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The serine protease prostasin is a negative regulator of lipopolysaccharide-induced inflammation and has a role in the regulation of cellular immunity. Prostasin expression in cancer cells inhibits migration and metastasis, and reduces epithelial-mesenchymal transition. Programmed death-ligand 1 (PD-L1) is a negative regulator of the immune response and its expression in cancer cells interferes with immune surveillance. The aim of the present study was to investigate if prostasin regulates PD-L1 expression. We established sublines overexpressing various forms of prostasin as well as a subline deficient for the prostasin gene from the Calu-3 human lung cancer cells. We report here that PD-L1 expression induced by interferon-γ (IFNγ) is further enhanced in cells overexpressing the wildtype membrane-anchored prostasin. The PD-L1 protein was localized on the cell surface and released into the culture medium in extracellular vesicles (EVs) with the protease-active prostasin. The epidermal growth factor-epidermal growth factor receptor (EGF-EGFR), protein kinase C (PKC), and mitogen-activated protein kinase (MAPK) participated in the prostasin-mediated up-regulation of PD-L1 expression. A Gene Set Enrichment Analysis (GSEA) of patient lung tumors in The Cancer Genome Atlas (TCGA) database revealed that prostasin and PD-L1 regulate common signaling pathways during tumorigenesis and tumor progression. [ABSTRACT FROM AUTHOR]

This study aimed at improving the understanding of speech characteristics of fricatives produced by five-year-old Mandarin-acquiring children with cerebral palsy (CP). Productions from nine CP children and nine gender-and-age-matched typically developing (TD) children were collected and analyzed. Results from transcription indicated that the CP group had lower production accuracy rates for all the five fricatives in Mandarin Chinese. Additionally, when the CP children failed to articulate the target fricative segments, they tended to delete them or convert them into non-continuant segments. Results from acoustic analyses indicated that the M2 values of the labiodental [f] and the M1 and M2 values of the alveolar [s] were higher among the CP children. The experimental results revealed that: (1) Observable differences were available once the age of the groups was properly controlled and acoustical measurements were adopted; (2) the lack of finer-grained speech motor control abilities among CP children were reflected in the M1 and M2 values; (3) for segments at the anterior places, the clinical group failed to extend the articulatory gestures to the desirable positions. It is suggested that future studies focusing on different age groups and children with different native languages would help to approach the nature of articulatory barriers among individuals with CP. [ABSTRACT FROM AUTHOR]

Objective: The type-II transmembrane extracellular serine protease matriptase was shown to cleave at Arg-102 in the amino-terminal region of the amyloid precursor protein (APP). In this study we determined matriptase cleavage sites in the amyloid-beta (Aβ) peptide region of APP (Asp-597 to Ala-638 in the APP695 isoform). A recombinant human matriptase protease domain was used to cleave a synthetic human Aβ1–42 peptide. The human APP695 or mutants at the candidate matriptase cleavage sites was co-expressed with the human matriptase or its protease-dead mutant in HEK293 cells to evaluate matriptase cleavage of APP. Overexpression of matriptase in the M17 human neuroblastoma cells was performed to determine the effect of matriptase expression on endogenous APP. Results: The human Aβ1–42 peptide can be cleaved by the matriptase serine protease domain, at Arg-5, Lys-16, and Lys-28, as determined by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Co-expression of matriptase but not its protease-dead mutant with APP695 resulted in site-specific cleavages of the latter. Replacement of Arg-601 (Arg-5 in Aβ1–42) by Ala in APP695 prevented matriptase cleavage at this site. Overexpression of matriptase but not its protease-dead mutant in the M17 cells resulted in a significant reduction of the endogenous APP quantity. [ABSTRACT FROM AUTHOR]

Purpose: This study explored the speech characteristics of Mandarin-speaking children with cerebral palsy (CP) and typically developing (TD) children to determine (a) how children in the 2 groups may differ in their speech patterns and (b) the variables correlated with speech intelligibility for words and sentences. Method: Data from 6 children with CP and a clinical diagnosis of moderate dysarthria were compared with data from 9 TD children using a multiple speech subsystems approach. Acoustic and perceptual variables reflecting 3 speech subsystems (articulatory-phonetic, phonatory, and prosodic), and speech intelligibility, were measured based on speech samples obtained from the Test of Children's Speech Intelligibility in Mandarin (developed in the lab for the purpose of this research). Results: The CP and TD children differed in several aspects of speech subsystem function. Speech intelligibility scores in children with CP were influenced by all 3 speech subsystems, but articulatory-phonetic variables had the highest correlation with word intelligibility. All 3 subsystems influenced sentence intelligibility. Conclusion: Children with CP demonstrated deficits in speech intelligibility and articulation compared with TD children. Better speech sound articulation influenced higher word intelligibility, but did not benefit sentence intelligibility. [ABSTRACT FROM AUTHOR]

Four new minor brominated indole related alkaloids (one indoles, 1 , one 1,3-dihydro-indole-2-one, 2 , one carbazole, 3 , and one 2-carbonylamino-benzoate, 4 ) were isolated and identified from Laurencia similis by extensive chromatographic and spectrometric methods. Among them, 1 and 2 were the first example of naturally occurring indole with 3-benzyl group and 1,3-dihydro-indole-2-one with 2-isopropylidene group, respectively, whereas 3 and 4 were the first carbazole alkaloids and 2-carbonylamino-benzoate, respectively, isolated from the genus Laurencia. Moreover, 1 showed the most potent antibacterial activity against seven bacterial strains with MIC values ranging from 2 to 8 μg/mL. [ABSTRACT FROM AUTHOR]

In Arabidopsis (Arabidopsis thaliana), the Shaker K+ channel AKT1 conducts K+ uptake in root cells, and its activity is regulated by CBL1/9-CIPK23 complexes as well as by the AtKC1 channel subunit. CIPK23 and AtKC1 are both involved in the AKT1-mediated low-K+ (LK) response; however, the relationship between them remains unclear. In this study, we screened suppressors of low-K+ sensitive [lks1 (cipk23)] and isolated the suppressor of lks1 (sls1) mutant, which suppressed the leaf chlorosis phenotype of lks1 under LK conditions. Map-based cloning revealed a point mutation in AtKC1 of sls1 that led to an amino acid substitution (G322D) in the S6 region of AtKC1. The G322D substitution generated a gain-of-function mutation, AtKC1D, that enhanced K+ uptake capacity and LK tolerance in Arabidopsis. Structural prediction suggested that glycine-322 is highly conserved in K+ channels and may function as the gating hinge of plant Shaker K+ channels. Electrophysiological analyses revealed that, compared with wild-type AtKC1, AtKC1D showed enhanced inhibition of AKT1 activity and strongly reduced K+ leakage through AKT1 under LK conditions. In addition, phenotype analysis revealed distinct phenotypes of lks1 and atkc1 mutants in different LK assays, but the lks1 atkc1 double mutant always showed a LK-sensitive phenotype similar to that of akt1. This study revealed a link between CIPK-mediated activation and AtKC1-mediated modification in AKT1 regulation. CIPK23 and AtKC1 exhibit distinct effects; however, they act synergistically and balance K+ uptake/leakage to modulate AKT1-mediated LK responses in Arabidopsis. [ABSTRACT FROM AUTHOR]

Isolation of human subtype H3N2 influenza viruses in embryonated chicken eggs yields viruses with amino acid substitutions in the hemagglutinin (HA) that often affect binding to sialic acid receptors. We used a glycan array approach to analyze the repertoire of sialylated glycans recognized by viruses from the same clinical specimen isolated in eggs or cell cultures. The binding profiles of whole virions to 85 sialoglycans on the microarray allowed the categorization of cell isolates into two groups. Group 1 cell isolates displayed binding to a restricted set of α2-6 and α2-3 sialoglycans, whereas group 2 cell isolates revealed receptor specificity broader than that of their egg counterparts. Egg isolates from group 1 showed binding specificities similar to those of cell isolates, whereas group 2 egg isolates showed a significantly reduced binding to α2-6- and α2-3-type receptors but retained substantial binding to specific O- and N-linked α2-3 glycans, including α2-3GalNAc and fucosylated α2-3 glycans (including sialyl Lewis x), both of which may be important receptors for H3N2 virus replication in eggs. These results revealed an unexpected diversity in receptor binding specificities among recent H3N2 viruses, with distinct patterns of amino acid substitution in the HA occurring upon isolation and/or propagation in eggs. These findings also suggest that clinical specimens containing viruses with group 1-like receptor binding profiles would be less prone to undergoing receptor binding or antigenic changes upon isolation in eggs. Screening cell isolates for appropriate receptor binding properties might help focus efforts to isolate the most suitable viruses in eggs for production of antigenically well-matched influenza vaccines. [ABSTRACT FROM AUTHOR]

The article discusses the factors of formaldehyde (HCHO) assimilation in transgenic plants that were introduced to bacterial ribulose monophosphate pathway genes. Researchers discovered that the pathway genes could be integrated as a bypass to the Calvin-Benson cycle in transgenic Arabidopsis thaliana and tobacco by genetic engineering. Results suggested a strategy for phytoremediation of HCHO pollution.

Factors influencing caregivers' decisions to place their older family members in nursing homes were explored. Data were based on interviews with primary caregivers, and patient records of a Geriatric Intermediate Care Facility during the eight-year data collection period. Findings show that over half of the primary caregivers (55%) decided to place their older family member in a nursing home due to their incapability of providing care rather than the older adult's declining physical or cognitive functioning. Although it is reported that the Japanese, whose culture is embedded in filial piety, stigmatize the notion of placing their older adults in nursing homes, results of this study suggest their attitudes may be changing. Practice implications for healthcare professionals based on findings are provided. [ABSTRACT FROM AUTHOR]

The segmented structure of the influenza virus genome plays a pivotal role in its adaptation to new hosts and the emergence of pandemics. Despite concerns about the pandemic threat posed by highly pathogenic avian influenza H5N1 viruses, little is known about the biological properties of H5N1 viruses that may emerge following reassortment with contemporary human influenza viruses. In this study, we used reverse genetics to generate the 63 possible virus reassortants derived from H5N1 and H3N2 viruses, containing the H5N1 surface protein genes, and analyzed their viability, replication efficiency, and mouse virulence. Specific constellations of avian-human viral genes proved deleterious for viral replication in cell culture, possibly due to disruption of molecular interaction networks. In particular, striking phenotypes were noted with heterologous polymerase subunits, as well as NP and M, or NS. However, nearly one-half of the reassortants replicated with high efficiency in vitro, revealing a high degree of compatibility between avian and human virus genes. Thirteen reassortants displayed virulent phenotypes in mice and may pose the greatest threat for mammalian hosts. Interestingly, one of the most pathogenic reassortants contained avian PB1, resembling the 1957 and 1968 pandemic viruses. Our results reveal the broad spectrum of phenotypes associated with H5N1/H3N2 reassortment and a possible role for the avian PB1 in the emergence of pandemic influenza. These observations have important implications for risk assessment of H5N1 reassortant viruses detected in surveillance programs. [ABSTRACT FROM AUTHOR]

Prostasin is a glycosylphosphatidylinositol-anchored serine protease, with epithelial sodium channel activation and tumor invasion suppression activities. We identified the bladder as an expression site of prostasin. In the mouse, prostasin mRNA expression was detected by reverse transcription and real-time polymerase chain reaction in the bladder, and the prostasin protein was localized by immunohistochemistry in the urothelial cells. In mice injected intraperitoneally with bacterial lipopolysaccharide (LPS), bladder prostasin mRNA expression was downregulated, whereas the expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), interferon-γ (IFN-γ), TNF-α, IL-1β, and IL-6 was upregulated. Viral promoter-driven expression of the human prostasin homolog in the bladder of transgenic mice attenuated the LPS induction of iNOS but did not abolish the induction. LPS induction of COX-2, TNF-α, IL-1β, and IL-6 expression, however, was not reduced by prostasin transgene expression. Liposome-mediated delivery of prostasin-expressing plasmid into mouse bladder produced similar attenuation effects on LPS-induced iNOS expression, while not affecting COX-2 or cytokine induction. Mice receiving plasmid expressing a catalytic mutant prostasin did not manifest the iNOS induction attenuation phenotype. We propose a proteolytic mechanism for prostasin to intercept cytokine signaling during LPS-induced bladder inflammation. [ABSTRACT FROM AUTHOR]

Avian influenza A H5N1 viruses continue to spread globally among birds, resulting in occasional transmission of virus from infected poultry to humans. Probable human-to-human transmission has been documented rarely, but H5N1 viruses have not yet acquired the ability to transmit efficiently among humans, an essential property of a pandemic virus. The pandemics of 1957 and 1968 were caused by avian-human reassortant influenza viruses that had acquired human virus-like receptor binding properties. However, the relative contribution of human internal protein genes or other molecular changes to the efficient transmission of influenza viruses among humans remains poorly understood. Here, we report on a comparative ferret model that parallels the efficient transmission of H3N2 human viruses and the poor transmission of H5N1 avian viruses in humans. In this model, an H3N2 reassortant virus with avian virus internal protein genes exhibited efficient replication but inefficient transmission, whereas HSN1 reassortant viruses with four or six human virus internal protein genes exhibited reduced replication and no transmission. These findings indicate that the human virus H3N2 surface protein genes alone did not confer efficient transmissibility and that acquisition of human virus internal protein genes alone was insufficient for this 1997 H5N1 virus to develop pandemic capabilities, even after serial passages in a mammalian host. These results highlight the complexity of the genetic basis of influenza virus transmissibility and suggest that H5N1 viruses may require further adaptation to acquire this essential pandemic trait. [ABSTRACT FROM AUTHOR]

A novel mutant of bovine viral diarrhea virus (BVDV) was found with a virion assembly phenotype attributable to an insertion into the NS5B polymerase locus. This mutant, termed 5B-741, was engineered by reverse genetics to express NS5B with a C-terminal peptide tag of 22 amino acids. Electroporation of bovine cells with genomic RNA from this mutant showed levels RNA synthesis which were regarded as sufficient for infectivity, yet infectious virions were not produced. Pseudorevertants of mutant 5B-741 that released infectious virions and formed plaques revealed a single nucleotide change (T12369C). This change resulted in a leucine-to-proline substitution within the NS5B tag (L726P). Genetic analysis revealed that indeed a single nucleotide change encoding proline at NS5B position 726 in the pseudorevertant polyprotein mediated recovery of virion assembly function without improving genomic RNA accumulation levels. A subgenomic BVDV reporter replicon (rNS3-5B) was used to analyze the consequences of alterations of the genomic region encoding the NS5B C terminus on replication and assembly. Interestingly, rNS3-5B-L726P (revertant) replicated with the same efficiency as the rNS3-5B-741 mutant but produced 10 times more virions in a trans-packaging assay. These results indicated that impairment of assembly function in 5B-741 was independent of RNA accumulation levels and agreed with the observations from the full-length mutant and revertant genomes. Finally, we recapitulated the packaging defect of 5B-741 with a vaccinia virus expression system to eliminate possible unwanted interactions between the helper virus and the packaged replicon. Taken together, these studies revealed an unexpected role of NS5B in infectious virion assembly. [ABSTRACT FROM AUTHOR]

The surfactin production genetic locus (sfp) is responsible for the ability of Bacillus subtilis to produce the lipopeptide biosurfactant, surfactin. This report demonstrates the utility of using PCR of the sfp gene as a means of identifying Bacillus species that produce surfactin. We carried out a hemolysis zone assay, quantitative HPLC and NMR in parallel to ensure that the PCR provided correct results. PCR analyses were performed for the sfp gene on 15 standard strains and 20 field-collected Bacillus spp. isolates native to Taiwan. Among the 15 standard strains, surfactin was produced by seven strains of B. subtilis and two closely related species, B. amyloliquefaciens B128 and B. circulans ATCC 4513. Of the 20 field-collected Bacillus spp. isolates; 16 strains yielded surfactin- positive results with PCR and HPLC. A good correlation was observed. Within the 16 field isolates, B. amyloliquefaciens S13 (452.5 mg/L) and B. subtilis S15 (125.6 mg/L) had high productivity of surfactin. The technique is valuable for finding out potential good yields of surfactin-producing strains. The PCR method we used could also be used to find different species or genera containing homologous genes. This is the first report of the detection of surfactin production by B. amyloliquefaciens and B. circulans based on PCR screening. RID="" ID="" Correspondence to: S.-S. Kao; email: sskao@tactri.gov.tw [ABSTRACT FROM AUTHOR]

Nearly all human immunodeficiency virus (HIV) infections are acquired mucosally, and the gut-associated lymphoid tissues are important sites for early virus replication. Thus, vaccine strategies designed to prime virus-specific cytotoxic T lymphocyte (CTL) responses that home to mucosal compartments may be particularly effective at preventing or containing HIV infection. The Salmonella type III secretion system has been shown to be an effective approach for stimulating mucosal CTL responses in mice. We therefore tested ΔphoP-phoQ attenuated strains of Salmonella enterica serovar Typhimurium and S. enterica serovar Typhi expressing fragments of the simian immunodeficiency virus (SIV) Gag protein fused to the type III-secreted SopE protein for the ability to prime virus-specific CTL responses in rhesus macaques. Mamu-A[sup *]01[sup +] macaques were inoculated with three oral doses of recombinant Salmonella, followed by a peripheral boost with modified vaccinia virus Ankara expressing SIV Gag (MVA Gag). Transient low-level CTL responses to the Mamu-A[sup *]01 Gag[sub 181-189] epitope were detected following each dose of Salmonella. After boosting with MVA Gag, strong Gag-specific CTL responses were consistently detected, and tetramer staining revealed the expansion of Gag[sub 181-189]-specific CD8[sup +] T-cell responses in peripheral blood. A significant percentage of the Gag[sub 181-189]specific T-cell population in each animal also expressed the intestinal homing receptor α4β7. Additionally, Gag[sub 181-189]-specific CD8[sup +] T cells were detected in lymphocytes isolated from the colon. Yet, despite these responses, Salmonella-primed/MVA-boosted animals did not exhibit improved control of virus replication following a rectal challenge with SIVmac239. Nevertheless, this study demonstrates the potential of mucosal priming by the Salmonella type III secretion system to direct SIV-specific cellular immune responses to the gastrointestinal mucosa... [ABSTRACT FROM AUTHOR]

A gene for phosphoenolpyruvate carboxylase (PEPC) was isolated from a thermophilic cyanobacterium, Synechococcus vulcanus, by screening a genomic DNA library using the coding region of Anacystis nidulans 6301 PEPC as a probe. The S. vulcanus PEPC gene (SvPEPC) had an open reading frame for a polypeptide of 1,011 amino acid residues with a calculated molecular mass of 116.4 kDa. SvPEPC was expressed in E. coli BL21 Codonplus (DE3), using pET32a as a vector. The purified recombinant SvPEPC protein with a tag showed a single band of 120 kDa on SDS-PAGE. The enzyme forms homotetramer as judged by gel filtration. SvPEPC retained full activity even after incubation at 50°C for 60 min or exposure to 0.5 M guanidine-HCl at 30°C for 20 h, being more stable than C4-form PEPC from Zea mays (ZmPEPC(C4)). SvPEPC activity showed a sharp optimum temperature of 42°C at pH 7.5 and an optimum pH of 9.0 at 30°C. The enzyme, unlike most plant PEPCs, was predominantly activated by fructose 1,6-bisphosphate (Fruc-1,6-P2), and slightly stimulated by 3-phosphoglycerate (3-PGA), glucose 6-phosphate (Gluc-6-P), glucose 1-phosphate, Glu and Gln. Acetyl-CoA known as a strong activator of most bacterial PEPCs but not of plant PEPCs, showed no effect on the enzyme activity. SvPEPC was more sensitive to the inhibition by Asp at higher pH (9.0) than lower pH (7.0), contrary to Coccochloris peniocystis PEPC and plant PEPCs. I0.5 for Asp was increased about 2-fold by Gluc-6-P while markedly decreased by Fruc-1,6-P2, Glu and Gln about 3- to 4-fold. The regulation mechanism of SvPEPC is not readily interpretable by conventional allosteric models. [ABSTRACT FROM AUTHOR]

Discusses a study that presented a receiver design from signal processing viewpoint for direct-sequence code-division multiple access systems under multipath fading channels. Signal model; Robust Kalman Channel estimation; Simulation results.

Objectives: Substantially increased funding for health care services occurred in Taiwan after the implementation of a national health insurance plan in 1995. This study attempts to examine the impact of this national health insurance plan on the utilization of prenatal and intrapartum care services. Methods: Nationally representative surveys of all pregnant women in Taiwan in 1989 (1,662 participants) and in 1996 (3,626 participants) were included in the analysis. We first compared the distribution of birth characteristics between the two surveys. We then calculated the rate of utilization of various prenatal and intrapartum care services in the two surveys in the overall sample and in subsamples, stratified by maternal education, age, and parity. Results: The utilization of most prenatal and intrapartum care services, especially the complicated laboratory tests, increased in 1996 compared to 1989. For example, the proportion of women who received amniocentesis increased from 1.62% in 1989 to 5.60% in 1996 and German measles testing increased from 5.96% to 27.11%. By contrast, the proportion of women who received consultation services was stable over time, or for family planning, consultation declined from 33.21% to 27.00%. These changes in utilization over time were consistently observed across different maternal education, age, and parity groups. Conclusions: The utilization of prenatal and intrapartum care services, especially for the more expensive services, has substantially increased in Taiwan since the implementation of the national health insurance. For countries considering similar national health insurance plan, it may be helpful to consider cost-containing measures before the implementation of such a plan. [ABSTRACT FROM AUTHOR]

Examines the developmental control of endocytosis in dendritic cells (DC) by Cdc42. Reactivation of endocytosis with microbial delivery of a Cdc42 nucleotide exchange factor; Downregulation of endocytosis during DC development; Significance of endocytosis downregulation for DC function in immune response.

Toxicodendron vernicifluum (Stokes) F.A. Barkley (syn. Rhus verniciflua or vernicifera Stokes, Anacardiaceae) (RVS), the lacquer tree, also known as sumac, has been used in China, Japan and South Korea for thousands of years as a highly durable coating material and a traditional herbal medicine, which contains medicinal ingredients with anti-tumor, anti-inflammatory, antiviral, and anti-rheumatic activities. This review intends to provide a comprehensive and critical appraisal of RVS, including its phytochemical data, botanical and pharmacological literature that support its therapeutic potential in treatment on human diseases, with emphasis on the isolation of natural occurring compounds and detailed pharmacological investigations. Specific information of RVS was collected by using the key words " Toxicodendron vernicifluum ", " Rhus verniciflua Stokes", " Rhus vernicifera Stokes" and "Lacquer tree" through published scientific materials (including PubMed, ScienceDirect, Wiley, ACS, CNKI, Scifinder, Springer, Web of Science, Google Scholar, and Baidu Scholar) and other literature sources. The major phytoconstituents, 175 of which are presented in this review, including flavonoids, urushiols, terpenes, phenolic acids and other types of compounds, of which flavonoids and urushiols are main components. The extracts and isolates purified from RVS showed a wide range of in vitro and in vivo pharmacological effects, such as anti-cancer, anti-oxidation, anti-inflammatory, antimicrobial, tyrosinase inhibition and so on. The modern pharmacological research of RVS mainly focus on the pharmacological effects of crude extract or active constituents, of which the flavonoids are widely studied. However, there are few reports on the relationship between pharmacological effects and their structures. And at present, there is still a lack of researches that are of both effective and in-depth. Meanwhile, there is little research on quality control. Apart from the wood and lacquer, other botanical parts also need to be explored further. In addition to phenolic compounds, the study on other types of components in T. vernicifluum would start more sparks for the discovery of new bioactive principles. Image 1 • Toxicodendron vernicifluum (Stokes) F.A. Barkley is a traditional herbal medicine, which is commonly used in China, Japan and south Korea to treat various diseases. • In the past five years, there have been many reports about the constituents and pharmacological effects of T. vernicifluum , and it is urgent to conduct a comprehensive evaluation of this plant. • This review intends to provide a comprehensive and critical appraisal of T. vernicifluum , including its phytochemical data, botanical and pharmacological literatures that support its therapeutic potential in treatment to human diseases, with emphasis on the isolation of additional compounds and detailed pharmacological investigations. [ABSTRACT FROM AUTHOR]

Phytochemical investigations on the fruit stalks and seeds of the plant Hovenia dulcis Thunb. led to the isolation of twenty-one compounds, including three triterpenes (1 – 3), two sterols (4–5), five flavonoids (6 – 10), two sesquiterpenes (11 – 12), one lignan (13), two phenylpropanoids (14 – 15), four benzoic acid derivatives (16 – 19), one acid amide (20) and one cerebroside (21). The structures of these compounds were elucidated on the basis of spectroscopic analysis and comparison with previous literatures. Among them, ten compounds (4 , 11 – 12 , 14 – 20) were isolated from familiy Rhamnaceae, two (13 , 21) from the genus Hovenia , and three (5 , 8 , 10) from the species Hovenia dulcis Thunb. for the first time, respectively. The chemotaxonomic significance of these isolates was also discussed. • Twenty-one compounds were isolated from Hovenia dulcis Thunb. • Three compounds (5 , 8 , 10) were isolated from the species Hovenia dulcis Thunb. for the first time. • Two compounds (13 , 21) were isolated from the genus Hovenia for the first time. • Ten compounds (4 , 11 – 12 , 14 – 20) were isolated from familiy Rhamnaceae firstly. • The chemotaxonomic significance of these isolates was discussed. [ABSTRACT FROM AUTHOR]

A phytochemical investigation of the heartwood of Toxicodendron vernicifluum (Stokes) F.A. Barkley led to the isolation of twenty-five compounds, including sixteen flavonoids (1 – 15), one flavonolignan (16), one chromone (17), four benzoic acid derivatives (18 – 21), two triterpenes (22 – 23) and two sterols (24 – 25). The structures of these compounds were elucidated on the basis of spectroscopic analysis and comparison with previous literature data. Among them, 8 and 21 were isolated from the genus Toxicodendron and 5 , 9 , 13 , 17 – 20 , 22 from the family Anacardiaceae for the first time. Furthermore, 21 had been identified only by LC/MS before. 20 was a synthetic product used as anthelmintics, newly identified as a natural product. Furthermore, the chemotaxonomic value of these isolates was also discussed in detail. Image 1 • 5 , 9 , 13 , 17 – 20 , 22 were isolated from the family Anacardiaceae for the first time. • 8 and 21 were isolated from the genus Toxicodendron for the first time. • NMR data of one compound which was first reported from natural source was reported. • The chemotaxonomic significance of the isolated compounds is discussed. [ABSTRACT FROM AUTHOR]

Purpose: The aim of this study was to conduct a scoping review of research on oral and laryngeal diadochokinesis (DDK) in children and adults, either typically developing/developed or with a clinical diagnosis. Method: Searches were conducted with PubMed/MEDLINE, Google Scholar, CINAHL, and legacy sources in retrieved articles. Search terms included the following: DDK, alternating motion rate, maximum repetition rate, sequential motion rate, and syllable repetition rate. Results: Three hundred sixty articles were retrieved and included in the review. Data source tables for children and adults list the number and ages of study participants, DDK task, and language(s) spoken. Cross-sectional data for typically developing children and typically developed adults are compiled for the monosyllables /pʌ/, /tʌ/, and /kʌ/; the trisyllable /pʌtʌkʌ/; and laryngeal DDK. In addition, DDK results are summarized for 26 disorders or conditions. Discussion: A growing number of multidisciplinary reports on DDK affirm its role in clinical practice and research across the world. Atypical DDK is not a well-defined singular entity but rather a label for a collection of disturbances associated with diverse etiologies, including motoric, structural, sensory, and cognitive. The clinical value of DDK can be optimized by consideration of task parameters, analysis method, and population of interest. [ABSTRACT FROM AUTHOR]

Presents a study on the maternal mortality rate in Taiwan during 1984-1988. Statistics on deaths for this time; How study was conducted; Trends in mortality; Results and conclusions of study; Discussion of topic.

Among the four new alkaloids, indole (I) shows the most potent antibacterial activity against seven bacterial strains. [ABSTRACT FROM AUTHOR]

Abstract Zeasesquiterpene A-E (1–5), five new sesquiterpenes with two cyclohexanes, were isolated from the roots of Zea mays. Their structures were elucidated on the basis of 1D and 2D NMR spectral data and ECD analysis. A plausible biosynthetic pathway for the compounds (1–6) were hypothesized. All isolated compounds were screened for cytotoxicities against five human cancer cell lines (A549, MDA-MB-231, SK-Hep-1, SNU638 and HCT116) in vitro by MTT assay. Compound 4 showed potential cytotoxic activities against A549 (14.3 μΜ) and SNU638 (9.7 μΜ). By contrast, compound 1 exhibited moderate cytotoxicity to the four human cancer cell lines (A549, SK-Hep-1, SUN638 and HCT116), and the IC 50 values are from 19.5 μΜ to 22.5 μΜ. Graphical abstract Unlabelled Image [ABSTRACT FROM AUTHOR]

The objective of this case study is to present our experience of a surgical approach for vaginal agenesis using an acellular porcine small intestinal submucosa (SIS) graft. The present report involved 2 patients diagnosed as having vaginal agenesis due to Meyer-von-Rokitansky-Küster-Hauser syndrome. The operation procedure involved the creation of a neovaginal tunnel and then a mold wrapped with the SIS graft was placed in the neovagina. The duration of surgery was less than 45 min with minimal blood loss and no operative and postoperative complications. Epithelialization of the neovagina was achieved within 2 months after surgery. The neovagina created with this procedure was the same as that of a normal adult vagina histologically and physiologically. In conclusion, the creation of a neovagina using a SIS graft resulted in a favorable outcome and this approach may be a potential alternative to the management of vaginal agenesis. [ABSTRACT FROM AUTHOR]

We recently demonstrated that tissue kallikrein (TK) promotes keratinocyte migration through activation of protease-activated receptor-1 (PAR1) and transactivation of the epi-dermal growth factor receptor (EGFR). In this study, we investigated the potential role of PAR1 in mediating the effect of TK on cancer cell migration, invasion and proliferation. Our results show that TK promotes DU145 prostate cancer cell migration in a concentration-dependent manner, but has no effect on A549 lung cancer cells. Active TK markedly increases DU145 cell migration and invasion, which are blocked by aprotinin but minimally affected by icatibant; kinin treatment has little effect. TK-induced cell migration and invasion are abolished by inhibition of PAR1 using a pharmacological inhibitor or RNA interference. The effect of TK on cell migration and invasion are also blocked by inhibitors of protein kinase C, c-Src, matrix metalloproteinase, EGFR and extracellular signal-regulated kinase (ERK). Moreover, TK stimulates ERK phosphorylation, which is inhibited by an EGFR antagonist. Additionally, TK but not kinin stimulates DU145 cell proliferation through activation of the kinin B2 receptor, but not PAR1 and EGFR. These results indicate differential signaling pathways mediated by TK in promoting prostate cancer cell migration and invasion via PAR1 activation, and proliferation via kinin B2 receptor stimulation. [ABSTRACT FROM AUTHOR]

This study investigated whether transmissible H5 subtype human-avian reassortant viruses could be generated in vivo. To this end, ferrets were coinfected with recent avian H5N1 (A/Thailand/16/04) and human H3N2 (A/Wyoming/3/03) viruses. Genotype analyses of plaque-purified viruses from nasal secretions of coinfected ferrets revealed that approximately 9% of recovered viruses contained genes from both progenitor viruses. H5 and H3 subtype viruses, including reassortants, were found in airways extending toward and in the upper respiratory tract of ferrets. However, only parental H5N1 genotype viruses were found in lung tissue. Approximately 34% of the recovered reassortant viruses possessed the H5 hemagglutinin (HA) gene, with five unique H5 subtypes recovered. These H5 reassortants were selected for further studies to examine their growth and transmissibility characteristics. Five H5 viruses with representative reassortant genotypes showed reduced titers in nasal secretions of infected ferrets compared to the parental H5N1 virus. No transmission by direct contact between infected and naïve ferrets was observed. These studies indicate that reassortment between H5N1 avian influenza and H3N2 human viruses occurred readily in vivo and furthermore that reassortment between these two viral subtypes is likely to occur in ferret upper airways. Given the relatively high incidence of reassortant viruses from tissues of the ferret upper airway, it is reasonable to conclude that continued exposure of humans and animals to H5N1 alongside seasonal influenza viruses increases the risk of generating H5 subtype reassortant viruses that may be shed from upper airway secretions. [ABSTRACT FROM AUTHOR]

Avian H7 influenza viruses from both the Eurasian and North American lineage have caused outbreaks in poultry since 2002, with confirmed human infection occurring during outbreaks in The Netherlands, British Columbia, and the United Kingdom. The majority of H7 infections have resulted in self-limiting conjunctivitis, whereas probable human-to-human transmission has been rare. Here, we used glycan microarray technology to determine the receptor-binding preference, of Eurasian and North American lineage H7 influenza viruses and their transmissibility in the ferret model. We found that highly pathogenic H7N7 viruses from The Netherlands in 2003 maintained the classic avian-binding preference for α2-3-linked sialic acids (SA) and are not readily transmissible in ferrets, as observed previously for highly pathogenic H5N1 viruses. However, H7N3 viruses isolated from Canada in 2004 and H7N2 viruses from the northeastern United States isolated in 2002-2003 possessed an HA with increased affinity toward α2-6-linked SA, the linkage type found prominently on human tracheal epithelial cells. We identified a low pathogenic H7N2 virus isolated from a man in New York in 2003, A/NY/107/03, which replicated efficiently in the upper respiratory tract of ferrets and was capable of transmission in this species by direct contact. These results indicate that H7 influenza viruses from the North American lineage have acquired sialic acid-binding properties that more closely resemble those of human influenza viruses and have the potential to spread to naïve animals. [ABSTRACT FROM AUTHOR]

This article describes a Delphi study to identify research priorities of gerontological social work practitioners and to compare these priorities with those of social work academic researchers. A national expert panel of 52 gerontological social work practitioners completed questionnaires to delimit a set of high-priority research topics. Findings were compared with a similar Delphi study conducted with academic social work researchers. The researcher panel and the practitioner panel endorsed a need for intervention research. Practitioners also identified several unique priorities, including income security and long-term care policies, decision making, and planning for later life. The authors suggest four substantive areas (housing and transitions in living arrangements, family caregiving, health and mental health, and workforce) and four crosscutting themes (intervention research, social policy, service delivery, and capacity building) as a potential organizing framework for a research agenda for gerontological social work. [ABSTRACT FROM AUTHOR]

Purpose: An increasingly important task for all disciplines involved in aging research is to identify and prioritize areas for investigation. This article reports the results of a national Delphi study on setting research priorities for gerontological social work. Design and Methods: Delphi methodology, a structured process for eliciting and correlating informed opinions from a panel of experts on a specific topic, was used. A national expert panel of 46 gerontological social workers completed three successive Webbased questionnaires with controlled feedback to delimit a set of high-priority research topics. Results: There were 49 independent research topics identified, 16 of which attained high or highest priority and high or moderate consensus ratings. The top-priority topic was developing and testing psychosocial interventions across specific populations and conditions. Three additional topics on intervention research achieved similar ratings, as did all four topics on services research. Implications: The research priorities identified by expert panelists in this study represent critical knowledge needs for the social work profession in aging, and they overlap and complement the current research agendas of the National Research Council and the National Institute on Aging. They are thus expected to help guide the development and prioritization of social work and interdisciplinary research to improve practice and policies affecting older adults and their families. [ABSTRACT FROM AUTHOR]

Studies the cellular localization and expression of tissue kallikrein-kinin system in human adrenal gland. Localization of kallikrein transcript; Riboprobe and tissue preparation; Primers used in Southern blot analysis; Detection of the expression of kininogen in chromaffin cells of the adrenal medulla.

Influenza A(H1) viruses circulating in swine represent an emerging virus threat, as zoonotic infections occur sporadically following exposure to swine. A fatal infection caused by an H1N1 variant (H1N1v) virus was detected in a patient with reported exposure to swine and who presented with pneumonia, respiratory failure, and cardiac arrest. To understand the genetic and phenotypic characteristics of the virus, genome sequence analysis, antigenic characterization, and ferret pathogenesis and transmissibility experiments were performed. Antigenic analysis of the virus isolated from the fatal case, A/Ohio/09/2015, demonstrated significant antigenic drift away from the classical swine H1N1 variant viruses and H1N1 pandemic 2009 viruses. A substitution in the H1 hemagglutinin (G155E) was identified that likely impacted antigenicity, and reverse genetics was employed to understand the molecular mechanism of antibody escape. Reversion of the substitution to 155G, in a reverse genetics A/Ohio/09/2015 virus, showed that this residue was central to the loss of hemagglutination inhibition by ferret antisera raised against a prototypical H1N1 pandemic 2009 virus (A/California/07/2009), as well as gamma lineage classical swine H1N1 viruses, demonstrating the importance of this residue for antibody recognition of this H1 lineage. When analyzed in the ferret model, A/Ohio/09/2015 and another H1N1v virus, A/Iowa/39/2015, as well as A/California/07/2009, replicated efficiently in the respiratory tract of ferrets. The two H1N1v viruses transmitted efficiently among cohoused ferrets, but respiratory droplet transmission studies showed that A/California/07/2009 transmitted through the air more efficiently. Preexisting immunity to A/California/07/2009 did not fully protect ferrets from challenge with A/Ohio/09/2015. IMPORTANCE Human infections with classical swine influenza A(H1N1) viruses that circulate in pigs continue to occur in the United States following exposure to swine. To understand the genetic and virologic characteristics of a virus (A/Ohio/09/2015) associated with a fatal infection and a virus associated with a nonfatal infection (A/ Iowa/39/2015), we performed genome sequence analysis, antigenic testing, and pathogenicity and transmission studies in a ferret model. Reverse genetics was employed to identify a single antigenic site substitution (HA G155E) responsible for antigenic variation of A/Ohio/09/2015 compared to related classical swine influenza A(H1N1) viruses. Ferrets with preexisting immunity to the pandemic A(H1N1) virus were challenged with A/Ohio/09/2015, demonstrating decreased protection. These data illustrate the potential for currently circulating swine influenza viruses to infect and cause illness in humans with preexisting immunity to H1N1 pandemic 2009 viruses and a need for ongoing risk assessment and development of candidate vaccine viruses for improved pandemic preparedness. [ABSTRACT FROM AUTHOR]

The study was designed to investigate the possible mechanisms of hepatic microRNAs (miRs) in regulating local thyroid hormone (TH) action and ultimately different propensities to high-fat diet (HFD)-induced obesity. When obesity-prone (OP) and obesity-resistant (OR) mice were fed HFD for 7 weeks, OP mice showed apparent hepatic steatosis, with significantly higher body weight and lower hepatic TH receptor b (TRb) expression and type 1 deiodinase (DIO1) activity than OR mice. Next-generation sequencing technology revealed that 13 miRs in liver were dysregulated between the two phenotypes, of which 8 miRs were predicted to target on Dio1 or TRb. When mice were fed for 17 weeks, OR mice had mild hepatic steatosis and increased Dio1 and TRb expression than OP mice, with downregulation of T3 target genes (including Srebp1c, Acc1, Scd1 and Fasn) and upregulation of Cpt1α, Atp5c1, Cox7c and Cyp7a1. A stemloop qRT-PCR analysis confirmed that the levels of miR-383, miR-34a and miR-146b were inversely correlated with those of DIO1 or TRb. Down-regulated expression of miR-383 or miR-146b by miR-383 inhibitor (anti-miR-383) or miR-146b inhibitor (anti-miR-146b) in free fatty acid-treated primary mouse hepatocytes led to increased DIO1 and TRb expressions, respectively, and subsequently decreased cellular lipid accumulation, while miR-34a inhibitor (anti-miR-34a) transfection had on effects on TRb expression. Luciferase reporter assay illustrated that miR-146b could directly target TRb 3'untranslated region (3'UTR). These findings suggested that miR-383 and miR-146b might play critical roles in different propensities to diet-induced obesity via targeting on Dio1 and TRb, respectively. [ABSTRACT FROM AUTHOR]

Background: Calcineurin (CaN) is a Ca2+ - and calmodulin (CaM)-dependent serine/threonine phosphatase. Previous studies have found that CaN is involved in the regulation of the stress responses. Results: In this study, the growth of Cryptococcus humicola was inhibited by the CaN inhibitor tacrolimus (FK506) under aluminum (Al) stress. The expression of CNA encoding a catalytic subunit A (CNA) and its interaction with CaM were upregulated when the concentration of Al was increased. A CaM-binding domain and key amino acids responsible for interaction with CaM were identified. ΔCNAb with a deletion from S454 to A639 was detected to bind to CaM, while ΔCNAa with a deletion from R436 to A639 showed no binding to CaM. The binding affinities of CNA1 and CNA2, in which I439 or I443 were replaced by Ala, were decreased relative to wild-type CNA. The phosphatase activities of ΔCNAa, CNA1 and CNA2 were lower than the wild-type protein. These results suggest that the region between R436 and S454 is essential for the interaction with CaM and I439, I443 are key amino acids in this region. The ability of the CNA transgenic yeast to develop resistance to Al was significantly higher than that of control yeast. Residual Al in the CNA transgenic yeast culture media was significantly lower than the amount of Al originally added to the media or the residual Al remaining in the control yeast culture media. These findings suggest that CNA confers Al tolerance, and the mechanism of Al tolerance may involve absorption of active Al. Conclusions: Al stress up-regulated the expression of CNA. CaM-binding domain and key amino acids responsible for interaction with CaM were identified and both are required for phosphatase activities. CNA conferred yeast Al resistance indicating that the gene has a potential to improve Al-tolerance through gene engineering. [ABSTRACT FROM AUTHOR]

This study sought to improve an existing live attenuated influenza vaccine (LAIV) by including nucleoprotein (NP) from wild-type virus rather than master donor virus (MDV). H7N9 LAIV reassortants with 6:2 (NP from MDV) and 5:3 (NP from wild-type virus) genome compositions were compared with regard to their growth characteristics, induction of humoral and cellular immune responses in mice, and ability to protect mice against homologous and heterologous challenge viruses. Although, in general, the 6:2 reassortant induced greater cellmediated immunity in C57BL6 mice than the 5:3 vaccine, mice immunized with the 5:3 LAIV were better protected against heterologous challenge. The 5:3 LAIV-induced CTLs also had better in vivo killing activity against target cells loaded with the NP366 epitope of recent influenza viruses. Modification of the genome of reassortant vaccine viruses by incorporating the NP gene from wild-type viruses represents a simple strategy to improve the immunogenicity and cross-protection of influenza vaccines. [ABSTRACT FROM AUTHOR]

Bat influenza virus H17N10 represents a distinct lineage of influenza A viruses with gene segments coding for proteins that are homologs of the surface antigens, hemagglutinin (HA) and neuraminidase (NA). Our recent study of the N10 NA homolog revealed an NA-like structure, but with a highly divergent putative active site exhibiting little or no NA activity, and provided strong motivation for performing equivalent structural and functional analyses of the H17 HA protein. The overall structure of the H17 HA homolog from A/little yellow-shouldered bat/Guatemala/060/2010 at 3.18 Å resolution is very similar to other influenza HAs, with a putative receptor-binding site containing some conserved aromatic residues that form the base of the sialic acid binding site. However, the rest of the H17 receptor-binding site differs substantially from the other HA subtypes, including substitution of other conserved residues associated with receptor binding. Significantly, electrostatic potential analyses reveal that this putative receptor-binding site is highly acidic, making it unfavorable to bind any negatively charged sialylated receptors, consistent with the recombinant H17 protein exhibiting no detectable binding to sialylated glycans. Furthermore, the fusion mechanism is also distinct; trypsin digestion with recombinant H17 protein, when exposed to pH 4.0, did not degrade the HA1 and HA2, in contrast to other HAs. These distinct structural features and functional differences suggest that the H17 HA behaves very differently compared with other influenza HAs. [ABSTRACT FROM AUTHOR]

Cell sheet-mediated tissue regeneration is a promising approach for corneal reconstruction. However, the fragility of bioengineered corneal endothelial cell (CEC) monolayers allows us to take advantage of cross-linked porous gelatin hydrogels as cell sheet carriers for intraocular delivery. The aim of this study was to further investigate the effects of biopolymer concentrations (5-15 wt%) on the characteristic and safety of hydrogel discs fabricated by a simple stirring process combined with freeze-drying method. Results of scanning electron microscopy, porosity measurements, and ninhydrin assays showed that, with increasing solid content, the pore size, porosity, and cross-linking index of carbodiimide treated samples significantly decreased from 508±30 to 292±42 µm, 59.8±1.1 to 33.2±1.9%, and 56.2±1.6 to 34.3±1.8%, respectively. The variation in biopolymer concentrations and degrees of cross-linking greatly affects the Young's modulus and swelling ratio of the gelatin carriers. Differential scanning calorimetry measurements and glucose permeation studies indicated that for the samples with a highest solid content, the highest pore wall thickness and the lowest fraction of mobile water may inhibit solute transport. When the biopolymer concentration is in the range of 5-10 wt%, the hydrogels have high freezable water content (0.89-0.93) and concentration of permeated glucose (591.3-615.5 µg/ml). These features are beneficial to the in vitro cultivation of CECs without limiting proliferation and changing expression of ion channel and pump genes such as ATP1A1, VDAC2, and AQP1. In vivo studies by analyzing the rabbit CEC morphology and count also demonstrate that the implanted gelatin discs with the highest solid content may cause unfavorable tissue-material interactions. It is concluded that the characteristics of cross-linked porous gelatin hydrogel carriers and their triggered biological responses are in relation to biopolymer concentration effects. [ABSTRACT FROM AUTHOR]

Recently, we reported a unique influenza A virus subtype H17N10 from little yellow-shouldered bats. Its neuraminidase (NA) gene encodes a protein that appears to be highly divergent from all known influenza NAs and was assigned as a new subtype N10. To provide structural and functional insights on the bat H17N10 virus, X-ray structures were determined for N10 NA proteins from influenza A viruses A/little yellow-shouldered bat/Guatemala/164/ 2009 (GUO9-164) in two crystal forms at 1.95 Å and 2.5 Å resolution and A/little yellow-shouldered bat/Guatemala/060/2010 (GU10060) at 2.0 Å. The overall N10 structures are similar to each other and to other known influenza NA structures, with a single highly conserved calcium binding site in each monomer. However, the region corresponding to the highly conserved active site of influenza A N1-N9 NA subtypes and influenza B NA differs substantially. In particular, most of the amino acid residues required for NA activity are substituted, and the putative active site is much wider because of displacement of the iso-loop and 430-loop. These structural features and the fact that the recombinant N10 protein exhibits no, or extremely low, NA activity suggest that it may have a different function than the NA proteins of other influenza viruses. Accordingly, we propose that the N10 protein be termed an NA-like protein until its function is elucidated. [ABSTRACT FROM AUTHOR]

The article presents the results of the analytical evaluation of available influenza diagnostic tests (RIDTs) that detect the influenza virus nucleoprotein (NP) antigen cleared by the U.S. Food and Drug Administration (FDA). Sixteen influenza A and seven influenza B viruses were provided by the Centers for Disease Control and Prevention to evaluate the commercially available RIDTs during the 2011-2012 influenza season. It describes the limitations of RIDTs.

Maternal cigarette smoking has adverse effects on pregnancy outcomes. The granulocyte-macrophage colony-stimulating factor (GM-CSF) is an essential cytokine for a normal pregnancy. We investigated the impact of cigarette smoke extract (CSE) on GM-CSF expression in human cytotrophoblast cells and suggested a cellular mechanism underlying the CSE-induced GM-CSF expression. An immortalized normal human trophoblast cell line (B6Tert-1) was treated with CSE. The viability and proliferation of the CSE-treated B6Tert-1 cells were evaluated, and the expression of GM-CSF in these cells was quantified at the mRNA and the protein levels by means of reverse-transcription and quantitative polymerase chain reaction (RT-qPCR); and enzyme-linked immunosorbent assay (ELISA), respectively. Human trophoblast cells treated with CSE had an increased expression of GM-CSF at both the mRNA and the protein levels. The CSE-induced GM-CSF expression was synergistically enhanced by the addition of the proteasome inhibitor MG-132, but inhibited by AG-1478, an inhibitor of the epidermal growth factor receptor (EGFR) kinase. Furthermore, CSE treatment increased the phosphorylation of the extracellular-signal regulated kinases (ERK1/2) in the trophoblast cells. The expression of other growth factors such as heparin- binding epidermal growth factor-like growth factor (HB-EGF) and vascular endothelial growth factor (VEGF) was also evaluated. Our data suggested that cigarette smoking and proteasome inhibition synergistically up-regulate GM-CSF cytokine expression by activating the EGFR signaling pathway. [ABSTRACT FROM AUTHOR]

Influenza A virus reservoirs in animals have provided novel genetic elements leading to the emergence of global pandemics in humans. Most influenza A viruses circulate in waterfowl, but those that infect mammalian hosts are thought to pose the greatest risk for zoonotic spread to humans and the generation of pandemic or panzootic viruses. We have identified an influenza A virus from little yellow-shouldered bats captured at two locations in Guatemala. It is significantly divergent from known influenza A viruses. The HA of the bat virus was estimated to have diverged at roughly the same time as the known subtypes of HA and was designated as H17. The neuraminidase (NA) gene is highly divergent from all known influenza NAs, and the internal genes from the bat virus diverged from those of known influenza A viruses before the estimated divergence of the known influenza A internal gene lineages. Attempts to propagate this virus in cell cultures and chicken embryos were unsuccessful, suggesting distinct requirements compared with known influenza viruses. Despite its divergence from known influenza A viruses, the bat virus is compatible for genetic exchange with human influenza viruses in human cells, suggesting the potential capability for reassortment and contributions to new pandemic or panzootic influenza A viruses. [ABSTRACT FROM AUTHOR]

Continued H5N1 virus infection in humans highlights the need for vaccine strategies that provide cross-clade protection against this rapidly evolving virus. We report a comparative evaluation in ferrets of the immunogenicity and cross-protective efficacy of isogenic mammalian cell-grown, live attenuated influenza vaccine (LAIV) and adjuvanted, whole-virus, inactivated influenza vaccine (IIV), produced from a clade 1 H5N1 6:2 reassortant vaccine candidate (caVN1203-Len17rg) based on the cold-adapted A/Leningrad/134/17/57 (H2N2) master donor virus. Two doses of LAIV or IIV provided complete protection against lethal homologous H5N1 virus challenge and a reduction in virus shedding and disease severity after heterologous clade 2.2.1 H5N1 virus challenge and increased virus-specific serum and nasal wash antibody levels. Although both vaccines demonstrated cross-protective efficacy, LAIV induced higher levels of nasal wash IgA and reduction of heterologous virus shedding, compared with IIV. Thus, enhanced respiratory tract antibody responses elicited by LAIV were associated with improved cross-clade protection. [ABSTRACT FROM AUTHOR]

We recently reported that HIV-1 infection can be inhibited by innate antimicrobial components of human seminal plasma (SP). Conversely, naturally occurring peptidic fragments from the SP-derived prostatic acid phosphatase (PAP) have been reported to form amyloid fibrils called "SEVI" and enhance HIV-1 infection in vitro. In order to understand the biological consequence of this proviral effect, we extended these studies in the presence of human SP. PAP-derived peptides were agitated to form SEVI and incubated in the presence or absence of SP. While PAP-derived peptides and SEVI alone were proviral, the presence of 1% SP ablated their proviral activity in several different anti-HIV-1 assays. The anti-HIV-1 activity of SP was concentration dependent and was reduced following filtration. Supraphysiological concentrations of PAP peptides and SEVI incubated with diluted SP were degraded within hours, with SP exhibiting proteolytic activity at dilutions as high as 1:200. Sub-physiological concentrations of two prominent proteases of SP, prostate-specific antigen (PSA) and matriptase, could degrade physiological and supraphysiological concentrations of PAP peptides and SEVI. While human SP is a complex biological fluid, containing both antiviral and proviral factors, our results suggest that PAP peptides and SEVI may be subject to naturally occurring proteolytic components capable of reducing their proviral activity. [ABSTRACT FROM AUTHOR]

BACKGROUND: Prostasin is a glycosylphosphatidylinositol-anchored extracellular serine protease with a role in epidermal growth factor receptor (EGFR) signal modulation. EGFR signaling has been shown to be important for regulating cytotrophoblast (CT) cell proliferation in human placenta. We investigated the impact of prostasin expression regulation on this cellular function as well as the molecular mechanisms involved in human cytotrophoblastic cells. [ABSTRACT FROM PUBLISHER]

Controlled invasion of the uterine wall by the trophoblast cells is pivotal for the successful pregnancy, and various kinds of protease are involved in this process. Serine protease prostasin has been shown to participate in the proteolytic activation of epithelial sodium channel as well as cleavage of epidermal growth factor receptor extracellular domain in human epithelial cells. Its physiological significance in human placentation has been suggested but not validated. In the present study, we found that prostasin was expressed at a relatively high level in human placenta trophoblasts in early pregnant weeks. In the in vitro cultured human choriocarcinomal JEG-3 cells, treatment with functional antibody against prostasin led to promotion in cell invasion capability, as well as increase in the production of MMP-2, MMP-26, TIMP-1, and TIMP-4. Our data indicated that this serine protease may function as an invasion suppressor in human trophoblast, participating in the invasion-restrictive regulation of trophoblasts to avoid their over-penetration into the uterine wall. [ABSTRACT FROM AUTHOR]

Background: Recurrent outbreaks of highly pathogenic H5N1 avian influenza virus pose a threat of eventually causing a pandemic. Early vaccination of the population would be the single most effective measure for the control of an emerging influenza pandemic. Methodology/Principal Findings: Influenza virus-like particles (VLPs) produced in insect cell-culture substrates do not depend on the availability of fertile eggs for vaccine manufacturing. We produced VLPs containing influenza A/Viet Nam1203/04 (H5N1) hemagglutinin, neuraminidase, and matrix proteins, and investigated their preclinical immunogenicity and protective efficacy. Mice immunized intranasally with H5N1 VLPs developed high levels of H5N1 specific antibodies and were 100% protected against a high dose of homologous H5N1 virus infection at 30 weeks after immunization. Protection is likely to be correlated with humoral and cellular immunologic memory at systemic and mucosal sites as evidenced by rapid anamnestic responses to re-stimulation with viral antigen in vivo and in vitro. Conclusions/Significance: These results provide support for clinical evaluation of H5N1 VLP vaccination as a public health intervention to mitigate a possible pandemic of H5N1 influenza. [ABSTRACT FROM AUTHOR]