CRISPRi knockdown of the cyabrB1 gene induces the divergently transcribed icfG and sll1783 operons related to carbon metabolism in the cyanobacterium Synechocystis sp. PCC 6803.

Most cyanobacterial genomes possess more than two copies of genes encoding cyAbrBs (cyanobacterial AbrB-like proteins) having an AbrB-like DNA-binding domain at their C-terminal region. Accumulating data suggest that a wide variety of metabolic and physiologic processes are regulated by cyAbrBs. In this study, we investigated the function of the essential gene cyabrB1 (sll0359) in Synechocystis sp. PCC 6803 by using CRISPR interference technology. The conditional knockdown of cyabrB1 caused increases of cyAbrB2 transcript and protein levels. However, the effect of cyabrB1 knockdown on global gene expression profile was quite limited compared to the previously reported profound effect of knockout of cyabrB2. Among 24 up-regulated genes, 16 genes were members of the divergently transcribed icfG and sll1783 operons related to carbon metabolism. The results of this and previous studies indicate the different contributions of two cyAbrBs to transcriptional regulation of genes related to carbon, hydrogen and nitrogen metabolism. Possession of a pair of cyAbrBs has been highly conserved during the course of evolution of the cyanobacterial phylum, suggesting physiological significance of transcriptional regulation attained by their interaction.

Case Report: Use of endobronchial Watanabe spigot and coagulation factor XIII supplementation in the treatment of persistent pneumothorax due to pneumocystis pneumonia with human immunodeficiency virus infection.

Pneumocystis jiroveceii pneumonia is one of the most common opportunistic infections associated with human immunodeficiency virus. Endobronchial Watanabe spigot has been recommended for refractory pneumothorax, even with persistant air leak despite continuous negative pressure control via thoracic drainage. Moreover, coagulation factor XIII is considered effective in wound healing.

Case Report: Massive Hemoptysis From a Spontaneously Regression Inflammatory Bronchial Polyp.

Background: Bronchial inflammatory polyps are usually treated by surgical operation or with steroids and/or antibiotics, and it is quite rare that such polys spontaneously disappear without any treatment. This report shows a rare case with a bronchial inflammatory polyp which caused massive hemoptysis but spontaneously disappeared without any treatment. Case Presentation: A 66-year-old man with type 2 diabetes mellitus and a history of cough and asthma suddenly developed massive hemoptysis while smoking and was brought to an emergency room in our institution. In bronchoscopy on admission, a polypoidal elevated lesion was observed in the left upper lobe bifurcation. Pulsatile hemorrhage from a polypoidal elevated lesion was observed upon stimulation of passage of the bronchoscope. Bronchoscopy performed 25 days after discharge showed no evidence of active bleeding and a tendency toward reduction of the elevated lesion. In bronchoscopy performed 106 days after the initial hospitalization, the bronchial inflammatory polyp completely disappeared. Conclusions: We should bear in mind the possibility of spontaneous disappearance of bronchial inflammatory polyps causing some serious symptoms such as massive hemoptysis and repeated bloody sputum. Finally, we should select the best therapy for bronchial inflammatory polys based on each patient's background and conditions in clinical practice.

Case Report: A Patient With Neuroleptic Malignant Syndrome, Water Intoxication and Hyponatremia Associated With Severe Cerebral Edema and Coma.

Background: Water intoxication is typically caused by primary or psychogenic polydipsia that potentially may lead to fatal disturbance in brain functions. Neuroleptic malignant syndrome (NMS) is a serious complication induced by administration of antipsychotics and other psychotropic drugs. The combination of inappropriate secretion of antidiuretic hormone (SIDAH), NMS and rhabdomyolysis have been rarely reported. Our patient also developed severe water intoxication. Case presentation: Herein we report a comatose case of NMS complicated with water intoxication, syndrome of SIADH and rhabdomyolysis. This patient had severe cerebral edema and hyponatremia that were improved rapidly by the correction of hyponatremia within a couple of days. Conclusions: Malignant neuroleptic syndrome water intoxication, SIADH and rhabdomyolysis can occur simultaneously. Comatose conditions induced by cerebral edema and hyponatremia can be successfully treated by meticulous fluid management and the correction of hyponatremia.

Temperature sensitivity of Notch signaling underlies species-specific developmental plasticity and robustness in amniote brains.

Ambient temperature significantly affects developmental timing in animals. The temperature sensitivity of embryogenesis is generally believed to be a consequence of the thermal dependency of cellular metabolism. However, the adaptive molecular mechanisms that respond to variations in temperature remain unclear. Here, we report species-specific thermal sensitivity of Notch signaling in the developing amniote brain. Transient hypothermic conditions increase canonical Notch activity and reduce neurogenesis in chick neural progenitors. Increased biosynthesis of phosphatidylethanolamine, a major glycerophospholipid components of the plasma membrane, mediates hypothermia-induced Notch activation. Furthermore, the species-specific thermal dependency of Notch signaling is associated with developmental robustness to altered Notch signaling. Our results reveal unique regulatory mechanisms for temperature-dependent neurogenic potentials that underlie developmental and evolutionary adaptations to a range of ambient temperatures in amniotes.

Animal models of chronic and recurrent Pseudomonas aeruginosa lung infection: significance of macrolide treatment.

Animal models of human diseases are invaluable and inevitable elements in identifying and testing novel treatments for serious diseases, including severe infections. Planning and conducting investigator-initiated human trials are generally accepted as being enormously challenging. In contrast, it is often underestimated how much planning, including background and modifying experiments, is needed to establish a relevant infectious disease animal model. However, representative animal infectious models, well designed to test generated hypotheses, are useful to improve our understanding of pathogenesis, virulence factors and host response and to identify novel treatment candidates and therapeutic strategies. Such results can subsequently proceed to clinical testing if suitable. The present review aims at presenting all the pulmonary Pseudomonas aeruginosa infectious models we have knowledge of and the detailed descriptions of established animal models in our laboratory focusing on macrolide therapy are presented.

Rapidly Exacerbating Autoimmune Hemolytic Anemia Together With Marked Cytokine Storm Triggered by Pneumonia Infection: A Case Report.

Background: Autoimmune hemolytic anemia (AIHA) is caused by hemolysis induced by the reaction of autoantibodies with red blood cells. AIHA is usually classified as either warm antibody or cold antibody-mediated AIHA. In addition, AIHA caused by infection is classified as secondary AIHA. It is well-known that alteration of various cytokine levels is closely associated with a variety of disorders such as infectious diseases. In addition, it is known that IL-10/ IL-12 imbalance is an indicator of Th2-dominat conditions and a progressive marker of AIHA. Case presentation: A 82-year-old Japanese man was brought to the emergency room with pneumonia and heart failure. After admission, we started antibiotics therapy. Next day, he had symptoms of jaundice and his total bilirubin level was elevated. Five days after admission, his inflammation markers were further elevated and he had hemolytic anemia. We finally diagnosed him as severe warm-type AIHA associated with pneumoniae infection. Seven days after admission, his severe leucocytosis was further aggravated, and then he suddenly had cardiac arrest and respiratory failure, and finally died of multiple organ failure. Unfortunately, we failed to rescue him from severe AIHA induced by pneumonia infection. Conclusions: Herein, we report a subject with pneumonia-triggered warm-type AIHA together with marked cytokine storm. IL-18 levels were markedly elevated without elevation of IL-12 levels. In addition, IL-10/IL-12 imbalance was observed. These data suggest that once marked cytokine storm is induced, infection-induced AIHA is exacerbated very rapidly.

Thoracoscopic Findings in IgG4-related Pleuritis.

A 46-year-old Japanese man was admitted to our hospital with a 1-year history of dyspnea and persistent right-dominant bilateral pleural effusions. Chest and abdominal computed tomography (CT) revealed no notable findings apart from the bilateral pleural effusions. 2-deoxy-2-[18F]-fluoro-D-glucose (FDG) positron emission tomography-CT showed no accumulation of FDG in the thorax and abdomen. Thoracoscopy revealed numerous small (approximately 2-3 mm in size), blister-like nodules on the left parietal pleura extending from the lower third of the chest wall to the diaphragm. A pathological examination revealed lymphocyte and plasma cell infiltrates with increasing numbers of IgG4-positive plasma cells in the fibrotic pleura, indicating IgG4-related pleuritis.

Elevated levels of periostin and TGF-ß1 in the bronchoalveolar lavage fluid of patients with idiopathic eosinophilic pneumonia.

BACKGROUND: Periostin is induced in bronchial epithelial cells and fibroblasts by various stimuli including interleukin (IL)13 and transforming growth factor (TGF)-ß1, and is involved in allergic diseases such as asthma and atopic dermatitis, playing an important role in tissue remodeling and fibrosis. The role of periostin in the pathogenesis of eosinophilic lung diseases, however, is unclear. OBJECTIVE: To examine the contribution of periostin to eosinophilic inflammation of the lung in humans, we evaluated periostin, IL-13, and TGF-ß1 levels in the bronchoalveolar lavage fluid (BALF) of patients with eosinophilic pneumonia (EP). METHODS: Periostin, IL-13, and TGF-ß1 concentrations in the BALF were measured by enzyme-linked immunosorbent assay in patients with acute EP, chronic EP, idiopathic pulmonary fibrosis (IPF), and sarcoidosis. Further, we analyzed the relationship between periostin, IL-13, and TGFß-1, levels and the number of inflammatory cells in the BALF. RESULTS: The absolute number of eosinophils, and the periostin, IL-13, and TGF-ß1 levels in the BALF were significantly higher in patients with EP than in patients with IPF and sarcoidosis. Concentrations of periostin significantly correlated with the concentrations of TGF-ß1, but not those of IL-13, in the BALF of patients with EP. Periostin levels also significantly correlated with the absolute number of eosinophils in the BALF of patients with IPF, but not EP. CONCLUSIONS: Our findings suggest that TGF-ß1 might increase the production of periostin in the lungs of patients with EP. Periostin might contribute the pathogenesis of not only EP, but also IPF.

Deciduoid type malignant pleural mesothelioma: a case report.

Here, we report a patient with deciduoid type malignant pleural mesothelioma (MPM), which rapidly progressed. A 55-year-old man who might have been exposed to asbestos a few decades ago had severe back pain. The chest X-ray scanning and computed tomography (CT) revealed pleural thickness on his right thoracic space, without the presence of a lung mass. A pleural biopsy was performed and the patient was histologically diagnosed with deciduoid type MPM. Although he received two cycles of chemotherapy, his disease rapidly progressed and he died within two months of the diagnosis of deciduoid type MPM.

Evaluation of Bioequivalence Between the New Procaterol Hydrochloride Hydrate Dry Powder Inhaler and the Approved Dry Powder Inhaler in Patients With Asthma in a Randomized, Double-Blind, Double-Dummy, Crossover Comparison Study: A Phase 3 Study.

Procaterol hydrochloride hydrate (procaterol) is a ß2 -adrenergic receptor agonist that induces a strong bronchodilatory effect. The procaterol dry powder inhaler (DPI) has been frequently used in patients with bronchial asthma or chronic obstructive pulmonary disease. We evaluated the bioequivalence and safety between the new procaterol DPI (new DPI) and the approved procaterol DPI (approved DPI). This study was a randomized, double-blind, double-dummy, crossover comparison to evaluate the pharmacodynamic equivalence of the new DPI and the approved DPI in patients with bronchial asthma. Primary efficacy variables were area under the concentration-time curve (AUC) forced expiratory volume in the first second (FEV1 )/h and maximum FEV1 during the 480-minute measurement period. Patients were divided into 2 groups, New-DPI-First (n = 8) and Approved-DPI-First (n = 8), according to the investigational medical product that was administered first. Patients inhaled 20 µg of procaterol in each period. FEV1 was measured by a spirometer at predose and at 15, 30, 60, 90, 120, 180, 240, 360, and 480 minutes after each investigational medical product administration. Equivalence was evaluated by confirming that the 2-sided 90%CIs for the difference between the new and the approved DPI in means of AUC (FEV1 )/h and maximum FEV1 were within the acceptance criteria of -0.15 to 0.15 L. The difference in means of AUC (FEV1 )/h and maximum FEV1 was 0.041 L and 0.033 L, respectively, and the 90%CI was 0.004 to 0.078 L and -0.008 to 0.074 L, respectively. These CIs were both within the acceptance criteria. The new DPI was assessed as being bioequivalent to the approved DPI.

Biomarkers for differentiation of patients with asthma and chronic obstructive pulmonary disease.

OBJECTIVE: Asthma and chronic obstructive pulmonary disease (COPD) are airflow limitation diseases with similar clinical manifestations but different pathophysiologic mechanisms. To implement the appropriate treatment, it is important to distinguish between asthma and COPD which sometimes might result difficult in clinical practice. We evaluated biomarkers to distinguish between asthma and COPD. METHODS: Blood eosinophil counts and fractional exhaled nitric oxide (FeNO) levels were analyzed. Serum periostin, interleukin-25 (IL-25), and immunoglobulin E (IgE) concentrations were compared between patients with asthma (n = 60), including atopic-asthma (n = 30) and non-atopic asthma (n = 30), and patients with COPD (n = 30). RESULTS: Significantly higher peripheral blood eosinophil counts (p < 0.001), FeNO levels (p < 0.001), and total serum IgE (P = 0.003) concentrations, but not serum periostin (p = 0.584) or serum IL-25 (p = 0.085) concentrations, were detected in patients with asthma compared to patients with COPD. Serum periostin and IgE concentrations were increased in patients with atopic-asthma compared with those with non-atopic asthma and COPD (p < 0.05). The FeNO levels were significantly correlated with the peripheral blood eosinophil counts (r = 0.430, p = 0.001) and serum IL-25 concentrations (r = 0.338, p = 0.009) in patients with asthma. The serum periostin concentrations were also correlated with the serum IgE concentrations (r = 0.375, p = 0.003)and FeNO levels (r = 0.291, p = 0.024) in patients with asthma. Asthma patients were effectively differentiated from COPD patients based on the FeNO levels (p < 0.001) and peripheral blood eosinophil counts (p < 0.001). CONCLUSIONS: FeNO levels and peripheral blood eosinophil counts were useful biomarkers for distinguishing between patients with asthma and COPD. Serum periostin and IgE concentrations could be biomarkers for atopic asthma.

Antimicrobial effect of titanium dioxide after ultraviolet irradiation against periodontal pathogen.

We focused on the antimicrobial effects of titanium dioxide (TiO2) after stopping ultraviolet (UV) irradiation as an adjunctive treatment for peri-implantitis in this study. The aim was to determine the continuous photocatalytic effects of TiO2 after UV irradiation and its antimicrobial activity against periodontal pathogen. The continuous photocatalytic effects of TiO2 after UV irradiation were determined by electron spin resonance (ESR) spectroscopy using TiO2 particles of various sizes with various UV irradiation times. In addition, antimicrobial activity against Porphyromonas gingivalis was investigated by quantitation of colony-forming units (CFUs). The results showed that the ESR signal ratio for the UV-irradiated TiO2 was significantly higher than that of the non-irradiated TiO2. UV-irradiated TiO2 significantly reduced the number of P. gingivalis when compared with non-irradiated controls. These results suggest that TiO2 has a continuous photocatalytic effect even after stopping UV irradiation and that it showed antimicrobial activity against periodontal pathogen.

Cloning of the lanosterol 14-α-demethylase ( ERG11 ) gene in Trichosporon asahii: a possible association between G453R amino acid substitution and azole resistance in T. asahii.

Lanosterol 14-α-demethylase ( Erg11 protein; Erg11p ), encoded by the ERG11 gene, is the primary target of azoles. Recently, a change in affinity of this enzyme for azoles has been reported as a resistance mechanism in several fungal species. Trichosporon asahii ( T. asahii) is susceptible to fluconazole (FLC). This report identified the ERG11 gene of T. asahii (NCBI accession; HQ176415). The Erg11p of T. asahii, presumed from the DNA sequence, was closely related to the Erg11p of Cryptococcus neoformans. Furthermore, a FLC-susceptible strain was cultured in medium containing FLC at concentrations from 4.0 to 16 µg mL(-1) in order to analyze the development of FLC resistance in T. asahii. The degree of resistance was related to the FLC concentration of the growth medium. One highly resistant strain that was cultured in the medium containing 16 µg mL(-1) FLC contained 1 point mutation (G1357C) that caused a single amino acid substitution at G453R. This amino acid is highly conserved among major fungal pathogens, and it is in a region very close to the heme-binding domain, which is characteristic of the cytochrome P450 superfamily, the primary target for the azole class of antifungal agents. This amino acid substitution may have caused the high resistance to azoles in T. asahii.

A case of eosinophilic pneumonia in a tobacco harvester.

BACKGROUND: Eosinophilic pneumonia comprises a group of lung diseases in which eosinophils appear in increased numbers in the lungs. The distinct etiology of eosinophilic pneumonia is unknown, although the previous case series have indicated a relationship between acute eosinophilic pneumonia and the exposure to exogenous substances including the constituents of cigarettes. CASE SUMMARY: A 60-year-old nonsmoking female, who had started to harvest and sort tobacco leaves two months before presentation, was admitted because of persistent coughing, breathlessness, and general malaise. Her laboratory findings revealed eosinophilia. Chest computed tomography showed nonsegmental airspace consolidations bilaterally. A bronchoalveolar lavage fluid analysis also revealed that the numbers of total cells and eosinophils had increased. Although the urine level of cotinine was within the normal range, positive findings were found in the skin scratch-patch tests using tobacco leaf and its extracts, and a biopsy specimen obtained from the positive site demonstrated infiltration of eosinophils in the dermis. The patient was successfully treated with corticosteroids. DISCUSSION: Green tobacco sickness, a type of nicotine poisoning caused by the dermal absorption of nicotine, is a well known occupational illness of tobacco harvesters. Although it is unclear whether the present case could be identified as a subtype of green tobacco sickness, this is the first report of eosinophilic pneumonia occurred in a tobacco harvester which was possibly induced by tobacco leaf exposure.

Clinical features of healthcare-associated pneumonia (HCAP) in a Japanese community hospital: comparisons among nursing home-acquired pneumonia (NHAP), HCAP other than NHAP, and community-acquired pneumonia.

BACKGROUND AND OBJECTIVE: More than 100000 Japanese die of pneumonia every year. The number of people residing in nursing homes is increasing with the ageing of the population. In 2005, the American Thoracic Society/Infectious Diseases Society of America (ATS/IDSA) published important guidelines for the management of healthcare-associated pneumonia (HCAP). In Japan, however, the optimum strategy for management of HCAP is still unclear. The purpose of this study was to clarify the clinical features of patients with HCAP. METHODS: Patients (n = 202) who were consecutively admitted with a diagnosis of acute pneumonia between October 2007 and September 2009 were retrospectively evaluated. Using the ATS/IDSA guidelines, patients were divided into three groups: a community-acquired pneumonia (CAP) group (n = 123), a nursing home-acquired pneumonia (NHAP) group (n = 46) and a HCAP other than NHAP (O-HCAP) group (n = 33). These groups were then compared with respect to laboratory data, microbiological findings and mortality. RESULTS: Thirty-day mortality in the NHAP group (10.9%) tended to be higher than that in the CAP group (3.3%) or the O-HCAP group (0%). The pathogens most frequently identified were Streptococcus pneumoniae and Haemophilus influenzae in the CAP group, methicillin-resistant Staphylococcus aureus and Klebsiella pneumoniae in the NHAP group, and S. pneumoniae and K. pneumoniae in the O-HCAP group. CONCLUSIONS: The NHAP group was clinically different from the O-HCAP group, based on bacteriological examination and mortality rates. In order to accurately diagnose, and formulate optimum treatment strategies for Japanese patients, the categories of HCAP, as specified in the ATS/IDSA guidelines, should not be applied directly either to patients with NHAP or those with O-HCAP.

[Two cases, of pulmonary sclerosing hemangioma, and peripheral lung carcinoid, in which the diagnoses were difficult by intraoperative frozen section examinations].

Case 1: A 38-year-old man was referred to our hospital because of a chest nodular shadow found on a medical check-up. Chest CT showed a mass about 2 cm in diameter with a sharp margin in the right S6 segment. Right lower lobectomy was performed by video-assisted thoracoscopic surgery, because the mass was thought to be a peripheral lung carcinoid by intraoperative frozen section examination. However, the postoperative histopathological diagnosis was pulmonary sclerosing hemangioma with lymph node metastasis. Case 2: An 81-year-old woman was referred to our hospital because of a chest nodular shadow found on a medical check-up. Chest CT showed a mass about 1.5 cm in diameter with a sharp margin in the right S. Partial lung resection was performed by video-assisted thoracoscopic surgery, because the mass was thought to be an inflammatory lymph node on intraoperative frozen section examination. However, the postoperative histopathological diagnosis was peripheral lung carcinoid. Then, a right middle lobectomy was performed. These cases suggest that it may be difficult to diagnose peripheral lung carcinoid or pulmonary sclerosing hemangioma by intraoperative frozen section examination because of their pathological diversity.

A retrospective study of the patients with positive ImmunoCard Mycoplasma test on an outpatient clinic basis.

The simplified ImmunoCard Mycoplasma test for rapidly detecting Mycoplasma pneumoniae-specific IgM antibodies has been widely used. We examined the usefulness of this test for outpatient practices as well as for the association with bronchial asthma. Among 196 patients whose ImmunoCard showed positive, we targeted 57 cases in which we were able to perform this test multiple times. We evaluated the degree of inflammatory response in the laboratory findings at the time of the test, the period until negative seroconversion for antibodies, and the course of their pulmonary function findings. The number of days from the onset of pyrexia until the test was a median of 10 (range 2-70) days, wherein the inflammatory response did not have a significant effect on that number of days. In 35 cases in which we observed the seroconversion to negative for antibodies, the period of the positive phase was 180 (21-421) days, wherein it was significantly longer for smokers. We observed exacerbation in the majority of the asthma patients, and some of the non-asthmatic patients showed either asthma or asthma-like clinical conditions. For ImmunoCard-positive individuals, it is desirable to ascertain the history of respiratory infection retrospectively to about half a year, although this test is known to suffer from lack of both of sensitivity and specificity. Additionally, it is necessary for ImmunoCard-positive patients to pay attention to the exacerbation of asthma and the development of asthma thereafter.

Low expression of T-cell co-stimulatory molecules in bone marrow-derived dendritic cells in a mouse model of chronic respiratory infection with Pseudomonas aeruginosa.

Pseudomonas (P.) aeruginosa frequently colonizes the respiratory tract of patients with chronic respiratory tract infections such as diffuse panbronchiolitis (DPB). The number of dendritic cells (DCs) that play a central role in immune functions as antigen-presenting cells is reportedly increased in the bronchiolar tissues of patients with DPB. However, the functions of DCs in chronic P. aeruginosa respiratory tract infection have not been defined. Here, we assessed the functions of DCs and the effect of macrolide antibiotics that are therapeutic agents for DPB, in a murine model of DPB caused by P. aeruginosa. Mice were intubated with either P. aeruginosa- or saline-precoated tubes for 80 days. Thereafter, the expression of T-cell co-stimulatory molecules (CD40, CD80, and CD86) and cytokine secretion (interleukin (IL)-10, IL-6, IL-12p40, and tumor necrosis factor (TNF)-alpha) on bone marrow-derived DCs stimulated by lipopolysaccharide were examined by flow cytometry and enzyme-linked immunosorbent assays. The expression of co-stimulatory molecules was significantly decreased in mice infected with P. aeruginosa compared to the saline-treated control mice, but production of these cytokines did not significantly differ between the two groups. Pretreatment with clarithromycin ex vivo decreased CD40 expression on DCs obtained from P. aeruginosa-infected mice and also decreased the production of IL-6, IL-12p40 and TNF-alpha by DCs. These findings suggest that chronic P. aeruginosa infection alters DC functions and that macrolides function as anti-inflammatory agents by modulating the functions of DCs in chronic P. aeruginosa infection.