Complete genome sequence of a novel lytic phage of Xanthomonas oryzae pv. oryzae, the bacterial leaf blight pathogen in rice.

Bacteriophage L522, which infects Xanthomonas oryzae pv. oryzae, was isolated from a paddy leaf sample collected in Long An province, Vietnam. The phage shows myovirus morphology based on transmission electron microscopy. It displays a latent period and burst size of approximately 3 h and 63 new virions per infected cell (PFU/infected cell), respectively. The genome of L522 is 44,497 bp in length, with 52% GC content. Of the 63 genes identified, functions were predicted for 26. No virulence or antibiotic-resistance genes were detected. The results of a BLASTn search showed similarity to a previously reported Xanthomonas phage, with 85% average nucleotide sequence identity and 87.15% query coverage. Thus, this L522 is a representative of a new species in the genus Xipdecavirus.

Biological Characterization and Genomic Analysis of Novel Phages DLDT_So2 and BHDT_So9 Against Pseudomonas solanacearum, an Infectious Agent in Tomato in Vietnam.

Tomato (Solanum lycopersicum L.) is an important grown vegetable in Vietnam. Bacterial wilt caused by Pseudomonas solanacearum has been considered to be an important disease resulting in a harvest loss up to 90% and significant economic loss to farmers. In this study, two bacteriophages DLDT_So2 and BHDT_So9 specific to P. solanacearum were isolated. Morphological analysis indicated that DLDT_So2 and BHDT_So9 had podovirus morphology and were classified into Autographiviridae family. The latent period and burst size of DLDT_So2 was found to be approximately 120 min and 20.0 ± 2.4 virions per infected cell. Meanwhile, the latent period of BHDT_So9 was 140 min with a burst size of 11.5 ± 2.8 virions per infected cell. Of the 23 bacterial strains tested, the phages infected 7/11 strains of P. solanacearum and none of the other bacteria tested were susceptible to the phages. Stability of the phages at different temperatures, pHs, solvents was also investigated. The genomes of DLDT_So2 and BHDT_So9 are 41,341 bp and 41,296 bp and long with a total GC content of 63%, contains 48 and 46 predicted protein-encoding CDSs. No virulence or antibiotic resistance genes were found in the genomes, suggesting they would be useful biocontrol agents against P. solanacearum. Classification of the phage using average nucleotide identity, phylogenetic analysis was also carried out. The two phages represented new species when they had overall average nucleotide identity of < 95%. This is first report of the isolation and characterization of P. solanacearum-specific phages from tomato farms in Vietnam. Supplementary Information: The online version contains supplementary material available at 10.1007/s12088-023-01090-9.

Demonstration of Single-Shot Picosecond Time-Resolved MeV Electron Imaging Using a Compact Permanent Magnet Quadrupole Based Lens.

We present the results of an experiment where a short focal length (∼1.3 cm), permanent magnet electron lens is used to image micron-size features (of a metal sample) with a single shot from an ultrahigh brightness picosecond-long 4 MeV electron beam emitted by a radio-frequency photoinjector. Magnification ratios in excess of 30× were obtained using a triplet of compact, small gap (3.5 mm), Halbach-style permanent magnet quadrupoles with nearly 600 T/m field gradients. These results pave the way towards single-shot time-resolved electron microscopy and open new opportunities in the applications of high brightness electron beams.

Association of the clinical efficacy of vancomycin with the novel pharmacokinetic parameter area under the trough level (AUTL) in elderly patients with hospital-acquired pneumonia.

WHAT IS KNOWN AND OBJECTIVE: The pharmacokinetic-pharmacodynamic parameter that best predicts the efficacy of vancomycin is the ratio of the area under the concentration versus time curve (AUC) to the minimum inhibitory concentration (MIC). A 24-h AUC (AUC24 )/MIC ratio ≥ 400 was recommended in an American consensus review, but vancomycin treatment occasionally fails despite maintenance of AUC24 /MIC ≥ 400. We evaluated the association between clinical efficacy of vancomycin and two novel pharmacokinetic parameters, the 'area under the trough level' (AUTL) and the 'area above the trough level' (AATL), in hospitalized elderly patients with methicillin-resistant Staphylococcus aureus (MRSA) pneumonia. METHODS: The subjects were hospitalized elderly patients who were administered vancomycin for treatment of MRSA pneumonia between 2006 and 2012 at Sasebo Chuo Hospital (Nagasaki, Japan). Pharmacokinetic parameters of vancomycin were estimated for each patient by Bayesian analysis using population pharmacokinetic parameters for Japanese patients. Based on the patient-specific parameters thus obtained, AUC24 values were calculated as the vancomycin dosage divided by vancomycin clearance. AUTL was calculated as the trough serum concentration multiplied by 24 h, whereas AATL was calculated by subtracting AUTL from AUC24 . RESULTS AND DISCUSSION: Logistic regression analysis demonstrated that efficacy of vancomycin was more strongly associated with AUTL than AUC24 . The optimal cut-off value of AUTL was 331 µg∙h/mL, which means that the optimal cut-off value of the trough serum concentration was 13·8 µg/mL. WHAT IS NEW AND CONCLUSION: Efficacy of vancomycin was associated with AUTL, a novel pharmacokinetic parameter. Determining the target AUTL or trough concentration may enhance the efficacy of vancomycin therapy in elderly patients with MRSA pneumonia. Given that nephrotoxicity may increase with a Ctrough in excess of 15 µg/mL, this level should ideally not be exceeded.

Surface-plasmon resonance-enhanced multiphoton emission of high-brightness electron beams from a nanostructured copper cathode.

We experimentally investigate surface-plasmon assisted photoemission to enhance the efficiency of metallic photocathodes for high-brightness electron sources. A nanohole array-based copper surface was designed to exhibit a plasmonic response at 800 nm, fabricated using the focused ion beam milling technique, optically characterized and tested as a photocathode in a high power radio frequency photoinjector. Because of the larger absorption and localization of the optical field intensity, the charge yield observed under ultrashort laser pulse illumination is increased by more than 100 times compared to a flat surface. We also present the first beam characterization results (intrinsic emittance and bunch length) from a nanostructured photocathode.

Changes in Generalized and Particularized Trust and Their Link to Depressive Symptoms: Exploring the Roles of Changes in Self-Rated Health, Family Support and Life Satisfaction Among Chinese Older Adults.

This study aimed to examine whether changes in generalized trust (GT) and particularized trust (PT) predict changes in depressive symptoms (CES-D 8), and whether changes in self-rated health (SRH), family support (FS), and life satisfaction (LS) mediate the relationship between changes in the two types of trust and depressive symptoms in Chinese older adults. Structural equation modeling was employed to analyze two-wave data on 3770 participants aged 65 and over. Our results showed that in a context where GT was low and PT was high, an increase in GT was associated with more depressive symptoms, while an increase in PT was associated with fewer depressive symptoms. As such, GT cannot be viewed as protective against depression in older adults in a given context. LS partially mediated the relationship between changes in PT and depressive symptoms. The findings support psychosocial processes rather than health-problem and support pathways.

Bacteriophage PVN06 protected catfish Pangasianodon hypophthalmus from Edwardsiella ictaluri infection.

Bacillary necrosis of pangasius (BNP) is a disease caused by Edwardsiella ictaluri bacteria in striped catfish Pangasianodon hypophthalmus that results in high mortality rates. To control this disease, bacteriophages have been considered as alternatives to antibiotics. In this study, we applied the lytic bacteriophage PVN06 in striped catfish fingerlings to prevent E. ictaluri infection. In an experimental trial, the phage was administered to fish by feeding phage-coated feed with doses of 7.17±0.09, 8.17±0.09 and 9.17±0.09 log PFU/g feed per day before bacterial infection. Fish were infected by bacteria once with concentrations ranging from 3.01 to 7.01 log CFU/ml tank water. A day after infection, phage treatment resumed at a rate of once per day until the end of the trial. The results of the trial show that bacterial infection caused typical symptoms of BNP in fish with the cumulative fish death rate of 36.7±2.9 to 75.0±5.0%, depending on the bacterial concentration used for infection. Phage treatment with 9.17±0.09 log PFU/g significantly reduced the mortality rate, while treatments with 8.17±0.09 and 7.17±0.09 log PFU/g did not. This phage dose resulted in a 61.7-fold reduction in the toxicity of the bacterial pathogen and the survival rate of 15-23.3% in fish. Our study has demonstrated that the bacteriophage PVN06 protected striped catfish from BNP.

Diverse Bacteriophages Infecting the Bacterial Striped Catfish Pathogen Edwardsiella ictaluri.

Bacteriophages infecting Edwardsiella ictaluri have been less investigated, although the host bacterium is one of the most important fish pathogens causing enteric septicemia of catfish (ESC). We present here two distinctly novel bacteriophages vB_EiM_PVN06 and vB_EiA_PVN09 infecting Edwardsiella ictaluri E1, with their geographical origins from the Mekong Delta, Vietnam. Bacteriophage vB_EiM_PVN06 native to a mud sample reveals complete differences of biological properties with the phage vB_EiA_PVN09 originated from a viscus of a healthy catfish (Pangasianodon hypophthalmus) cultured in the same area. Morphological analyses combined with genomic data indicate that phage vB_EiM_PVN06 is classified to Myoviridae family and shares high similarity with E. ictaluri phage PEi21 genome, while vB_EiA_PVN09 is a member of Teseptimavirus genus, Autographiviridae family, and mostly closes to phage vB_EcoP_IME390. The vB_EiA_PVN09 is a T7-like bacteriophage, which has been firstly found infecting to E. ictaluri, and host range analysis also evidences for the cross-infection of this phage to Escherichia coli K12 and Escherichia coli DH5α. Together, our research highlights the diversity of bacteriophages infecting the pathogen E. ictaluri and suggests further explorations of lytic phages in environmental niches, to be exploited in feasible strategies of phage therapy in ESC disease control.

Inactivation of the retinoblastoma susceptibility gene in human breast cancers.

Mutational inactivation of the retinoblastoma susceptibility (RB) gene, a recessive cancer gene, has been implicated in the genesis of retinoblastoma and certain other human neoplasms. This gene is now shown to be inactivated in two of nine human breast cancer cell lines examined. The RB gene of one cell line had a homozygous internal duplication of a 5-kilobase region containing exons 5 and 6. The RB messenger RNA transcript was correspondingly lengthened, and its translation was probably terminated prematurely due to a shifted reading frame. The other cell line had a homozygous deletion of the RB gene that removed the entire gene beyond exon 2. The RB gene product, pp110RB, was not detectable in either cell line by immuno-precipitation with specific antibodies. These findings are significant in relation to proposed genetic mechanisms of breast cancer formation.

Tumor suppressor activity of RB and p53 genes in human breast carcinoma cells.

Breast cancer is the most common cancer of women in Western countries. Various genetic alterations have been implicated in its development. Two tumor suppressor genes, the retinoblastoma susceptibility gene (RB) and the gene encoding the p53 protein, are frequently found to be deleted or mutated in breast cancer cell lines and primary tumor samples. Breast carcinoma cell lines MDA-MB468 and BT549 both harbor partial RB gene deletions as well as point mutations of their p53 genes, thus providing an excellent model system for testing the roles played by these two genes in the oncogenesis of breast cancer. Single copies of wild-type RB or p53 were delivered to these cells by retrovirus-mediated gene transfer. Restoration of RB expression in cells reduced their ability to grow in soft agar and their tumorigenicity in nude mice, although no significant alteration of growth rate in culture could be detected. Introduction of wild-type p53 into these cells exerted a similar effect on the neoplastic phenotypes. This effect occurred even in the presence of their endogenous mutated p53 alleles, which argues for the phenotypical dominance of the wild-type p53 over mutated p53 during human oncogenesis. The ability of RB and p53 genes to suppress the tumorigenicity of breast carcinoma cells provides functional evidence that deletion or mutational inactivation of tumor suppressor genes represents an important step in the genesis of breast cancer.

Induction of c-fos by prostaglandin F2 alpha in human ciliary smooth muscle cells.

PURPOSE: To evaluate the induction of the proto-oncogene c-fos in cultured human ciliary muscle cells by prostaglandin F2 alpha (PGF2 alpha) by 17-phenyltrinor-PGF 2 alpha. METHOD: Human ciliary muscle cells were grown to confluency in monolayer cell culture, placed in medium containing 1% fetal bovine serum for 5 days, treated by addition of PGF2 alpha or the trinor derivative, fixed, and then immunocytochemically stained using an antibody to c-Fos, the protein product of the translation of c-fos. RESULTS: After treatment with either agonist, the mean induction score (proportion of brightly immunostained nuclei) increased to a maximal level during the first hour and returned to basal levels 4 to 8 hours after treatment. Increasing the agonist concentration increased the maximal level, but had no effect on the time course of the response. The dose responses after 1 hour of treatment with PGF2 alpha or 17-phenyltrinor-PGF2 alpha increased similarly between 1.6 x 10(-9) M and 2 x 10(-7) M. When treated with 1 x 10(-6) M of either agonist, however, the induction was half that obtained at 2 x 10(-7) M. CONCLUSION: Exposure of ciliary smooth muscle cells to either PGF2 alpha or 17-phenyltrinor-PGF2 alpha induces an immediate early gene expression response that is similar to c-Fos induction in other cell systems. These results establish the basis for future investigations evaluating the potential role of c-fos induction in mediating the effects of PGF2 alpha on uveoscleral outflow.

Population-based investigation of valproic acid relative clearance using nonlinear mixed effects modeling: influence of drug-drug interaction and patient characteristics.

Nonlinear mixed effects modeling (NONMEM) was used to estimate the effects of drug-drug interaction on valproic acid relative clearance values using 792 serum levels gathered from 400 pediatric and adult patients with epilepsy (age range, 0.3-54.8 years) during their clinical routine care. Patients received valproic acid as monopharmacy or in combination with either the antiepileptic drugs, phenobarbital, or carbamazepine. The final model describing valproic acid relative clearance was CL (mL/hr/kg) = 15.6.TBW (kg)-0.252.DOSE (mg/kg/day)0.183.0.898GEN.COPB.COCBZ, where COPB equals 1.10 if the patient is treated with phenobarbital, a value of unity otherwise, and COCBZ equals 0.769.DOSE (mg/kg/day)0.179 if the patient is treated with carbamazepine, a value of unity otherwise. Valproic acid relative clearance was highest in the very young and decreased in a weight-related fashion in children, with minimal changes observed in adults. This pattern was consistent whether valproic acid was administered alone or coadministered with phenobarbital or carbamazepine. When valproic acid was coadministered with phenobarbital or carbamazepine, valproic acid relative clearance increased as compared with that in monopharmacy. Its magnitude in the presence of carbamazepine increased in a valproic acid daily dose-related fashion. Concomitant administration of phenobarbital and valproic acid resulted in a 10% increase on valproic acid relative clearance. The clearance in female patients was approximately 10% less than that in male patients.

Molecular cloning of an immunogenic and acid-induced isocitrate dehydrogenase gene from Coxiella burnetii.

The Coxiella burnetii icd gene encoding an immunogenic dimeric NADP(+)-dependent isocitrate dehydrogenase (IDH) was cloned by screening a C. burnetii genomic library with a human positive serum and sequenced. The predicted gene product consists of 427 amino acids (M(r) = 46,600) and showed high identity to the IDHs of Escherichia coli (74%), Salmonella enterica (73%) and IDH-I of Vibrio sp. (71%). The cloned gene complemented an icd-defective E. coli mutant producing a recombinant IDH that had the same biochemical properties as the enzyme from purified C. burnetii. Unlike the homologs from other bacteria, the cloned enzyme was expressed to the highest level in low pH conditions. This distinct property of the cloned IDH suggests that C. burnetii icd gene may have a role in the adaptation of the organism to the harsh acidic environment of the eucaryotic phagolysosomes.

Cisplatin-induced vomiting depends on circadian timing.

We examined whether the clock time of cisplatin plus antiemetic and diuretic administration affects the amount of cisplatin-associated emesis and severity of renal toxicity. We treated 22 patients with urogenital cancer with two courses of chemotherapy containing 70 mg/m2 of cisplatin. Cisplatin together with furosemide was administered in the morning (05:00) or evening (17:00) during two courses 1 month apart in a crossover fashion. Ondansetron was given either before or after cisplatin to control nausea and vomiting. The number of vomiting episodes, serum creatinine, serum urea nitrogen (BUN), creatinine clearance, and urinary beta-N-acetyl glucosamidase (NAG) concentration were evaluated before and after each treatment course. Regardless of the timing of ondansetron, morning compared to evening cisplatin was always associated with greater vomiting in the first treatment course. However, prophylactic administration of ondansetron markedly diminished the impact of the clock time of cisplatin administration. Serum creatinine transiently decreased rather than increased 14 days after cisplatin and furosemide administration, while NAG excretion increased 3 days after cisplatin and furosemide administration. In the first course, serum creatinine levels were similar regardless of the clock time of cisplatin and furosemide administration. However, in the second course, serum creatinine rose in patients given evening cisplatin and furosemide, while it remained unchanged in those given morning cisplatin and furosemide. Moreover, the first course morning cisplatin and furosemide treatment was associated with less change in NAG excretion (less kidney toxicity) than the first course of evening cisplatin and furosemide treatment. The second course evening cisplatin and furosemide treatment was associated with an increase in NAG excretion compared to the first course of treatment, while morning cisplatin and furosemide treatment in the second course showed less change in NAG excretion compared to the first course. The clock time of cisplatin administration had an impact on the frequency of emesis. Prophylactic ondansetron, however, diminished the time-of-day dependency of cisplatin-induced vomiting. Administration of cisplatin and furosemide in the morning rather than evening appears to cause less renal damage, and this damage may be further reduced with aggressive hydration and routine administration of furosemide.

Effects of dosing time and schedule on cisplatin-induced nephrotoxicity in rats.

Renal dysfunction induced by a single injection of cisplatin depends on the timing of the dose. However, the effects of repeated administration of cisplatin on time-dependent toxicity have not been evaluated despite the fact that in clinical practice high doses are repeatedly injected at intervals or low doses are administered daily. We studied chrononephrotoxicity in rats after weekly or daily cisplatin injections. Weekly high doses (5 mg kg(-1)) or daily low doses (1.2 mg kg(-1)) of cisplatin were injected at four time points (3, 9, 15 and 21 h after the light was turned on (HALO)) for 3 weeks. Changes in body weight after weekly cisplatin administration were independent of the timing of the doses. In the group that received daily cisplatin, the loss in body weight in the 3 HALO group was smaller than in animals receiving injections at 15 and 21 HALO (P < 0.05 and 0.001, respectively). The blood urea nitrogen and serum creatinine levels in the control rats showed a significant circadian change (peak at 15 HALO and trough at 9 HALO), but these parameters were markedly elevated in both trials and their circadian variations were disturbed. In conclusion, cisplatin-induced nephrotoxicity was the lowest at 3HALO compared with other time points of both dose regimens.

Time-dependent nephrotoxicity associated with daily administration of cisplatin in mice.

The chronopharmacokinetics and chronopharmacodynamics of cisplatin were studied in a mouse model to reveal the mechanisms of dosing time-dependent nephrotoxicity induced by daily administration. Chronotoxicity was tested by daily intraperitoneal injections of cisplatin (6mg kg(-1)) for 5 days at four time points (04:00, 10:00, 16:00 and 22:00h) in BALB/c mice (n = 6 in each group). After following the changes in body weight, serum concentrations of blood urea nitrogen (BUN) and creatinine obtained on day 6 were compared. The results showed diurnal variations in cisplatin toxicity, with the 04:00 and 16:00h time points the best and the worst, respectively. We then measured platinum concentrations in blood, liver and kidney and compared the results of the 04:00 and 16:00 h groups (n = 4 in each group). Kidney sensitivity to cisplatin alone, lipopolysaccharide (LPS) alone, cisplatin with LPS and saline (control) were also measured using a tissue culture system (a measurement system of interleukin-6 (IL-6) production) between the 04:00 and the 16:00 h groups (n = 4 in each group). These results showed no significant difference in platinum accumulation between the two groups. IL-6 production was higher in the 16:00 h group than in the 04:00 h group after saline injection alone (P < 0.05). Cisplatin treatment alone did not increase IL-6 production. However, IL-6 levels were markedly augmented by cisplatin with LPS. In conclusion, chrononephrotoxicity induced by daily cisplatin administration does not only depend on cisplatin accumulation, but might also depend on kidney sensitivity to diurnal variations in inflammatory reaction without direct cisplatin toxicity.

Advances in the understanding of Coxiella burnetii infection in Japan.

Q fever is a zoonotic disease caused by a rickettsia Coxiella burnetii. Since its first description in 1937, the disease has been found to be present in most countries of the world. Serological evidences of Q fever in humans and coxiellosis in animals were reported in Japan in the 1950s, however, systematic studies of the disease did not begin until the report of isolation of C. burnetii from an acute Q fever patient in 1989. In addition to the extensive information about epidemiology of the disease, the understanding of Japanese isolates of C. burnetii is increasing rapidly in recent years. In this review, the epidemiology of the disease along with some characteristics of isolates of C. burnetii in Japan is summarized in five sections, i.e., coxiellosis, Q fever, modes of spread of the infection, laboratory diagnosis of the infection and some characteristics of Japanese isolates. This review includes some recent, unpublished data from our and other groups.

Antigenic characteristic of the lipopolysaccharides of Coxiella burnetii isolates.

Lipopolysaccharides (LPSs) of 8 isolates of Coxiella burnetii from a variety of clinical and geographical sources could be divided into four groups based on molecular heterogeneity in silver-stained sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) profiles in the region of the 10 to 17 kDa. The lipopolysaccharide of group 1 was identified on isolates from acute Q fever patient, milk and tick. The three remaining groups were primarily found on isolates from human cases of chronic Q fever. These LPSs shared many antigenic epitopes, as determined by immunoblotting with mouse anti-C. burnetii antisera.

Prevalence of Coxiella burnetii infection in dairy cattle with reproductive disorders.

The prevalence of Coxiella burnetii infection in 207 cattle with reproductive disorders was studied by using an indirect immunofluorescence (IF) test, nested polymerase chain reaction (PCR) and isolation. IF antibodies to phase I and phase II antigens of C. burnetii were found in 122 (58.9%) and 125 (60.4%) of the sera, respectively, and PCR-positives were found in 8 (3.9%) of the sera and in 51 (24.6%) of the milk samples. In addition, C. burnetii was isolated from 51 (24.6%) of the milk samples by inoculating laboratory mice. The results indicate that the IF test plus PCR are useful in the diagnosis of bovine coxiellosis. It is difficult to deny that dairy cattle with reproductive disorders would be one of the important reservoirs of C. burnetii responsible for infection in both animal and human populations in Japan.

Sequence comparison of the VP2 variable region of infectious bursal disease virus isolates from Vietnam.

The variable region in the VP2 gene of twenty-three infectious bursal disease virus (IBDV) isolates, collected in Vietnam in 1997 and 1998, was amplified as cDNA by using the reverse transcription-polymerase chain reaction and sequenced. Analysis of amino acid substitutions and phylogenetic relationships of the deduced amino acid sequences (residues 206-350) showed that the nineteen Vietnamese vv IBDVs clustered with the European vv IBDVs, Japanese vv IBDVs and Chinese vv strains, and that the four vietnamese virulent strains were closely related to European virulent strain 52/70. These results suggest that Vietnamese vv IBDVs, European vv IBDVs, Japanese vv IBDVs and Chinese vv strains have the same origin.