Discovery and bioinspired total syntheses of unprecedented sesquiterpenoid dimers unveiled bifurcating [4 + 2] cycloaddition and target differentiation of enantiomers.

[4 + 2] cycloaddition has led to diverse polycyclic chiral architectures, serving as novel sources for organic synthesis and biological exploration. Here, an unprecedented class of cadinane sesquiterpene [4 + 2] dimers, henryinins A-E (1-5), with a unique 6/6/6/6/6-fused pentacyclic system, were isolated from Schisandra henryi. The divergent total syntheses of compounds 1-5 and their enantiomers (6-10) were concisely accomplished in eight linear steps using a protection-free approach. Mechanistic studies illustrated the origin of selectivity in the key [4 + 2] cycloaddition as well as the inhibition of reaction pathway bifurcation via desymmetrization. The chemical proteomics results showed that a pair of enantiomers shared common targets (PRDX5 C100 and BLMH C73) and had unique targets (USP45 C588 for 4 and COG7 C419 for 9). This work provides experimental evidence for the discovery of unprecedented cadinane dimers from selective Diels-Alder reaction and a powerful strategy to explore the biological properties of natural products.

Harnessing Natural Products by a Pharmacophore-Oriented Semisynthesis Approach for the Discovery of Potential Anti-SARS-CoV-2 Agents.

Natural products possessing unique scaffolds may have antiviral activity but their complex structures hinder facile synthesis. A pharmacophore-oriented semisynthesis approach was applied to (-)-maoelactone A (1) and oridonin (2) for the discovery of anti-SARS-CoV-2 agents. The Wolff rearrangement/lactonization cascade (WRLC) reaction was developed to construct the unprecedented maoelactone-type scaffold during semisynthesis of 1. Further mechanistic study suggested a concerted mechanism for Wolff rearrangement and a water-assisted stepwise process for lactonization. The WRLC reaction then enabled the creation of a novel family by assembly of the maoelactone-type scaffold and the pharmacophore of 2, whereby one derivative inhibited SARS-CoV-2 replication in HPA EpiC cells with a low EC50 value (19±1 nM) and a high TI value (>1000), both values better than those of remdesivir.

3-Hydroxy-4-methyldecanoic Acid-Containing Cyclotetradepsipeptides from an Endolichenic Beauveria sp.

An investigation of an endolichenic Beauveria sp. led to the discovery of seven new cyclotetradepsipeptides, beauveamides A-G (2-8), along with the known beauverolide Ka (1). All incorporate a 3-hydroxy-4-methyldecanoic acid (HMDA) moiety in their structures. Their configuration was determined through Marfey's, J-based configuration analysis, and NMR computational methods, representing the first time that the stereostructures of HMDA-moiety-containing cyclotetradepsipeptides have been established. Compounds 1 and 2 exhibited protecting effects on HEI-OC1 cells at 10 µM, while 1, 4, and 5 could stimulate glucose uptake in cultured rat L6 myoblasts at 50 µM. Compound 1 showed dose-dependent activity in both L6 myoblasts and myotubes.

Chaetolactam A, an Azaphilone Derivative from the Endophytic Fungus Chaetomium sp. g1.

Chaetolactam A (1), an unprecedented azaphilone derivative bearing a unique 9-oxa-7-azabicyclo[4.2.1]octan-8-onering system, together with two new compounds, 11-epi-chaetomugilide B (2) and chaetomugilide D (3) was isolated from an endophytic fungus, Chaetomium sp. g1. Notably, extensive NMR data analyses, NMR calculations with DP4 and DP4+ analyses, ECD calculations, and the RDC method were employed to establish the structure of 1. Furthermore, 2 exhibited potent apoptosis induction activity by mediating caspase-3 activation and PARP degradation at 3 µM in HL-60.

Logistic regression analysis of the related factors in discarded semen of boars in Southern China.

This study aimed to investigate factors affecting the discarded semen of boars. A total of 176,368 ejaculates of boars from nine AI centers were collected from January 2013 to May 2016 in Southern China. The criteria for determining whether their semen was abnormal included cytoplasmic droplets, coiling tail, sperm agglutination, impurity, poor motility, oligozoospermia, necrozoospermia, azoospermia, and hemospermia. The cause of discarded semen was evaluated with a Chi-square test, and the effects of housing type, breed, age at collection, season identified in the northern hemisphere, and age at herd entry of the discarded semen of boars were analyzed with a logistic regression model. Results indicated the proportion of the discarded semen (PDS) in the nine AI centers was 13.09%. Chi-square test showed the greatest PDS among all causes was found in semen discarded due to cytoplasmic droplets (31.60%), followed by impurity (25.96%), sperm agglutination (20.31%), coiling tail (17.72%), oligozoospermia (10.86%), and others (6.78%; P < 0.0001). Logistic regression analysis revealed the PDS was affected by all these five factors (P < 0.0001). The PDS of boars raised individually in stalls was greater than that of boars raised individually in pens (OR: 1.657; 95% CI: 1.607 to 1.709). The PDSs of Duroc boars (OR: 1.130; 95% CI: 1.093 to 1.167) and Yorkshire boars (OR: 1.432; 95% CI: 1.380 to 1.486) were greater than that of Landrace boars. The PDSs of adult boars (aged from 13 to 24 mo, from 25 to 36 mo, and more than 37 mo with OR: 0.800, 0.941, and 0.838, respectively; 95% CI: 0.771 to 0.831, 0.902 to 0.983, and 0.790 to 0.889, respectively) were lower than those of young boars (aged less than 12 mo). The PDSs of semen collected in summer (OR: 1.367; 95% CI: 1.314 to 1.422), autumn (OR: 1.185; 95% CI: 1.138 to 1.234), and winter (OR: 1.159; 95% CI: 1.115 to 1.206) were greater than those of semen obtained in spring. The PDSs of boars introduced at ages of 5-7 mo (OR: 1.432; 95% CI: 1.380 to 1.486) and 10-12 mo (OR: 1.432; 95% CI: 1.380 to 1.486) were greater than those of boars introduced at an age of 8 and 9 mo. In conclusion, logistic regression analysis reveals discarded semen is affected by housing type, breed, age at collection, season, and age at herd entry. More importantly, cytoplasmic droplets is the primary reason for discarding boar semen, and 8 months at herd entry is the most suitable age for boar introduction.

(+)- and (-)-Alternarilactone A: Enantiomers with a Diepoxy-Cage-like Scaffold from an Endophytic Alternaria sp.

The enantiomers (+)- and (-)-alternarilactone A (1), the first examples of dibenzo-α-pyrones bearing a diepoxy-cage-like moiety, were isolated from the endophytic fungus Alternaria sp. hh930. The deficiency in 1H-1H COSY and HMBC correlations caused by the highly oxidized caged system of 1 and the deceptive and ambiguous signals such as "W" couplings in NMR data increased the risk of structure misassignment of 1. By performing a quantum chemical calculation of the NMR chemical shifts together with a DP4+ probability analysis and single-crystal X-ray crystallographic experiment, their structures were unambiguously determined, and their absolute configurations were determined by ECD calculations.

Serum and Seminal Plasma Element Concentrations in Relation to Semen Quality in Duroc Boars.

Element concentrations in serum and seminal plasma were studied in Duroc boars with different semen quality characteristics. Based on the utilization rate of 2174 ejaculates from June to August in 2016, a total of 166 Duroc boars were allocated into three groups: low utilization rate group (LG, 0 to 60% utilization rate), medium utilization rate group (MG, 60 to 80%), and high utilization rate group (HG, 80 to 100%). Serum and seminal plasma samples were collected, and element levels were analyzed using inductively coupled plasma mass spectrometry. The results showed that LG boars had higher concentrations of serum copper and seminal plasma cadmium compared with MG and HG boars (P < 0.05), and serum copper and seminal plasma cadmium were negatively correlated with sperm motility, while positively correlated with the abnormal sperm rate. We observed the abnormal sperm rate increased by approximately 4.53% with serum copper increasing from 1.63 to 2.44 mg/L, while sperm motility decreased by approximately 2.85% with seminal plasma cadmium increasing from 0 to 0.82 µg/L. Moreover, serum iron and manganese levels in the LG group were significantly reduced compared with the HG boars (P < 0.05), and the two elements were negatively correlated with the abnormal sperm rate (P < 0.05). In conclusion, excessive copper and absence of iron and manganese in serum as well as higher seminal plasma cadmium may reduce the utilization rate of semen by impairing sperm motility and morphology, indicating the importance of adding and monitoring microelements in boar diet.

Secondary Metabolites from the Endophytic Fungus Xylaria sp. hg1009.

A detailed chemical investigation of the secondary metabolites produced by the endophytic fungus Xylaria sp. isolated from the stems of Isodon sculponeatus afforded six new compounds, xylariahgins A-F (1-6), two new natural products (7 and 8), along with two known compounds (9 and 10) (Fig. 1). The structures of all compounds were unambiguously established by analyzing their spectroscopic data or referring to pertinent literature. Compounds 1-8 were tested for their cytotoxic activity against five human tumor cell lines.

Multi-level mixed models for evaluating factors affecting the mortality and weaning weight of piglets in large-scale commercial farms in central China.

This study investigated the factors affecting piglet mortality (square root of mortality, SQRM) and average weaning weight (AWW) in commercial farms in central China. Information on sow diets, management and climate from 2478 weaning batches completed in 16 pig farms was collected from 2009 to 2013. Multi-level mixed models, which included batch level (level 1) and farm level (level 2), were used to analyze the factors associated with SQRM and AWW. The mean values of SQRM and AWW were 2.52% (SD = 0.96) and 7.31 (SD = 0.77), respectively. Lactation sow diets supplemented with oregano essential oils (OEOs) decreased the SQRM (P < 0.05) and increased the AWW of piglets (P < 0.01). The SQRM was lower in period 2 (June to September, hot) than in period 1 (February to May, warm) and period 3 (October to January, cold; P < 0.05 and 0.001, respectively). The AWW was lower in periods 2 and 3 than in period 1 (P < 0.01). In conclusion, supplying OEOs to lactation diets can increase the weaning weight and reduce the mortality of piglets. The sources of variations in SQRM and AWW are of greater concern in the warm season than in the hot season.

Analysis of influencing factors of boar claw lesion and lameness.

This study aimed to investigate the factors affecting boar claw lesions and lameness. A total of 1299 boars were examined for claw lesions and lameness, including 788 boars reared in individual pens with solid concreted floor (IPS) and 511 boars raised in individual stalls with slatted floor (ISS). Flooring type showed significant impacts on all claw lesion types (P < 0.01). Except for swelling ankle, boar age had significant effects on all other claw lesion types (P < 0.01). In addition, only heel overgrowth and erosion, cracked wall horizontal, heel-sole crack, dew claws, and toes were significantly related to boar breeds (P < 0.05). Furthermore, IPS lame boars had higher prevalence of lameness in the hind limb (P < 0.05), whereas in ISS lame boars, there were no significant differences in prevalence of lameness between the fore and hind limbs (P > 0.05). Boar lameness was moderately correlated with swelling ankle (Φ = 0.5571). In conclusion, claw lesions can be influenced by flooring type, boar age and breed, and could serve as a predictor for boar lameness.

Gypmacrophin A, a Rare Pentacyclic Sesterterpenoid, Together with Three Depsides, Functioned as New Chemical Evidence for Gypsoplaca macrophylla (Zahlbr.) Timdal Identification.

The phytochemical investigation on 1 g of materials from Gypsoplacamacrophylla (Zahlbr.) Timdal resulted in the discovery of gypmacrophin A, a rare pentacyclic sesterterpenoid; brialmontin III, a new polysubstituted depside and two known ones, brialmontins I and II. The structure and absolute configurations of gypmacrophin A were elucidated by spectroscopic analyses and computational methods. Gypmacrophin A showed weak inhibition of AchE with an IC50 value of 32.03 µM. The four compounds provided new chemical evidence for G. macrophylla identification.

Linear model analysis of the influencing factors of boar longevity in Southern China.

This study aimed to investigate the factors influencing the boar herd life month (BHLM) in Southern China. A total of 1630 records of culling boars from nine artificial insemination centers were collected from January 2013 to May 2016. A logistic regression model and two linear models were used to analyze the effects of breed, housing type, age at herd entry, and seed stock herd on boar removal reason and BHLM, respectively. Boar breed and the age at herd entry had significant effects on the removal reasons (P < 0.001). Results of the two linear models (with or without removal reason including) showed boars raised individually in stalls exhibited shorter BHLM than those raised in pens (P < 0.001). Boars aged 5 and 6 months at herd entry (44.6%) showed shorter BHLM than those aged 8 and 9 months at herd entry (P < 0.05). Approximately 95% boars were culled for different reasons other than old age, and the BHLM of these boars was at least 12.3 months longer than that of boars culled for other reasons (P < 0.001). In conclusion, abnormal elimination in boars is serious and it had a negative effect on boar BHLM. Boar removal reason and BHLM can be affected by breed, housing type, and seed stock herd. Importantly, 8 months is suggested as the most suitable age for boar introduction.

Eicosapentaenoic acid abolishes inhibition of insulin-induced mTOR phosphorylation by LPS via PTP1B downregulation in skeletal muscle.

Dietary n-3 polyunsaturated fatty acids (n-3 PUFAs) increase insulin signaling in skeletal muscle. In the current study, we investigated the effect of eicosapentaenoic acid (EPA) on insulin-induced mammalian target of rapamycin (mTOR) phosphorylation in myotubes. We showed that EPA did not affect basal and insulin-induced mTOR phosphorylation in myotubes. However, EPA abolished lipopolysaccharide (LPS) -induced deficiency in insulin signaling (P < 0.05). Pre-incubation of nuclear factor κB (NF-κΒ) and c-Jun N-terminal kinases (JNK) inhibitors prevented the decreased insulin-induced mTOR phosphorylation elicited by LPS (P < 0.05). In addition, in protein tyrosine phosphatase-1B (PTP1B) knockdown myotubes, LPS failed to decrease insulin-induced mammalian target of rapamycin (mTOR) phosphorylation in myotubes (P > 0.05). In myotubes, LPS stimulated PTP1B expression via NF-κB and activation protein-1 (AP1). Pre-incubation of 50 µM EPA prevented the LPS-induced activation of AP1 and NF-κΒ as well as PTP1B expression (P < 0.05). Interestingly, incubation of peroxisome proliferator-activated receptor γ (PPARγ) antagonist (GW9662) prior to EPA treatment, the effect of EPA on insulin-induced mTOR phosphorylation was blocked. Accordingly, EPA did not inhibit the LPS-induced activation of AP1 or NF-κΒ as well as PTP1B expression when incubation of GW9662 prior to EPA treatment. The in vivo study showed that EPA prevented LPS-induced PTPT1B expression and a decrease in insulin-induced mTOR phosphorylation in muscle of mice. In summary, EPA abolished LPS inhibition of insulin-induced mTOR phosphorylation in myotubes, and one of the key mechanisms was to inhibit AP1 and NF-κB activation and PTP1B transcription.

Effects of dietary oregano essential oil and vitamin E supplementation on meat quality, stress response and intestinal morphology in pigs following transport stress.

This study investigates the effects of dietary oregano essential oil (OEO) and vitamin E (Vit E) supplementation on meat quality, stress response and intestinal morphology in pigs following transport stress. A total of 288 finishing pigs were randomly assigned to three groups: a basal diet or a basal diet supplemented either with 200 mg/kg Vit E or 25 mg/kg OEO. After a 28-day feeding trial, total of 132 finishing pigs according diet and transport stress were assigned to one of four treatment groups: 1) control treatment without transport stress (Control group), 2) control treatment with 5-hr transport stress (Negative group), 3) Vit E treatment with 5-hr transport stress and 4) OEO treatment with 5-hr transport stress. Transport stress pigs had lower muscle 45 min pH (pHi) and higher drip loss than control pigs. Dietary OEO and Vit E supplementation significantly increased 45min pH under transport stress, and the OEO groups produced lower 24-hr drip loss values (P<0.05) than that of pigs from the negative group. The OEO-supplemented pigs showed decreased serum levels of creatine kinase (CK) and cortisol (P<0.05), and decreased Hsp 27 (heat shock protein 27) and Hsp 70 (heat shock protein 70) mRNA expression in the muscle (P<0.05). Additionally, histological analysis revealed intestinal epithelial damage in transport stress pigs that was reversed by dietary supplementation with OEO. In conclusion, supplementation with dietary OEO may be superior to supplementation with dietary Vit E in alleviating the meat quality, stress response and intestinal morphology of pigs after challenge due to transportation stress.

Dietary n-6:n-3 ratio and Vitamin E improve motility characteristics in association with membrane properties of boar spermatozoa.

This study was aimed to evaluate the effects of dietary n-6:n-3 ratio and Vitamin E on the membrane properties and motility characteristics of spermatozoa in boars. Forty Duroc boars were randomly distributed in a 2 × 2 factorial design with two n-6:n-3 ratios (14.4 and 6.6) and two Vitamin E levels (200 and 400 mg kg-1 ). During 16 weeks of treatment, fresh semen was collected at weeks 0, 8, 12, and 16 for measurements of motility characteristics, contents of fatty acids, membrane properties (membrane fluidity and membrane integrity), and lipid peroxidation of the spermatozoa. The semen was diluted in Beltsville Thawing Solution (BTS) extender and stored at 17°C, and the sperm motility was assessed at 12, 36, 72, and 120 h of storage. The 6.6 n-6:n-3 dietary ratio increased the contents of n-3 polyunsaturated fatty acids (PUFAs) and docosahexaenoic acid (DHA) and improved the membrane integrity and membrane fluidity of the spermatozoa, resulting in notably increased total motility, sperm progressive motility, and velocity parameters of fresh semen. Feeding diet with Vitamin E (400 mg kg-1 ) prevented sperm lipid peroxidation, and resulted in higher total motility and sperm progressive motility in fresh and liquid stored semen. In conclusion, the adjustment of n-6:n-3 ratio (6.6) and supply of Vitamin E (400 mg kg-1 ) successfully improved sperm motility characteristics and thus may be beneficial to the fertility of boars, which might be due to the modification of the physical and functional properties of spermatozoa membrane in response to dietary supplementation.

Protective effect of quercetin on pig intestinal integrity after transport stress is associated with regulation oxidative status and inflammation.

This experiment was conducted to evaluate the effects of quercetin supplementation on intestinal integrity, intestinal reactive oxygen species (ROS) levels and intestinal inflammation in pigs under transport stress. A total of 170 finishing pigs were randomly assigned into two groups. Animals in the control group consumed a basal diet, while those in the treatment group consumed the same diet supplemented with 25 mg quercetin per kg feed. After a 4-week period, pigs were transported for 5 hr. The quercetin-supplemented pigs showed decreased serum levels of endotoxin (P<0.05), increased height of jejunum villi (P<0.05), and increased occludin and zonula occudens-1 (ZO-1) mRNA expression in the jejunum (P<0.05). These parameters are associated with intestinal health and were markedly improved by quercetin supplementation. Pigs consuming the quercetin-supplemented diet had lower intestinal levels of ROS and malondialdehyde (MDA) compared with the control group (P<0.05). This finding coincided with greater inhibition of the innate immune system (P<0.05), including mitogen-activated protein kinase (MAPK), protein kinase B (Akt) and nuclear factor κB (NF-κB) signaling pathways, as well as decreased expression of inflammatory cytokines in the jejunum. These results indicate that quercetin alleviates intestinal injury in pigs during transport, probably through modulation of intestinal oxidative status and inflammation.

[Morphological characteristics and gene mapping of a novel bent pedicel branch (bpb1) mutant in rice].

Rice pedicels are tightly associated with the yield of grain. In the present study, a novel and stable pedicel mutant bpb1 (bent pedicel branch 1) was obtained from the wild type "Zhenong 7" after 60Co γ-ray treatment. The mutant had the typical phenotype of bent pedicel branches with multiple abnormal phenotypes, such as longer pedicels, short panicles, and dwarfism. Detail examination using scanning electron microscopy revealed that the pedicel epidermal hairs and stomas in the mutant were smaller than those in the wild type. The epidermal and sclerenchymatous cells were arranged irregularly, and the cells in the bend region of pedicels became smaller and arranged closely. The transverse observation of the mutant pedicel branches showed that the small vascular bundles arranged differently from those of the wild type. Genetic analysis indicated that the abnormal phenotypes were controlled by a single recessive gene. Using the F2 mapping population from the bpb1 mutant crossed with the japonica rice variety "Zhenongda 104", the bpb1 gene was mapped in a 343 kb region between two SSR markers, RM21537 and RM21552, at the long arm of chromosome 7. Because no homologous gene was found in this region until now, bpb1 might be a novel gene related to the pedicel development and growth. This study could be beneficial to future cloning and functional analysis of the bpb1 gene.

Fine mapping and candidate gene prediction of the photoperiod and thermo-sensitive genic male sterile gene pms1(t) in rice.

Pei'ai64S, an indica sterile variety with photoperiod and thermo-sensitive genic male sterile (PTGMS) genes, has been widely exploited for commercial seed production for "two-line" hybrid rice in China. One PTGMS gene from Pei'ai64S, pms1(t), was mapped by a strategy of bulked-extreme and recessive-class approach with simple sequence repeat (SSR) and insert and deletion (In-Del) markers. Using linkage analysis for the F(2) mapping population consisting of 320 completely male sterile individuals derived from a cross between Pei'ai64S and 93-11 (indica restorer) lines, the pms1(t) gene was delimited to the region between the RM21242 (0.2 cM) and YF11 (0.2 cM) markers on the short arm of chromosome 7. The interval containing the pms1(t) locus, which was co-segregated with RM6776, is a 101.1 kb region based on the Nipponbare rice genome. Fourteen predicted loci were found in this region by the Institute for Genomic Research (TIGR) Genomic Annotation. Based on the function of the locus LOC_Os07g12130 by bioinformatics analysis, it is predicted to encode a protein containing a Myb-like DNA-binding domain, and may process the transcript with thermosensory response. The reverse transcription-polymerase chain reaction (RT-PCR) results revealed that the mRNA levels of LOC_Os07g12130 were altered in different photoperiod and temperature treatments. Thus, the LOC_Os07g12130 locus is the most likely candidate gene for pms1(t). These results may facilitate not only using the molecular marker assisted selection of PTGMS genes, but also cloning of the pms1(t) gene itself.

[Gene cloning and structure characterization of CMS/Rf system in plants].

It has achieved great success in plant heterosis by using cytoplasmic male sterility and fertility restorer (CMS/Rf) system. In order to elucidate the action mechanism of Rf gene in this system, many researchers have been contributing themselves to map and clone fertility restorer genes in recent years. Four Rf genes were recently cloned successfully. Here we presented the overview of plant CMS fertility restorer gene mapping, cloning and its molecular mechanism. It was also discussed on utilization of fertility restorer genes in the molecular breeding in plants.