Novel MEI1 mutations cause chromosomal and DNA methylation abnormalities leading to embryonic arrest and implantation failure.

This study presents a case of a female infertile patient suffering from embryonic arrest and recurrent implantation failure. The primary objective was to assess the copy number variations (CNVs) and DNA methylation of her embryos. Genetic diagnosis was conducted by whole-exome sequencing and validated through Sanger sequencing. CNV evaluation of two cleavage stage embryos was performed using whole-genome sequencing, while DNA methylation and CNV assessment of two blastocysts were carried out using whole-genome bisulfite sequencing. We identified two novel pathogenic frameshift variants in the MEI1 gene (NM_152513.3, c.3002delC, c.2264_2268 + 11delGTGAGGTATGGACCAC) in the proband. These two variants were inherited from her heterozygous parents, consistent with autosomal recessive genetic transmission. Notably, two Day 3 embryos and two Day 6 blastocysts were all aneuploid, with numerous monosomy and trisomy events. Moreover, global methylation levels greatly deviated from the optimized window of 0.25-0.27, measuring 0.344 and 0.168 for the respective blastocysts. This study expands the mutational spectrum of MEI1 and is the first to document both aneuploidy and abnormal methylation levels in embryos from a MEI1-affected female patient presenting with embryonic arrest. Given that females affected by MEI1 mutations might experience either embryonic arrest or monospermic androgenetic hydatidiform moles due to the extrusion of all maternal chromosomes, the genetic makeup of the arrested embryos of MEI1 patients provides important clues for understanding the different disease mechanisms of the two phenotypes.

Pregnancy outcomes of patients with positive anticentromere antibodies receiving in vitro fertilization-embryo transfer. / æŠ—ç€ä¸ç‚¹æŠ—ä½“é˜³æ€§æ‚£è€ ä½“å¤–å—ç²¾ç»“å±€åˆ†æž.

OBJECTIVES: To analyze the pregnancy outcomes in patients with positive anti-centromere antibodies (ACA) receiving in vitro fertilization (IVF)-embryo transfer (ET) and natural conception. METHODS: A case-control study was used to retrospectively analyze the clinical data of 3955 patients who received IVF-ET therapy and had the results of antinuclear antibody (ANA) spectrum at Zhejiang Provincial People's Hospital from June 2016 to June 2023. Patients with positive ACA and negative ACA were matched at a ratio of 1∶3 using propensity score matching. Embryo outcomes of IVF were compared between the two groups, and the impact of different fertilization methods and the use of immunosuppressants on pregnancy outcomes were analyzed using self-matching. The natural conception and disease progress were followed up for ACA-positive patients after IVF failure. RESULTS: The ACA-positive patients accounted for 0.86% of all IVF patients (34/3955) and 2.51% of total ANA-positive IVF patients. Regardless of whether patients received conventional IVF (c-IVF) or intracytoplasmic sperm injection (ICSI), the ACA-positive group exhibited significant differences in oocyte maturity and fertilization compared to the ACA-negative group (both P<0.01). Moreover, the ACA-positive group had a decreased number of D3 suboptimal embryos and D3 optimal embryos (both P<0.05). In 5 cases of ACA-positive patients who underwent ICSI cycles, the two pronucleus (2PN) rate did not increase compared to c-IVF cycles (P>0.05), and there was a decrease in the number of D3 high-quality embryos and D3 suboptimal embryos (both P<0.05). After 1-2 months of immuno-suppressant treatment, 12 ACA-positive patients underwent c-IVF/ICSI again, and there were no changes in egg retrieval and fertilization before and after medication (both P>0.05), but there was an improvement in the 2PN embryo cleavage rate (P<0.05). The number of embryos transferred was similar between the ACA-positive and negative groups, but the ACA-positive group had significantly lower embryo implantation rate and clinical pregnancy rate compared to the ACA-negative group (both P<0.05), with no significant differences in the miscarriage rate between the two groups (P>0.05). Twenty-seven ACA-positive patients attempted natural conception or artificial insemination after IVF failure, resulting in a total of 7 cases of clinical pregnancy. CONCLUSIONS: Serum ACA positivity may disrupt oocyte maturation and normal fertilization processes, with no improvement observed with ICSI and immunosuppressant use. However, ACA-positive patients may still achieve natural pregnancy.

Physiological mechanisms by which gypsum increases the growth and yield of Lentinula edodes.

Lentinula edodes is one of the most important commercially cultivated edible mushrooms. It is well known that gypsum (CaSO4·2H2O) supplementation in sawdust medium increases the yield of L. edodes, while the physiological mechanisms remain unclear. Our previous study showed that the acidification of the medium to pH 3.5-4.0 was essential for the growth of L. edodes. In this study, it was found that the oxalic acid excreted by L. edodes was responsible for the acidification of the medium. The biosynthesis of oxalic acid was regulated by the ambient pH and buffer capacity of the medium. To acidify the sawdust medium, the concentrations of total and soluble oxalate were 51.1 mmol/kg and 10.8 mmol/kg, respectively. However, when the concentration of soluble oxalate was 8.0 mmol/kg, the mycelial growth rate decreased by 29% compared with the control. Soluble oxalate was toxic to L. edodes, while soluble sulfate was nontoxic. CaSO4 reacted with soluble oxalate to form nontoxic insoluble CaC2O4 and the strong acid H2SO4. When the CaSO4 supplemented in sawdust medium was more than 25 mmol/kg, the soluble oxalate decreased to less than 1 mmol/kg, and the mycelial growth rate increased by 32% compared with the control. In conclusion, gypsum improved the growth and yield by relieving the toxicity of oxalate and facilitating the acidification of sawdust medium. KEY POINTS: • L. edodes excretes oxalic acid to acidify the ambient environment for growth. • Soluble oxalate is toxic to L. edodes. • Gypsum increases growth by reacting with oxalate to relieve its toxicity.

The outcomes of sequential embryo transfer in patients undergoing in vitro fertilization with frozen-thawed embryos: A retrospective study.

AIM: To explore whether sequential embryo transfer benefits patients with repeated implantation failure (RIF) undergoing frozen-thawed embryo transfer (FET) cycles. METHODS: We included 311 patients with a history of RIF in this retrospective study. We did sequential transfers with a cleavage embryo on day 3 and a blastocyst on day 5 in 77 patients; blastocyst transfers with two blastocysts on day 5 in 80 patients; and cleavage embryo transfers with two cleavage embryos on day 3 in 154 patients. We compared clinical outcomes between the three groups. RESULTS: The clinical pregnancy rate was comparable between the blastocyst transfer group (48.8%), the sequential transfer group(48.1%) and the cleavage embryo transfer group (48.1%). There was no statistically significant difference found (p > 0.05). The ongoing pregnancy and multipregnancy rates were also comparable between the three groups (p > 0.05). The early miscarriage rate was significantly higher in the sequential transfer group (32.4%) compared with the blastocyst group (12.8%) and the cleavage embryo group (12.2%) (p < 0.05). However, after adjusting for confounders, there was no significant difference in early miscarriage rates in the sequential transfer group compared with the blastocyst group (odds ratio [OR], 2.97; 95% confidence interval [CI], 0.85-9.24; p = 0.07) and the cleavage embryo group (OR, 3.03; 95% CI, 0.94-8.06; p = 0.08). CONCLUSIONS: Sequential embryo transfer failed to improve clinical outcomes for patients with RIF.

Nitric Oxide Improves the Tolerance of Pleurotus ostreatus to Heat Stress by Inhibiting Mitochondrial Aconitase.

Pleurotus ostreatus is widely cultivated in China. However, its cultivation is strongly affected by seasonal temperature changes, especially the high temperatures of summer. Nitric oxide (NO) was previously reported to alleviate oxidative damage to mycelia by regulating trehalose. In this study, we found that NO alleviated oxidative damage to P. ostreatus mycelia by inhibiting the protein and gene expression of aconitase (ACO), and additional studies found that the overexpression and interference of aco could affect the content of citric acid (CA). Furthermore, the addition of exogenous CA can induce alternative oxidase (aox) gene expression under heat stress, reduce the content of H2O2 in mycelium, and consequently protect the mycelia under heat stress. An additional analysis focused on the function of the aox gene in the heat stress response of mycelia. The results show that the colony diameter of the aox overexpression (OE-aox) strains was significantly larger than that of the wild-type (WT) strain under heat stress (32°C). In addition, the mycelia of OE-aox strains showed significantly enhanced tolerance to H2O2 In conclusion, this study demonstrates that NO can affect CA accumulation by regulating aco gene and ACO protein expression and that CA can induce aox gene expression and thereby be a response to heat stress.IMPORTANCE Heat stress is one of the abiotic stresses that affect the growth and development of edible fungi. Our previous study found that exogenous NO had a protective effect on mycelia under heat stress. However, its regulatory mechanism had not been elucidated. In this study, we found that NO altered the respiratory pathway of mycelia under heat stress by regulating aco The results have enhanced our understanding of NO signaling pathways in P. ostreatus.

Lactic acid accumulation under heat stress related to accelerated glycolysis and mitochondrial dysfunction inhibits the mycelial growth of Pleurotus ostreatus.

High temperature is a major threat to Pleurotus ostreatus cultivation. In this study, a potential mechanism by which P. ostreatus mycelia growth is inhibited under heat stress was explored. Lactate, as a microbial fermentation product, was found unexpectedly in the mycelia of P. ostreatus under heat stress, and the time-dependent accumulation and corresponding inhibitory effect of lactate on mycelial growth was further confirmed. The addition of a glycolysis inhibitor, 2-deoxy-D-glucose (2DG), reduced the lactate content in mycelia and slightly restored mycelial growth under high-temperature conditions, which indicated the accumulation of lactate can be inhibited by glycolysis inhibition. Further data revealed mitochondrial dysfunction under high-temperature conditions, with evidence of decreased oxygen consumption and adenosine triphosphate (ATP) synthesis and increased reactive oxygen species (ROS). The removal of ROS with ascorbic acid decreased the lactate content, and mycelial growth recovered to a certain extent, indicating lactate accumulation could be affected by the mitochondrial ROS. Moreover, metabolic data showed that glycolysis and the tricarboxylic acid cycle were enhanced. This study reported the accumulation of lactate in P. ostreatus mycelia under heat stress and the inhibitory effect of lactate on the growth of mycelia, which might provide further insights into the stress response mechanism of edible fungi. Key Points • Lactate can accumulate in Pleurotus ostreatus mycelia under heat stress and inhibit its growth. • The accumulation of lactate may be due to the acceleration of glycolysis and the dysfunction of mitochondria of P. ostreatus mycelia under high-temperature stress. • The glycolysis and tricarboxylic acid cycle of P. ostreatus mycelia were accelerated under high-temperature stress.

Expression patterns of two pal genes of Pleurotus ostreatus across developmental stages and under heat stress.

BACKGROUND: Phenylalanine ammonia-lyase (PAL, EC 4.3.1.24) is the first key enzyme in the phenylpropanoid pathway. The pal gene has been widely studied in plants and participates in plant growth, development and defense systems. However, in Pleurotus ostreatus, the biological functions of pal during organismal development and exposure to abiotic stress have not been reported. RESULTS: In this study, we cloned and characterized the pal1 (2232 bp) and pal2 (2244 bp) genes from the basidiomycete P. ostreatus CCMSSC 00389. The pal1 and pal2 genes are interrupted by 6 and 10 introns, respectively, and encode proteins of 743 and 747 amino acids, respectively. Furthermore, prokaryotic expression experiments showed that PAL enzymes catalyzed the conversion of L-phenylalanine to trans-cinnamic acid. The function of pal1 and pal2 was determined by constructing overexpression (OE) and RNA interference (RNAi) strains. The results showed that the two pal genes had similar expression patterns during different developmental stages. The expression of pal genes was higher in the reproductive growth stage than in the vegetative growth stage. And the interference of pal1 and pal2 delayed the formation of primordia. The results of heat stress assays showed that the RNAi-pal1 strains had enhanced mycelial tolerance to high temperature, while the RNAi-pal2 strains had enhanced mycelial resistance to H2O2. CONCLUSIONS: These results indicate that two pal genes may play a similar role in the development of P. ostreatus fruiting bodies, but may alleviate stress through different regulatory pathways under heat stress.

Trehalose induced by reactive oxygen species relieved the radial growth defects of Pleurotus ostreatus under heat stress.

Trehalose is a nonreducing disaccharide, and it plays an intracellular protective role in organisms under various stress conditions. In this study, the trehalose synthesis and its protective role in Pleurotus ostreatus were investigated. As a signal in metabolic regulation, reactive oxygen species (ROS) accumulated in the mycelia of P. ostreatus under heat stress (HS). Furthermore, mycelial growth was significantly inhibited, and the malondialdehyde (MDA) level significantly increased under HS. First, exogenous addition of H2O2 inhibited mycelial growth and elevated the MDA level, while N-acetyl cysteine (NAC) and vitamin C (VC) reduced the MDA level and recovered mycelial growth under HS by scavenging ROS. These results indicated that the mycelial radial growth defect under HS might be partly caused by ROS accumulation. Second, adding NAC and VC to the media resulted in rescued trehalose accumulation, which indicated that ROS has an effect on inducing trehalose synthesis. Third, the mycelial growth was recovered by addition of trehalose to the media after HS, and the MDA level was reduced. This effect was further verified by the overexpression of genes for trehalose-6-phosphate synthase (TPS) and neutral trehalase (NTH), which led to increased and reduced trehalose content, respectively. In addition, adding validamycin A (NTH inhibitor) to the media promoted trehalose accumulation and the recovered mycelial growth after HS. In conclusion, trehalose production was partly induced by ROS accumulation in the mycelia under HS, and the accumulated trehalose could promote the recovery of growth after HS, partly by reducing the MDA level in the mycelia.

A Partnership and Coping Enhancement Program for Couples Undergoing In Vitro Fertilization Treatment: An Intervention Study.

This is a feasibility study to examine the effects of a partnership and coping enhancement program (PCEP) on improving the psychological well-being and marital functions of couples undergoing in vitro fertilization treatment. A total of 100 couples were recruited consecutively and assigned to a PCEP intervention group or a routine care control group. The couples in the PCEP group received an additional face-to-face, couple-based, 90-minute session on enhancing partnership and coping on the day of the embryo transfer (ET). The outcome measures were assessed at baseline (T0), 10 days after the ET (T1), and one month after the ET (T2). The level of anxiety of the women was lower in the intervention than in the control group at T1. Significant improvements in partnership and dyadic coping were seen in women at T2. The men of infertile couples reported a significant improvement in the scores for partnership at T2. The PCEP had no significant effects on marital satisfaction and marital adjustment for both the females and males of the infertile couples. The findings indicated that the PCEP is feasible and acceptable. Before a larger simple-size randomized controlled trial with participants drawn from multiple reproductive medical centers is conducted to further confirm its effectiveness.

High temperature induced disruption of the cell wall integrity and structure in Pleurotus ostreatus mycelia.

Fungal cells are surrounded by a tight cell wall to protect them from harmful environmental conditions and to resist lysis. The synthesis and assembly determine the shape, structure, and integrity of the cell wall during the process of mycelial growth and development. High temperature is an important abiotic stress, which affects the synthesis and assembly of cell walls. In the present study, the chitin and ß-1,3-glucan concentrations in the cell wall of Pleurotus ostreatus mycelia were changed after high-temperature treatment. Significantly higher chitin and ß-1,3-glucan concentrations were detected at 36 °C than those incubated at 28 °C. With the increased temperature, many aberrant chitin deposition patches occurred, and the distribution of chitin in the cell wall was uneven. Moreover, high temperature disrupts the cell wall integrity, and P. ostreatus mycelia became hypersensitive to cell wall-perturbing agents at 36 °C. The cell wall structure tended to shrink or distorted after high temperature. The cell walls were observed to be thicker and looser by using transmission electron microscopy. High temperature can decrease the mannose content in the cell wall and increase the relative cell wall porosity. According to infrared absorption spectrum, high temperature broke or decreased the glycosidic linkages. Finally, P. ostreatus mycelial cell wall was easily degraded by lysing enzymes after high-temperature treatment. In other words, the cell wall destruction caused by high temperature may be a breakthrough for P. ostreatus to be easily infected by Trichoderma.

Genome-Wide Characterization and Expression Analyses of Pleurotus ostreatus MYB Transcription Factors during Developmental Stages and under Heat Stress Based on de novo Sequenced Genome.

Pleurotus ostreatus is a commercially grown mushroom species in China. However, studies on the mechanisms of the fruiting body development and stress response of P. ostreatus are still at a primary stage. In this study, we report the entire genome sequence of P. ostreatus CCMSSC03989. Then, we performed comprehensive genome-wide characterization and expression analysis of the MYB transcription factor family during a series of developmental stages and under the condition of heat stress. A 34.76 Mb genome was obtained through next-generation sequencing (NGS) and Bionano optical mapping approaches. The genome has a scaffold N50 of 1.1 Mb and contains 10.11% repeats, and 10,936 gene models were predicted. A total of 20 MYB genes (PoMYB) were identified across the genome, and the full-length open reading frames were isolated. The PoMYBs were classified into 1 repeat (1R), 2R, and 3R-MYB groups according to their MYB domain repeat numbers, and 3R-MYBs possessed relatively more introns than 1R and 2R-MYBs. Based on phylogenetic analysis, the PoMYBs were divided into four groups and showed close relationships with the MYB genes of plants and fungi. RNA-sequencing (RNA-Seq) and quantitative PCR (qPCR) analyses revealed that PoMYB expression showed stage-specific patterns in reproductive stages and could be induced by heat stress. The P. ostreatus draft genome will promote genome-wide analysis, and our study of PoMYBs will promote further functional analysis of MYB genes in mushrooms.

Endurance with partnership: a preliminary conceptual framework for couples undergoing in vitro fertilisation treatment.

BACKGROUND/OBJECTIVES: Infertility affects both women and men in the physical, emotional, existential, and interpersonal realms. When couples seek in vitro fertilisation (IVF) treatment, they further suffer from the difficulties of the treatment and the uncertainty of its outcome. The aim of this study was to develop a preliminary conceptual framework for couples undergoing IVF treatment to give health professionals a better understanding of the experiences of such couples, and to guide the development of an intervention. METHODS: The process of identifying frameworks adopted in intervention studies confirmed that there is no established framework for infertile couples undergoing IVF treatment. A skeletal framework identified from previous studies provides an internal structure for the proposed framework for couples undergoing IVF treatment, filled out with concepts drawn from a concept analysis and a qualitative study, knitting the structure together. RESULTS: This preliminary framework is the Endurance with Partnership Conceptual Framework (P-EPCF). It consists of four domains: the impacts of infertility and stressors, dyadic mediators, dyadic moderators and dyadic outcomes. According to the P-EPCF, the impacts of infertility and IVF treatment can be mediated by the couples' partnership and dyadic coping. Improvements in the psychological well-being and marital functioning of IVF couples can then be expected. CONCLUSIONS: The P-EPCF would be potentially valuable in guiding the development of a complex, couple-based intervention, which could focus on enhancing the partnership of couples and their coping strategies.

Association between Methionine Synthase Reductase A66G Polymorphism and Male Infertility: A Meta-Analysis.

The aim of this meta-analysis was to assess the methionine synthase reductase (MTRR) gene 66A>G polymorphism and male infertility susceptibility. Studies were identified in PubMed and Embase databases. An odds ratio (OR) was used to assess the relationship between MTRR 66A>G polymorphism and male infertility risk. A total of seven case-control studies containing 1438 patients and 1363 controls were enrolled in the meta-analysis. Our results showed that no association exists between the MTRR 66A>G polymorphism and male infertility risk in the total population (GG vs. AA: OR = 1.29, 95% confidence interval [CI] = 1.00-1.66; GA vs. AA: OR = 1.09, 95% CI = 0.92-1.30; dominant model: OR = 1.13, 95% CI = 0.96-1.34; recessive model: OR = 0.83, 95% CI = 0.67-1.03). In a subgroup analysis by nationality, the 66A>G polymorphism was not associated with male infertility in both Asians and Caucasians. In a subgroup analysis stratified by male infertility type, significant association was found with oligoasthenozoospermia (OAT) (GG vs. AA: OR = 1.83, 95% CI = 1.24-269; GA vs. AA: OR = 1.36, 95% CI = 0.88-2.11; dominant model: OR = 1.51, 95% CI = 1.13-2.01; recessive model: OR = 0.62, 95% CI = 0.46-0.85). In summary, the present meta-analysis suggests that MTRR 66A>G polymorphism may be associated with OAT risk.

The Partnership and Coping Enhancement Programme for couples undergoing in vitro fertilization treatment: the development of a complex intervention in China.

PURPOSE: Couples as dyads suffer from the diagnosis of infertility and related treatment. These couples commonly experience emotional and physical pain and tension in their marital lives. The purpose of this study is to report on the process of developing a potentially feasible and effective complex intervention for couples undergoing in vitro fertilization treatment in China. METHODS: The Medical Research Council (MRC) framework for developing and evaluating the complex intervention was adopted to guide the development of the Partnership and Coping Enhancement Programme (PCEP). In developing the PCEP, three steps were taken, namely, (1) identifying evidence by conducting literature reviews, a concept analysis and a qualitative study; (2) identifying/developing a theory-in this case, a preliminary Endurance with Partnership Conceptual Framework (P-EPCF) was proposed; and (3) modelling the process and outcomes of the PCEP. RESULTS: The PCEP that was developed is targeted mainly at the domains of the partnership mediators of stress in the P-EPCF. It consists of two sections-partnership and coping-and will be delivered to infertile couples on the day of embryo transfer. The main focuses of the programme are to facilitate mutual sharing and support in infertile couples, and to improve their individual and dyadic coping strategies while undergoing IVF treatment, especially in the period when they are waiting for the results of a pregnancy test and after the disclosure of a negative treatment outcome. The programme is couple-based, consisting of experience sharing, psychoeducation, meditation exercise, skill practise and supplemental written materials. CONCLUSIONS: The Partnership and Coping Enhancement Programme (PCEP) for couples undergoing in vitro fertilization treatment was developed according to the guideline of the MRC framework. It is recommended that a pilot study be conducted to evaluate its feasibility and to model the process and outcomes of the programme.

Gene cloning, expression, and characterization of trehalose-6-phosphate synthase from Pleurotus ostreatus.

Trehalose-6-phosphate synthase (TPS; EC2.4.1.15) catalyzes the first step in trehalose synthesis, which involves transfer of glucose from uridine diphosphate glucose (UDPG) to glucose 6-phosphate (G6P) to form trehalose-6-phosphate. To determine the gene and enzymatic characteristics of TPS in Pleurotus ostreatus, we cloned and sequenced the cDNA of PoTPS1, which contains a 1665 bp open reading frame that encodes a 554-amino acid protein with a predicted molecular weight of 62.01 kDa. This gene was expressed in Escherichia coli BL21 and then the recombinant protein was purified and characterized. Results showed that the optimum pH and temperature for the recombinant PoTPS1 were 7.4 and 30 °C, respectively; the Km value against G6P and UDPG were 0.14 and 0.17 mM, respectively, and the Vmax and Kcat values were 91.86 nkat/g and 5.89 s-1 , respectively. Trehalose content was as high as 158.88 mg g-1 dry weight after heat treatment at 40 °C for 15 h, which was consistent with highest TPS1 activity at that time point. This result indicated that PoTPS1 was responsible for trehalose synthesis in P. ostreatus.

Establishment of an efficient transformation system for Pleurotus ostreatus.

Pleurotus ostreatus is widely cultivated worldwide, but the lack of an efficient transformation system regarding its use restricts its genetic research. The present study developed an improved and efficient Agrobacterium tumefaciens-mediated transformation method in P. ostreatus. Four parameters were optimized to obtain the most efficient transformation method. The strain LBA4404 was the most suitable for the transformation of P. ostreatus. A bacteria-to-protoplast ratio of 100:1, an acetosyringone (AS) concentration of 0.1 mM, and 18 h of co-culture showed the best transformation efficiency. The hygromycin B phosphotransferase gene (HPH) was used as the selective marker, and EGFP was used as the reporter gene in this study. Southern blot analysis combined with EGFP fluorescence assay showed positive results, and mitotic stability assay showed that more than 75% transformants were stable after five generations. These results showed that our transformation method is effective and stable and may facilitate future genetic studies in P. ostreatus.

Heat stress induces apoptotic-like cell death in two Pleurotus species.

High temperature is an important environmental factor that affects the growth and development of most edible fungi, however, the mechanism(s) for resistance to high temperature remains elusive. Nitric oxide is known to be able to effectively alleviate oxidative damage and plays an important role in the regulation of trehalose accumulation during heat stress in mycelia of Pleurotus eryngii var. tuoliensis. In this paper, we investigated whether heat stress can activate apoptosis-like cell death in mycelia of Pleurotus. Two Pleurotus species were used to detect morphological features characteristic of apoptosis including nuclear condensation, reactive oxygen species accumulation, and DNA fragmentation when exposed to heat stress (42 °C). The results showed that these classical apoptosis markers were apparent in Pleurotus strains after heat treatment. The heat-induced apoptosis-like cell death in Pleurotus was further probed using oligomycin and N-acetylcysteine, both of which were shown to block processes leading to apoptosis. This is the first report that apoptosis-like cell death occurs in Pleurotus species as a result of abiotic stress, and that this process can be inhibited with chemicals that block mitochondrial-induced apoptotic pathways and/or with ROS-scavenging compounds.

A novel laccase with inhibitory activity towards HIV-I reverse transcriptase and antiproliferative effects on tumor cells from the fermentation broth of mushroom Pleurotus cornucopiae.

A novel laccase with a molecular mass of 67 kDa was isolated from the fermentation broth of Pleurotus cornucopiae through ion exchange chromatography and gel filtration. The optimal pH and temperature for the laccase was pH 4.2 and 30°C, respectively. The laccase activity was remarkably inhibited by Fe(3+) and Hg(2+) , while it was stimulated by Cu(2+) and Pb(2+) . It inhibited proliferation of the hepatoma cells HepG2 and the breast cancer cells MCF-7, and the activity of HIV-I reverse transcriptase with IC50 values of 3.9, 7.6 and 3.7 µM, respectively.

[Association of menopausal specific fat distribution changes with metabolic risk factors].

OBJECTIVE: To investigate the association of menopausal specific fat distribution with metabolic risk factors. METHODS: Two hundred Chinese women including 110 premenopausal women aged 39.7±9.2 y and 90 postmenopausal women aged 55.4±4.6 years were enrolled. Total and regional fat depots including android fat and gynoid fat were measured by dual-energy X-ray absorptiometry. The differences of regional fat depot between pre-and post menopausal women were compared by covariate analysis. The association of regional fat changes with metabolic risk factors were analyzed by logistic models. RESULTS: Gynoid fat was significantly decreased in postmenopausal women after accounting for total fat changes. After adjusting covariates (including age, BMI, and %BF), the inverse associations of gynoid fat with metabolic risk factors still remained (OR=1.8-3.7,P<0.05). Furthermore, the interaction between menopause and gynoid fat was significantly in all logistic models (P<0.05). CONCLUSION: Android fat was increased whereas gynoid fat was decreased after menopause. Opposite to the detrimental effects of android fat, gynoid fat was inversely associated with metabolic risk factors, especially in postmenopausal women.