blaTEM-1/105, and aph(6)-Id, aph(3″)-Ib) conferring resistance to certain antibiotics, whereas G2M6U had a unique heat-stable enterotoxin gene (astA) and 7192 single nucleotide polymorphisms. Furthermore, there were 43 and 111 unique genes identified in G2M6U and G6M1F genomes, respectively. These results indicate distinct differences in the genomic characteristics of E. coli strain G2M6U and G6M1F that might have important implications in the pathophysiology of mammalian mastitis, and treatment strategies for mastitis in dairy animals.

Insight into SARS-CoV-2 Omicron variant immune escape possibility and variant independent potential therapeutic opportunities.

The Omicron, the latest variant of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), was first detected in November 2021 in Botswana, South Africa. Compared to other variants of SARS-CoV-2, the Omicron is the most highly mutated, with 50 mutations throughout the genome, most of which are in the spike (S) protein. These mutations may help the Omicron to evade host immunity against the vaccine. Epidemiological studies suggest that Omicron is highly infectious and spreads rapidly, but causes significantly less severe disease than the wild-type strain and the other variants of SARS-CoV-2. With the increased transmissibility and a higher rate of re-infection, Omicron has now become a dominant variant worldwide and is predicted to be able to evade vaccine-induced immunity. Several clinical studies using plasma samples from individuals receiving two doses of US Food and Drugs Administration (FDA)-approved COVID-19 vaccines have shown reduced humoral immune response against Omicron infection, but T cell-mediated immunity was well preserved. In fact, T cell-mediated immunity protects against severe disease, and thus the disease caused by Omicron remains mild. In this review, I surveyed the current status of Omicron variant mutations and mechanisms of immune response in the context of immune escape from COVID-19 vaccines. I also discuss the potential implications of therapeutic opportunities that are independent of SARS-CoV-2 variants, including Omicron. A better understanding of vaccine-induced immune responses and variant-independent therapeutic interventions that include potent antiviral, antioxidant, and anti-cytokine activities may pave the way to reducing Omicron-related COVID-19 complications, severity, and mortality. Collectively, these insights point to potential research gaps and will aid in the development of new-generation COVID-19 vaccines and antiviral drugs to combat Omicron, its sublineages, or upcoming new variants of SARS-CoV-2.

Spirulina supplementation to alleviate negative effects of lead in layer chicken.

Objectives: Lead (Pb), a toxic heavy metal, is a serious concern for poultry that negatively affects their productivity and health. To combat those issues efficiently, it is necessary to include feed supplements that have rich antioxidant properties for satisfactory health and productivity. Spirulina platensis (Sp), a microalgae, is a compound that provides several health benefits for humans and animals. This study explores that supplementation of Sp in diet as well as in water reduces the burden of Pb in different tissues, improves hematology, and improves the productive performance of advanced-age laying hens. Materials and methods: Forty birds were separated into four groups: the control (C), Spirulina (Sp), Pb, and (Pb + Sp) groups. The Pb group was given Pb acetate at a dose of 2 gm/l in water ad libitum for 4 weeks. Sp group was fed Sp at a dose of 4 gm/kg feed. The Pb + Sp group was given Pb and Sp as in the previous groups. Results: Productive performance and hematology such as hemoglobin (Hb), packed cell volume, red blood cell, mean corpuscular volume, mean corpuscular Hb (MCH) concentration, and MCH were significantly (p < 0.05) decreased in Pb-treated groups compared to controls. The distribution of Pb concentration was highest in the bones and lowest in the gizzard. However, Sp treatment significantly (p < 0.05) increased the productive performance and the hematological parameters. Moreover, Pb concentration in different organs significantly decreased in the group treated with Sp. Conclusion: This study indicates that Sp can possibly be used as a natural and powerful dietary additive to mitigate heavy metal intoxication in chickens, thereby being efficient and effective for production.

N-acetylcysteine reduces severity and mortality in COVID-19 patients: A systematic review and meta-analysis.

Objectives: Recent clinical studies suggest that oxidative stress is one of the key players in the pathogenesis of coronavirus disease 2019 (COVID-19), and N-acetylcysteine (NAC), a potent antioxidant, has been shown to improve clinical outcomes in COVID-19 patients. We conducted a systematic review and meta-analysis of the literature published on the therapeutic intervention of NAC on COVID-19 infection. Methods: We searched PubMed, Google Scholar, and Science Direct. We identified and screened eight studies with 20,503 participants, including 2,852 in the NAC-treated group and 17,651 in the placebo group, which reported the effect of NAC on COVID-19 infection. A meta-analysis was performed using forest plots under fixed effect estimates based on the standardized mean difference (SMD) and risk ratio (RR). Results: Pooled analysis showed that NAC was associated with lower mortality in patients with COVID-19 compared with the placebo group [RR, 0.65; (95% CI: 0.56 to 0.75); p < 0.0001]. Similarly, C-reactive protein (CRP) [SMD, -0.32; (95% CI: -56 to -0.09); p = 0.0070] and D-dimer [SMD, -0.35, (95% CI: -0.59 to -0.10; p = 0.0062] levels were significantly decreased, and the oxygenation marker, PaO2/FiO2 ratio, was increased in the NAC-treated group compared with the placebo group [SMD, 0.76; (95% CI: 0.48 to 1.03); p < 0.0001]. Conclusion: Although the number of included studies was minimal, this meta-analysis suggests that NAC may have a positive effect on COVID-19 outcomes, specifically, a significant decrease in CRP and D-dimer levels and a significant increase in oxygen saturation, which decreased mortality. We have also presented a comprehensive review of the role and mechanisms of NAC in patients with COVID-19.

Calcium and L-glutamate present the opposite role in managing arsenic in barley.

Arsenic contamination in agricultural soils has posed tremendous threat to sustainable crop production and human health via food chain. Calcium and Glutamate have been well-documented in metal(loid)s detoxification, but it is poorly understood how they regulate arsenic-induced toxicity to plants. In this study, the effect of glutamate and calcium at high concentration on arsenic toxicity and accumulation in barley seedling was accessed in terms of plant growth, photosynthetic efficacy, arsenic uptake, translocation and accumulation, antioxidant defense, nutrient uptake and the expression of As transporters. Our results have demonstrated that calcium could effectively ameliorate arsenic toxicity to barley seedlings, which is mainly attributed to its beneficial effect on increasing nutrient uptake, reducing the aboveground arsenic accumulation and enhancing antioxidative defense capacity. However, it is unexpected that glutamate considerably exacerbated the arsenic toxicity to barley seedlings. More importantly, for the first time, glutamate was observed to tremendously facilitate the root-to-shoot translocation of arsenic by 41.8- to 60.8-fold, leading to 90% of the total amount of As accumulating in barley shoots. The reason of this phenomenon can be well explained by the glutamate-triggered enormous upregulation of genes involved in arsenic uptake (HvPHT1;1, HvPHR2 and HvNIP3;2), reduction (HvHAC1;1), translocation (HvABCC7, HvNIP1;1 and HvNIP3;3) and intracellular sequestration (HvABCC1). These findings suggest that calcium and glutamate function as the opposite player in managing arsenic, with calcium being an effective alleviator of arsenic stress to ensure the safe production of crops; while glutamate being a highly efficient phytoextraction agent for phytoremediation of arsenate-contaminated soils.

SARS-CoV-2 infection and oxidative stress: Pathophysiological insight into thrombosis and therapeutic opportunities.

The current coronavirus disease 2019 (COVID-19) pandemic has presented unprecedented challenges to global health. Although the majority of COVID-19 patients exhibit mild-to-no symptoms, many patients develop severe disease and need immediate hospitalization, with most severe infections associated with a dysregulated immune response attributed to a cytokine storm. Epidemiological studies suggest that overall COVID-19 severity and morbidity correlate with underlying comorbidities, including diabetes, obesity, cardiovascular diseases, and immunosuppressive conditions. Patients with such comorbidities exhibit elevated levels of reactive oxygen species (ROS) and oxidative stress caused by an increased accumulation of angiotensin II and by activation of the NADPH oxidase pathway. Moreover, accumulating evidence suggests that oxidative stress coupled with the cytokine storm contribute to COVID-19 pathogenesis and immunopathogenesis by causing endotheliitis and endothelial cell dysfunction and by activating the blood clotting cascade that results in blood coagulation and microvascular thrombosis. In this review, we survey the mechanisms of how severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) induces oxidative stress and the consequences of this stress on patient health. We further shed light on aspects of the host immunity that are crucial to prevent the disease during the early phase of infection. A better understanding of the disease pathophysiology as well as preventive measures aimed at lowering ROS levels may pave the way to mitigate SARS-CoV-2-induced complications and decrease mortality.

CRISPR/Cas9 Mediated Knockout of the OsbHLH024 Transcription Factor Improves Salt Stress Resistance in Rice (Oryza sativa L.).

Salinity stress is one of the most prominent abiotic stresses that negatively affect crop production. Transcription factors (TFs) are involved in the absorption, transport, or compartmentation of sodium (Na+) or potassium (K+) to resist salt stress. The basic helix-loop-helix (bHLH) is a TF gene family critical for plant growth and stress responses, including salinity. Herein, we used the CRISPR/Cas9 strategy to generate the gene editing mutant to investigate the role of OsbHLH024 in rice under salt stress. The A nucleotide base deletion was identified in the osbhlh024 mutant (A91). Exposure of the A91 under salt stress resulted in a significant increase in the shoot weight, the total chlorophyll content, and the chlorophyll fluorescence. Moreover, high antioxidant activities coincided with less reactive oxygen species (ROS) and stabilized levels of MDA in the A91. This better control of oxidative stress was accompanied by fewer Na+ but more K+, and a balanced level of Ca2+, Zn2+, and Mg2+ in the shoot and root of the A91, allowing it to withstand salt stress. Furthermore, the A91 also presented a significantly up-regulated expression of the ion transporter genes (OsHKT1;3, OsHAK7, and OsSOS1) in the shoot when exposed to salt stress. These findings imply that the OsbHLH024 might play the role of a negative regulator of salt stress, which will help to understand better the molecular basis of rice production improvement under salt stress.

Loss-of-function mutations of OsbHLH044 transcription factor lead to salinity sensitivity and a greater chalkiness in rice (Oryza sativa L.).

The most hazardous abiotic stress, salinity, restricted the world crop production, and grain chalkiness affected the grain quality to limit consumers' acceptance. The basic helix-loop-helix (bHLH) proteins modulate massive biological processes in plants. Here the CRISPR/Cas9 gene editing mutants were obtained to detect the function of OsbHLH044. The loss-of-function of OsbHLH044 mutants showed numerous altered plant phenotypes. Notably, the osbhlh044 mutants resulted in prominently reduced morphological and physiological parameters under salt stress. Lower antioxidant activities and higher lipid peroxidation and hydrogen peroxide (H2O2) accumulation in the osbhlh044 mutants caused salinity sensitivity due to elevated reactive oxygen species (ROS). Under salt stress, both shoots and roots of the osbhlh044 mutants acquired higher Na+. Moreover, the expression of ion homeostasis-related genes (OsHKTs, OsHAK, OsSOSs, and OsNHX) and ABA-responsive gene (OsLEA3) was significantly altered in the osbhlh044 mutants after salt stress. The expression levels of genes coding for starch (OsAGPL1, OsSSIIa, OsWx, and OsFLO2) and seed storage proteins (GluA1 and Globulin 1) were significantly decreased, indicating that they synthesize less store starch and proteins, resulting in grain chalkiness in the osbhlh044 mutants. Yeast one Hybrid (Y1H) showed that OsbHLH044 could activate salt- (OsHKT1;3, OsHAK7, OsSOS1, OsSOS2, OsNHX2, and OsLEA3 but not OsHKT2;1), and starch-related genes (OsSSIIa, OsWx, and OsFLO2) by binding to the G-boxes of their promoters. Therefore, the OsbHLH044 gene editing mutants revealed multiple functions, specifically a positive regulator of salt stress and grain quality, which might bring new insights into the breeding of rice varieties.

OsLHY is involved in regulating flowering through the Hd1- and Ehd1- mediated pathways in rice (Oryza sativa L.).

Flowering time (or heading date in crops) is a critical agronomic trait for rice reproduction and adaptation. The circadian clock is an endogenous oscillator that is involved in controlling photoperiodic flowering. The rice LATE ELONGATED HYPOCOTYL (OsLHY), the core oscillator component of circadian clock, is a homolog of the LHY/CCA1 in Arabidopsis. Here we showed that CRISPR/Cas9-engineered mutations in OsLHY caused late flowering in rice only under natural long-day (nLD) and short-day (nSD) conditions, but not artificial SD (10 h light/14 h dark) conditions. In the oslhy mutant, the diurnal expression of circadian clock-related genes was seriously affected under both LD and SD conditions. Furthermore, the expression of the flowering activators Ehd1, Hd3a and RFT1 was down-regulated and flowering repressors Hd1 and Ghd7 was up-regulated in the oslhy mutant under LD conditions. While the transcripts of flowering-related genes were not dramatically influenced under SD conditions. Dual-luciferase assays showed that OsLHY repressed the transcription of OsGI, Hd1, Ghd7, Hd3a, RFT1 and OsELF3, and activated the transcription of Ehd1. Moreover, the yeast one hybrid assay and electrophoretic mobility shift assay confirmed that OsLHY directly repressed OsGI, RFT1 and OsELF3 by binding to their promoters, which is consistent with that in Arabidopsis. These results suggested that the OsLHY can promote rice flowering mainly through regulating Hd1 and Ehd1.

Efficient and Fast Immuno-Labeling of Clarified Tissues Using Low-Field Enhanced Diffusion.

OBJECTIVE: To alleviate the severe limitation of the prohibitively long process of immune-fluorescence labeling on the routine applications of revolutionary intact tissue clearing techniques in diverse biomedical arenas. METHODS: We proposed an easily adaptable approach, electro-enhanced rapid staining (EERS), for highly efficient and fast immuno-labeling of thick clarified tissues. In EERS, an optimized and precisely controlled weak external electric field is engineered into a compact device to enable efficient and uniform transport of antibodies into clarified tissues while minimizing the detrimental effect of macromolecular crowding at the tissue-solution interface. RESULTS AND CONCLUSIONS: The experimental results show that, with EERS, a current density of only ∼0.2 mA mm-2 is sufficient to achieve uniform labeling of clarified tissues of several millimeters thick in a few hours without detectable tissue damage. In addition, the amount of antibodies required is also several-fold lower than conventional immuno-labeling assays under comparable conditions. SIGNIFICANCE: It is expected that the implementation of EERS in most laboratories should significantly expedite the application of tissue clearing in a broad range of research explorations, both basic and clinical.

Extensively Current Activity of Transposable Elements in Natural Rice Accessions Revealed by Singleton Insertions.

Active transposable elements (TEs) have drawn more attention as they continue to create new insertions and contribute to genetic diversity of the genome. However, only a few have been discovered in rice up to now, and their activities are mostly induced by artificial treatments (e.g., tissue culture, hybridization etc.) rather than under normal growth conditions. To systematically survey the current activity of TEs in natural rice accessions and identify rice accessions carrying highly active TEs, the transposon insertion polymorphisms (TIPs) profile was used to identify singleton insertions, which were unique to a single accession and represented the new insertion of TEs in the genome. As a result, 10,924 high-confidence singletons from 251 TE families were obtained, covering all investigated TE types. The number of singletons varied substantially among different superfamilies/families, perhaps reflecting distinct current activity. Particularly, eight TE families maintained potentially higher activity in 3,000 natural rice accessions. Sixty percent of rice accessions were detected to contain singletons, indicating the extensive activity of TEs in natural rice accessions. Thirty-five TE families exhibited potentially high activity in at least one rice accession, and the majority of them showed variable activity among different rice groups/subgroups. These naturally active TEs would be ideal candidates for elucidating the molecular mechanisms underlying the transposition and activation of TEs, as well as investigating the interactions between TEs and the host genome.

Induction of spermatogenic cell apoptosis in prepubertal rat testes irrespective of testicular steroidogenesis: a possible estrogenic effect of di(n-butyl) phthalate.

Although di(n-butyl) phthalate (DBP), a suspected endocrine disruptor, induces testicular atrophy in prepubertal male rats, whether it exerts estrogenic activity in vivo remains a matter of debate. In the present study, we explored the estrogenic potency of DBP using 3-week-old male rats, and then examined the relationship between estrogen-induced spermatogenic cell apoptosis and testicular steroidogenesis. Daily exposure to DBP for 7 days caused testicular atrophy due to loss of spermatogenic cells, whereas testicular steroidogenesis was almost the same with the control values. A single exposure of DBP decreased testicular steroidogenesis in addition to decreasing the level of serum LH at 3 h after DBP treatment, with an extremely high incidence of apoptotic spermatogenic cells at 6 h after administration. To elucidate the estrogenic activity of DBP, we carried out an inhibition study using pure antiestrogen ICI 182,780 (ICI) in a model of spermatogenic cell apoptosis induced by DBP or estradial-3-benzoate (EB). Although both the DBP- and EB-treated groups showed a significant increase in spermatogenic cell apoptosis, ICI pretreatment significantly decreased the number of apoptotic spermatogenic cells in these two groups. In contrast, testicular steroidogenesis and serum FSH were significantly reduced in all the treated groups, even in the DBP+ICI and EB+ICI groups. Taken together, these findings led us to conclude that estrogenic compounds such as DBP and EB induce spermatogenic cell apoptosis in prepubertal rats, probably by activating estrogen receptors in testis, and that reduction in testicular steroidogenic function induced by estrogenic compounds is not associated with spermatogenic cell apoptosis.

Effects of di-iso-butyl phthalate on testes of prepubertal rats and mice.

Di-iso-butyl phthalate (DiBP), a special plasticizer, is used as a substitute for di(n-butyl) phthalate (DBP). The effects of DiBP on testes in prepubertal rodents still remain to be obscure. Testicular toxicity of DiBP was investigated in 21-day-old Sprague-Dawley rats and C57BL/6N mice, using with in situ TUNEL method. For an acute exposure experiment, animals were once given DiBP at various concentrations by oral gavage. For a subchronic exposure experiment, they were daily given DiBP at various concentrations for consecutive 7 days. Controls were treated with corn oil under the same condition. For a recovery experiment, rats were once given DiBP (1000 mg/kg), and were sacrificed at day 1 to 8 after administration. Furthermore, the disorder of vimentin filaments in Sertoli cells after daily administration of DiBP (500 mg/kg) for consecutive 7 days in rats also identified by immunohistochemistry using anti-vimentin antibody. As a result, the present study demonstrated that DiBP can induce testicular atrophy in rats due to the increase of TUNEL-positive spermatogenic cells in both acute and subchronic exposure experiments. At the same time, the disorder of vimentin filaments in Sertoli cells was recognized. However, no such damages could be found in mouse testis. For the recovery experiment, the testis weight and testicular morphology returned to normal at day 6 after administration. In conclusion, the present study indicates that DiBP causes the significant increase of TUNEL-positive spermatogenic cells and the disorder of vimentin filaments in Sertoli cells in rats and that DiBP shows a species-specific toxicity.