Genetic ablation of Saposin-D in Krabbe disease eliminates psychosine accumulation but does not significantly improve demyelination.

Krabbe disease is an inherited demyelinating disease caused by a genetic deficiency of the lysosomal enzyme galactosylceramide (GalCer) ß-galactosidase (GALC). The Twitcher (Twi) mouse is a naturally occurring, genetically and enzymatically authentic mouse model that mimics infantile-onset Krabbe disease. The major substrate for GALC is the myelin lipid GalCer. However, the pathogenesis of Krabbe disease has long been explained by the accumulation of psychosine, a lyso-derivative of GalCer. Two metabolic pathways have been proposed for the accumulation of psychosine: a synthetic pathway in which galactose is transferred to sphingosine and a degradation pathway in which GalCer is deacylated by acid ceramidase (ACDase). Saposin-D (Sap-D) is essential for the degradation of ceramide by ACDase in lysosome. In this study, we generated Twi mice with a Sap-D deficiency (Twi/Sap-D KO), which are genetically deficient in both GALC and Sap-D and found that very little psychosine accumulated in the CNS or PNS of the mouse. As expected, demyelination with the infiltration of multinucleated macrophages (globoid cells) characteristic of Krabbe disease was milder in Twi/Sap-D KO mice than in Twi mice both in the CNS and PNS during the early disease stage. However, at the later disease stage, qualitatively and quantitatively comparable demyelination occurred in Twi/Sap-D KO mice, particularly in the PNS, and the lifespans of Twi/Sap-D KO mice were even shorter than that of Twi mice. Bone marrow-derived macrophages from both Twi and Twi/Sap-D KO mice produced significant amounts of TNF-α upon exposure to GalCer and were transformed into globoid cells. These results indicate that psychosine in Krabbe disease is mainly produced via the deacylation of GalCer by ACDase. The demyelination observed in Twi/Sap-D KO mice may be mediated by a psychosine-independent, Sap-D-dependent mechanism. GalCer-induced activation of Sap-D-deficient macrophages/microglia may play an important role in the neuroinflammation and demyelination in Twi/Sap-D KO mice.

Factors of influenza vaccine inoculation and non-inoculation behavior of community-dwelling residents in Japan: Suggestions for vaccine policy and public health ethics after COVID-19.

The aim of this study is to provide basic information that contributes to vaccine inoculation policy after COVID-19. We used the secondary data of the influenza vaccine inoculation behavior survey for community-dwelling adults conducted in 2011, before the COVID-19 pandemic, but after the 2009 novel influenza A (H1N1) pdm 09 pandemic. All factors such as socio-demographic characteristics, health-related behaviors, family environment, physical and social environment, and area of residence were adjusted, and factors related to vaccine inoculation behavior were analyzed. Those living with pregnant women had a significantly higher odds ratio of inoculation; this was self-evident in that those people considered infection to their family. Regarding the social environment, those aged 20-64 years with a significantly higher adjusted odds ratio of inoculation were those with "at least five people with which they interacted in the neighborhood". This result can be interpreted in two ways relating to altruism in Japan. Finally, we indicated the importance of learning from the past, including the case of 2009.

Plasma membrane proteomic changes of Arabidopsis DRP1E during cold acclimation in association with the enhancement of freezing tolerance.

The freezing tolerance of plants that live in cold regions increases after exposure to low temperature, a process termed cold acclimation (CA). During CA, restructuring of the plasma membrane (PM) is important to enhance freezing tolerance. We have previously shown that the function of DYNAMIN-RELATED PROTEIN 1 E (DRP1E), which regulates endocytosis by pinching vesicles from the PM, is associated with the enhancement of freezing tolerance during CA in Arabidopsis. DRP1E is predicted to play a role in reconstituting the PM composition during CA. In this study, to test the validity of this hypothesis, we studied the changes in PM proteome patterns induced by drp1e mutation. In a detailed physiological analysis, after 3 days of CA, only young leaves showed significantly less increase in freezing tolerance in the mutant than in the wild type (WT). Using nano-liquid chromatography-tandem mass spectrometry, 496 PM proteins were identified. Among these proteins, 81 or 71 proteins were specifically altered in the WT or the mutant, respectively, in response to CA. Principal component analysis showed that the proteomic pattern differed between the WT and the mutant upon cold acclimation (CA), suggesting that DRP1E contributes to reconstruction of the PM during CA. Cluster analysis revealed that proteins that were significantly increased in the mutant after CA were biased toward glycosylphosphatidylinositol-anchored proteins, such as fasciclin-like arabinogalactan proteins. Thus, a primary target of DRP1E-associated PM reconstruction during CA is considered to be glycosylphosphatidylinositol-anchored proteins, which may be removed from the PM by DRP1E in young leaves after 3 days of CA.

Old plasma dilution reduces human biological age: a clinical study.

This work extrapolates to humans the previous animal studies on blood heterochronicity and establishes a novel direct measurement of biological age. Our results support the hypothesis that, similar to mice, human aging is driven by age-imposed systemic molecular excess, the attenuation of which reverses biological age, defined in our work as a deregulation (noise) of 10 novel protein biomarkers. The results on biological age are strongly supported by the data, which demonstrates that rounds of therapeutic plasma exchange (TPE) promote a global shift to a younger systemic proteome, including youthfully restored pro-regenerative, anticancer, and apoptotic regulators and a youthful profile of myeloid/lymphoid markers in circulating cells, which have reduced cellular senescence and lower DNA damage. Mechanistically, the circulatory regulators of the JAK-STAT, MAPK, TGF-beta, NF-κB, and Toll-like receptor signaling pathways become more youthfully balanced through normalization of TLR4, which we define as a nodal point of this molecular rejuvenation. The significance of our findings is confirmed through big-data gene expression studies.

Mechanisms and Minimization of False Discovery of Metabolic Bioorthogonal Noncanonical Amino Acid Proteomics.

Metabolic proteomics has been widely used to characterize dynamic protein networks in many areas of biomedicine, including in the arena of tissue aging and rejuvenation. Bioorthogonal noncanonical amino acid tagging (BONCAT) is based on mutant methionine-tRNA synthases (MetRS) that incorporates metabolic tags, for example, azidonorleucine [ANL], into newly synthesized proteins. BONCAT revolutionizes metabolic proteomics, because mutant MetRS transgene allows one to identify cell type-specific proteomes in mixed biological environments. This is not possible with other methods, such as stable isotope labeling with amino acids in cell culture, isobaric tags for relative and absolute quantitation and tandem mass tags. At the same time, an inherent weakness of BONCAT is that after click chemistry-based enrichment, all identified proteins are assumed to have been metabolically tagged, but there is no confirmation in mass spectrometry data that only tagged proteins are detected. As we show here, such assumption is incorrect and accurate negative controls uncover a surprisingly high degree of false positives in BONCAT proteomics. We show not only how to reveal the false discovery and thus improve the accuracy of the analyses and conclusions but also approaches for avoiding it through minimizing nonspecific detection of biotin, biotin-independent direct detection of metabolic tags, and improvement of signal to noise ratio through machine learning algorithms.

Case Report: Therapeutic and immunomodulatory effects of plasmapheresis in long-haul COVID.

Many patients with COVID-19 experience a range of debilitating symptoms months after being infected, a syndrome termed long-haul COVID. A 68-year-old male presented with lung opacity, fatigue, physical and cognitive weaknesses, loss of smell and lymphocytopenia. After rounds of therapeutic plasma exchange (TPE), the patient returned to normal activities and work. Mechanistically in the patient's peripheral blood mononuclear cells (PBMCs), markers of inflammatory macrophages diminished and markers of lymphocytes, including natural killer (NK) cells and cytotoxic CD8 T-cells, increased. Circulating inflammatory proteins diminished, while positive regulators of tissue repair increased. This case study suggests that TPE has the capacity to treat long-haul COVID.

Autonomous Continuation of Community Health Workers' Activities in Thegon Township, Bago Region, Myanmar.

Background: Few studies have reported on the autonomous continuation of Community Health Worker (CHW) activities after external supervision and support have ended. Objective: The study reports CHW activity continuation in Thegon Township, Bago Region, Myanmar, observed after the supervision by the external organization is completed. Method: Following the completion of a child nutrition program in Thegon Township, CHWs were left unsupervised and uninformed of a follow-up at 10, 23, and 40 months from the end of the program survey due to unforeseen circumstances. In a follow-up survey in 2014, data on the activity implementation status from CHWs and activity attendance from caregivers of the target children were collected. Focus group discussions were held with caregivers concerning their information sources on child nutrition and health. Results: On average, CHWs were found to have continued with 2.6 of the four core activities, often with modifications, irrespective of the time since completion of the non-profit-organization-led program. Meanwhile, caregiver attendance decreased over time. Caregivers recognized CHWs as information sources. Discussion: Although unsupervised, CHWs ambitiously continued with their activities, but sorted through and modified them, which may have been unrelated to the local acceptance of the program, as caregiver attendance decreased even as CHWs continued the activities. The observation may highlight the importance of proactive engagement and thus, the autonomy of CHWs in their activity continuation.

AtUBL5 regulates growth and development through pre-mRNA splicing in Arabidopsis thaliana.

Ubiquitin-like proteins play important roles in the regulation of many biological processes. UBL5 (Ubiquitin-like protein 5)/Hub1 (Homologous to ubiquitin 1), a member of the ubiquitin family, acts as a ubiquitin-like modifier on a specific target, the spliceosomal protein Snu66, in yeast and human cells. The 22nd aspartic acid (Asp22) is involved in the attachment of Hub1 to the Hub1 interaction domain (HIND) of Snu66 in yeast to modulate spliceosomal activity. Hub1 differs from other modifiers which interact covalently with their targets. It modulates pre-mRNA splicing by binding to Snu66 non-covalently in both yeast and human cells. However, the molecular mechanisms of Hub1-mediated pre-mRNA splicing in plant systems remains unclear. To better understand the function of Hub1 in plants, we examined the role of this ubiquitin-like modifier in Arabidopsis thaliana, which has two Hub1 homologues. Arabidopsis UBL5/Hub1(UBL5) is highly conserved at the amino acid level, compared to eukaryotic homologues in both plants and animals. In this study, phenotypic analysis of A. thaliana with reduced UBL5 gene expression, generated by RNA interference of AtUBL5a and AtUBL5b were performed. Interestingly, knock down plants of AtUBL5 showed abnormalities in root elongation, plant development, and auxin response. AtUBL5b is highly expressed in the vascular tissue of the leaf, stem, and root tissue. Yeast two-hybrid analysis revealed that AtUBL5a and AtUBL5b interact with the putative splicing factor AtPRP38 through its C-terminal domain (AtPRP38C). Knock down of AtUBL5b resulted in a pattern of insufficient pre-mRNA splicing in several introns of AtCDC2, and in introns of IAA1, IAA4, and IAA5. Defects of pre-mRNA splicing in an AtPRP38 mutant resulted in an insufficient pre-mRNA splicing pattern in the intron of IAA1. Based on these results, we showed that AtUBL5b positively regulates plant root elongation and development through pre-mRNA splicing with AtPRP38C in A. thaliana.

Novel neuroblastoma amplified sequence (NBAS) mutations in a Japanese boy with fever-triggered recurrent acute liver failure.

Biallelic mutations in the neuroblastoma amplified sequence (NBAS) gene have been reported to cause two different clinical spectra: short stature with optic nerve atrophy and Pelger-Huët anomaly (SOPH) syndrome and infantile liver failure syndrome 2 (ILFS2). Here, we describe a case of a 3-year-old Japanese boy who presented with fever-triggered recurrent acute liver failure (ALF). The clinical characteristics were considerable elevation of liver enzymes, severe coagulopathy, and acute renal failure. In addition to the liver phenotype, he had short stature and Pelger-Huët anomaly in the peripheral granulocytes. Whole-exome and Sanger sequencing of the patient and his parents revealed that he carried novel compound heterozygous missense mutations in NBAS, c.1018G>C (p.Gly340Arg) and c.2674 G>T (p.Val892Phe). Both mutations affect evolutionarily conserved amino acid residues and are predicted to be highly damaging. Immunoblot analysis of the patient's skin fibroblasts showed a normal NBAS protein level but a reduced protein level of its interaction partner, p31, involved in Golgi-to-endoplasmic reticulum retrograde vesicular trafficking. We recommend NBAS gene analysis in children with unexplained fever-triggered recurrent ALF or liver dysfunction. Early antipyretic therapy may prevent further episodes of ALF.

Influence of abdominal obesity and habitual behaviors on incident atrial fibrillation in Japanese.

BACKGROUND: As atrial fibrillation (AF) increases with the aging of the population, it is urgently required to clarify modifiable factors to prevent AF. However, evidence regarding the independent influence of abdominal-obesity and habitual behaviors on incident AF is limited among Japanese. METHODS AND RESULTS: Those aged 40-79 years undergoing periodic health checkups during 2008-2014 were followed-up in 2015 (n=96,841) and the independent risk of incident AF was estimated using multivariate Cox proportional hazards regression models after adjustment for potential covariates. Participants were classified into four groups according to the baseline body mass index (BMI: kg/m2) (normal-BMI or overweight: < or ≥25) and waist circumference (WC: cm) (normal-WC or abdominal-obesity: < or ≥85 for male, ≥90 for female). Baseline habitual behaviors, smoking status, alcohol intake, and physical activity, were also included as modifiable factors. Among 65,984 eligible participants, 349 developed AF over mean follow-up of 5.5±1.6 years. Increase of both BMI and WC significantly elevated the risk of AF. Compared to the normal-BMI and normal-WC group, the normal-BMI but abdominal-obesity and the overweight and abdominal-obesity groups in males and the overweight and abdominal-obesity group in females had significantly elevated risk of AF. Among modifiable behavioral factors including abdominal-obesity, alcohol intake (≥40g/day) and abdominal-obesity significantly elevated the risk of AF in males, and abdominal-obesity was the strongest risk factor in both sexes, but smoking and physical activity were not significant. However, an aggregation of these four behavioral factors increased the risk of AF more than 2.5 times in both sexes. CONCLUSIONS: Abdominal-obesity could be a crucial risk factor in prediction of AF in Japanese, and an aggregation of four behavioral factors increased the risk of AF almost three times. To prevent incident AF, practicing healthy habitual behaviors is recommended.

HSP90 stabilizes auxin receptor TIR1 and ensures plasticity of auxin responses.

Heat shock protein 90 (HSP90) is a highly conserved molecular chaperone that facilitates the maturation of target proteins. Here, we report that the auxin receptor TIR1 is a target of cytosolic HSP90 and that HSP90 and TIR1 form a complex. Inhibition of HSP90 compromised the nuclear localization of TIR1, and abrogated plant responses to the hormone auxin. Our findings suggest that HSP90 positively regulates auxin receptor function. We also propose that HSP90 buffers or hides phenotypic variations in animals and plants by masking mutations in some of its target proteins. Support for this proposal comes from the tir1-1 mutant of Arabidopsis, which showed a root growth defect that was only seen after inhibition of HSP90. We have developed a model in which cytosolic HSP90 works like a capacitor for auxin-related phenotypic variation via regulation of the auxin receptor in response to environmentally and genetically induced perturbations.

How do health behaviours relate to dietary practice patterns among Japanese adults?

BACKGROUND AND OBJECTIVES: To identify dietary practice patterns for Japanese adults and investigate the links between health behaviours and these patterns. METHODS AND STUDY DESIGN: A random sample, stratified according to area, sex, and age, of 4570 adults aged 20-80 years completed a survey conducted in 2011 in a city, in Yamagata Prefecture, Northeast Japan. RESULTS: Cluster analysis of 16 dietary practice items revealed four patterns labelled as: low fat, sugar, or salt; emphasis on nutrition; regular breakfast and staples; and meals not snacks. Findings from multiple linear regression analyses showed that those not engaged in habitual physical exercise had lower scores on low fat, sugar, or salt (beta coefficient -0.22: 95% confidence intervals -0.30, -0.14); emphasis on nutrition (-0.17: -0.25, -0.09); meals not snacks; (-0.38: -0.46, -0.3) that other participants. Current smokers had lower scores than never smokers on low fat, sugar, or salt (-0.23: -0.32, -0.14); emphasis on nutrition (-0.28: -0.37, -0.19); regular breakfast and staples (-0.42: -0.51, -0.33) patterns. Compared with nondrinkers, those who had reduced their consumption of alcohol had higher scores on low fat, sugar, or salt (0.19: 0.09, 0.29) and emphasis on nutrition (0.17: 0.07, 0.27). These relationships were adjusted for other dietary practice patterns, sociodemographic factors, body mass index, and the presence of major illness or pain. CONCLUSIONS: Findings support an integrated and targeted approach as part of public health policy by considering links between dietary practices and other health behaviours, such as habitual exercise and smoking behaviour that may facilitate changes in dietary practices.

Clustering patterns of obesity-related multiple lifestyle behaviours and their associations with overweight and family environments: a cross-sectional study in Japanese preschool children.

OBJECTIVES: The purpose of this study is (1) to identify obesity-related lifestyle behaviour patterns of diet, physical activity, sedentary and sleep behaviours in preschool children, (2) to examine the association between identified behaviour clusters and overweight/obesity and (3) to investigate differences in children's family environments according to clusters. DESIGN SETTING AND PARTICIPANTS: A cross-sectional study on 2114 preschool children aged 3-6 years who attended childcare facilities (24 nursery schools and 10 kindergartens) in Tsuruoka city, Japan in April 2003 was conducted. MAIN OUTCOME MEASURES: Children's principal caregivers completed a questionnaire on children's lifestyle behaviours (dinner timing, outside playtime, screen time and night-time sleep duration), family environment (family members, maternal employment, mealtime regularity and parents' habitual exercise and screen time) and measurements of weight and height. Cluster analysis was performed using children's 4 lifestyle behaviours based on those non-missing values (n=1545). The χ2 tests and analysis of variance (ANOVA) estimated cluster differences in overweight/obesity and family environments. RESULTS: 6 clusters were identified. Children's overweight/obesity varied across clusters (p=0.007). The cluster with the most screen time, shorter night-time sleep duration, average dinner timing and outside playtime had the highest overweight/obesity prevalence (15.1%), while the cluster with the least screen time, the longest sleep duration, the earliest dinner timing and average outside playtime had the lowest prevalence (4.0%). Family environments regarding mealtime regularity and both parents' screen time also significantly varied across clusters. The cluster having the highest overweight/obesity prevalence had the highest proportion of irregular mealtimes and the most screen time for both parents. CONCLUSIONS: This study suggests that public health approaches to prevent children's overweight/obesity should focus on decreasing screen time and increasing night-time sleep duration. To shape those behaviours, regular mealtimes and decreasing parents' screen time within family environments need to be targeted among family members.

HSP90 Stabilizes Auxin-Responsive Phenotypes by Masking a Mutation in the Auxin Receptor TIR1.

Heat shock protein 90 (HSP90) is a molecular chaperone that is required for the function of various substrate proteins, also known as client proteins. It is proposed that HSP90 buffers or hides phenotypic variations in animals and plants by masking mutations in some of its client proteins. However, none of the client proteins with cryptic mutations has been identified to date. Here, we identify the first client protein example by which HSP90 buffers a mutation: the auxin receptor transport inhibitor response 1 (TIR1). TIR1 interacts with HSP90 in the nucleus. An HSP90-specific inhibitor abolished the nuclear localization of TIR1 and the auxin-induced degradation of a TIR1-substrate, indicating that TIR1 is an HSP90 client protein. Plants with a null mutation in the TIR1 gene had a defect in auxin response, whereas plants with a point mutation in the TIR1 gene responded to auxin treatment in young seedlings, but a cryptic defect in its auxin response was exposed with HSP90 inhibitor treatment. These results demonstrate that HSP90 masks a point mutation in the auxin receptor TIR1 and thereby buffers auxin-responsive phenotypes.

Combined eating behaviors and overweight: Eating quickly, late evening meals, and skipping breakfast.

OBJECTIVE: Various eating behaviors have been linked with body weight management. However, combined effects of major eating behaviors are not fully understood. This study aimed to clarify the association of the combination of eating quickly (EQ), late evening meals (LEM), and skipping breakfast (SB) with being overweight. METHOD: A cross-sectional study with standardized questions for EQ, LEM, and SB was conducted. Stratified random sampling of 5% of residents aged 20 to 80years was surveyed in a city in northeast Japan in 2011, and 4249 (84.9%) residents were analyzed. Association of combinations of eating behaviors on being overweight (BMI (kg/m(2)≥25.0)) was estimated by using logistic analysis, and odds ratio (OR) and 95% confidential interval were calculated after adjustment for potential covariates. RESULTS: LEM, SB, or a combination of LEM and SB was not significantly associated with being overweight. However, the combination of EQ or only EQ was significantly associated with being overweight. As the number of eating behavior practices increased, there was a linear increase in OR for being overweight. The OR of all three combined eating behaviors was higher than that of any combined two behaviors or of each behavior. DISCUSSION: This study result supports the evidence that EQ increases the risk of being overweight whether by itself or in combinations with LEM and/or SB. However, only LEM or only SB did not increase the risk of being overweight.

Joining characteristics of titanium-based orthodontic wires connected by laser and electrical welding methods.

This study investigated the possibility of electrical and laser welding to connect titanium-based alloy (beta-titanium and nickel-titanium) wires and stainless-steel or cobalt-chromium alloy wires for fabrication of combination arch-wires. Four kinds of straight orthodontic rectangular wires (0.017 × 0.025 inch) were used: stainless-steel (S-S), cobalt-chromium (Co-Cr), beta-titanium alloy (ß-Ti), and nickel-titanium (Ni-Ti). Homogeneous and heterogeneous end-to-end joints (15 mm long each) were made by electrical welding and laser welding. Non-welded wires (30 mm long) were also used as a control. Maximum loads at fracture (N) and elongation (%) were measured by conducting tensile test. The data (n = 10) were statistically analyzed using analysis of variance/Tukey test (P < 0.05).The S-S/S-S and Co-Cr/Co-Cr specimens showed significantly higher values of the maximum load (ML) at fracture and elongation (EL) than those of the Ni-Ti/Ni-Ti and ß-Ti/ß-Ti specimens for electrical welding and those of the S-S/S-S and Co-Cr/Co-Cr specimens welded by laser. On the other hand, the laser-welded Ni-Ti/Ni-Ti and ß-Ti/ß-Ti specimens exhibited higher values of the ML and EL compared to those of the corresponding specimens welded by electrical method. In the heterogeneously welded combinations, the electrically welded Ni-Ti/S-S, ß-Ti/S-S and ß-Ti/Co-Cr specimens showed significantly (P < 0.05) higher ML and EL than those of the corresponding specimens welded by laser. Electrical welding exhibited the higher values of maximum load at fracture and elongation for heterogeneously welded combinations than laser-welding.

Consolidation with a twisted appearance along the airways: a report of five cases of interstitial pneumonia.

High-resolution CT showed areas of airspace consolidation with a twisted appearance of the airways, along with areas of peribronchial ground-glass attenuation and traction bronchiectasis, in five patients with interstitial pneumonia. These areas of airspace consolidation were termed "twisted consolidation" (TwC). The five patients included two patients receiving treatment for rheumatoid arthritis (RA), one patient with newly diagnosed RA, and one patient who subsequently showed RA. Three patients showed improvement after steroid administration. An association of TwC with RA is suspected, but further studies are necessary.

Association between changes in 12 lifestyle behaviors and the development of metabolic syndrome during 1 year among workers in the Tokyo metropolitan area.

BACKGROUND: Practicing healthy lifestyle behaviors is a means to prevent metabolic syndrome (MetS), but the effect of changes of various behaviors over a short period is not fully understood. The purpose of this study was to elucidate the influence of changes in 12 behaviors on the development of MetS during 1 year. METHODS AND RESULTS: Of 10,442 workers who received a periodic health checkup in a health center in Tokyo in 2008, 3,137 workers aged 30-69, without MetS, who received another health checkup in 2009 were analyzed. Smoking, amounts and frequency of alcohol drinking, sleeping, exercise, walking duration and speed, late-night dinners, bedtime snacking, breakfast, eating speed, and weight control were classified into 4 groups according to change from 2008 to 2009. To examine the influence of behavioral changes on developing MetS, multiple logistic analysis was conducted after adjustment for sex, baseline age and MetS components. Changes from healthy to unhealthy behaviors in exercise, walking duration and speed, daily drinking, and weight control were significant in developing MetS compared with maintaining healthy behaviors. Those risks were higher than keeping unhealthy behaviors. Unhealthy to healthy behavior in smoking increased the risk while healthy to unhealthy behavior in eating speed decreased the risk of developing MetS. CONCLUSIONS: To prevent developing MetS during 1 year, healthy behaviors regarding physical activity, drinking, and weight management should be maintained.

[Study on pulmonary lesions in which nontuberculous mycobacteria were detected by percutaneous aspiration--a proposal to add "culture positivity of percutaneous aspiration material" to the bacteriological diagnostic criteria of pulmonary nontuberculous mycobacterial diseases].

OBJECTIVES: Culture positivity of percutaneous aspiration material" is not included in the current bacteriological criteria for diagnosis of pulmonary nontuberculous mycobacterial (NTM) diseases, which were published by the Infectious Diseases Society of America (IDSA)/American Thoracic Society (ATS) in 2007 or those released by the Japanese Society for Tuberculosis in 2008. However, percutaneous aspiration is a reliable technique for the detection of causative microorganisms isolated from the focus of infection. We discuss the benefits of including positive culture of percutaneous aspiration material in the bacteriological diagnostic criteria of pulmonary NTM diseases. METHODS: We reviewed the radiological images and clinical courses of pulmonary diseases in which NTM cultures were obtained from percutaneously aspirated materials at our hospital from 1991 to 2011. Aspiration was carried out under local anesthesia, usually with fluoroscopic guidance. After percutaneous insertion of a 22-gauge needle attached to a 20-mL syringe containing about 3 mL of saline, the lesion specimen was withdrawn together with the saline. After the needle was pulled out, the aspirated material and saline were transferred to test tubes for cytological and microbiological examinations. In patients with thin-walled cavitary lesions, saline was injected into the cavity and then aspirated. RESULTS: Percutaneous aspiration was performed in 2,742 patients and NTM disease was detected in 51 patients. Of these 51 patients, 12 had solitary nodular lesions, and in many of these patients, no NTM bacilli could be detected in the sputa or bronchial washing specimens. Mycobacterium avium was identified in 10 of the 12 cases. Four of these 10 patients were followed up after their diagnosis without any treatment: 3 showed spontaneous reduction in lesion size, while 1 patient's condition remained unchanged. Four of the remaining 6 cases were treated with anti-NTM medications, and lesion size reduced in 2 cases, while no change or deterioration was seen in the other 2. Aspiration from solitary small cavitary lesions showed a relatively high number of NTM colonies. Pneumothorax was the only complication of the aspiration procedure. DISCUSSION: If the diagnostic criteria for pulmonary NTM diseases include positive culture in percutaneous aspiration material, the diagnosis of solitary nodular NTM lesions would become easier; at present, these lesions are often diagnosed only upon surgical resection. Further, clinical studies on the possibility of spontaneous shrinkage of the solitary lesion and the value of its medical treatment would be promoted. Aspiration can easily differentiate NTM disease from pulmonary abscess or fungal infection in patients with a solitary lesion or small cavity.