Identification of an atypical monocyte and committed progenitor involved in fibrosis.

Monocytes and macrophages comprise a variety of subsets with diverse functions. It is thought that these cells play a crucial role in homeostasis of peripheral organs, key immunological processes and development of various diseases. Among these diseases, fibrosis is a life-threatening disease of unknown aetiology. Its pathogenesis is poorly understood, and there are few effective therapies. The development of fibrosis is associated with activation of monocytes and macrophages. However, the specific subtypes of monocytes and macrophages that are involved in fibrosis have not yet been identified. Here we show that Ceacam1+Msr1+Ly6C-F4/80-Mac1+ monocytes, which we term segregated-nucleus-containing atypical monocytes (SatM), share granulocyte characteristics, are regulated by CCAAT/enhancer binding protein ß (C/EBPß), and are critical for fibrosis. Cebpb deficiency results in a complete lack of SatM. Furthermore, the development of bleomycin-induced fibrosis, but not inflammation, was prevented in chimaeric mice with Cebpb-/- haematopoietic cells. Adoptive transfer of SatM into Cebpb-/- mice resulted in fibrosis. Notably, SatM are derived from Ly6C-FcεRI+ granulocyte/macrophage progenitors, and a newly identified SatM progenitor downstream of Ly6C-FcεRI+ granulocyte/macrophage progenitors, but not from macrophage/dendritic-cell progenitors. Our results show that SatM are critical for fibrosis and that C/EBPß licenses differentiation of SatM from their committed progenitor.

Be careful of splenic rupture caused by hit by a pitch during a baseball game: a case report.

BACKGROUND: To the best of our knowledge, splenic rupture caused by hit by a pitch (HBP) has not been previously reported. We present a patient who underwent emergency laparotomy for splenic rupture after being HBP during a baseball game. CASE PRESENTATION: A 41-year-old male was HBP in the left abdomen during his first at-bat during a baseball game. During the operation, vascular injury of the splenic hilum and a deeply extending parenchymal injury were observed, and splenectomy was performed. Histologic findings were consistent with splenic rupture. CONCLUSIONS: The patient's postoperative course was uneventful. Although extremely rare, the possibility of intra-abdominal organ injury should be considered in batters who are hit in the abdomen by a pitched baseball, as illustrated by our patient.

Schlafen-8 is essential for lymphatic endothelial cell activation in experimental autoimmune encephalomyelitis.

Schlafen-8 (Slfn8) is a member of the Schlafen family of proteins, which harbor helicase domains and are induced by LPS and interferons. It has been reported that the Schlafen family are involved in various cellular functions, including proliferation, differentiation and regulation of virus replication. Slfn8 has been implicated in T-cell differentiation in the thymus. However, the roles of Slfn8 in the immune system remains unclear. In this study, we generated Slfn8 knockout mice (Slfn8-/-) and investigated the immunological role of Slfn8 using the T-cell-mediated autoimmune model experimental autoimmune encephalomyelitis (EAE). We found that the clinical score was reduced in Slfn8-/- mice. IL-6 and IL-17A cytokine production, which are associated with EAE onset and progression, were decreased in the lymph nodes of Slfn8-/- mice. Immune cell populations in Slfn8-/- mice, including macrophages, neutrophils, T cells and B cells, did not reveal significant differences compared with wild-type mice. In vitro activation of Slfn8-/- T cells in response to TCR stimulation also did not reveal significant differences. To confirm the involvement of non-hematopoietic cells, we isolated CD45- CD31+ endothelial cells and CD45-CD31- gp38+ fibroblastic reticular cells by FACS sorting. We showed that the levels of IL-6 and Slfn8 mRNA in CD45- CD31+ endothelial cells were increased after EAE induction. In contrast, the level of IL-6 mRNA after EAE induction was markedly decreased in CD31+ endothelial cells from Slfn8-/- mice. These results indicate that Slfn8 may play a role in EAE by regulating inflammation in endothelial cells.

Critical role of Trib1 in differentiation of tissue-resident M2-like macrophages.

Macrophages consist of at least two subgroups, M1 and M2 (refs 1-3). Whereas M1 macrophages are proinflammatory and have a central role in host defence against bacterial and viral infections, M2 macrophages are associated with responses to anti-inflammatory reactions, helminth infection, tissue remodelling, fibrosis and tumour progression. Trib1 is an adaptor protein involved in protein degradation by interacting with COP1 ubiquitin ligase. Genome-wide association studies in humans have implicated TRIB1 in lipid metabolism. Here we show that Trib1 is critical for the differentiation of F4/80(+)MR(+) tissue-resident macrophages--that share characteristics with M2 macrophages (which we term M2-like macrophages)--and eosinophils but not for the differentiation of M1 myeloid cells. Trib1 deficiency results in a severe reduction of M2-like macrophages in various organs, including bone marrow, spleen, lung and adipose tissues. Aberrant expression of C/EBPα in Trib1-deficient bone marrow cells is responsible for the defects in macrophage differentiation. Unexpectedly, mice lacking Trib1 in haematopoietic cells show diminished adipose tissue mass accompanied by evidence of increased lipolysis, even when fed a normal diet. Supplementation of M2-like macrophages rescues the pathophysiology, indicating that a lack of these macrophages is the cause of lipolysis. In response to a high-fat diet, mice lacking Trib1 in haematopoietic cells develop hypertriglyceridaemia and insulin resistance, together with increased proinflammatory cytokine gene induction. Collectively, these results demonstrate that Trib1 is critical for adipose tissue maintenance and suppression of metabolic disorders by controlling the differentiation of tissue-resident M2-like macrophages.

Simultaneous Screening of Major Flame Retardants and Plasticizers in Polymer Materials Using Pyrolyzer/Thermal Desorption Gas Chromatography Mass Spectrometry (Py/TD-GC-MS).

This study was conducted with the aim of achieving the simultaneous screening of various additives in polymer materials by utilizing a solvent-free pyrolyzer/thermal desorption gas chromatography mass spectrometry (Py/TD-GC-MS) method. As a first step to achieve this goal, simultaneous screening has been examined by selecting major substances representing plasticizers and flame retardants, such as short chain chlorinated paraffins (SCCPs), decabromodiphenyl ether (DecaBDE), hexabromocyclododecane (HBCDD), and di(2-ethylhexyl) phthalate (DEHP). A quantitative MS analysis was performed to check for the peak areas and sensitivities. Since Py/TD-GC-MS is fraught with the risk of thermal degradation of the sample, temperatures during the analytical process were finely tuned for securing reliable results. The instrumental sensitivity was confirmed by the S/N ratio on each component. The detection limits of all components were less than 50 mg/kg, which are sufficiently lower than the regulatory criteria. With regard to reproducibility, a relative standard deviation (RSD) of about 5% was confirmed by employing a spike recovery test on a polystyrene polymer solution containing mixed standard solution (ca. 1000 mg/kg). In conclusion, the results obtained in this study indicate that Py/TD-GC-MS is applicable for the screening of major flame retardants and plasticizers in real samples with sufficient reproducibility at regulatory levels.

Chemical Synthesis of O-Glycosylated Human Interleukin-2 by the Reverse Polarity Protection Strategy.

The chemical synthesis of human interleukin-2 (IL-2) , having a core 1 sugar, by a ligation method is reported. Although IL-2 is a globular glycoprotein, its C-terminal region, in particular (99-133), is extremely insoluble when synthesized by solid-phase method. To overcome this problem, the side-chain carboxylic acid of the Glu residues was protected by a picolyl ester, thus reversing its polarity from negative to positive. This reverse polarity protection significantly increased the isoelectric point of the peptide segment and made it positive under acidic conditions and facilitated the purification. An efficient method to prepare the prolyl peptide thioester required for the synthesis of the (28-65) segment was also developed. These efforts resulted in the total synthesis of the glycosylated IL-2 having full biological activity.

Exploratory Feasibility Study of Cerebral Cooling by Transpulmonary Cooling During Cardiac Arrest in a Swine Cardiac Arrest Model.

Studies on targeted temperature management for postcardiac arrest syndrome have shown no difference in outcomes between normothermia and hypothermia in patients with postcardiac arrest brain injury. Therefore, further development of therapeutic methods for temperature control in cardiac arrest patients is desirable. Although animal studies have shown that inducing hypothermia during cardiac arrest improves outcomes, no clinically effective method has yet been reported. We investigated whether intra-arrest lung cooling (IALC) effectively lowers brain temperature. A device capable of cooling oxygen was developed. The pigs were subjected to cardiac arrest using the device, ventilated, cooled during cardiopulmonary resuscitation, and resuscitated for 1 hour, with changes in brain temperature closely monitored. A device capable of cooling oxygen to -30°C was used to cool the lungs during cardiac arrest. Through this approach, IALC successfully reduced the brain temperature. Optimal cooling efficiency was observed when chest compressions and ventilation were synchronized at a ratio of 5:1, resulting in an approximate brain temperature reduction of 1.5°C/h. Our successful development of an oxygen-cooling device underscores the potential for lowering brain temperature through IALC using inhaled oxygen cooling.

Undernutrition Scored Using the CONUT Score with Hypoglycemic Status in ICU-Admitted Elderly Patients with Sepsis Shows Increased ICU Mortality.

This study aimed to clarify whether the influence of undernutrition status and the degree of glycemic disorders affected the prognosis of patients with sepsis. A total of 307 adult patients with sepsis were retrospectively enrolled and analyzed. Characteristics, including nutrition status, calculated according to the Controlling Nutritional Status (CONUT) score of survivors and non-survivors, were examined. The independent prognostic factors of these patients with sepsis were extracted using multivariable logistic regression analysis. The CONUT scores in three glycemic categories were compared. Most patients with sepsis (94.8%) in the study had an undernutrition status according to their CONUT scores. High CONUT scores (odds ratio, 1.214; p = 0.002), indicating a poor nutritional status, were associated with high mortality. The CONUT scores in the hypoglycemic group were significantly higher than those in other groups with an undernutrition status (vs. hyperglycemic, p < 0.001; vs. intermediate glycemic, p = 0.006). The undernutrition statuses of patients with sepsis in the study scored using the CONUT were independent predictors of prognostic factors.

Relationship between brain volume reduction during the acute phase of sepsis and activities of daily living in elderly patients: A prospective cohort study.

Brain damage in acute sepsis may be associated with poor long-term outcomes that impair reintegration into society. We aimed to clarify whether brain volume reduction occurs during the acute phase of sepsis in patients with acute brain damage. In this prospective, noninterventional observational study, brain volume reduction was evaluated by comparing head computed tomography findings at admission with those obtained during hospitalization. We examined the association between brain volume reduction and performance of the activities of daily living in 85 consecutive patients (mean age, 77 ± 12.7 years) with sepsis or septic shock. The bicaudate ratio increased in 38/58 (65.5%) patients, Evans index increased in 35/58 (60.3%) patients, and brain volume by volumetry decreased in 46/58 (79.3%) patients from the first to the second measurement, with significant increases in the bicaudate ratio (P < 0.0001) and Evans index (P = 0.0005) and a significant decrease in the brain volume by volumetry (P < 0.0001). The change rate for brain volume by volumetry was significantly correlated with the Katz index (ρ = -0.3790, P = 0.0094). In the acute phase of sepsis in this sample of older patients, 60-79% of patients showed decreased brain volumes. This was associated with a decreased capacity for performing activities of daily living.

The Characteristics of Patients with Severe COVID-19 Pneumonia Treated with Direct Hemoperfusion Using Polymyxin B-Immobilized Fiber Column (PMX-DHP).

Purpose: The characteristics of patients with severe COVID-19 pneumonia who underwent direct hemoperfusion using polymyxin B-immobilized fiber column (PMX-DHP), in addition to steroids and immunomodulators, remain unclear. Patients and Methods: We conducted a retrospective observational study on 31 patients with severe COVID-19 pneumonia treated with PMX-DHP in an intensive care unit (ICU) from December 2020 to September 2021. Results: Outcomes 28 days after admission to the ICU were 20 in the survival group and 11 in the death group. Parameters significantly different between the survival and death group before PMX-DHP were percentage of invasive mechanical ventilation (25% vs 72.7%, P = 0.0209), PaO2/FIO2 (P/F) ratio (104.5 vs 75, P = 0.0317), and sequential organ failure assessment (SOFA) score (2 vs 3, P = 0.0356). Invasive mechanical ventilation avoidance rate was significantly different between the survival (100%) and death group (0%) (P = 0.0012). P/F ratio, respiratory ratio (RR), and lymphocyte counts improved significantly after PMX-DHP for all patients. The lymphocyte counts changed significantly in the survival (P < 0.0001), but not the death group (P = 0.7927). Conclusion: PMX-DHP, in addition to steroids and immunomodulators, may improve oxygenation and alleviate tachypnea by modulating the lymphocyte numbers and levels of various mediator against severe COVID-19 pneumonia. It may be better to perform PMX-DHP before multi organ dysfunction and lung injury has progressed. Furthermore, the early increase in lymphocyte counts after PMX-DHP might be an indicate a positive outcome.

Development of a screening method for phthalate esters in polymers using a quantitative database in combination with pyrolyzer/thermal desorption gas chromatography mass spectrometry.

Seven phthalate esters (di-isobutyl phthalate (DIBP), di-n-butyl phthalate (DBP), benzylbutyl phthalate (BBP), di-(2-ethylhexyl) phthalate (DEHP), di-n-octyl phthalate (DNOP), di-isononyl phthalate (DINP) and di-isodecyl phthalate (DIDP)) were analyzed by pyrolyzer/thermal desorption-gas chromatography/mass spectrometry (Py/TD-GC/MS), the retention index and relative response factor (RRF) relative to DEHP was calculated for each compound and used to construct a quantitative database (qDB). This qDB enables normalization of the retention time and response factor of each phthalate ester between any laboratory simply by analyzing an n-alkane solution and DEHP standard material. This allows for easy calculation of the phthalate ester content of samples without preparation of calibration curves. The efficacy of this qDB method was verified by performing a quantitative analysis of phthalate esters at 4 different laboratories that showed actual retention times were within ±0.012 min of the estimated retention times for all compounds at all laboratories. Similarly, the mean recovery rate (n = 6) at each laboratory was within 79-113%. Quantitative analysis was also performed on 30 real samples using both the qDB method and the Py/TD-GC/MS method set forth in IEC62321-8, which involves the preparation of 1-point calibrations to perform quantitative analysis. The difference in quantitative results between the methods was approximately within ±200 mg/kg for compounds in the concentration region of <2000 mg/kg.

Improvement of sample throughput using fast gas chromatography mass-spectrometry for biochemical diagnosis of organic acid disorders.

BACKGROUND: Gas chromatograph mass-spectrometric (GC/MS) method of analysis for urinary organic acids is used for the diagnosis of a variety of metabolic disorders. The method is time-consuming and does not allow for improvements in sample throughput. Although the sample preparation and the data processing have been improved, the long GC/MS analysis time still remains to be problematic. METHODS: The fast-GC/MS method, which utilizes a short microbore capillary GC column and fast temperature programming, was applied to the analysis for urinary organic acids. Urine samples obtained from 15 patients with 9 different disorders and 16 healthy controls were analyzed using conventional GC/MS and fast-GC/MS. RESULTS: Analysis cycle time was shortened from 1 h to 15 min. The automated data system uses retention indices determined by conventional-GC/MS for the identification of 134 organic acids. These retention indices can also be used in data obtained by fast-GC/MS. New fast-GC/MS method with the automated data system gave the same diagnostic results as conventional-GC/MS except for 1 healthy control. CONCLUSIONS: The combined system of fast-GC/MS and the automated data system will be powerful tools in clinical laboratories due to increased sample throughput and reduced analysis costs.

A Simple and Robust Method for Determination of Alkylmercury in Seawater and Industrial Wastewater by Phenylation Pretreatment Combined with GC-MS.

For determination of methylmercury (MeHg) and ethylmercury (EtHg) in seawater and industrial wastewater, a simple and robust analytical method was developed based on phenylation and solvent extraction followed by GC-MS measurement. Alkylmercury compounds were directly phenylated with sodium tetraphenylborate in water and extracted into toluene. The method detection limits obtained for MeHg and EtHg in pure water were 53.3 and 33.5 ng Hg L-1, respectively, which are almost 10 times lower than the environmental quality standards for water pollution in Japan (EQSJ): 0.5 µg Hg L-1. The recoveries of alkylmercury compounds from seawater and four kinds of industrial wastewater except for EtHg from treated wastewater of an optic lens factory were satisfactory (>90%) at 1- or 4-fold concentrations of the EQSJ. Contrarily, the low recovery of EtHg from the treated wastewater (75.4 ± 4.7%) was found to be caused by the rapid decomposition of EtHg into inorganic mercury.

Surgically removed thoracolithiasis: report of two cases.

Thoracolithiasis is a rare condition with only 12 cases of surgically removed nodules reported in the literature. We report 2 additional cases. Case 1: A 19-year-old male admitted with an abnormal shadow on a chest X-ray. Computed tomography (CT) revealed a nodule in the right lower lung lobe. The material extirpated by thoracoscopy was milky white, glossy, and 1.6 cm in diameter. Histopathologically, it consisted of fatty necrotic tissue covered with hyalinized fibrous tissue. Case 2: A 78-year-old female, with a past history of breast cancer, admitted with an abnormal shadow on chest X-ray. CT revealed a nodule in the left lung S(1+2) segment, of which transbronchial biopsy findings indicated primary lung adenocarcinoma. Exploratory thoracoscopy incidentally revealed some pearly material, 0.4 cm in diameter, in the thoracic cavity. They were extirpated during left upper lobectomy for lung cancer; all of them demonstrated concentric hyalinized fibrous tissue. Thoracic surgeons should consider this condition in the differential diagnosis of a peripheral pulmonary nodule.

Pulmonary suture abscess with false-positive 18F-fluorodeoxyglucose positron emission scan mimicking lung cancer recurrence.

We present the case of a 57-year-old woman with pulmonary suture abscess. She had undergone right S3 segmentectomy for early lung adenocarcinoma 7 years before and right breast-conserving surgery for invasive ductal carcinoma 5 months previously, followed by irradiation plus endocrine therapy. Chest radiography and computed tomography revealed an irregular mass (3.5 cm in diameter) between the residual S1 segment and the middle lobe, neighboring the staple line of the segmentectomy. 18F-fluorodeoxyglucose uptake into the mass increased, seen by positron emission scans. Therefore, we could not rule out the possibility of local recurrence of lung cancer and resected it. Pathologically and microbiologically, the mass was a suture abscess arising around the nylon suture of the previous segmentectomy. This lesion was the result of a foreign-body reaction, as confirmed by polarized microscopy. Moreover, titanium staples at the segmentectomy and breast-conserving surgery may also have contributed to this condition.

Ectopic cervico-mediastinal thymoma confirmed by flow cytometric analysis of tumor-derived lymphocytes.

Ectopic cervical or cervico-mediastinal thymomas are very rare and most of them are asymptomatic, except for the presence of a cervical mass. We present the case of a 71-year-old man with an ectopic cervico-mediastinal thymoma threatening superior vena cava syndrome. He had a slight headache and presented with venous dilatation on the chest wall. A computed tomographic scan and magnetic resonance imaging of the chest demonstrated a mass extending from the right neck to the hilum, that indented the trachea and compressed and displaced the brachiocephalic veins anteriorly. Under a right hemicollar incision and median sternotomy, the mass was resected en bloc together with the thymus. The resected specimen was an encapsulated mass measuring 11x7x4 cm. The pathological diagnosis was type AB, non-invasive thymoma, confirmed by 3-color flow cytometry of tumor-derived lymphocytes. Flow cytometry using biopsy material may contribute to the preoperative diagnosis of ectopic thymoma.

[Prostate small cell carcinoma: report of two cases that differed in treatment responsiveness].

Case 1 : A 76-year-old man with a chief complaint of dysuria had an elevated prostate specific antigen (PSA) level of 24.9. He underwent a transperineal needle biopsy of the prostate, and the histopathological diagnosis was prostatic small cell carcinoma. The cancer was clinically diagnosed as T3bN1M1 with multiple lung metastases. He started receiving hormonal therapy. After three months of hormonal therapy, the multiple lung metastases disappeared. Thereafter, the serum PSA level and the tumor volume increased and he died 12 months from the start of therapy. Case 2: A 79-year-old man was referred to our hospital with a chief complaint of dysuria. The serum level of PSA was elevated to 10.4. Transperineal prostate biopsy revealed prostatic small cell carcinoma. The cancer was clinically diagnosed as T3bN1M1, and hormonal therapy was started. Subsequently, although his serum PSA level declined, his condition worsened rapidly and he died five months after the start of therapy.

[An elderly patient with non-resectable lung cancer responding to TS-1].

A 76-year-old man had complained of hoarseness, productive cough and left chest pain in June 2005. Chest radiograph and computed tomography showed a tumor 7.5x5 cm in size which contacted the aortic arch, some nodules in left upper lobe and right lower lobe of lung and left pleural effusion. The broncho-fiberscopic biopsy histologically proved the tumor to be moderately differentiated squamous cell carcinoma in the left upper lobe of lung. The patient was diagnosed as a non-resectable lung cancer with clinical stage IV (c-T4N2M1). In July 2005, he was treated with oral administration of TS-1 (120 mg/day) and Jyu-zen daiho toh (7.5 g/day). After one week, he complained of grade 2 leukopenia and grade 1 skin rash. Then TS-1 was reduced to 100 mg/day. After this modulation of the treatment, he could take TS-1 and Jyu-zen daiho toh days 1-14, every 3 weeks. The patient then received 17 courses until June 2006. The left pleural effusion and left chest pain disappeared only 2 weeks after taking TS-1. In June of 2006, central necrosis of the left lung tumor was confirmed by positron emission computed tomography. No serious adverse effect was observed, and the patient maintained good quality of life. This case suggests that TS-1 with Jyu-zen daiho toh may be an effective treatment for elderly advanced lung cancer.

Hepatoma-derived growth factor as a prognostic marker in completely resected non-small-cell lung cancer.

Hepatoma-derived growth factor (HDGF), unrelated to hepatocyte growth factor, is a heparin-binding protein originally purified from human hepatoma HuH-7 cells. HDGF exhibits mitogenic activities for certain hepatoma cells, fibroblasts and vascular smooth muscle cells, and angiogenic activities through nuclear targeting. Recently, HDGF was found to be a mitogen for lung epithelial cells in vitro and in vivo. This suggests that HDGF may play a critical role in the development and progression of lung cancer. We investigated, immunohistochemically, the relationship between HDGF expression and clinicopathological variables, and the prognostic significance of HDGF in 102 patients with completely resected non-small-cell lung cancer (NSCLC: 70 adenocarcinomas and 32 squamous cell carcinomas). To address the mechanism of action of HDGF, we evaluated the contribution of HDGF to tumor cell proliferation and intratumor angiogenesis using anti-Ki-67 and anti-CD31 antibodies, respectively. HDGF expression was strongly detected in the nucleus of cancer cells; the HDGF-labeling index (LI) was 20-95% (median 64.5%). There was no significant association between HDGF-expression level and clinicopathological variables. Patients with NSCLC showing a high HDGF-LI (> or =65%) had significantly worse overall and disease-free survivals than those with NSCLC showing a low HDGF-LI. Multivariate analysis revealed that HDGF is a significant independent prognostic factor, more powerful than pathological stage. Moreover, HDGF expression correlated with Ki-67-LI and intratumor microvessel density. We consider HDGF as a useful prognostic marker for patients with completely resected NSCLC and it may play a critical role in the pathobiology of lung cancer through its mitogenic and angiogenic activities.

Novel gas chromatography-mass spectrometry database for automatic identification and quantification of micropollutants.

A novel gas chromatography-mass spectroscopy (GC-MS) database for identification and quantification of micropollutants in environmental and food samples is reported. GC retention times, calibration curves, and mass spectra of nearly 700 chemicals were registered in the database, and the GC retention times of registered chemicals in actual samples were predicted from the retention times of n-alkanes measured before sample analysis. Differences between predicted and actual retention times were less than 3 s, an accuracy that is nearly identical to that obtained by analysis of standard substances. After the retention times were predicted, a calibration file for the GC-MS instrument was created from the predicted retention times, calibration curves, and mass spectra of the registered chemicals. With the resulting calibration file, automated identification of all the chemicals in actual samples was possible without the use of standards, and the identification method was as reliable as conventional methods. When the GC inlet, column, and tuning conditions were adjusted using GC-MS performance check standards, relative standard deviations of 20% or less for determination values could be obtained. More than 90% of the chemicals in the database could be detected at a sensitivity sufficient for all practical purposes (100 pg or less). Because each chemical in the database, to which new substances can easily be added, can be determined in 1 h, micropollutants in samples can be analyzed efficiently and inexpensively.