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Chlamydomonas reinhardtii is a model organism with three sequenced genomes capable of genetic transformation. C. reinhardtii has the advantages of being low cost, non-toxic, and having a post-translational modification system that ensures the recombinant proteins have the same activity as natural proteins, thus making it a great platform for application in molecular biology and other fields. In this review, we summarize the existing methods for nuclear transformation of C. reinhardtii, genes for selection, examples of foreign protein expression, and factors affecting transformation efficiency, to provide insights into effective strategies for the nuclear transformation of C. reinhardtii.

CYP27A1, CYP2R1 and CYP27B1 hydroxylases are involved in the synthesis of 1, 25-hydroxyvitamin D3, which plays a role in the immune regulation and pathogenesis of hepatitis C virus (HCV) infection. The aim of the present study was to investigate the relationships between polymorphisms in vitamin D pathway genes and HCV infection outcomes in a Chinese population. Nine single-nucleotide polymorphisms (SNPs) of CYP27A1, CYP2R1 and CYP27B1 were genotyped in a high-risk Chinese population. The distributions of these SNPs were compared among groups with different outcomes of HCV infection, including 863 cases of persistent HCV infection, 524 cases of spontaneous clearance, and 1079 uninfected controls. The results showed that the CYP2R1 rs12794714-G, rs10741657-A, rs1562902-C, and rs10766197-G alleles were significantly associated with increased susceptibility to HCV infection (all PFDR

Vitamin D binding protein (VDBP) plays an important role in the immune modulation and pathogenesis of hepatitis C viral (HCV) infection by influencing serum vitamin D levels. The present study aims to evaluate the association of VDBP genetic polymorphisms with susceptibility to and chronicity of HCV infection in a high-risk Chinese population. Seven genetic variants in the VDBP gene were genotyped in a case-control study of 886 patients with HCV persistent infection, 539 subjects with spontaneous clearance, and 1081 uninfected controls. Logistic regression analysis was used to assess the effects of these variants on HCV infection outcomes. The results showed that two variants rs7041-G and rs3733359-T alleles were significantly associated with increased susceptibility of HCV infection, and the combined effect of the two unfavorable alleles was related to an elevated risk of HCV infection in a locus-dosage manner (Ptrend = 8.16 × 10−4). Interaction analysis manifested that rs7041-GT/GG and rs3733359-CT/TT jointly increased risk of HCV infection. Moreover, haplotype analysis suggested that compared with the most frequent TC haplotype, the haplotype carrying GT indicated a risk effect of HCV infection [odds ratio (OR) = 1.464]. However, no significant associations were observed for the other five variants. These findings implied that VDBP rs7041-G and rs3733359-T variants may contribute to increased susceptibility to HCV infection in a high-risk Chinese population.

BackgroundChaperonin-containing TCP-1 (TRiC or CCT) was demonstrated to be involved in oncogenesis of cancers carcinogenesis and development of various malignancies. Increasing experimental evidence indicated that dysregulation of TRiC was implicated in the tumor progression of breast cancer (BCa). However, few definitive studies have addressed the diverse expression patterns and prognostic values of eight TRiC subunits. Thus, we aimed to investigate the clinical significance of TRiC subunit expression and prognostic values for their possible implications in diagnosis and treatment of BCa.MethodsBased on updated public resources and comprehensive bioinformatics analysis, we used some online databases (e.g., UALCAN, GEPIA, cBioPortal, TIMER, BC-GenExMiner, metascape, and GeneMANIA) to comprehensively explore the expression levels and the prognostic effects of eight TRiC subunits in patients with BCa.ResultsThe transcriptional levels of most subunits of the Chaperonin TRiC (CCT2, CCT3, CCT4, CCT5, CCT6A, and CCT7) were significantly elevated compared with normal breast tissues, whereas TCP1, CCT4, and CCT6B were lower in BCa tissues than in normal tissues. Besides, copy-number alterations (CNA) of eight TRiC subunits positively regulated their mRNA expressions. Furthermore, high mRNA expression of TCP1/CCT2/CCT4/CCT5/CCT6A/CCT7/CCT8 was significantly associated with poor overall survival (OS) in BCa patients. The eight subunits of the chaperonin TRiC was related to tumor purity and immune infiltration levels of BCa. Co-expression analysis showed CCT6B was negatively associated with other subunits of TRiC and other subunits of TRiC were positively correlated with each other. Additionally, TRiC and their interactive proteins were correlated with positive regulation of biological process, localization, and biological regulation.ConclusionThis study systematically illustrated the expression profiles and distinct prognostic values of chaperonin TRiC in BCa, providing insights for further investigation of subunits of the chaperonin TRiC as novel therapeutic targets and potential prognostic biomarkers in BCa.

Circular RNAs (circRNAs) are a unique family of noncoding RNAs that could regulate multiple biological processes, which play a crucial role in carcinogenesis, progression and chemotherapy resistance of cancers. Growing studies have demonstrated that circRNAs act as novel biomarkers and therapeutic targets for cancers by sponging microRNAs (miRNAs). Up to date, another function of circRNAs, combining with RNA-binding proteins (RBPs), was uncovered. However, there is limit studies illustrating the underlying mechanism of circRNAs-RBPs interactions, as well as showing its roles in diverse types of cancers. In this review, we collected the biogenesis, properties of circRNAs, and then synthesize the connection between circRNAs and RBPs, and try to clarify its molecular mechanisms involving in the pathogenesis and progression of several common cancers, aiming to provide a brand-new insight to the prognosis and treatment strategy for cancers.

Sleep disorder (SD) has a high incidence and seriously affects quality of life, mental health and even the manifestation of physical diseases. The combination of Pinellia ternata (Chinese name: banxia) and Prunella vulgaris (Chinese name: xiakucao), known as the Banxia–Xiakucao Chinese herb pair (BXHP), is a proven Chinese herbal medicine that has been used to treat SD for thousands of years due to its significant clinical effects. However, its active pharmacological components and sedative–hypnotic mechanisms have not been fully elucidated. Thus, the present study used a systematic pharmacological approach to develop pharmacokinetic screens and target predictions via construction of a protein–protein interaction network and annotation database for SD-related and putative BXHP-related targets. Visualization, screening and integrated discovery enrichment analyses were conducted. The BXHP chemical database contains 166 compounds between the two herbal ingredients, and of these, 22 potential active molecules were screened by pharmacokinetic evaluation. The targets of 114 of the active molecules were predicted, and 34 were selected for further analysis. Finally, gene ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses suggested that BXHP can reduce inflammatory responses. and mediate immune-related and central nervous system neurotransmitters via regulation of multiple targets and pathways. The use of a systematic pharmacology-based approach in the present study further elucidated the mechanisms of action underlying BXHP for the treatment of SD from a holistic perspective and sheds light on the systemic mechanisms of action of Chinese herbal medicines in general.

A esterase gene was characterized from a halophilic bacterium Chromohalobacter canadensis which was originally isolated from a salt well mine. Sequence analysis showed that the esterase, named as EstSHJ2, contained active site serine encompassed by a conserved pentapeptide motif (GSSMG). The EstSHJ2 was classified into a new lipase/esterase family by phylogenetic association analysis. Molecular weight of EstSHJ2 was 26 kDa and the preferred substrate was p-NP butyrate. The EstSHJ2 exhibited a maximum activity at 2.5 M NaCl concentration. Intriguingly, the optimum temperature, pH and stability of EstSHJ2 were related to NaCl concentration. At 2.5 M NaCl concentration, the optimum temperature and pH of EstSHJ2 were 65 ℃ and pH 9.0, and enzyme remained 81% active after 80 ℃ treatment for 2 h. Additionally, the EstSHJ2 showed strong tolerance to metal ions and organic solvents. Among these, 10 mM K+, Ca2+ , Mg2+ and 30% hexane, benzene, toluene has significantly improved activity of EstSHJ2. The EstSHJ2 was the first reported esterase from Chromohalobacter canadensis, and may carry considerable potential for industrial applications under extreme conditions.

Exogenous H2 S donor, sodium hydrosulfide (NaHS), can influence the bleomycin-induced pulmonary fibrosis by attenuating the epithelial-mesenchymal transition (EMT) of alveolar epithelial cells, but whether NaHS affects paraquat (PQ)-induced EMT and the molecular mechanisms remain unclarified. The aim of the present study is to examine the effect of exogenous NaHS on PQ-induced EMT in human alveolar epithelial cells (A549 cells) and assess if this effect occurs through regulating transforming growth factor (TGF)-β1/Smad2/3 signaling pathway. The expressions of endogenous H2 S producing enzymes, namely cystathionine β-synthase, cystathionine γ-lyase and 3-mercaptopyruvate sulfur transferase, were detected by reverse transcription-polymerase chain reaction and western blotting. The induced EMT was assessed by morphological and phenotypic characterizations, and the protein level of E-cadherin and vimentin were detected by western blotting. To investigate the effect of NaHS on PQ-induced EMT and potential mechanism, A549 cells were pretreated with NaHS before incubating with PQ and then evaluated by morphological changes, cell migration ability, the expression of EMT markers and TGF-β1/Smad2/3 signaling pathway related proteins. PQ significantly downregulated the expression levels of cystathionine β-synthase and cystathionine γ-lyase, but not 3-mercaptopyruvate sulfur transferase, in a time-dependent manner in A549 cells. Exogenous NaHS could significantly retard PQ-induced morphological changes and cell migration ability. Furthermore, exogenous NaHS significantly upregulated the expression of E-cadherin, whereas it downregulated the expression of vimentin. In addition, exogenous NaHS could also significantly attenuates PQ-induced TGF-β1, phosphorylated Smad2/3 proteins expression, which induced by PQ in a time-dependent manner. This study provides the first evidence that exogenous NaHS attenuates PQ-induced EMT and migration of human alveolar epithelial cells through regulating the TGF-β1/Smad2/3 signaling pathway.

Peptide-based T cell vaccines targeting the conserved epitopes of influenza virus can provide cross-protection against distantly related strains, but they are generally not immunogenic. Foreign antigen-specific regulatory T (Treg) cells are induced under subimmunogenic conditions peripherally, although their development and role in vaccine-mediated antiviral immunity is unclear. Here, we demonstrated primary vaccination with peptides alone significantly induced antigen-specific Foxp3+ Treg cells, which were further expanded by repeated vaccination with unadjuvanted peptides. Certain adjuvants, including CpG, suppressed the induction and expansion of antigen-specific Treg cells by peptide vaccination. Interestingly, secondary influenza virus infection significantly increased the frequency of preexisting antigen-specific Treg cells, although primary infection barely induced them. Importantly, specific depletion of vaccine-induced antigen-specific Treg cells promoted influenza viral clearance, indicating their inhibitory role in vivo. Immunization with CpG-adjuvanted peptides by the subcutaneous prime-intranasal-boost strategy restricted the recruitment and accumulation of antigen-specific Treg cells in lung, and stimulated robust T cell immunity. Finally, subcutaneous prime-intranasal-boost immunization with CpG-adjuvanted peptides or whole-inactivated influenza vaccines protected mice from heterosubtypic influenza virus infection. In conclusion, antigen-specific Treg cells induced by peptide vaccines attenuate the antiviral immunity against influenza virus infection. CpG-adjuvanted peptide vaccines provide heterosubtypic influenza protection probably by inhibiting Treg development and enhancing T cell immunity.

Mutations within the Shank3 gene, which encodes a key postsynaptic density (PSD) protein at glutamatergic synapses, contribute to the genetic etiology of defined autism spectrum disorders (ASDs), including Phelan-McDermid syndrome (PMS) and intellectual disabilities (ID). Although there are a series of genetic mouse models to study Shank3 gene in ASDs, there are few rat models with species-specific advantages. In this study, we established and characterized a novel rat model with a deletion spanning exons 11–21 of Shank3, leading to a complete loss of the major SHANK3 isoforms. Synaptic function and plasticity of Shank3-deficient rats were impaired detected by biochemical and electrophysiological analyses. Shank3-depleted rats showed impaired social memory but not impaired social interaction behaviors. In addition, impaired learning and memory, increased anxiety-like behavior, increased mechanical pain threshold and decreased thermal sensation were observed in Shank3-deficient rats. It is worth to note that Shank3-deficient rats had nearly normal levels of the endogenous social neurohormones oxytocin (OXT) and arginine-vasopressin (AVP). This new rat model will help to further investigate the etiology and assess potential therapeutic target and strategy for Shank3-related neurodevelopmental disorders.

Genetic variation of related genes in Vitamin D (VD) metabolic pathway played an important role in antiviral immune response and chronic hepatitis C virus (HCV) infection. Retinoid X receptor (RXR) is one of the key genes in the metabolism pathway of VD. This study aims to investigate the effect of single nucleotide polymorphisms (SNPs) in RXR on the outcomes of HCV infection. Three SNPs (RXRɑ-rs4842194, rs1045570 and RXRβ-rs2076310) were genotyped using Sequenom MassARRAY platform in 515 spontaneous clearance subjects, 830 persistent infection subjects, and 1062 uninfected subjects. Multivariate stepwise regression analyss was used to identify the prediction factors for HCV infection outcomes. The USCS Brower and RNAfold web serves were performed to further explore the potential biological functions of positive SNPs. The results of logistic regression analysis after adjusting for age, gender and types of high-risk population showed that subjects with RXRβ rs2076310-T (recessive model: adjusted OR = 1.598, 95%CI = 1.126-2.267, P = 0.009; additive model: adjusted OR = 1.196, 95%CI = 1.011-1.416, P = 0.037) had a significantly increased possibility of HCV infection chronicity. Rs2076310, age, types of high-risk population and aspartate aminotransferase were independent predictors of chronic HCV infection (P < 0.05). And the area under the receiver operating characteristic curve of combined effects of these factors was 0.679. Bioinformatics analysis indicated that rs2076310 could affect the gene expression level by affecting the transcriptional regulatory activity of the corresponding gene region. These findings indicated that genetic variation of RXRβ was associated with the risk of HCV infection chronicity among a high-risk Chinese population.

Background and aim Irritable bowel syndrome is characterized by abdominal pain and altered bowel habits and may occur following stressful events or infectious gastroenteritis such as giardiasis. Recent findings revealed a link between cholecystokinin (CCK), neurotrophin synthesis, and intestinal hyperalgesia. The aim was to investigate the role of CCK in visceral hypersensitivity using mouse models challenged with a bout of infection with Giardia lamblia or psychological stress, either alone or in combination. Methods Abdominal pain was evaluated by visceromoter response to colorectal distension. Nerve fibers in intestinal tissues were stained using immunohistochemistry (PGP9.5). Human neuroblastoma SH-SY5Y cells incubated with bacterial-free mouse gut supernatant or recombinant CCK-8S were assessed for neurite outgrowth and nerve growth factor (NGF) production. Results Intestinal hypersensitivity was induced by either stress or Giardia infection, and a trend of increased pain was seen following dual stimuli. Increased CCK levels and PGP9.5 immunoreactivity were found in colonic mucosa of mice following stress and/or infection. Inhibitors to the CCK-A receptor (L-364718) or CCK-B receptor (L-365260) blocked visceral hypersensitivity caused by stress, but not when induced by giardiasis. Nerve fiber elongation and NGF synthesis were observed in SH-SY5Y cells after incubation with colonic supernatants from mice given the dual stimuli, or after treatment with CCK-8S. Increased nerve fiber length by colonic supernatant and CCK-8S was attenuated by L-365260 or neutralizing anti-NGF. Conclusions This new model successfully recapitulates intestinal hypernociception induced by stress or Giardia. Colonic CCK contributes to visceral hypersensitivity caused by stress, but not by Giardia, partly via NGF-dependent neurite outgrowth.

Nickel compounds are known to be common environmental and occupational carcinogens which also promote the migration of lung cancer cells. However, the molecular mechanism yet remains to be clarified. Hydrogen sulfide (H2S) is involved in cancer biological processes. However, the exact effect and functionality of H2S on nickel, towards the promotion of the migration ability of lung cancer cells, remains to be unknown. In this study, we have found that the nickel chloride (NiCl2) treatment significantly downregulates the protein levels of endogenous H2S enzyme cystathionine β-synthase (CBS), cystathionine γ-lyase (CSE) and 3-Mercaptopyruvate sulfurtransferase (3-MST). A correlation between NiCl2-induced epithelial-mesenchymal transition (EMT) and the migration ability of lung cancer A549 cells has been observed. Exogenous H2S donor, sodium hydrogen sulfide (NaHS) (100 μmol/L), can reverse NiCl2-induced EMT as well as the migration ability of A549 cells. NiCl2 treatment is able to upregulate the protein level of transforming growth factor-β1 (TGF-β1), p-Smad2, p-Smad3, p-JNK, p-ERK and p-P38 in a time-dependent fashion, indicating that both TGF-β1/Smad2/Smad3 and mitogen-activated protein kinase (MAPK) signaling cascades (a non-Smad pathway) may play essential roles in NiCl2-dependent EMT as well as cell migration of human lung cancer cells. Furthermore, exogenous NaHS alleviates the NiCl2-induced EMT and the migration ability of A549 cells only by regulating TGF-β1/Smad2/Smad3, rather than the MAPK, signaling pathway. These results indicate that the exogenous administration of NaHS might be a potential therapeutic strategy against nickel-induced lung cancer progression.

The human induced pluripotent stem cell (iPSC) line YAHKMUi001-A was derived from the dermal fibroblasts of a patient with Tetralogy of Fallot (TOF), with a mutation in the TBX1 gene (c.928G > A). The skin fibroblasts were obtained from a 4-year-old boy, and were infected with Sendai virus expressing the Yamanaka factors. The YAHKMUi001-A iPSC line expresses pluripotent stem cell markers, displays a normal karyotype, and has the capacity to differentiate into 3 germ layers. This cell line model can be a good tool to study the pathological mechanism of the TBX1 gene mutations associated with TOF.

The corpora allata in adult Loreyi leafworms Leucania loreyi (Lepidoptera: Noctuidae) exhibit sexual dimorphism. The male possesses right and left corpora allata of about the same size. Each gland is composed of a cluster of approximately 40 semi-transparent, spherical, isolated cells held together by fine tracheae and nerve fibers. The largest cell diameter found in male glands was 203 pm. In contrast, the female gland cells and clusters are much smaller. The largest dimensions of one whole female gland cluster were 452 μm in length and 280 μm in width. Using bilateral and unilateral larval allatectomy, we confirmed that the adult isolated cell type glands develop ontogenetically from larval capsular type glands. Ultrastructural study re- vealed many similarities between the subcellular structures of the isolated cell type glands of L. loreyi and the more common capsular gland reported by others. These similarities include very large numbers of mitochondria, abun- dant whorled smooth endoplasmic reticulum, irregularly shaped nuclei, Golgi bodies, and free ribosomes. Compared with the corpora allata of 3- to 9-day-old adults, the glands of 1-day-old adults possessed much less smooth endoplasmic reticulum. The gland cells in females usually have more neurosecretory nerve endings, less-abundant stacked smooth endoplasmic reticulum, and less- defined interdigitations than the gland cells in males. © 1995 Wiley-Liss, Inc.

Vitamin D has been considered as an immune modulator, and exerted the effect through the vitamin D receptor (VDR). This study investigated the associations of single-nucleotide polymorphisms (SNPs) of VDR with the outcomes of Hepatitis C virus (HCV) infection. Three SNPs (rs2228570, rs757343 and rs739837) were genotyped by TaqMan assay among Chinese population, including 538 HCV spontaneous clearance subjects, 834 persistent infection subjects and 1030 uninfected subjects. Binary logistic analyses were used to control the effects of confounding factors. The results showed that subjects with the rs757343 A allele and rs739837 A allele had the significantly reduced risk of HCV susceptibility (all PBonferroni

Human leukocyte antigen (HLA) class II molecule influences host antigen presentation and anti-viral immune response. The aim of this study was to investigate whether single nucleotide polymorphisms (SNPs) within HLA class II gene were associated with different clinical outcomes of hepatitis C virus (HCV) infection. Three HLA class II SNPs (rs3077, rs2395309 and rs2856718) were genotyped by TaqMan assay among Chinese population, including 350 persistent HCV infection patients, 194 spontaneous viral clearance subjects and 973 HCV-uninfected control subjects. After logistic regression analysis, the results indicated that the rs2856718 TC genotype was significantly associated with the protective effect of the HCV natural susceptibility (adjusted OR: 0.712, 95% CI: 0.554–0.914) when compared with reference TT genotype, and this remained significant after false discovery rate (FDR) correction (p = 0.024). Moreover, the protective effect of rs2856718 was observed in dominant genetic models (adjusted OR: 0.726, 95% CI: 0.574–0.920), and this remained significant after FDR correction (p = 0.024). In stratified analysis, a significant decreased risk was found in rs2856718C allele in the male subgroup (adjusted OR: 0.778, 95% CI: 0.627–0.966) and hemodialysis subgroup (adjusted OR: 0.713, 95% CI: 0.552–0.921). Our results indicated that the genetic variations of rs2856718 within the HLA-DQ gene are associated with the natural susceptibility to HCV infection among the Chinese population.

The unique 75 K γ-secalins encoded by Sec2 loci in Secale species is composed of almost half rye storage proteins. The chromosomal location of Sec2 loci in wild Secale species, Secale africanum, was carried out by the wheat—S. africanum derivatives, which were identified by genomic in situ hybridization and multi-color fluorescence in situ hybridization. The Sec2 gene-specific PCR analysis indicated that the S. cereale Sec2 was located on chromosome 2R, while the S. africanum Sec2 was localized on chromosome 6Rafr of S. africanum. A total of 38 Sec2 gene sequences were isolated from S. africanum, S. cereale and S. sylvestre by PCR-based cloning. Phylogenetic analysis showed that S. africanum Sec2 diverged from S. cereale Sec2 approximately 2–3 million years ago. The illegitimate recombination of chromosome 2R–6R involving the Sec2 loci region may accelerate sequence variation during evolutionary process from wild to cultivated Secale species.

Objective: To study the distribution and patterns of resistance to antimicrobial agents of normal conjunctival bacteria. Materials and Methods: Conjunctival specimens were collected from 8,224 patients and then cultured, which underwent antimicrobial susceptibility test following standard methods. Patients with infectious symptoms such as erythema or oedema and those using systemic or topical antibiotics within one month were excluded. Results: In this study, the incidence of isolated bacteria was 24.2%. The middle aged group of 41-65 years presented the lowest rate of bacterial isolation which was 19.4%, while the highest isolation rate (83.1%) was found in patients in the age range of 0-6 years. In every age group, the incidence of bacterial isolation in men was higher than that in women. The top 3 most commonly isolated micro-organisms were Staphylococcus epidermidis (39.7%), Streptococcus pneumoniae (4.5%) and Staphylococcus aureus (2.7%), of which about 83.1% Staphylococcus aureus were isolated in the group of 0-6 years. We found that coagulase-negative Staphylococcus (CONS) were more resistant to penicillin, macrolides, clindamycin and sulfonamides with the rate ranging from 57.9% to 90.8%, which were highly susceptible to vancomycin, linezolid, rifampin, tetracyclines and aminoglycosides. Contrasting to CONS, the general resistance rate of Staphylococcus aureus (S.aureus) was significantly lower. Additionally, Streptococcus was susceptible well to the majority of antimicrobial agents, while highly resistant to macrolides and tetracyclines with the rate greater than 80%. Conclusions: In conclusion, our study revealed the incidence and antimicrobial sensitivity profiles of normal conjunctiva bacterial flora in the central area of China, which could be useful in the prevention of ocular infections. Importantly, our data could be used to guide the selection of appropriate prophylactic agents.

The interleukin (IL)-6/gp130 family of cytokines (e.g., IL-6, IL-11, leukemia inhibitory factor (LIF) and oncostatin M (OSM)) play important roles in the central nervous system (CNS) during neuroinflammation and neurodevelopment. However, little is known regarding the responses by astroglia and microglia to this family of cytokines. Here the expression of the IL-6/gp130 cytokine receptors and subsequent signal pathway activation was examined in murine astrocytes and microglia in vitro. Astrocytes had high levels of OSMR mRNA while lower levels of IL-6R, LIFR and IL-11R mRNAs were also present. In comparison, in microglia there was no detectable OSMR mRNA, higher levels of IL-6R mRNA and lower levels of the LIFR and IL-11R mRNAs. The OSMR protein was present in astrocytes but was undetectable in microglia. Conversely, the IL-6R protein was present in microglia but not detectable in astrocytes. In astrocytes but not microglia, phosphorylation of STAT1 and STAT3 occurred in response to OSM, whereas both microglia and astrocytes responded to hyper-IL-6 (IL-6 linked to the soluble IL-6 receptor). Finally, in both microglia and astrocytes, OSM failed to activate NFκB or induce iNOS and nitrite production. We conclude: (1) notable differences exist in the expression of receptors utilized by the IL-6/gp130 family of cytokines in astrocytes and microglia, and (2) the findings provide a molecular basis for the differential response to OSM by astrocytes versus microglia and demonstrate a fundamental means for achieving cellular specificity in the response of these glial cells to this cytokine.

Aims Our main purposes were to determine the effects of Vigna radiata, a nitrogen fixing plant, on growth, water balance and gas exchange of the intercropping Juglans regia seedlings and to investigate the hydraulic mechanism involved in photosynthesis and growth. Methods We measured growth, hydraulic characteristics, and rate of gas exchange in J. regia seedlings, and analyzed the effects of the intercropping nitrogen fixing V. radiata on xylem anatomic structure, water balance and photosynthetic characteristics of J. regia seedlings under conditions of nitrogen deprivation and enrichment. Important findings Under conditions of nitrogen deficiency, the nitrogen fixing V. radiata facilitated the growth of the intercropping J. regia seedlings by improving xylem development, water transport and hydraulic characteristics in high transpiration demand. However, with nitrogen addition, the occurrence of V. radiata inhibited the growth of J. regia seedlings, likely by competing for water and other elements.

Aims Our main purposes were to explore effects of phosphorus deficiency and soil acidification on growth, water balance and phosphorus nutrition in Juglans regia seedlings and to investigate the hydraulic mechanism involved in reduced photosynthesis and growth. Methods We measured growth, hydraulic characteristics, and gas exchanges in J. regia seedlings. The effects of phosphorus deficiency and soil acidification on xylem anatomic structure, water balance and photosynthetic characteristics were also analyzed. Important findings Soil acidification inhibited the phosphorous uptake and utilization efficiency in J. regia seedlings. A combination of soil acidification and phosphorous deficiency interrupted water balance, and reduced photosynthesis and growth of J. regia seedlings. Under soil acidification and phosphorus deficiency, non-stomatal factors also played a role in inhibiting photosynthesis.

The wild species, Secale africanum Stapf., serves as a valuable germplasm resource for increasing the diversity of cultivated rye (Secale cereale L.) and providing novel genes for wheat improvement. The objective of this study was to identify new wheat-S. africanum chromosome 2Rafr derivatives by molecular markers and in situ hybridization, and to test the effects of centric translocations of S. africanum chromosome 2Rafr on disease resistance and agronomic performance of wheat. The T2RafrS.2DL and T2DS.2RafrL translocations, and other S. africanum chromosome introgressions such as a 2Rafr substitution as well as monotelo-2RafrS and 2RafrL addition lines, were identified by the genomic in situ hybridization and sequential fluorescence in situ hybridization (FISH). Twenty-three molecular markers were localized on chromosome 2Rafr arms which will facilitate future identification of their introgressions into wheat. A comparison of FISH patterns and the molecular marker distribution between the S. africanum chromosome 2Rafr and cultivated rye chromosome 2R indicates differentiation between the wild and cultivated Secale genomes. Tests of disease resistance and agronomic performance of wheat-S. africanum chromosome 2Rafr derivatives indicated that chromosome 2RafrL carried gene(s) for dwarfing and stripe rust resistance, while the 2RafrS translocation line was the most favorable for agronomic performance when compared with the substitution and 2RafrL translocation lines. These results encourage the continued use of novel wheat-S. africanum 2Rafr derivative lines in wheat improvement.

The grasshoppers are ideal materials to study various meiotic stages of spermatogenesis due to their easy availability, fairly large chromosomes, and fewer numbers of chromosomes. It is easy to make temporary squash preparation of grasshopper testes; however, it is usually difficult for the beginners to differentiate between stages of meiosis. In view of this, we demonstrated the method of identification of meiotic stages by chromosome number and chromosome conformation, taking spermatogonial meiosis of Locusta migratoria manilensis as an example. We described briefly the mitosis of spermatogonia and the spermatogenesis of this species as well.

Vitamin D and vitamin D receptor (VDR) are involved in multiple immune-mediated disorders including chronic hepatitis C virus (HCV) infection. The aim of this study was to determine the association between plasma vitamin D level, VDR genetic polymorphisms and risk of HCV infection susceptibility and chronicity. Seven single nucleotide polymorphisms (SNPs) in VDR gene were genotyped and plasma 25-hydroxyvitamin D [25(OH)D] levels were measured in a Han Chinese population of 898 HCV persistent infection cases, 558 spontaneous clearance subjects and 1136 uninfected controls with high risk of HCV infection. In this case-control study, the average plasma 25(OH)D level in persistent infection patients was significantly lower than that in spontaneous clearance cases (P=0.039) and controls (P=0.005). Logistic analyses indicated that rs7975232-C, rs2239185-T and rs11574129-T alleles were significantly associated with a decreased risk of HCV infection susceptibility (all PBonferroni

Trees and crops in agroforestry system can make the best use of water,nutrient,radiance and heat.However,competition may occur when resources are scarce,especially for the water competition in the warm temperate semi-arid region.Due to arid climate,less rainfall and poor irrigation condition,water is one of the major factors limiting the development of agriculture and forestry in the low hilly area of North China.Stable carbon isotope ratio(δ13C) of walnut(Juglans regia L.)-wheat(Triticum aestivum) intercropping system and monoculture wheat as well as sap flow of walnut tree at seedling establishment,jointing,filling and maturing growth period of winter wheat in the low hilly area of North China were researched.Water use efficiency(WUE) and water use(WU) were calculated by using the δ13C value,crop biomass,walnut sap flow and meteorological data.The results showed that the WUE of intercropping walnut,monoculture wheat and intercropping wheat were 10.771—21.233,9.946—19.149 and 9.878—18.431 mmol C/mol H2O,respectively.The WUE of intercropping wheat were significantly greater than that in monoculture wheat at wheat jointing growth period.The total water use of the walnut-wheat intercropping system was 1755.19 t/hm2,which was 30.85% less than that in the monoculture wheat system(2538.13 t/hm2).Water consumption of the intercropping walnut accounted for 36.34% in the walnut-wheat intercropping system,respectively 62.09%,30.80%,35.58% and 27.24% at sowing-seedling establishment,jointing,flowering-filling and filling-maturing growth period of winter wheat.The walnut tree consumed the greatest amount of water at earlier growth period of winter wheat,and the less at vigorous growth stages of winter wheat.Contrary to the walnut,the water use of wheat was greater at its later growth period.Intercropping walnut and wheat staggered water demand periods and avoided temporal competition for water.Soil volumetric water content in the intercropping system was the greater the closer to the walnut tree in the shallow soil layer,and the less the closer to the walnut tree in the deep soil,which suggested that the walnut tree mainly absorbed deep water,while the intercropping wheat mainly absorb shallow water,it avoided spatial competition for water.The grain yield,total biomass and water use of monoculture wheat were respectively 26.79%,27.12% and 36.30%(P=0.001、P=0.033、P=0.050) higher than that of the intercropping wheat.The average fruit yield of intercropping and monoculture walnut were respectively 0.88 kg and 0.94 kg per tree(P=0.829).However,yield LER(Land equivalent ratio) and WUEe(WUE of economics) of the walnut-wheat intercropping system respectively reached 1.67 and 25.92RMB · mm-1 · hm-2,improved obviously comparing with monoculture system.The walnut-wheat intercropping system utilized water with high efficiency and exhibited both production advantage and economic advantage at the same time.

Trees,especially big ones,have huge and complicated vascular systems which make the long-distance water transport difficult to study.We used the heat dissipation method and deuterated water as tracers to characterize water transport and storage properties of Quercus variabilis trees under the different water stress treatment.The transit time for D2O transport from the trunk base to the upper branches and the tracer residence time were determined by measuring hydrogen isotope ratios in water extracted from branches sampled in regular intervals.The result indicated that the maxima δD of full irrigation(FI),mild water stress(LWS) and severe water stress(SWS) respectively increased to 586.67‰、997.33‰ and 1364.89‰ after injecting deuterium tracer.There were significant differences in maxima δD among the three treatments.The tracer velocities with LWS and SWS were significantly lower than FI treatment but the tracer half-life and residence time were longer than FI treatment.The branch PLC,sapflow rate,water potential and transpiration rate were also observed.The branch PLC treated with LWS and SWS was significantly higher than FI treatment and the sapflow rate,water potential and transpiration rate were lower.There significantly positive correlations between tracer velocity and leaf transpiration rate(P0.01,n=27),and between tracer velocity and sapflow rate(P0.01,n=9).The results indicated that water transport was determined by transpiration rate.The PLC and branch water potential were inversely correlated with tracer velocity(P0.01,n=27),suggesting that drought stress-induced embolism and lower water potential should increase resistance to water transport.Tracer half-life and residence time increased as water stress increased.Tracer half-life and residence time were significant exponential functions with the branch PLC(P0.01,n=27),but power functions with the leaf transpiration rate.The times needed for transporting the same volume tracer were increased,indicating that water transport efficiencies were reduced because of PLC,transpiration and storage water variations under water-stress condition.Analyses of sapflow showed that the calculative fluxes of drought treatments within tracer half-life and residence time were significantly higher than FI treatment.The amounts of water carrying the same volume tracer were increased as stress degree increased.The results indicated that the increased water was possibly stored in the sapwood and re-exchanged to transpiration stream when transpiration was strong in the daytime.Quercus variabilis trees should reduce the water transport velocity and efficiency to adapt water stress environment by xylem embolism and storage-water exchange.

In the present study, we report on a new triticale (×Triticosecale Wittm.) ZH-1, derived from the progeny of common wheat (Triticum aestivum L.) cv. MY15 × Chinese Weining rye (Secale cereale L.). ZH-1, exhibiting shorter plant height and higher tillering ability compared to MY15, is immune to both powdery mildew and stripe rust and has stable fertility. Genomic in situ hybridization (GISH), fluorescence in situ hybridization (FISH) and C-banding revealed that the chromosome composition of triticale ZH-1 was 14 A-genome (1A-7A), 12 B-genome (1B-2B, 4B-7B), 12 R-genome (1R, 3R-7R), chromosomes 6D and T2DS.2DL-?R. Moreover, the PCR results of PLUG and EST-SSR markers also strongly supported the above stated chromosome composition of triticale ZH-1. In addition, the physical mapping of chromosome T2DS.2DL-?R showed a minute chromosomal fragment derived from rye was attached at the distal end of 2DL. The new triticale ZH-1 could be a valuable source for wheat improvement, especially for resistance to disease.

Wheat yellow rust resistance gene Yr17 was originated from the wheat-Aegilops ventricosa introgression, and still effective on the adult plant in Southern China. The previous studies located the gene Yr17 on the translocation of 2NS-2AS using the molecular and cytological markers. In the present study, we screened new PCR-based markers to map the gene Yr17 region from the investigation of a segregating 120 F2 population. All markers including four EST-PCR markers, a SCAR (sequence characterized amplified region) and a PLUG (PCR based landmark unique gene) marker specific to Yr17 gene were mapped on the chromosome 2AS, and located on the chromosomal deletion bin 2AS5-0.8–1.00 region. Based on the wheat-rice collinearity, we found that the sequences of the Yr17 gene linked markers were comparatively matched at rice chromosome 4 and chromosome 7. However, the identified closely linked genomic sequence of Yr17 gene is most likely collinear with genomic region of rice chromosome 4. The newly produced PCR based markers closely linked to Yr17 gene will be useful for the marker-assisted selection in wheat breeding for rust resistance.

Two amphiploids, AF-1(Triticum aestivum L. cv. Anyuepaideng–Secale africanum Stapf.) and BF-1 (T. turgidum ssp. carthlicum–S. africanum), were evaluated by chromosomal banding and in situ hybridization. The individual S. africanum chromosomes were identified in the BF-1 background by sequential C-banding and genomic in situ hybridization (GISH), and were distinguishable from those of S. cereale, because they exhibited less terminal heterochromatin. Fluorescence in situ hybridization (FISH) using the tandem repeat pSc250 as a probe indicated that only 6Ra of S. africanum contained a significant hybrid signal, whereas S. cereale displayed strong hybridization at the telomeres or subtelomeres in all seven pairs of chromosomes. Extensive wheat–S. africanum non-Robertsonian translocations were observed in both AF-1 and BF-1 plants, suggesting a frequent occurrence of chromosomal recombination between wheat and S. africanum. Moreover, introgression lines selected from the progeny of wheat/AF-1 crosses were resistant when field tested with widely virulent strains of Puccinia striiformis f. sp. tritici. Three highly resistant lines were selected. GISH and C-banding revealed that resistant line L9-15 carried a pair of 1BL.1RS translocated chromosomes. This new type of S. africanum derived wheat–Secale translocation line with resistance to Yr9-virulent strains will broaden the genetic diversity of 1BL.1RS for wheat breeding.

Genome in situ hybridization (GISH) analysis of wheat-Secale africanum amphiploid revealed that the S. afri-canum genome displayed significant divergence to the Secale cereale genome. It is thus valuable to deploy genes from S. africanum. We performed the PCR analysis on S. africanum, wheat-S. afticanum amphiploid, T. eastivum cv. Anyuepaideng and other genetic stocks by 100 ISSR primers. A specific segment of 561 bp, named pSaUBC815561, was obtained from S. africanum using primer UBC815. This segment was not amplified from the control wheat lines. Primer UBC815 also am-plified fragments from wild species of genus Secale, including S. vavilovii, S. sylvestre, and other cultivated ryes. Based on the sequence of pSaUBC815561, a pair of special primers U815-F and U815-R was designed and was used to amplify the DNA of wheat related species in Triticeae aimed at validating the specificity of pSaUBC815561. PCR analysis indicated that this specific DNA fragment was amplified not only from a set of Chinese Spring wheat-Imperial rye chromosome addition lines but also from certain wheat-rye introgression lines. Therefore, pSaUBC815561 can be used as a specific marker for detection of chromosomes of Secale genome in wheat.

Summary Understandinghowtraitsareintegratedattheorganismal level remains a fundamental problem at the interface of developmental and evolutionary biology. Hormones, regulatory signaling molecules that coordinate multiple developmental and physiological processes, are major determinants underlying phenotypic integration. The probably best example for this is the lipid-like juvenile hormone (JH) in insects. Here we review the manifold effects of JH, the most versatile animal hormone, with an emphasis on the fruit fly Drosophila melanogaster ,a n organism amenable to both genetics and endocrinology. JH affects a remarkable number of processes and traits in Drosophila development and life history, including metamorphosis, behavior, reproduction, diapause, stress resistance and aging. While many molecular details underlying JH signaling remain unknown, we argue that studying ‘‘hormonal pleiotropy’’ offers intriguing insights into phenotypic integration and the mechanisms underlying life history evolution. In particular, we illustrate the role of JH as a key mediator of life history trade-offs. BioEssays 27:999–1010, 2005. 2005 Wiley Periodicals, Inc.

In Drosophila melanogaster, ageing is slowed when insulin-like signalling is reduced: life expectancy is extended by more than 50% when the insulin-like receptor (InR) or its receptor substrate (chico) are mutated, or when insulin-producing cells are ablated. But we have yet to resolve when insulin affects ageing, or whether insulin signals regulate ageing directly or indirectly through secondary hormones. Caenorhabditis elegans lifespan is also extended when insulin signalling is inhibited in certain tissues, or when repressed in adult worms, and this requires the forkhead transcription factor (FOXO) encoded by daf-16 (ref. 6). The D. melanogaster insulin-like receptor mediates phosphorylation of dFOXO, the equivalent of nematode daf-16 and mammalian FOXO3a. We demonstrate here that dFOXO regulates D. melanogaster ageing when activated in the adult pericerebral fat body. We further show that this limited activation of dFOXO reduces expression of the Drosophila insulin-like peptide dilp-2 synthesized in neurons, and represses endogenous insulin-dependent signalling in peripheral fat body. These findings suggest that autonomous and non-autonomous roles of insulin signalling combine to control ageing.

Mutations of the insulin signal pathway in Drosophila melanogaster produce long-lived adults with many correlated phenotypes. Homozygotes of insulin-like receptor (InR) and insulin-like receptor substrate (chico) delay time to eclosion, reduce body size, decrease reproduction and increase life span. Because these mutations are expressed through all life stages it is unclear when insulin signals must be reduced to increase life span. As a first analysis of this problem in D. melanogaster we have manipulated the larval diet to determine if changes in metabolic regulation at this stage are sufficient to slow aging. We controlled the dietary yeast fed to third instar larvae and studied the size, mortality, fecundity and hormones of the resulting adults, which were fed a normal, yeast-replete diet. Adults from yeast-deprived larvae phenocopied many traits of InR and chico mutants: small body size, delayed eclosion, reduced ovariole number and reduced age-specific fecundity. But unlike constitutive mutants of the insulin/IGF system, adults from yeast-deprived larvae had normal patterns of demographic senescence, and this was accompanied by normal insulin-like peptide and juvenile hormone syntheses. Surprisingly, the normal aging in these adults was also associated with greatly reduced fecundity. Although nutritional conditions of the larvae can affect the subsequent body size and fecundity of adults, these are not sufficient to slow aging.

Hypomorphic mutants affecting the Drosophila insulin/IGF signal pathway are reported to increase longevity in females but not in males. To understand this sex-difference, we conducted a large-scale demographic study with three new isogenic strains of alleles at chico, the insulin-receptor substrate homologue. We verify that female dwarf homozygotes (ch1/ch1) and normal-sized heterozygotes (ch1/+) are long-lived, as originally reported. We find for the first time that male heterozygotes are long-lived relative to wildtype, by about 50%. The life span of male ch1/ch1 is similar to that of wildtype but these dwarf males age at a slow demographic rate. The levels of demographic frailty and of age-independent mortality are elevated in ch1/ch1 males, counteracting the effect of slow aging upon life expectancy. Mortality deceleration occurs amongst the oldest-old wildtype adults, as seen in many organisms. Remarkably, in similarly sized cohorts of male and female ch1/ch1 and of male ch1/+ mortality deceleration is absent. Mortality deceleration is a phenotype of chico.

The Drosophila melanogaster gene insulin-like receptor ( InR ) is homologous to mammalian insulin receptors as well as to Caenorhabditis elegans daf-2 , a signal transducer regulating worm dauer formation and adult longevity. We describe a heteroallelic, hypomorphic genotype of mutant InR , which yields dwarf females with up to an 85% extension of adult longevity and dwarf males with reduced late age-specific mortality. Treatment of the long-lived InR dwarfs with a juvenile hormone analog restores life expectancy toward that of wild-type controls. We conclude that juvenile hormone deficiency, which results from InR signal pathway mutation, is sufficient to extend life-span, and that in flies, insulin-like ligands nonautonomously mediate aging through retardation of growth or activation of specific endocrine tissue.

The gp130 cytokines include IL-6, IL-11, LIF and OSM. These cytokines have important roles in neuroinflammation and neurodevelopment. However, the nature of any cell-specific responses of astrocytes and microglia to this cytokine family and their molecular basis is not well understood. Here, the gp130 family cytokine signal pathway activation and global transcriptional responses were examined in murine astrocytes and microglia in vitro. In astrocytes, OSM-induced STAT1 phosphorylation was significantly greater than for hyperIL-6 (IL-6 linked to the soluble IL-6 receptor) or LIF. However, STAT3 phosphorylation was induced to similar levels for all three cytokines. Conversely, in microglia which lacked the OSMR, hyperIL-6 and LIF but not OSM, induced phosphorylation of STAT3 and only minimal phosphorylation of STAT1. In STAT1 KO astrocytes, OSM conferred a stronger activation of STAT3 compared with WT astrocytes. Transcriptomic analysis of WT and STAT1 KO astrocyte response to OSM, hyperIL-6 or LIF revealed up-regulation and downregulation of many transcripts with OSM > IL-6 > LIF. A dominant IFN-like response to OSM was largely abolished in STAT1 KO astrocytes, suggesting that a large portion of OSM-regulated genes is STAT1-dependent. The gene expression profile of OSM- and hyperIL-6 treated astrocytes was somewhat similar while that for LIF differed the most. Despite signalling via common gp130 pathways, our findings show that specificity is achieved through multiple mechanisms including cell-restricted expression of some receptors, varying strength of signal pathway activation and differential activation of STAT1 and STAT3, which mediate differential gene expression and potential functions by the gp130 cytokines in astrocytes and microglia.

Wild Secale species, Secale africanum Stapf., serve as a valuable source for increasing the diversity of cultivated rye (Secale cereale L.) and provide novel genes for wheat improvement. New wheat – S. africanum chromosome 1Rafr addition, 1Rafr(1D) substitution, 1BL.1RafrS and 1DS.1RafrL translocation, and 1RafrL monotelocentric addition lines were identified by chromosome banding and in situ hybridization. Disease resistance screening revealed that chromosome 1RafrS carries resistance gene(s) to new stripe rust races. Twenty-nine molecular markers were localized on S. africanum chromosome 1Rafr by the wheat – S. africanum introgression lines. Twenty markers can also identically amplify other reported wheat – S. cereale chromosome 1R derivative lines, indicating that there is high conservation between the wild and cultivated Secale chromosome 1R. Nine markers displayed polymorphic amplification between S. africanum and S. cereale chromosome 1Rafr derivatives. The comparison of the nucleotide sequences of these polymorphic markers suggested that gene duplication and sequence divergence may have occurred among Secale species during its evolution and domestication.

The in vitro release of juvenile hormones (JH) by female, and of JH acids (JHA) by male corpora allata (CA) ofLeucania loreyi was identified by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). Separation and quantification were accomplished by HPLC and GC, respectively. JH II and JH III were the major components released by CA of females. Four JHA analogues were identified as the release products of male CA, i.e. JHA III, Iso-JHA II, JHA II and JHA I. JHA III and Iso-JHA II were reported for the first time as the major release products of CA of adult male Lepidoptera. Iso-JHA II is a new member of the insect juvenile hormone analogue family.

A stable, highly fertile wheat Secale africanum substitution line LF24, derived from the F7 generation of a cross between Mianyang11 (MY11) and Triticum durum, S. africanum amphiploid (YF) was identified through molecular cytogenetic analysis. Application of C-banding, in situ hybridization and molecular markers analysis showed that LF24 was a wheat S. africanum 2R(a)(2D) substitution line. When inoculated with stripe rust isolates, T. durum and MY11 were highly susceptible, while S. africanum, YF and LF24 were immune. It is confirmed through molecular cytogenetic analysis that the stripe rust resistance of LF24 was derived from S. africanum chromosome 2R(a). We compared the banding patterns and disease resistance of reported chromosomes 2R from different S. cereale introduced into wheat background, and found that there was new stripe rust resistance gene(s) on S. africanum 2R(a). LF24 is a new substitution line which can be used as stripe rust resistant source in wheat improvement.

Two cytologically stable wheat-Dasypyrum breviarisatatum addition lines, Y93-1-6-6 and Y93-1-A6-4, were identified by integrated molecular and cytogenetic techniques. C-banding and genomic in situ hybridization (GISH) showed that Y93-1-6-6 and Y93-1-A6-4 were different wheat-D. breviaristatum additions. A total of 51 markers (primer/enzyme combinations), including 6 PCR-based Landmark Unique Gene (PLUG) markers and 45 Sequence-Tagged-Site (STS) markers, were selected from 3,774 primer/enzyme combinations to further characterize these two additions. Marker haploytpes suggested that both D. breviaristatum chromosomes in Y93-1-6-6 and Y93-1-A6-4 were rearranged. Stem rust resistance screening indicated that both additions were highly resistant to race RKQQC, whereas only Y93-1-6-6 was resistant to race TTKSK (Ug99). Powdery mildew resistance screening showed that only Y93-1-6-6 was resistant. Pedigree analysis suggested that the stem rust and powdery mildew resistance of Y93-1-6-6 was derived from D. breviaristatum, indicating that the D. breviaristatum chromosomes in Y93-1-6-6 possess a new powdery mildew resistance gene(s), and new stem rust resistance gene(s). These two additions could be used as stem rust or powdery mildew resistance sources in wheat breeding programs.

Publisher Summary This chapter reviews hormonal effects on aging in Drosophila melanogaster . The insect juvenile hormone (JH) is a sesquiterpenoid compound produced by the corpora allata (CA), a pair of endocrine glands with nervous connections to the brain. In pre-adult development and metamorphosis, JH functions as a “status quo” hormone, allowing continued growth after ecdysteroid-induced molting. Metamorphosis can only take place when the ecdysteroids act in the absence of JH. During the final larval instar, the JH titer declines because of a cessation of synthesis and increased degradation in the hemolymph and target tissues. Steroid hormones, such as ecdysteroids, are another class of vital hormones in insects. In pre-adult flies, 20E is produced in the larval prothoracic gland, which makes part of the larval ring gland in dipterans. Both JH and 20E play major antagonistic or synergistic roles in regulating Drosophila development. Application of commonly used JH inhibitors, such as precocene or fluvastatin, can reduce or inhibit JH synthesis in the CA and may thus be used to study the effects of JH deficiency on life span.

In Phormia regina, the rate of juvenile hormone (JH) synthesis rises rapidly after the ingestion of an adequate protein meal. In a previous publication we have localized the neurons containing Manduca sexta allatotropin (Mas-AT)-like substances in the brain of P. regina and demonstrated the allatotropic effect of synthetic Mas-AT in sugar-fed flies in vitro. In this current study, we examined the possible role of the brain of P. regina after the fly received a protein meal. In vitro studies showed that the brain releases, at 8 h after a protein meal, a factor(s) with a strong allatotropic effect. This factor(s) stimulates the corpus allatum (CA) to produce 6.9 times more juvenile hormones (JHs) than the control CA. Time course studies showed that the release of this allatotropic factor(s) is temporally controlled. Only the brains collected from flies at 6 and 8 h after the onset of a liver meal release allatotropic factor(s). Injection of anti-Mas-AT antiserum partially suppressed the fly follicle development in vivo. Presence of anti-Mas-AT antiserum decreased the allatotropic effect of the brain released allatotropic factor(s) in vitro. In addition to a Mas-AT-like substance, it is possible that the brains of liver-fed P. regina females may synthesize other allatotropic factors that are chemically unrelated or partially related to Mas-AT, which cannot be recognized/neutralized by our anti-Mas-AT antiserum.

The gp130 cytokines, including IL-6, IL-11, LIF and OSM, have important roles in neuroinflammation and neurodevelopment. However, the nature of any cell-specific responses of astrocytes and microglia to this family of cytokines and their molecular basis are not well understood. Here, the expression of the gp130 family cytokine receptors and subsequent signal pathway activation were examined in murine astrocytes and microglia in vitro . Astrocytes had high levels of OSMR mRNA and lower levels of IL-6R, LIFR and IL-11R mRNAs. In contrast, microglia expressed higher levels of IL-6R mRNA, similar levels of LIFR and IL-11R mRNAs, and no detectable OSMR mRNA. The OSMR protein was present in astrocytes but was undetectable in microglia. Conversely, the IL-6R protein was present in microglia but not detectable in astrocytes. In astrocytes, OSM induced STAT1 and STAT3 phosphorylation to a greater extent than hyperIL-6 (IL-6 linked to the soluble IL-6 receptor) and LIF. Conversely, hyperIL-6 and LIF but not OSM, induced phosphorylation of STAT1 and STAT3 in microglia. Upregulation of OSMR occurred in astrocytes, in response to a number of inflammatory mediators (e.g. lipo-polysaccharide) but remained undetectable in microglia. In contrast, the OSM mRNA was induced by the same inflammatory mediators in microglia but not astrocytes. Furthermore, the same inflammatory mediators also differentially induced other gp130 cytokines and their receptors between the two cell types. These findings show: (1) differences exist in the receptor expression utilized by the gp130 family of cytokines in astrocytes and microglia, (2) a molecular basis for the differential response to OSM by astrocytes versus microglia, and (3) a potential regulatory network exists between astrocytes and microglia in the central nervous system involving OSM and other gp130 cytokines.

Insect allatotropin upregulates the biosynthesis of juvenile hormones by the corpus allatum. We raised two rabbit antisera against the allatotropin of Manduca sexta (Mas AT) using a synthetic, multiple-antigenic-peptide that contains a branching heptalysine core and eight Mas AT molecules. Both antisera recognized specifically the same neurons in the larval brain, frontal ganglion and terminal abdominal ganglion of M. sexta as previously reported by others. Immunoassay showed reactivity specific to the Mas AT. Very low or nearly no cross-reactivity was found for two Mas AT-like peptides, a myotropin from Locusta migratoria and a Mas AT-like peptide deduced from the DNA sequence of Aedes aegypti, respectively. Immunopositive neurons also were identified in adult Phormia regina, Dacus dorsalis, Oncopeltus fasciatus, and Mythimna loreyi, and in larval M. loreyi, Bombyx mori, and Andraca bipunctata. At 20 pmol per 25 μl incubation medium (i.e. 8×10−7 M), synthetic Mas AT significantly stimulated in vitro juvenile hormone biosynthesis by the corpus allatum of adult, sugar-fed females of P. regina to 2.64-fold that of controls. Thus, this study provides the first demonstration that at the higher end of the physiological concentration range, the Mas AT has allatotropic effect in vitro to CA of non-lepidopterans. However, in vivo functions of Mas AT and/or Mas AT-like peptide in P. regina remain to be defined.