Cavidine Ameliorates Lipopolysaccharide-Induced Acute Lung Injury <Emphasis Type="Italic">via</Emphasis> NF-κB Signaling Pathway <Emphasis Type="Italic">in vivo</Emphasis> and <Emphasis Type="Italic">in vitro</Emphasis>

Acute lung injury (ALI) is characterized by widespread inflammation in the lungs and alveolar-capillary destruction, causing high morbidity and mortality. Cavidine, isolated from Corydalis impatiens, have been exhibited to have potent anti-inflammatory effects in previous studies. The purpose of this study was to evaluate the protective effect of cavidine on lipopolysaccharide (LPS)-induced ALI and to enunciate the underlying in vivo and in vitro mechanisms. Mice were intraperitoneally administrated with cavidine (1, 3, or 10 mg/kg) at 1 and 12 h, prior to the induction of ALI by intranasal administration of LPS (30 mg/kg). Blood samples, lung tissues, and bronchoalveolar lavage fluid (BALF) were harvested after LPS challenge. Furthermore, we used LPS-induced lung epithelial cells A549 to examine the mechanism of cavidine to lung injury. The results showed that pretreatment with cavidine significantly decreased lung wet-to-dry weight (W/D) ratio, reduced pro-inflammatory cytokine levels including TNF-α and IL-6 in BALF and serum from LPS-stimulated mice, and attenuated lung histopathological changes. In addition, western blot results showed that cavidine inhibited the phosphorylation of nuclear factor-kappaB (NF-κB) p65 and IκBα induced by LPS. In conclusion, our results demonstrate that cavidine protects against LPS-induced acute lung injury in mice via inhibiting of pro-inflammatory cytokine TNF-α and IL-6 production and NF-κB signaling pathway activation. Taken together, cavidine may be useful for the prevention and treatment of pulmonary inflammatory diseases, such as ALI.     

Effects of Human Defensin-α<Subscript>1</Subscript> on <Emphasis Type="Italic">Trypanosoma cruzi</Emphasis> Trypomastigotes <Emphasis Type="Italic">in Vitro</Emphasis>

Human defensin-α₁ is a biologically active peptide exhibiting a dose-dependent trypanocidal effect in vitro against trypomastigotes and amastigotes of Trypanosoma cruzi line Tulahuen. This effect is determined by fragmentation of parasite DNA reducing the capacity of passaged T. cruzi to invade HeLa cells.     

<Emphasis Type="Italic">RNR4</Emphasis> mutant alleles <Emphasis Type="Italic">pso3-1</Emphasis> and <Emphasis Type="Italic">rnr4Δ</Emphasis> block induced mutation in <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis>

The PSO3 gene of Saccharomyces cerevisiae was molecularly cloned by complementing the cold-sensitivity phenotype of a pso3-1 mutant and was found to be allelic to RNR4, encoding one of the two DNA damage-inducible small subunits of the ribonucleotide reductase (RNR) complex. Compared to a rnr4Δ mutant that allows only very little mutation induction at very low doses of 254_nm ultraviolet light (UVC), the pso3-1 mutant allele confers leakiness in that it permits some DNA damage-induced mutagenesis at low doses of UVC. Similarly, the pso3 mutant is slightly less sensitive to UVC than an rnr4Δ mutant. Cloning and sequencing of the RNR4 locus of the pso3-1 mutant revealed that its intermediate phenotype is attributable to a G → A transition at nucleotide 352, leading to replacement of glycine by arginine [G118R] in the mutant’s protein. Both RNR4 mutant alleles confer significantly less sensitivity to UVC than mutant alleles of non-UVC-mutable REV3, indicating that, apart from nucleotide excision repair, RAD6-dependent error-free DNA repair may still be functional. The phenotype of a strongly reduced UVC-induced mutagenesis for rnr4 mutant alleles has not yet been described; it suggests the importance of this gene for a fully functional RNR providing correct amounts of DNA precursor molecules, thereby, allowing translesion synthesis (error-prone) of UVC-damaged DNA. Stationary phase cells of the rnr4Δ mutant, but not of the original pso3-1 mutant, are swollen with a fourfold to eightfold increase in volume. The central role of RNR in DNA precursor metabolism and its complex regulation allow for several modes of suppression that may influence the phenotypes of RNR4 mutants, especially those containing the leaky pso3-1 mutant allele.     

The mating type-specific homeodomain genes <Emphasis Type="Italic">SXI1α</Emphasis> and <Emphasis Type="Italic">SXI2a</Emphasis> coordinately control uniparental mitochondrial inheritance in <Emphasis Type="Italic">Cryptococcus neoformans</Emphasis>

In the great majority of sexual eukaryotes, mitochondrial genomes are inherited almost exclusively from a single parent. While many hypotheses have been proposed to explain this phenomenon, very little is known about the genetic elements controlling uniparental mitochondria inheritance. In the bipolar, isogamous basidiomycete yeast Cryptococcus neoformans, progeny from crosses between strains of mating type a (MATa) and mating type α (MATα) typically inherit mitochondrial DNA (mtDNA) from the MATa parent. We recently demonstrated that a mating type α (MATα)-specific gene SXI1α, controls mitochondrial inheritance in C. neoformans. Here, we show that another homeodomain gene SXI2a in the alternative mating type MATa is also required for uniparental mtDNA inheritance in this fungus. Disruption of SXI2a resulted in biparental mtDNA inheritance in the zygote population with significant numbers of progeny inheriting mtDNA from the MATa parent, the MATα parent, and both the MATa and the MATα parents. In addition, progeny from same-sex mating between MATα strains showed a biparental mitochondrial inheritance pattern. Our results suggest that SXI1α and SXI2a coordinately control uniparental mitochondrial inheritance in C. neoformans.     

What are the infectious larvae in <Emphasis Type="Italic">Ascaris suum</Emphasis> and <Emphasis Type="Italic">Trichuris muris?</Emphasis>

In the present study, larvae of Ascaris suum and Trichuris muris were investigated by light and electron microscopy after incubation in a hatching medium containing 89% phosphate-buffered saline (pH 7.4), 10% RPMI-1640 and 1% sodiumhypochlorite at 40 and 37 degrees C, respectively. The larvae were obtained from fertilised eggs of the worms during defined phases of development (A. suum, 36th-50th day of development; T. muris, once a week from week 16 to 20). Light and electron micrographs of the larvae gave evidence that the third larval stage of A. suum is probably the infectious stage. The first moult of the larvae had already taken place before the 36th day of incubation starting at day 1. After 36 days of incubation, only the second larval stage was found within eggs. Some of these larvae were coated by a separated sheath so that a second moult of the larvae is reasonable. On the other hand, no sheathed larvae of T. muris were found in the eggs incubated for 20 weeks in distilled water. No signs of moult were seen for 20 weeks neither on light nor on the electron micrographs. Therefore, in T. muris, the first larval stage is the infectious stage, which was proven by means of re-infections of mice 16, 18 or 20 weeks after incubation of the eggs.     

Chemopreventive potential of β-Sitosterol in experimental colon cancer model - an <Emphasis Type="Italic">In vitro</Emphasis> and <Emphasis Type="Italic">In vivo</Emphasis> study

Background  

Asclepias curassavica Linn. is a traditional medicinal plant used by tribal people in the western ghats, India, to treat piles, gonorrhoea, roundworm infestation and abdominal tumours. We have determined the protective effect of β-sitosterol isolated from A. curassavica in colon cancer, using in vitro and in vivo models.     

<Emphasis Type="Italic">Phlebotomus</Emphasis> (<Emphasis Type="Italic">Euphlebotomus</Emphasis>) <Emphasis Type="Italic">mascomai</Emphasis> n. sp. (Diptera–Psychodidae)

A new species of sandfly is described from limestone caves in Thailand. The inclusion of this species in the subgenus Euphlebotomus is justified on the basis of characters of the male genitalia (paramere, basal lobe). The male–female gathering in the same taxon is based on ecological (cavernicolous species), morphological (length of male genital filaments and female spermathecal ducts) and molecular (homology of cytochrome b mt DNA sequences) criteria. A differential diagnosis between Phlebotomus mascomai n. sp. and P. argentipes Annandale & Brunetti, the vector of Leishmania donovani (Laveran & Mesnil) in India, is proposed based on several morphological characters like antennal formula and genitalia.     

Interleukin-10 production and T cell-suppressive capacity in B cell subsets from atherosclerotic <Emphasis Type="Italic">apoE</Emphasis><Superscript><Emphasis Type="Italic">−/−</Emphasis></Superscript> mice

The evidence regarding the role of regulatory B cells (Breg) in atherosclerosis are scarce, and there are contradictory data about their atheroprotective properties. Due to the demonstrated protective function of Breg in different inflammatory diseases mainly through interleukin-10 (IL-10) production, the knowledge of their participation in atherosclerosis immunopathology would be very valuable. To further study which B cell subsets participate in IL-10 production and their regulatory role, splenocytes from apolipoprotein-E-deficient mice were evaluated by ex vivo and in vitro cultures. Atherosclerotic mice had increased frequency of IL-10⁺ B cells, which presented high CD1d, CD19, and IgM, but variable CD5, CD21, and CD23 expression. IL-10⁺ B cells were not enriched in B cell subsets previously reported as Breg. Increased frequency of IL-10⁺ B cells with transitional 1-like (T1-like) and follicular (FO) and reduced CD5⁺ and marginal zone (MZ) phenotypes were observed ex vivo. Increased frequency of IL-10⁺ B cells with T1-like and MZ, and decreased IL-10⁺ FO and T2 phenotypes were also observed in vitro. To determine regulatory capacity of B cells in the atherosclerotic model, each subset were co-cultured with CD4⁺CD25^? T cells. CD5⁺, FO, MZ, and T1-like cells from atherosclerotic mice exhibited regulation in an IL-10-dependent manner. However, only FO cells decreased both frequency of interferon gamma (IFN-γ)⁺ and tumor necrosis factor alpha (TNF-α)⁺ and proliferation of T cells. Finally, splenocytes showed increased frequency of IFN-γ⁺ and TNF-α⁺ cells only when FO-depleted B cells were evaluated. These results suggest that mainly FO B cells can modulate in some level the inflammatory responses observed in atherosclerosis.     

Genetic regulation of arrested development in nematodes: are <Emphasis Type="Italic">age</Emphasis>-1 and <Emphasis Type="Italic">daf</Emphasis>-gene orthologs present in <Emphasis Type="Italic">Dictyocaulus viviparus</Emphasis>?

In opposite to the free-living soil nematode Caenorhabditis elegans, the genetic regulation of hypobiosis or inhibited or arrested development in parasitic nematodes is completely unknown. In C. elegans, the daf-genes or the age-1 gene are of major importance in signaling pathways regulating arrested development. To investigate if orthologs of these genes are present in the bovine lungworm Dictyocaulus viviparus, a PCR analysis with gene-specific primer combinations was performed. No orthologs of the age-1 or daf-genes could be identified in D. viviparus. The possible differences in the role of the daf-genes concerning arrested development in parasitic and free-living nematodes will be discussed.     

The <Emphasis Type="Italic">rpl5</Emphasis>–<Emphasis Type="Italic">rps14</Emphasis> mitochondrial region: a hot spot for DNA rearrangements in <Emphasis Type="Italic">Solanum</Emphasis> spp. somatic hybrids

Southern analysis with rpl5 and rps14 mtDNA gene probes of Solanum tuberosum, S. commersonii and a sample of somatic hybrids detected polymorphisms between parents and the appearance of a novel restriction fragment in various hybrids. In one of them, detailed mtDNA analyses revealed various configurations of the rpl5–rps14 region present at different stoichiometries. Multiple inter-parental recombination events across homologous sequences were assumed to have caused these rearrangements. Sequence similarity searches detected one sequence putatively involved in the recombination upstream of the rpl5 gene. The presence of a second recombinogenic sequence was inferred. We propose two models to explain the mechanism responsible for obtaining the different rpl5–rps14 arrangements shown after somatic hybridization. Variability in the rpl5–rps14 region observed in both the parental species and their somatic hybrids suggests this region is a hot spot for mtDNA rearrangements in Solanum spp.Contribution no. 39 from the Institute of Plant Genetics, Research Division of Portici.Communicated by A. Brennicke     

Circulatory “Efficacy” during progressive aerobic exercise in children: insights from the <Emphasis Type="Italic">Q</Emphasis>: <Emphasis Type="Italic">V</Emphasis>O<Subscript>2</Subscript> relationship

The relationship between circulatory flow (Q) and oxygen uptake ( ) may provide insights into performance of peripheral mechanisms which govern blood flow during exercise (circulatory efficacy). This study evaluated the response of Q relative to during progressive upright cycle exercise in a group of 39 preadolescent boys (mean age 12.2 ± SD 0.5 years). The Q– relationship was curvilinear, best described by the cubic equation Q = 3.60()³ + 5.24()² + 2.40() − 0.94. Circulatory efficacy, defined as the %ΔQ/%Δ × 100, fell from 70.4% between the first two workloads to 49.7% at peak exercise. This decline in circulatory efficacy is consistent with other published data suggesting a decline in skeletal muscle pump function at high intensity workloads. The pattern of change in relationship of Q and during progressive exercise in these children is similar to that observed in studies of adults. This implies that performance of peripheral determinants of circulatory responses to exercise is not affected by biological maturation.     

Expression of the <Emphasis Type="Italic">Acremonium chrysogenum</Emphasis><Emphasis Type="Italic">cefT</Emphasis> gene in <Emphasis Type="Italic">Penicillum chrysogenum</Emphasis> indicates that it encodes an hydrophilic β-lactam transporter

The Acremonium chryrsogenum cefT gene encoding a membrane protein of the major facilitator superfamily implicated in the cephalosporin biosynthesis in A. chrysogenum was introduced into Penicillium chrysogenum Wisconsin 54-1255 (a benzylpenicillin producer), P. chrysogenum npe6 pyrG (-) (a derivative of Wisconsin 54-1255 lacking a functional penDE gene) and P. chrysogenum TA98 (a deacetylcephalosporin producer containing the cefD1, cefD2, cefEF and cefG genes from A. chrysogenum). RT-PCR analysis revealed that the cefT gene was expressed in P. chrysogenum strains. HPLC analysis of the culture broths of the TA98 transformants showed an increase in the secretion of deacetylcephalosporin C and hydrophilic penicillins (isopenicillin N and penicillin N). P. chrysogenum Wisconsin 54-1255 strain transformed with cefT showed increased secretion of the isopenicillin N intermediate and a drastic decrease in the benzylpenicillin production. Southern and northern blot analysis indicated that the untransformed P. chrysogenum strains contain an endogenous gene similar to cefT that may be involved in the well-known secretion of the isopenicillin N intermediate. In summary, the cefT transporter is a hydrophilic beta-lactam transporter that is involved in the secretion of hydrophilic beta-lactams containing alpha-aminoadipic acid side chain (isopenicillin N, penicillin N and deacetylcephalosporin C).     

Use of a <Emphasis Type="Italic">ura5</Emphasis><Superscript>+</Superscript>–<Emphasis Type="Italic">lys7</Emphasis><Superscript>+</Superscript> cassette to construct unmarked gene knock-ins in <Emphasis Type="Italic">Schizosaccharomyces pombe</Emphasis>

While the counterselectable Schizosaccharomyces pombe ura4 ⁺ gene can be used to prepare a site in the S. pombe genome to receive an unmarked mutant allele (loss of ura4 ⁺ confers 5FOA-resistant (5FOA^R) growth), the desired unmarked knock-in strains are generally outnumbered by spontaneously arising 5FOA^R mutants. Relative to the same approach using the homologous URA3 ⁺ gene in Saccharomyces cerevisiae, knock-ins in S. pombe are harder to identify due to a lower efficiency of homologous recombination and a relatively high background of spontaneous 5FOA^R colonies. To develop an improved method for identifying cells receiving unmarked mutant alleles, we first determined that 5FOA^R strains carry mutations in either of two genes; ura4 ⁺ and ura5 ⁺. We then cloned the S. pombe ura5 ⁺ orotate phosphoribosyltransferase gene and constructed a 2.1 kb cassette containing ura5 ⁺ together with the S. pombe lys7 ⁺ gene. Using this doubly marked cassette to disrupt the sck1 ⁺ kinase gene, we can distinguish between strains created by homologous knock-in of unmarked wild-type or kinase-dead alleles and spontaneously arising ura4 ⁻ and ura5 ⁻ mutants by screening 5FOA^R colonies for the loss of the lys7 ⁺ marker. The utility of this system, especially when the phenotype for the strain carrying the knock-in allele is indistinguishable from that of the disruption strain, is borne out by the fact that ~95% of 5FOA^R colonies in our studies arose from background ura4 ⁻ and ura5 ⁻ mutations.     

<Emphasis Type="Italic">Ex vivo</Emphasis> production of interferon-γ, tumor necrosis factor-α, and interleukin-6 by mouse macrophages during infection with <Emphasis Type="Italic">M. bovis</Emphasis> and <Emphasis Type="Italic">M. tuberculosis</Emphasis> H37Rv

Ex vivo production of IFN-γ, TNF-α, and IL-6 by mouse peritoneal macrophages was studied during successive infection with the vaccine strain M. bovis BCG and virulent strain M. tuberculosis H37Rv. The increase in the concentrations of TNF-α and IL-6 did not depend on the sequence of macrophage infection with the vaccine or virulent strain, but was related to the presence of the vaccine strain M. bovis BCG in the medium. IFN-γ production depended on infection of macrophages with the virulent strain M. tuberculosis H37Rv. The concentration of IFN-γ was maximum during primary infection with the virulent strain and did not increase after successive infection with the virulent and vaccine strain. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 144, No. 11, pp. 552–555, November, 2007     

First report of <Emphasis Type="Italic">Cryptosporidium</Emphasis><Emphasis Type="Italic">parvum</Emphasis> ‘ferret’ genotype in American mink (<Emphasis Type="Italic">Mustela vison</Emphasis> Shreber 1777)

A total of 51 faecal samples from wild and farmed mink were analysed by a direct immunofluorescence antibody test. Cryptosporidium oocysts were identified in eight, apparently healthy, farmed American mink (Mustela vison). The isolates were identified as Cryptosporidium parvum ‘ferret’ genotype by PCR-RFLP and sequencing analysis of a 341-base-pair fragment of the Cryptosporidium oocyst wall protein (COWP) gene. This is the first report of Cryptosporidium in American mink. Genbank accession numbers of the sequences used in this study C. parvum, ‘bovine’ genotype AF266273; C. hominis, AF266265; C. parvum, ‘mouse’ genotype AF266268, C. wrairi, U35027; C. parvum, ‘ferret’ genotype AF266267; C. meleagridis, AF266266; C. parvum, ‘marsupial’ genotype AF266269; C. parvum, ‘pig’ genotype AF266270; C. canis, AF266274; C. felis, AF266263; C. baileyi, AF266276; C. serpentis, AF266275; C. andersoni, AF266262 and C. muris, AF161579     

First description of natural <Emphasis Type="Italic">Echinococcus multilocularis</Emphasis> infections in chinchilla (<Emphasis Type="Italic">Chinchilla laniger</Emphasis>) and Prevost’s squirrel (<Emphasis Type="Italic">Callosciurus prevostii borneoensis</Emphasis>)

This report describes for the first time the occurrence of alveolar echinococcosis in two exotic rodent species in Europe. A pet chinchilla (Chinchilla laniger) was euthanized due to a painful enlargement of the abdominal cavity, and a Prevost's squirrel (Callosciurus prevostii borneoensis) was found dead in the enclosure of a zoo. At necropsy, extended liver lesions consisting of small vesicles and cysts were observed in the livers of both animals. Histological examination revealed that these cysts were composed of an outer, homogenous, eosinophilic layer and an inner, cellular germinal layer. The cysts from both animals contained numerous protoscolices. The morphological diagnosis of Echinococcus multilocularis metacestode infections was confirmed by molecular means.     

Prevalence and antibiotic susceptibility of <Emphasis Type="Italic">Ureaplasma urealyticum</Emphasis> and <Emphasis Type="Italic">Mycoplasma hominis</Emphasis> in Xi’an,China

This study analyzed the prevalence and antibiotic susceptibility of urogenital Ureaplasma urealyticum and Mycoplasma hominis isolated in Xi’an, China. A total of 2161 individuals from 2011 to 2015 were included, and antibiotic susceptibility tests were performed by using the Mycoplasma IST kit. Of the individuals studied, 1018 (47.11 %) were identified to be positive for urogenital mycoplasmas. The single U. urealyticum, single M. hominis, and dual U. urealyticum and M. hominis infections accounted for 772 (75.83 %), 66 (6.48 %), and 180 (17.68 %), respectively. The total positive rate was higher in females than in males (58.76 % vs. 28.86 %, p?<?0.001). The highest total positive rate (48.88 %) was observed in individuals aged 25 years to 30 years. In symptomatic and asymptomatic individuals, the positive rates were both higher in females than in males (67.36 % vs. 31.02 %, p?<?0.001 and 42.58 % vs. 7.69 %, p?<?0.001, respectively) and individuals aged 25 years to 30 years, and those aged 30 years to 35 years had the highest positive rates (54.35 and 57.14 %, respectively). The U. urealyticum and M. hominis identified from single or dual infections displayed low resistance rates to josamycin, doxycycline, and minocycline (<10 %) in both the symptomatic and asymptomatic groups. These results suggest that females and individuals with symptoms and younger age had higher mycoplasma infection rates and that josamycin, doxycycline, and minocycline may be recommended for the clinical treatment of patients infected with urogenital mycoplasmas, irrespective of the symptoms.     

Cerebral Imaging Markers of <Emphasis Type="Italic">GBA</Emphasis> and <Emphasis Type="Italic">LRRK2</Emphasis> Related Parkinson’s Disease and Their First-Degree Unaffected Relatives

Cerebral atrophy has been detected in patients with Parkinson’s disease (PD) both with and without dementia, however differentiation based on genetic status has thus far not yielded robust findings. We assessed cortical thickness and subcortical volumes in a cohort of PD patients and healthy controls carriers of the G2019S mutation in the LRRK2 gene and the common GBA mutations, in an attempt to determine whether genetic status influences structural indexes. Cortical thickness and subcortical volumes were computed and compared between six groups of participants; idiopathic PD, GBA-PD, LRRK2-PD, non-manifesting non-carriers (NMNC), GBA-non-manifesting carriers (NMC) and LRRK2-NMC utilizing the FreeSurfer software program. All participants were cognitively intact based on a computerized cognitive assessment battery. Fifty-seven idiopathic PD patients, 9 LRRK2-PD, 12 GBA-PD, 49 NMNC, 41 LRRK2-NMC and 14 GBA-NMC participated in this study. Lower volumes among patients with PD compared to unaffected participants were detected in bilateral hippocampus, nucleus accumbens, caudate, thalamus, putamen and amygdala and the right pallidum (p?=?0.016). PD patients demonstrated lower cortical thickness indexes in a majority of regions assessed compared with non-manifesting participants. No differences in cortical thickness and subcortical volumes were detected within each of the groups of participants based on genetic status. Mutations in the GBA and LRRK2 genes are not important determinants of cortical thickness and subcortical volumes in both patients with PD and non-manifesting participants. PD is associated with a general reduction in cortical thickness and sub-cortical atrophy even in cognitively intact patients.