Intravesical treatment of advanced urothelial bladder cancers with oncolytic HSV-1 co-regulated by differentially expressed microRNAs.

Urothelial bladder cancer is the most common malignancy of the urinary tract. Although most cases are initially diagnosed as non-muscle-invasive, more than 80% of patients will develop recurrent or metastatic tumors. No effective therapy exists currently for late-stage metastatic tumors. By intravesical application, local administration of oncolytic Herpes Simplex virus (oHSV-1) can provide a promising new therapy for this disease. However, its inherent neurotoxicity has been a perceived limitation for such application. In this study, we present a novel microRNA-regulatory approach to reduce HSV-1-induced neurotoxicity by suppressing viral replication in neurons while maintaining oncolytic selectivity toward urothelial tumors. Specifically, we designed a recombinant virus that utilizes differentially expressed endogenous microR143 (non-cancerous, ubiquitous) and microR124 (neural-specific) to regulate expression of ICP-4, a gene essential for HSV-1 replication. We found that expression of ICP-4 must be controlled by a combination of both miR143 and miR124 to achieve the most effective attenuation in HSV-1-induced toxicity while retaining maximal oncolytic capacity. These results suggest that interaction between miR143 and miR124 may be required to successfully regulate HSV-1 replication. Our resent study is the first proof-in-principle that miRNA combination can be exploited to fine-tune the replication of HSV-1 to treat human cancers.

Real-space observation of a two-dimensional skyrmion crystal.

Crystal order is not restricted to the periodic atomic array, but can also be found in electronic systems such as the Wigner crystal or in the form of orbital order, stripe order and magnetic order. In the case of magnetic order, spins align parallel to each other in ferromagnets and antiparallel in antiferromagnets. In other, less conventional, cases, spins can sometimes form highly nontrivial structures called spin textures. Among them is the unusual, topologically stable skyrmion spin texture, in which the spins point in all the directions wrapping a sphere. The skyrmion configuration in a magnetic solid is anticipated to produce unconventional spin-electronic phenomena such as the topological Hall effect. The crystallization of skyrmions as driven by thermal fluctuations has recently been confirmed in a narrow region of the temperature/magnetic field (T-B) phase diagram in neutron scattering studies of the three-dimensional helical magnets MnSi (ref. 17) and Fe(1-x)Co(x)Si (ref. 22). Here we report real-space imaging of a two-dimensional skyrmion lattice in a thin film of Fe(0.5)Co(0.5)Si using Lorentz transmission electron microscopy. With a magnetic field of 50-70 mT applied normal to the film, we observe skyrmions in the form of a hexagonal arrangement of swirling spin textures, with a lattice spacing of 90 nm. The related T-B phase diagram is found to be in good agreement with Monte Carlo simulations. In this two-dimensional case, the skyrmion crystal seems very stable and appears over a wide range of the phase diagram, including near zero temperature. Such a controlled nanometre-scale spin topology in a thin film may be useful in observing unconventional magneto-transport effects.

Azapirones for Attention Deficit Hyperactivity Disorder: A Systematic Review.

INTRODUCTION: No meta-analysis has evaluated azapirones (serotonin1A receptor partial agonists) as anxiolytics for attention deficit hyperactivity disorder (ADHD). METHODS: Randomized controlled trials (RCTs) and single-arm trials published before October 27, 2015 were retrieved from major healthcare databases and clinical trial registries. Relative risk and 95% confidence intervals were calculated. RESULTS: 5 RCTs (n=429) and 3 single-arm studies (n=70) were identified. 3 RCTs compared buspirone vs. methylphenidate in children/adolescents, one buspirone patches vs. placebo patches in children/adolescents, and one atomoxetine plus buspirone vs. atomoxetine vs. placebo in adults. The single-arm studies were buspirone trails in children/adolescents. All-cause discontinuation rates and adverse events did not differ between pooled buspirone and methylphenidate groups. No other meta-analyses of buspirone efficacy and safety vs. comparators were conducted due to insufficient data. 2 RCTs found no significant differences in parent and teacher ADHD-Rating Scale total scores between buspirone and methylphenidate, while one reported that methylphenidate improved parent and teacher ADHD-RS total scores vs. buspirone. DISCUSSION: It remains unclear whether buspirone use has benefit for ADHD patients and therefore further evidence is needed for better clinical use of buspirone in patients with ADHD.

Quasicrystal as a Catalyst for the Synthesis of Carbon Nanotubes.

The present report describes the catalytic activity of mechanically activated nano quasicrystalline Al65Cu20Fe15 and related nano crystalline Al50Cu28Fe22 for the synthesis of carbon nanotubes (CNTs). CNTs are synthesized by catalytic decomposition of ethanol through nano quasicrystalline Al65Cu20Fe15 and related crystalline Al50Cu28Fe22 alloys as a catalyst. The synthesized multi-walled CNTs exhibits tube diameter ranging from 5 to 25 nm. The synthesized CNTs are characterized by scanning and transmission electron microscopy. It is found that Al65Cu20Fe15 nanoquasicystal shows better catalytic behaviour as compared to nano-crystalline Al50Cu28Fe22 alloys for decomposition of ethanol during the synthesis of multi-walled CNTs.

Effect of the mass of immunoglobulin (Ig)G intake and age at first colostrum feeding on serum IgG concentration in Holstein calves.

Forty-four Holstein calves (19 male and 25 female) were used in this study of the relationships among age at first colostrum feeding, IgG intake, and apparent efficiency of IgG absorption. Time of birth was recorded for each calf and the calves were fed colostrum ad libitum after birth at either 0930 or 1630 h. Blood samples were collected immediately before and 24h after colostrum feeding. Data from calves were then categorized into 4 groups representing time from birth to colostrum feeding: A=fed within 1h (n=5); B=fed from 1 to 6h (n=10); C=fed from 6 to 12 h (n=21); and D=fed from 12 to 18 h (n=8) after birth. Average total intake of colostrum was 3.6 ± 0.1L. Over 80% of the calves consumed ≥3 L of colostrum. Apparent efficiency of IgG absorption declined remarkably 12 h after birth. Mean apparent efficiency of absorption of IgG in group D (15.8 ± 3.0%) was lower than that in groups A (30.5 ± 3.9%) and B (27.4 ± 2.8%). Serum IgG concentration in calves was positively correlated with IgG intake in all groups. The relationship between mass of IgG consumed and calf serum IgG at 24 h was different for each time of colostrum feeding, with only limited differences observed between groups A and B. We concluded that failure of transfer of passive immunity in newborn calves may be avoided if calves consume ≥3 L of colostrum with IgG concentration >40 mg/mL within 6 h after birth. These findings help define the opportunity to minimize failure of transfer of passive immunity to newborn calves under management programs similar to those used on commercial dairy farms.

Structural phase transition and magnetic-field effect on the modulated structure in GdBaCo2O5+δ (δ < 0.5).

We investigated the crystal structures of an ordered perovskite-type cobaltate, GdBaCo2O(5+δ) (δ < 0.5), at elevated temperatures by transmission electron microscopy. Above the magnetic ordering temperature, we observed a first-order structural phase transition between the low-temperature tetragonal 3a(p) × 3a(p) and high-temperature orthorhombic 1a(p) × 2a(p) superstructure phases (where a(p) is the perovskite-unit cell). Upon the application of a magnetic field, an incommensurate phase emerges around the structural phase-transition temperature, which indicates a magnetic-field-induced structural phase transition via no magnetic ordering in the ordered perovskite-type cobaltate.

Investigation of removal and inactivation efficiencies of human sapovirus in drinking water treatment processes by applying an in vitro cell-culture system.

Here, we examined the efficiencies of drinking water treatment processes for the removal and inactivation of human sapovirus (HuSaV). We applied a recently developed in vitro cell-culture system to produce purified solutions of HuSaV containing virus concentrations high enough to conduct virus-spiking experiments, to develop an integrated cell culture-polymerase chain reaction (ICC-PCR) assay to quantify the infectivity of HuSaV, and to conduct virus-spiking experiments. In virus-spiking coagulation-sedimentation-rapid sand filtration (CS-RSF) and coagulation-microfiltration (C-MF) experiments, HuSaV removals of 1.6-3.7-log10 and 1.2->4.3-log10, respectively, were observed. The removal ratios observed with CS-RSF were comparable and correlated with those of murine norovirus (MNV, a widely used surrogate for human noroviruses) and pepper mild mottle virus (PMMoV, a potential surrogate for human enteric viruses in physical and physicochemical drinking water treatment processes), and those observed with C-MF were higher than but still correlated with those of MNV and PMMoV, indicating that MNV and PMMoV are both potential surrogates for HuSaV in CS-RSF and C-MF. For astrovirus (AstV, a representative human enteric virus), removal ratios of 1.8-3.3-log10 and 1.1->4.0-log10 were observed with CS-RSF and C-MF, respectively. The removal ratios of AstV observed with CS-RSF were comparable and correlated with those of PMMoV, and those observed with C-MF were higher than but still correlated with those of PMMoV, indicating that PMMoV is a potential surrogate for AstV in CS-RSF and C-MF. When the efficacy of chlorine treatment was examined by using the developed ICC-PCR assay, 3.8-4.0-log10 inactivation of HuSaV was observed at a CT value (free-chlorine concentration [C] multiplied by contact time [T]) of 0.02 mg-Cl2·min/L. The infectivity reduction ratios of HuSaV were comparable with those of MNV. For AstV, 1.3-1.7-log10 and >3.4-log10 inactivation, as evaluated by ICC-PCR, was observed at CT values of 0.02 and 0.09 mg-Cl2·min/L, respectively. These results indicate that HuSaV and AstV are both highly sensitive to chlorine treatment and more sensitive than a chlorine-resistant virus, coxsackievirus B5 (1.3-log10 inactivation at a CT value of 0.4 mg-Cl2·min/L, as evaluated by the ICC-PCR assay).

Relationships among Grip Strength Measurement, Response Time, and Frailty Criteria.

BACKGROUND: Muscle response in older adults is believed to decrease with maximal muscle strength, although it has not been adequately assessed; further, the relationship between frailty and muscle response remains unexamined. OBJECTIVES: This study aimed to develop a practical method for measuring muscle response using grip strength in older adults and to clarify the relationship between frailty and grip strength response. DESIGN, SETTING, AND PARTICIPANTS: We performed a cross-sectional, clinical, observational study. A total of 248 patients (94 men and 154 women, mean age: 78.2 years) who visited the outpatient unit in the Integrated Healthy Aging Clinic of our Hospital for the first time were enrolled. MEASUREMENTS: Using a grip strength measuring device originally developed by us, we measured grip strength response indices, such as reaction time, time constant, rate of force development (response speed), and maximum grip strength. Grip strength response indices were compared among three groups (robust, pre-frail, and frail) according to the Fried and Kihon checklist assessments for frailty. RESULTS: Based on Fried's assessment, marked differences were found between groups not only in maximal grip strength but also in response time and response speed. Based on the Kihon checklist assessment, there was no significant difference in response time; however, a considerable difference in response speed for the left hand was observed. Moreover, according to the Kihon checklist assessment, some cases showed differences in muscle response although not in maximal muscle strength. CONCLUSIONS: The response speed of grip strength was suggested to decrease with frailty. The results suggest that measurement of grip strength response in both hands is useful to examine the relationship between frailty and grip strength response.

Activation of Lactate Receptor Positively Regulates Skeletal Muscle Mass in Mice.

G protein-coupled receptor 81 (GPR81), a selective receptor for lactate, expresses in skeletal muscle cells, but the physiological role of GPR81 in skeletal muscle has not been fully elucidated. As it has been reported that the lactate administration induces muscle hypertrophy, the stimulation of GPR81 has been suggested to mediate muscle hypertrophy. To clarify the contribution of GPR81 activation in skeletal muscle hypertrophy, in the present study, we investigated the effect of GPR81 agonist administration on skeletal muscle mass in mice. Male C57BL/6J mice were randomly divided into control group and GPR81 agonist-administered group that received oral administration of the specific GPR81 agonist 3-Chloro-5-hydroxybenzoic acid (CHBA). In both fast-twitch plantaris and slow-twitch soleus muscles of mice, the protein expression of GPR81 was observed. Oral administration of CHBA to mice significantly increased absolute muscle weight and muscle weight relative to body weight in the two muscles. Moreover, both absolute and relative muscle protein content in the two muscles were significantly increased by CHBA administration. CHBA administration also significantly upregulated the phosphorylation level of p42/44 extracellular signal-regulated kinase-1/2 (ERK1/2) and p90 ribosomal S6 kinase (p90RSK). These observations suggest that activation of GRP81 stimulates increased the mass of two types of skeletal muscle in mice in vivo. Lactate receptor GPR81 may positively affect skeletal muscle mass through activation of ERK pathway.

Near room-temperature formation of a skyrmion crystal in thin-films of the helimagnet FeGe.

The skyrmion, a vortex-like spin-swirling object, is anticipated to play a vital role in quantum magneto-transport processes such as the quantum Hall and topological Hall effects. The existence of the magnetic skyrmion crystal (SkX) state was recently verified experimentally for MnSi and Fe(0.5)Co(0.5)Si by means of small-angle neutron scattering and Lorentz transmission electron microscopy. However, to enable the application of such a SkX for spintronic function, materials problems such as a low crystallization temperature and low stability of SkX have to be overcome. Here we report the formation of SkX close to room temperature in thin-films of the helimagnet FeGe. In addition to the magnetic twin structure, we found a magnetic chirality inversion of the SkX across lattice twin boundaries. Furthermore, for thin crystal plates with thicknesses much smaller than the SkX lattice constant (as) the two-dimensional SkX is quite stable over a wide range of temperatures and magnetic fields, whereas for quasi-three-dimensional films with thicknesses over as the SkX is relatively unstable and observed only around the helical transition temperature. The room-temperature stable SkX state as promised by this study will pave a new path to designing quantum-effect devices based on the controllable skyrmion dynamics.

Relaxation dynamics of photoexcited excitons in rubrene single crystals using femtosecond absorption spectroscopy.

The relaxation dynamics of an exciton in rubrene was investigated by femtosecond absorption spectroscopy. Exciton relaxation to a self-trapped state occurs via the coherent oscillation with 78 cm(-1) due to a coupled mode of molecular deformations with phenyl-side-group motions and molecular displacements. From the temperature dependence of the decay time of excitons, the energy necessary for an exciton to escape from a self-trapped state is evaluated to be ~35 meV (~400 K). As a result, a self-trapped exciton is stable at low temperatures. At room temperature, excitons can escape from a self-trapped state and, subsequently, they are dissociated to charged species. The exciton dissociation mechanism is discussed on the basis of the results.

Anomalously large anisotropic magnetoresistance in a perovskite manganite.

The signature of correlated electron materials (CEMs) is the coupling between spin, charge, orbital and lattice resulting in exotic functionality. This complexity is directly responsible for their tunability. We demonstrate here that the broken symmetry, through cubic to orthorhombic distortion in the lattice structure in a prototype manganite single crystal, La(0.69)Ca(0.31)MnO(3), leads to an anisotropic magneto-elastic response to an external field, and consequently to remarkable magneto-transport behavior. An anomalous anisotropic magnetoresistance (AMR) effect occurs close to the metal-insulator transition (MIT) in the system, showing a direct correlation with the anisotropic field-tuned MIT in the system and can be understood by means of a simple phenomenological model. A small crystalline anisotropy stimulates a "colossal" AMR near the MIT phase boundary of the system, thus revealing the intimate interplay between magneto- and electronic-crystalline couplings.

Evaluation of reduction efficiencies of pepper mild mottle virus and human enteric viruses in full-scale drinking water treatment plants employing coagulation-sedimentation-rapid sand filtration or coagulation-microfiltration.

Here, we evaluated the reduction efficiencies of indigenous pepper mild mottle virus (PMMoV, a potential surrogate for human enteric viruses to assess virus removal by coagulation-sedimentation-rapid sand filtration [CS-RSF] and coagulation-microfiltration [C-MF]) and representative human enteric viruses in four full-scale drinking water treatment plants that use CS-RSF (Plants A and B) or C-MF (Plants C and D). First, we developed a virus concentration method by using an electropositive filter and a tangential-flow ultrafiltration membrane to effectively concentrate and recover PMMoV from large volumes of water: the recovery rates of PMMoV were 100% when 100-L samples of PMMoV-spiked dechlorinated tap water were concentrated to 20 mL; even when spiked water volume was 2000 L, recovery rates of >30% were maintained. The concentrations of indigenous PMMoV in raw and treated water samples determined by using this method were always above the quantification limit of the real-time polymerase chain reaction assay. We therefore were able to determine its reduction ratios: 0.9-2.7-log10 in full-scale CS-RSF and 0.7-2.9-log10 in full-scale C-MF. The PMMoV reduction ratios in C-MF at Plant C (1.0 ± 0.3-log10) were lower than those in CS-RSF at Plants A (1.7 ± 0.5-log10) and B (1.4 ± 0.7-log10), despite the higher ability of MF for particle separation in comparison with RSF owing to the small pore size in MF. Lab-scale virus-spiking C-MF experiments that mimicked full-scale C-MF revealed that a low dosage of coagulant (polyaluminum chloride [PACl]) applied in C-MF, which is determined mainly from the viewpoint of preventing membrane fouling, probably led to the low reduction ratios of PMMoV in C-MF. This implies that high virus reduction ratios (>4-log10) achieved in previous lab-scale virus-spiking C-MF studies are not necessarily achieved in full-scale C-MF. The PMMoV reduction ratios in C-MF at Plant D (2.2 ± 0.6-log10) were higher than those at Plant C, despite similar coagulant dosages. In lab-scale C-MF, the PMMoV reduction ratios increased from 1-log10 (with PACl [basicity 1.5], as at Plant C) to 2-4-log10 (with high-basicity PACl [basicity 2.1], as at Plant D), suggesting that the use of high-basicity PACl probably resulted in higher reduction ratios of PMMoV at Plant D than at Plant C. Finally, we compared the reduction ratios of indigenous PMMoV and representative human enteric viruses in full-scale CS-RSF and C-MF. At Plant D, the concentrations of human norovirus genogroup II (HuNoV GII) in raw water were sometimes above the quantification limit; however, whether its reduction ratios in C-MF were higher than those of PMMoV could not be judged since reduction ratios were >1.4-log10 for HuNoV GII and 2.3-2.9-log10 for PMMoV. At Plant B, the concentrations of enteroviruses (EVs) and HuNoV GII in raw water were above the quantification limit on one occasion, and the reduction ratios of EVs (>1.2-log10) and HuNoV GII (>1.5-log10) in CS-RSF were higher than that of PMMoV (0.9-log10). This finding supports the usefulness of PMMoV as a potential surrogate for human enteric viruses to assess virus removal by CS-RSF.

Physical Frailty and Hemoglobin-to-Red Cell Distribution Width Ratio in Japanese Older Outpatients.

The Frailty screening should be widely performed; however, simple and inexpensive biomarkers are missing. Biomarkers that can be routinely assessed in many patients are desirable. Recently, the hemoglobin-to-red cell distribution width ratio (Hb/RDW, HRR) has been suggested as a new prognostic marker and has been reported to be associated with inflammation, one of the factors contributing to frailty. Therefore, we aimed to address the role of HRR in frailty among 557 older outpatients (aged 65-96 years). Frailty was assessed using the Japanese version of the Cardiovascular Health Study criteria, and HRR was calculated from clinical records. Participants were classified into five groups based on a sex-stratified quintile of HRR (Q1-Q5). Of the participants, 20.3% were frail. Using multiple logistic regression models with the Q5 group as a reference, after adjusting for sex, age, body mass index, polypharmacy, pre-orthopedic surgery, and the use of iron medications, the multivariable-adjusted odds ratios (95% confidence intervals) of the Q4 to Q1 groups were 0.92 (0.58-1.47), 1.04 (0.67-1.61), 1.29 (0.84-1.96), and 1.85 (1.22-2.82), respectively, indicating that a lower HRR was significantly associated with frailty. The robustness of these results was also shown in the multiple imputation analysis. The results suggest that HRR measurement may be one of the indicators to identify frail older adults in routine practice.

Association between Sarcopenia and Fall Risk According to the Muscle Mass Adjustment Method in Japanese Older Outpatients.

We investigated whether the association between sarcopenia and fall risk (FR) differs according to the muscle mass adjustment method in 357 outpatients who were not disabled in the activities of daily living or indicated for orthopedic surgery. Sarcopenia was diagnosed by the Asian Working Group for Sarcopenia (AWGS) 2019 criteria, using adjusting methods of muscle mass by height squared (ht2), body mass index (BMI), or any of these (i.e., ht2-adjusted Sarc", "BMI-adjusted Sarc", or "ht2 and/or BMI-adjusted Sarc, respectively). FR was defined by FR index ≥10. There were 111, 105, and 157 participants with ht2-adjusted, BMI-adjusted, and ht2 and/or BMI-adjusted Sarc, respectively. After multivariable adjustment, ht2 and/or BMI-adjusted Sarc was the most closely associated with FR [odds ratio and 95% confidence interval: 2.94, 1.75-4.93]. Our data suggest that the sarcopenia definition using low ASM/ht2 and/or ASM/BMI muscle mass might better predict adverse outcomes in older patients.

Quantifying Muscle Mass by Adjusting for Body Mass Index Is the Best for Discriminating Low Strength and Function in Japanese Older Outpatients.

OBJECTIVES: To investigate the association of three muscle mass adjustment methods with low muscle strength (MS) and low physical function (PF) defined by the Asian Working Group for Sarcopenia (AWGS) 2019 criteria. DESIGN: Cross-sectional study. SETTING: Clinical setting. PARTICIPANTS: We included 361 outpatients (77.9 ± 5.9 years) without scheduled orthopedic surgery or activities of daily living disability. MEASUREMENTS: Appendicular skeletal muscle mass (ASM) was measured by dual-energy X-ray absorptiometry, then divided by height-square (ht2), body weight, and body mass index (BMI) to calculate the ASM indexes. We assessed grip strength, gait speed, short physical performance battery, and five-time chair stand test. Low MS and low PF were defined by the AWGS2019 criteria. To compare the association of three muscle mass adjustments with low MS and low PF, multiple logistic regression analysis was performed, adjusted for age in each sex. RESULTS: Participants with low MS was 31.5%, low PF was 50.1%. After adjustment, only ASM/BMI was significantly associated with all independent variables, such as low MS, low PF, and either of these, with the odds ratios of 2.09, 2.08, and 2.50 for males; and 1.87, 2.43, and 2.71 for females, respectively. CONCLUSION: Our findings suggest that ASM/BMI is best associated with low MS and low PF in older Japanese outpatients. Longitudinal outcome studies are needed to confirm our findings.

Evaluation of norovirus removal performance in a coagulation-ceramic microfiltration process by using recombinant norovirus virus-like particles.

Norovirus (NV) is a prototype strain of a group of human caliciviruses responsible for epidemic outbreaks of acute gastroenteritis worldwide. Because of the lack of a cell culture system or an animal model for this virus, studies on drinking water treatment such as separation and disinfection processes are still hampered. In the present study, we investigated NV removal performance as particles during a coagulation-ceramic microfiltration (MF) process by using recombinant NV virus-like particles (rNV-VLPs), which are morphologically and antigenically similar to native NV. We also experimentally investigated the behaviors of two widely accepted surrogates for pathogenic waterborne viruses, bacteriophages Qbeta and MS2, for comparison with the behavior of rNV-VLPs. More than 4-log removal was observed for rNV-VLPs with a 1.08 mg-Al/L dose of polyaluminium chloride in the coagulation-ceramic MF process. This high removal ratio of rNV-VLPs satisfies the U.S. Environmental Protection Agency requirement of 4-log removal or inactivation. In addition, the removal ratios of Qbeta and MS2 were approximately 2-log and 1-log, smaller than the ratio of rNV-VLPs. Accordingly, both bacteriophages have the potential to become appropriate surrogates for native NV in the coagulation-ceramic MF process, and, of the two, Qbeta is the more conservative surrogate.

Geosmin and 2-methylisoborneol adsorption on super-powdered activated carbon in the presence of natural organic matter.

Geosmin and 2-methylisoborneol (2-MIB) are naturally occurring compounds responsible for musty-earthy-odors in surface water supplies. They are a severe problem confronting utilities worldwide. Adsorption by powdered activated carbon (PAC) is a widely used process to control this problem, but it has low efficiency, which engenders large budget spending for utilities services. Super-powdered activated carbon (S-PAC) is activated carbon with much finer particles than those of PAC. Experiments on geosmin and 2-MIB adsorptions on S-PAC and PAC were conducted. Geosmin and 2-MIB adsorption capacities on S-PAC were not smaller than those on PAC although natural organic matter, which adversely impacted the adsorption capacity of geosmin and 2-MIB, was more adsorbed on S-PAC than on PAC, meaning that the adsorption competition is less severe for S-PAC than for PAC.

Suitability of pepper mild mottle virus as a human enteric virus surrogate for assessing the efficacy of thermal or free-chlorine disinfection processes by using infectivity assays and enhanced viability PCR.

Evaluating the efficacy of disinfection processes to inactivate human enteric viruses is important for the prevention and control of waterborne diseases caused by exposure to those viruses via drinking water. Here, we evaluated the inactivation of two representative human enteric viruses (adenovirus type 40 [AdV] and coxsackievirus B5 [CV]) by thermal or free-chlorine disinfection. In addition, we compared the infectivity reduction ratio of a plant virus (pepper mild mottle virus [PMMoV], a recently proposed novel surrogate for human enteric viruses for the assessment of virus removal by coagulation‒rapid sand filtration and membrane filtration) with that of the two human enteric viruses to assess the suitability of PMMoV as a human enteric virus surrogate for use in thermal and free-chlorine disinfection processes. Finally, we examined whether conventional or enhanced viability polymerase chain reaction (PCR) analysis using propidium monoazide (PMA) or improved PMA (PMAxx) with or without an enhancer could be used as alternatives to infectivity assays (i.e., plaque-forming unit method for AdV and CV; local lesion count assay for PMMoV) for evaluating virus inactivation by disinfection processes. We found that PMMoV was more resistant to heat treatment than AdV and CV, suggesting that PMMoV is a potential surrogate for these two enteric viruses with regard to thermal disinfection processes. However, PMMoV was much more resistant to chlorine treatment compared with AdV and CV (which is chlorine-resistant) (CT value for 4-log10 inactivation: PMMoV, 84.5 mg-Cl2·min/L; CV, 1.15-1.19 mg-Cl2·min/L), suggesting that PMMoV is not useful as a surrogate for these enteric viruses with regard to free-chlorine disinfection processes. For thermal disinfection, the magnitude of the signal reduction observed with PMAxx-Enhancer-PCR was comparable with the magnitude of reduction in infectivity, indicating that PMAxx-Enhancer-PCR is a potential alternative to infectivity assay. However, for free-chlorine disinfection, the magnitude of the signal reduction observed with PMAxx-Enhancer-PCR was smaller than the magnitude of the reduction in infectivity, indicating that PMAxx-Enhancer-PCR underestimated the efficacy of virus inactivation (i.e., overestimated the infectious virus concentration) by chlorine treatment. Nevertheless, among the PCR approaches examined in the present study (PCR alone, PMA-PCR or PMAxx-PCR either with or without enhancer), PMAxx-Enhancer-PCR provided the most accurate assessment of the efficacy of virus inactivation by thermal or free chlorine disinfection processes.

The first case of chronic pulmonary Mycobacterium shigaense infection in an immunocompetent patient.

Mycobacterium shigaense has recently been recognized as an emerging human pathogen, and is well known as a skin pathogen in immunocompromised individuals. In this report we describe the first case of chronic pulmonary infectious disease caused by M. shigaense in an immunocompetent individual.