Enhancing the heralded single-photon rate from a silicon nanowire by time and wavelength division multiplexing pump pulses.

Heralded single photons produced on a silicon chip represent an integrated photon source solution for scalable photonic quantum technologies. The key limitation of such sources is their non-deterministic nature introduced by the stochastic spontaneous four-wave mixing (SFWM) process. Active spatial and temporal multiplexing can improve this by enhancing the single-photon rate without degrading the quantum signal-to-noise ratio. Here, taking advantage of the broad bandwidth of SFWM in a silicon nanowire, we experimentally demonstrate heralded single-photon generation from a silicon nanowire pumped by time and wavelength division multiplexed pulses. We show a 90±5% enhancement on the heralded photon rate at the cost of only 14±2% reduction to the signal-to-noise ratio, close to the performance found using only time division multiplexed pulses. As single-photon events are distributed to multiple wavelength channels, this new scheme overcomes the saturation limit of avalanche single-photon detectors and will improve the ultimate performance of such photon sources.

Swimming exercise stimulates neuro-genesis in the subventricular zone via increase in synapsin I and nerve growth factor levels.

In this study, we investigated the effects of 8-weeks of swimming exercise on neurogenesis in the subventricular zone (SVZ) and on the levels of nerve growth factor (NGF) and synapsin I protein in the olfactory bulb (OB) of adult rats at a series of relevant time points (2 days, 1 week, 2 weeks, 4 weeks, 3 months, and 6 months). Ninety-six male Sprague Dawley rats were divided into 2 groups: (1) a control group (COG; n = 48, n = 8 for each time point) and (2) a swimming exercise group (SEG; total n = 48; n = 8 for each time point). SEG performed swimming exercise for 5 days per week over a period of 8 weeks. We found that the number of 5-bromo-2'-deoxyuridine-5'-monophosphate (BrdU)- and doublecortin (DCX)-positive cells was significantly higher in SEG than in COG at all time points (Day 2, Week 1, Week 2, Week 4, Month 3, and Month 6; p < 0.001). Furthermore, NGF and synapsin I protein levels were significantly higher in SEG on Day 2, and Weeks 1, 2, and 4 than in COG (p < 0.05 for each time point). Our findings suggest that regular swimming exercise in adult rats increases neurogenesis, neuronal survival, and neuronal maintenance in the SVZ; furthermore, swimming exercise increases the levels of NGF and synapsin I in the OB.

Presence of Porphyromonas gingivalis and plasma cell dominance in gingival tissues with periodontitis.

OBJECTIVE: Porphyromonas gingivalis can invade and survive within its host epithelial cells. The aim of this study was to test our hypothesis that persistent presence of intracellular periodontal pathogens in gingival tissue causes the chronic inflammation and that an inappropriate immune response is a risk factor for periodontitis. METHODS: Together with the presence of P. gingivalis, the distribution of B cells, plasma cells, and CD4(+), CD8(+), and FOXP3(+) regulatory T cells was evaluated in gingival tissues from healthy (n = 7) and periodontitis (n = 8) sites by in situ hybridization and immunohistochemistry, respectively. RESULTS: Porphyromonas gingivalis was detected in proximity to inflammatory infiltrates in three and seven biopsies from the healthy and periodontitis sites, respectively. Compared with healthy sites, periodontal lesions contained a significantly increased number of each immune cell studied with a relative dominance of plasma cells over T cells. CONCLUSIONS: Persistent bacterial invasion of gingival tissues in combination with a plasma cell-dominant immune response may be involved in the pathogenesis of periodontitis.

Association of a protease with polytene chromosomes of Drosophila melanogaster.

Incubation of Drosophila salivary glands with radioactive diisopropyl fluorophosphate results in the uniform labeling of polytene chromosomes. Extensive labeling is seen only when chromosome squashes are prepared by a formaldehyde fixation procedure and not by standard acetic acid techniques. The labeling is inhibited in the presence of tosylphenylalanine chloromethyl ketone and phenylmethane sulfonylfluoride but not by tosyllysine chloromethyl ketone, suggesting that a chymotrypsin-like serine protease is associated with the chromosomes. Protease inhibitors show no apparent effect on heat-shock specific puffing.

Expression of mucins and trefoil factor family protein-1 in the colon of pigs naturally infected with Salmonella typhimurium.

The expression patterns of different mucins (MUC1, MUC2, MUC4, MUC5AC, MUC5C and MUC6) and trefoil factor family protein-1 (TFF1) in the colon of healthy pigs and pigs naturally infected with Salmonella typhimurium is reported. Twenty infected pigs approximately 80-160 days of age from 20 different herds were studied. These animals had similar microscopical change in colonic tissue characterized by mucosal erosion or sloughing and acute inflammation. S. typhimurium was cultured from all lesions and the identity of each isolate was confirmed by serotyping. Immunohistochemical studies of colonic tissue revealed reduced expression of MUC4 on the surface of the cryptal epithelium of S. typhimurium-infected pigs compared with non-infected pigs (P<0.001). By contrast, colon from infected animals had increased expression of MUC5AC (P<0.0001) and TFF1 (P=0.0095) relative to controls and there was a significant positive correlation between expression of these two molecules (Spearman coefficient 0.64, P<0.0001). Further studies are needed to evaluate the functional relationship between altered expression of these molecules and inflammation in Salmonella-infected pigs.

Evidence of shedding of porcine circovirus type 2 in milk from experimentally infected sows.

Detection of porcine circovirus type 2 (PCV2) was to evaluate the milk from experimentally infected sows using polymerase chain reaction (PCR) and virus isolation. Six pregnant sows were inoculated intranasally with PCV2 at 93 days of gestation, and milk samples were collected from all sows at 1, 3, 6, 9, 12, 15, 18, 21, 24, and 27 days of lactation. PCV2 was detected in milk as early as day 1 of lactation in all six sows. Thereafter, all infected sows remained positive by PCR for PCV2 in milk until 27 days of lactation. In addition, PCV2 itself was isolated from milk collected from a virus-infected sows. These results suggest that PCV2 may be shed in milk following infection of pregnant sows by the virus.

Effects of a modified live CSFV vaccine on the development of PMWS in pigs infected experimentally with PCV-2.

The objective of this study was to determine the effect of vaccination against classical swine fever virus (CSFV) on the development of postweaning multisystemic wasting syndrome (PMWS) in conventional pigs infected experimentally with porcine circovirus type 2 (PCV-2). The pigs infected with PCV-2 and immunised with modified live CSFV developed mild to moderate PMWS, whereas none of the pigs infected with PCV-2 alone or immunised with modified live CSFV alone developed PMWS. Lesions histologically characteristic of PMWS were observed in lymph nodes from the pigs infected with PCV-2 and immunised with modified live CSFV vaccine, and extensive replication of PCV-2 was detected in the nodes by in situ hybridisation.

Detection by in-situ hybridization of Pasteurella multocida toxin (toxA) gene in the lungs of naturally infected pigs.

In-situ hybridization with a non-radioactive digoxigenin-labelled probe was used to detect the Pasteurella multocida toxin (PMT) gene in tissue sections of pneumonic lung from pigs naturally infected with toxigenic P. multocida. The morphology of host cells was preserved despite the relatively high temperature used in the incubation procedure. Pulmonary abscessation was observed in 13 pigs naturally infected with toxigenic P. multocida type A (three pigs) or D (10 pigs). In these 13 pigs a strong hybridization signal for PMT DNA was detected, mainly in degenerate leucocytes in abscesses. Occasionally, PMT DNA was detected in degenerate neutrophils and macrophages in alveolar spaces. Detection of hybridization signals for PMT DNA would seem to be a potential indicator of the production of PMT. The study suggested that PMT plays an important role in pulmonary abscessation caused by P. multocida.

A modified-live porcine reproductive and respiratory syndrome virus (PRRSV)-1 vaccine protects late-term pregnancy gilts against heterologous PRRSV-1 but not PRRSV-2 challenge.

The objective of this study was to determine the efficacy of a commercially available porcine reproductive and respiratory syndrome virus (PRRSV)-1 modified-live virus (MLV) vaccine against PRRSV-1 and PRRSV-2 challenge in late-term pregnancy gilts. Gilts were vaccinated with the PRRSV-1 MLV vaccine at 4 weeks prior to breeding and then challenged intranasally with PRRSV-1 or PRRSV-2 at 93 days of gestation. After PRRSV-1 challenge, vaccinated pregnant gilts had a significantly longer gestation period, significantly higher numbers of live-born and weaned piglets and a significantly lower number of stillborn piglets at birth compared to unvaccinated pregnant gilts. No significant improvement in reproductive performance was observed between vaccinated and unvaccinated pregnant gilts following PRRSV-2 challenge. Vaccinated pregnant gilts also exhibited a significantly improved reproductive performance after challenge with PRRSV-1 compared to vaccinated pregnant gilts following PRRSV-2 challenge. The PRRSV-1 MLV vaccine was able to reduce PRRSV-1 but not PRRSV-2 viremia in pregnant gilts. Vaccinated gilts also showed a significantly higher number of PRRSV-1-specific IFN-γ-secreting cells (IFN-γ-SC) compared to PRRSV-2-specific IFN-γ-SC. The data presented here suggest that the vaccination of pregnant gilts with a PRRSV-1 MLV vaccine provides good protection against PRRSV-1 but only limited protection against PRRSV-2 challenge in late-term pregnancy gilts based on improvement of reproductive performance, reduction in viremia and induction of IFN-γ-SC.

S-phase-specific transcription regulatory elements are present in a replication-independent testis-specific H2B histone gene.

The testis-specific H2B histone (TH2B) gene is expressed in pachytene spermatocytes during meiotic prophase I in the absence of any significant DNA synthesis. Unlike somatic histones, synthesis of testis-specific histones is not affected by inhibitors of DNA synthesis. A genomic rat TH2B gene was cloned by using a DNA fragment derived from TH2B cDNA as a probe. Expression of the cloned TH2B was investigated by gene transfer experiments. From these studies, we found that the 5' upstream region of the cloned TH2B gene contained S-phase-specific transcription elements which regulated expression of a reporter gene in an S-phase-specific manner. The S-phase-regulatory element was found to be located in two regions containing CCAAT elements between -153 and -110 base pairs (bp) and an octamer element (ATTTGCAT) between -109 and -84 bp. The two regions were required for a maximal stimulation of transcription of the cloned TH2B gene in S phase. On the other hand, only the octamer element was reported be important for the S-phase-specific transcription of human H2B gene. Since the synthesis of the TH2B histone is independent of DNA synthesis and specific for pachytene spermatocytes in vivo, the presence of the S-phase-specific transcription regulatory elements in the TH2B gene is surprising.

Demethylation of somatic and testis-specific histone H2A and H2B genes in F9 embryonal carcinoma cells.

In contrast to many other genes containing a CpG island, the testis-specific H2B (TH2B) histone gene exhibits tissue-specific methylation patterns in correlation with gene activity. Characterization of the methylation patterns within a 20-kb segment containing the TH2A and TH2B genes in comparison with that in a somatic histone cluster revealed that: (i) the germ cell-specific unmethylated domain of the TH2A and TH2B genes is defined as a small region surrounding the CpG islands of the TH2A and TH2B genes and (ii) somatic histone genes are unmethylated in both liver and germ cells, like other genes containing CpG islands, whereas flanking sequences are methylated. Transfection of in vitro-methylated TH2B, somatic H2B, and mouse metallothionein I constructs into F9 embryonal carcinoma cells revealed that the CpG islands of the TH2A and TH2B genes were demethylated like those of the somatic H2A and H2B genes and the metallothionein I gene. The demethylation of those CpG islands became significantly inefficient at a high number of integrated copies and a high density of methylated CpG dinucleotides. In contrast, three sites in the somatic histone cluster, of which two sites are located in the long terminal repeat of an endogenous retrovirus-like sequence, were efficiently demethylated even at a high copy number and a high density of methylated CpG dinucleotides. These results suggest two possible mechanisms for demethylation in F9 cells and methylation of CpG islands of the TH2A and TH2B genes at the postblastula stage during embryogenesis.

Specific regions of beta-globin RNA are resistant to nuclease digestion in RNA-protein complexes in chicken reticulocyte nuclei.

The interaction between beta-globin RNA and proteins in chicken reticulocyte nuclei was studied by determining the sequence of nuclease-resistant beta-globin RNA. Two types of nuclease-resistant RNAs were isolated for this study: endogenous nuclease-resistant RNA from 50S heterogeneous nuclear RNA-protein complexes and micrococcal nuclease-resistant nuclear RNA from whole nuclei. The nuclease-resistant regions were identified with the use of a RNA mapping method we recently developed (J.R. Patton and C.-B. Chae, J. Biol. Chem. 258:3991-3995, 1983). We found that beta-globin RNA is assembled into heterogeneous nuclear RNA-protein complexes in a specific manner. There are several regions of nuclease resistance in the first and third exons interrupted at regular intervals by sensitive regions. The second exon has only one nuclease-resistant region. The resistant regions range in size from 20 to 50 nucleotides. This organization may reflect a specific mode of assembly for heterogeneous nuclear RNA-protein complexes.

Characterization of the S-phase-specific transcription regulatory elements in a DNA replication-independent testis-specific H2B (TH2B) histone gene.

The testis-specific H2B histone (TH2B) gene is expressed in pachytene spermatocytes of meiotic prophase I during rat spermatogenesis. The TH2B RNA and histones are not synthesized in any other tissues, and the synthesis is independent of DNA replication. However, the cloned TH2B gene has two DNA sequence elements which stimulate transcription of the cloned gene in an S-phase-dependent manner when introduced into somatic cells. The factors interacting with the two elements, CCAAT at -127 base pairs and octamer ATTTGCAT at -93 base pairs, interact with each other to bring about a maximum stimulation of S-phase-dependent transcription. The level of CCAAT and octamer-binding proteins is unchanged during the cell cycle, and the S-phase-dependent transcription of TH2B and endogenous mouse H2B genes does not require synthesis of new proteins during the S phase. Cell cycle-specific posttranslational modification of regulatory proteins may be responsible for the S-phase-dependent transcription of H2B histone genes. The biological significance of the presence of S-phase-specific transcription regulatory elements in the DNA replication-independent and tissue-specific TH2B gene is not known.

A novel DNA-binding protein bound to the mitochondrial inner membrane restores the null mutation of mitochondrial histone Abf2p in Saccharomyces cerevisiae.

The yeast mitochondrial HMG-box protein, Abf2p, is essential for maintenance of the mitochondrial genome. To better understand the role of Abf2p in the maintenance of the mitochondrial chromosome, we have isolated a multicopy suppressor (YHM2) of the temperature-sensitive defect associated with an abf2 null mutation. The function of Yhm2p was characterized at the molecular level. Yhm2p has 314 amino acid residues, and the deduced amino acid sequence is similar to that of a family of mitochondrial carrier proteins. Yhm2p is localized in the mitochondrial inner membrane and is also associated with mitochondrial DNA in vivo. Yhm2p exhibits general DNA-binding activity in vitro. Thus, Yhm2p appears to be novel in that it is a membrane-bound DNA-binding protein. A sequence that is similar to the HMG DNA-binding domain is important for the DNA-binding activity of Yhm2p, and a mutation in this region abolishes the ability of YHM2 to suppress the temperature-sensitive defect of respiration of the abf2 null mutant. Disruption of YHM2 causes a significant growth defect in the presence of nonfermentable carbon sources such as glycerol and ethanol, and the cells have defects in respiration as determined by 2,3,5,-triphenyltetrazolium chloride staining. Yhm2p may function as a member of the protein machinery for the mitochondrial inner membrane attachment site of mitochondrial DNA during replication and segregation of mitochondrial genomes.

Prevalence and genotyping of hepatitis E virus in swine population in Korea between 1995 and 2004: a retrospective study.

Hepatitis E virus (HEV) infections have been reported in pigs throughout the world but have only recently been recorded in Korean pigs. The aim of this study was to investigate whether HEV was present in archived porcine hepatic tissues collected between 1995 and 2004 using RT-PCR and immunohistochemistry and, if so, to determine the genotype of the isolates. Swine HEV was identified in the liver tissue of 42 pigs of 388 submissions (four pigs every year on average). The isolates showed genetic homology with swine and human HEV isolates identified in the United States and Japan (92.5-97%) and phylogenetic tree analysis indicated they belonged to genotype III. The study indicates that HEV is not a newly emerging virus in Korean pigs, but a pathogen that has existed in the country since at least 1995.

Two Commercial Type 1 Porcine Reproductive and Respiratory Syndrome Virus (PRRSV)-Modified Live Vaccines Reduce Seminal Shedding of Type 1 PRRSV but not Type 2 PRRSV in Infected Boars.

The objective of this study was to compare the effects of two commercial type 1 porcine reproductive and respiratory syndrome virus (PRRSV)-modified live vaccines on type 1 and type 2 PRRSV shedding in the semen of experimentally infected boars. Upon challenge with PRRSV, unvaccinated boars exhibited an increase in daily rectal temperature (39.4-39.7°C). Vaccination of boars with type 1 PRRSV significantly reduced the amount of type 1 PRRSV load in blood and semen after challenge with type 1 PRRSV, but barely reduced the amount of type 2 PRRSV load in blood and semen after the type 2 PRRSV challenge. There were no significant differences in the reduction of viremia and seminal shedding of type 1 and type 2 PRRSV between the two commercial vaccines. The seminal shedding of PRRSV is independent of viremia. The reduction of type 1 PRRSV seminal shedding coincided with the appearance of type 1 PRRSV-specific interferon-γ secreting cells (IFN-γ-SC) in vaccinated type 1 PRRSV-challenged boars. The frequencies of type 1 PRRSV-specific IFN-γ-SC induced by type 1 PRRSV vaccine are relatively high compared to type 2 PRRSV-specific IFN-γ-SC induced by the same vaccine which may explain why type 1 PRRSV vaccine is more effective in reducing seminal shedding of type 1 PRRSV when compared to type 2 PRRSV in vaccinated challenged boars. These results provide clinical information on how to reduce seminal shedding of type 1 PRRSV in boars using type 1 PRRSV-modified live vaccine.

Expression of inflammatory cytokines in pigs experimentally infected with Mycoplasma hyopneumoniae.

The expression of interleukin-1 (IL-1), tumour necrosis factor-alpha (TNF-alpha) and IL-6 were studied over a period of 35 days in the lungs of pigs experimentally infected with Mycoplasma hyopneumoniae, by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR), morphometric analysis and in-situ hybridization. Fifteen colostrum-deprived pigs aged 14 days were inoculated intranasally with M. hyopneumoniae. IL-1, TNF-alpha and IL-6 were detected by RT-PCR in the lungs of the infected pigs from 7 days post-inoculation (dpi) onwards, but not in the uninfected control pigs. Concurrent expression of all three cytokines was always observed, in association with lung lesions. Inflammatory cytokine-positive cells were detected in the lungs at 7 dpi, their number increasing at 21dpi, and decreasing thereafter. The results suggest that such cytokines play a role in mediating and regulating inflammation in M. hyopneumoniae infection.

Decreased activity of brush border membrane-bound digestive enzymes in small intestines from pigs experimentally infected with porcine epidemic diarrhea virus.

Brush border membrane-bound digestive enzymes such as disaccharidases (lactase, sucrase, and maltase), leucine aminopeptidase N, and alkaline phosphatase were measured in jejunum from pigs experimentally infected with porcine epidemic diarrhea virus (PEDV). Three piglets from the infected and control groups were euthanized by electrocution and subjected to necropsy at 24, 36, 48, 60, and 72 hours post-inoculation (hpi). The infection of PEDV to jejunum resulted in significant decreases in brush border membrane-bound digestive enzymes such as disaccharidases (lactase, sucrase, and maltase), leucine aminopeptidase N, and alkaline phosphatase. PEDV replication results in massive destruction of villous enterocytes leading to a marked reduction of intestinal epithelial surface and brush border membrane-bound digestive enzyme activity. Reduced enzymatic activity and villous atrophy in the small intestine is thought to result in a maldigestive and malabsorptive diarrhea.

The effects of transplacental porcine circovirus type 2 infection on porcine epidemic diarrhoea virus-induced enteritis in preweaning piglets.

The effects of transplacental porcine circovirus type 2 (PCV2) infection on porcine epidemic diarrhoea virus (PEDV)-induced enteritis were examined in neonatal piglets. Six pregnant sows were randomly allocated to an infected (n=3) or control group (n=3). Three pregnant sows were inoculated intranasally with 6 mL of tissue culture fluid containing 1.2 x 10(5) tissue culture infective doses 50% (TCID(50))/mL of PCV2 strain SNUVR000470 three weeks before the expected farrowing date. Three control pregnant sows were similarly exposed to uninfected cell culture supernatants. Thirty piglets from PCV2-infected sows were randomly assigned to two groups (A and B) of 15 piglets each. Another 30 piglets from noninfected sows were randomly assigned to two groups (C and D) of 15 piglets each. The piglets in groups A and C were dosed orally at three days of age with 2mL of virus stock (1 x 10(6.5) TCID(50)/mL) of the PEDV strain, SNUVR971496, at the third passage. The mean villous height and crypt depth (VH:CD) ratio in PEDV-infected piglets from PCV2-infected sows (group A) were significantly different from those of the PEDV-infected piglets from PCV2 negative sows (group C) at 36, 48, and 72 h post-inoculation (hpi) (P<0.05). In PEDV-infected piglets from PCV2-infected sows (group A), significantly more PEDV nucleic acid was detected in the jejunal tissues (P<0.05) at 24 hpi than in the same tissues of the PEDV-infected piglets from PCV2 negative sows (group C). Thereafter, at 36, 48, 60, and 70 hpi significantly more PEDV nucleic acid (P<0.05) was detected in the jejunal tissues of the PEDV-infected piglets from PCV2 negative sows (group C) than those of the PEDV-infected piglets from the PCV2-infected sows (group A). It is concluded that the clinical course of PEDV disease was markedly affected by transplacental infection of PCV2.

Identification of porcine circovirus type 2 in retrospective cases of pigs naturally infected with porcine epidemic diarrhoea virus.

The identification of porcine circovirus type 2 (PCV2) was studied in fresh intestinal tissues by polymerase chain reaction (PCR) and in formalin-fixed, paraffin-wax-embedded intestinal tissues by in situ hybridisation. The tissues came from pigs naturally infected with porcine epidemic diarrhoea virus (PEDV). A total of 35 (32.7%) of 107 small intestinal samples from pigs naturally infected with PEDV were found to be positive using PCR. Positive signals for PCV2 were detected in 32 (29.9%) of 107 small intestinal samples from pigs naturally infected with PEDV by in situ hybridisation. The distribution of positive cells in the jejunum and ileum was multifocal or patchy. Distinct positive labelling was found throughout the lamina propria in the small intestines. The results of this study indicate that PCV2 is highly prevalent in pigs naturally infected with PEDV.