As the second most common occupational disease in China, occupational poisoning is one of the major public health problems that seriously affect workers' health. This study aimed to investigate the epidemiological and occupational characteristics of acute and chronic occupational poisoning cases in Zhejiang Province, so as to provide a scientific basis for proposing intervention measures and preventive strategies of occupational poisoning.The data on occupational poisoning cases in Zhejiang Province from 2006 to 2020 was derived from the National Occupational Disease Network Direct Report System. A descriptive statistical analysis was employed on this data utilizing R software.P < 0.05). Benzene and lead and its compounds (excluding tetraethyl lead) were the major toxicants causing occupational poisoning. More than 60% of occupational poisoning cases were reported in private enterprises. Meanwhile, over 90% of the cases were distributed in medium enterprises and small enterprises. The type of industry with the most occupational poisoning cases was the manufacturing industry.From 2006 to 2020, 1,008 occupational poisoning cases were reported in Zhejiang Province, with a downward trend since 2007. Of these cases, 81.94% were chronic poisoning and 18.06% were acute poisoning. Ningbo reported the most occupational poisoning cases among the 11 cities in Zhejiang Province, accounting for 20.34% of the total cases. Besides, the occupational poisoning cases in Wenzhou, Jiaxing, and Shaoxing also accounted for 18.15%, 18.06%, and 17.76% of the total number of cases, respectively. Occupational poisoning in male were 693 cases and in female 315 cases. Most of the occupational poisoning cases studied involved people aged between 40 and 49 years (38.19%). The length of work in chronic occupational poisoning cases was significantly higher than that of acute occupational poisoning cases (Although the cases of occupational poisoning in Zhejiang Province have declined, more comprehensive and effective prevention and control measures are still needed. More attention ought to be paid to the management of key points according to the epidemiological and occupational characteristics of occupational poisoning cases. Occupational poisoning, as the poisoning caused by exposure to toxic chemicals during work, is the occupational disease second only to pneumoconiosis in the Chinese occupational disease catalog , 2. In tShort-term high-concentration exposure of workers to toxicants in the workplace could lead to acute poisoning accidents, which can be fatal in severe cases , 7. MoreWhile occupational diseases have long been a concern, many countries lack official figures on occupational accidents and work-related illnesses due to the absence of proper recording and notification systems . In ChinThrough the analysis of occupational poisoning cases in Zhejiang Province from 2006 to 2020 collected on the direct reporting system, this study elucidates the incidence trend and characteristics of occupational poisoning in Zhejiang Province in recent 15 years, so as to provide a scientific basis for the prevention and control strategies of occupational poisoning. Many enterprises in Zhejiang Province may use or produce various chemicals in the process of product manufacturing, which greatly increases the risk of occupational poisoning incidents. Therefore, clarifying the epidemiological characteristics of occupational poisoning in Zhejiang Province in the past 15 years is of great significance for decision-makers to effectively prevent occupational poisoning.The study utilized data on occupational poisoning cases from 1 January 2006 to 31 December 2020 in Zhejiang Province via the Occupational Disease and Health Hazards Monitoring System which is part of the China information system for disease control and prevention. All data has been confirmed, reviewed, and reported by occupational disease diagnosis institutions.The descriptive analysis of occupational poisoning cases reported in Zhejiang Province from 2006 to 2020 was carried out based on factors such as general information, region, types of toxicants, enterprise ownership type and scale, industry, and occupation.Types of toxicants: 60 toxicants (including one open item) causing occupational poisoning are listed in the National Occupational Disease Classification and Catalog .Enterprise ownership type: state-owned, collectively owned, privately owned, foreign, Hong Kong/Macao/Taiwan, and others .Enterprise scale: Based on the Enterprise Scale Standard established by the State Statistical Bureau , enterprIndustry classification: The industry classification of the enterprises is based on the Industrial Classification for National Economic Activities , divided25, P75). For the categorical variables, frequencies (n), and proportions (%) were calculated. After descriptive analysis of the data, further comparison between groups was performed using Chi-square test, Student's t-test and Wilcoxon rank sum test, and differences were considered statistically significant at P < 0.05. Mann-Kendall trend test was applied to determine whether the number of occupational poisoning cases over the years has a monotonic upward or downward trend, and P < 0.01 indicated a significant trend change.The data on occupational poisoning in Zhejiang Province from 2006 to 2020 were introduced to Excel for sorting. Data management, analysis, and plotting were performed using R4.1.3. The continuous variables for the normal distribution were described as mean ± standard deviation (SD), and data with the skewed distribution were described using median and percentile , while cases involving acute poisoning were 182 (18.06%).2 = 49.24, P < 0.05). The number of occupational poisoning cases increased since 2006, and the growth rate from 2006 to 2007 reached 146.94%. From 2007 to 2010, the number of cases was generally at a high level, with a temporary decrease in 2009, after which the numbers rose again. From 2007 to 2020, although the number of occupational poisoning cases fluctuated, it showed a significant downward trend (P < 0.01). In particular, the trend of acute poisoning cases was relatively stable from 2006 to 2020, with a maximum of 22 cases in 2017. Chronic poisoning shows a similar trend to the overall occupational poisoning, with a maximum of 111 cases in 2007 and a significant downward trend since 2007 (P < 0.01).2 = 299.06, P < 0.05). Occupational poisoning exhibited different characteristics in different regions. Acute poisoning was reported the most in Quzhou with 48 cases (26.37%), followed by Taizhou with 28 cases (15.38%), Jiaxing , and Ningbo . Chronic poisoning was reported the most in Ningbo with 185 cases (22.40%), followed by Wenzhou , Shaoxing , and Jiaxing .From 2006 to 2020, occupational poisoning cases were reported in 11 cities in Zhejiang Province, of which Ningbo had the most with 205 cases (20.34%), followed by Wenzhou with 183 cases (18.15%), Jiaxing , Shaoxing , Zhoushan the least with only 1 case (0.10%) . The disAccording to the trend of occupational poisoning cases reported over the years, occupational poisoning in Shaoxing reached its peak in 2007 (74 cases), and then gradually declined and remained at a low level since 2010 . The num2 = 6.45, P < 0.05). The average age of poisoning cases was 38.83 ± 9.28, of which the age of acute poisoning cases and chronic poisoning cases were 40.51 ± 9.75 and 38.46 ± 9.15, respectively. A statistical difference was found between the age of acute and chronic poisoning cases . The acute and chronic occupational poisoning cases were both primarily found in the age group of 40–49 years, accounting for 39.01% and 38.01% of all acute and chronic poisoning cases, respectively  occupational poisoning cases in males and 315 (31.25%) in females were reported in Zhejiang Province, among which the number of acute poisoning cases and chronic poisoning cases in males was 3.33 times and 2.03 times that of females, respectively. The number of male and female poisoning cases differed in a statistically significant manner ] at the onset time of occupational poisoning cases in Zhejiang Province was 3.25  years. The length of work in chronic occupational poisoning cases was 3.50  years, which was significantly higher than that of acute occupational poisoning cases with 1.06  years (P < 0.05). The acute occupational poisoning cases were mainly distributed in the group of work length at 0–1 years with 87 cases (47.80%), while the group of work length with the most chronic occupational poisoning cases was 1–4 years (336 cases), accounting for 40.68% of the total number of chronic poisoning cases , where the major toxicant that caused occupational poisoning were benzene with 309 cases (30.65%), and lead and its compounds (excluding tetraethyl lead) with 280 cases (27.78%). As shown in From 2006 to 2020, there were differences between the number of cases of different toxicants , with most of the occupational poisoning cases occurring in private enterprises and foreign enterprises, whose were 683 (67.76%) and 196 (19.44%), respectively. The enterprises with other ownership types reported 129 cases (12.80%)  . There w2 = 70.85, P < 0.05). Among the occupational poisoning cases, there were 42 cases (4.17%) in large enterprises, 551 cases (54.66%) in medium enterprises, 399 cases (39.58%) in small enterprises, 2 cases (0.20%) in micro enterprises, and 14 cases (1.39%) in enterprises with unknown size. Among them, the small enterprises had the most acute poisoning cases ; the medium enterprises had the most chronic poisoning cases   .The curve of poisoning cases from medium enterprises and small enterprises fluctuated between 2006 and 2020 . Medium 2 = 41.54, P < 0.05), with cases mainly distributed in the manufacturing industry, accounting for more than 90%  were the major toxicants of chronic poisoning, and they were also the toxicants that caused the most occupational poisoning cases, accounting for more than 50% of the total occupational poisoning cases. A study in 2018 by Wang et al. . AccordiIn recent years, the occurrence of lead and benzene occupational poisoning was severe in Zhejiang Province. Previous studies have shown that most organic solvents have chronic toxic effects, among which benzene is a known carcinogen in humans , 30. RegThere are many types of enterprises in Zhejiang Province, and many types and quantities of chemicals are used in production . This stA wide range of toxic chemicals is in heavy use, and numerous new technologies, new materials, and new products are continuously introduced in Zhejiang Province in recent years . BesidesIndustry distribution showed that occupational poisoning cases in Zhejiang Province were mainly concentrated in the manufacturing industry. Zhejiang Province is one of the earliest developing provinces of manufacturing industry in China. Since the late 1970s, after decades of rapid development, the manufacturing industry in Zhejiang Province has now developed into an important domestic manufacturing agglomeration and export base for industrial manufactured products . The sumOccupational chemical poisoning is a category of occupational diseases with high fatality rate and great harm to workers. In this study, epidemiological and occupational characteristics of occupational poisoning cases in Zhejiang Province from 2006 to 2020 were described. The results exhibited that occupational poisoning cases reported in Zhejiang Province were predominantly chronic poisoning, and acute poisoning occasionally occurred. According to the characteristics of occupational poisoning cases, classified prevention and control strategies should be implemented, and the key points should be highlighted. Lead and benzene poisoning should be listed as key occupational diseases for surveillance, and the control of these toxicants should be strengthened. Great importance should be attached to private small and medium-sized enterprises in regions with a high incidence of occupational poisoning, and the prevention and treatment of occupational poisoning shall be carried out according to the characteristics of industry distribution.The original contributions presented in the study are included in the article/The studies involving human participants were reviewed and approved by Ethics Committee of Zhejiang Provincial Center for Disease Control and Prevention (2022-026-01). The patients/participants provided their written informed consent to participate in this study.LZ: conceptualization, investigation, formal analysis, and writing original draft. FW: data curation, methodology, and formal analysis. XF: data curation and funding acquisition. YZ: formal analysis. YH: data curation and supervision. XL: conceptualization and supervision. PX: conceptualization, data curation, visualization, and writing—review and editing. HZ: methodology, writing—review and editing, funding acquisition, and supervision. All authors contributed to the article and approved the submitted version.This work was supported by the Medical Health Technology Project by the Health Commission of Zhejiang (Grant Number: 2019KY056), 2021 Zhejiang Provincial Center for Disease Control and Prevention for the Technology Talent Incubation Project.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Post-transplant lymphoproliferative disease (PTLD) is a serious complication occurring as a consequence of immunosuppression in the setting of allogeneic hematopoietic stem cell transplantation  or solid organ transplantation (SOT). The majority of PTLD arises from B-cells, and Epstein–Barr virus (EBV) infection is present in 60–80% of the cases, revealing the central role played by the latent infection in the pathogenesis of the disease. Therefore, EBV serological status is considered the most important risk factor associated with PTLDs, together with the depth of T-cell immunosuppression pre- and post-transplant. However, despite the advances in pathogenesis understanding and the introduction of novel treatment options, PTLD arising after alloHSCT remains a particularly challenging disease, and there is a need for consensus on how to treat rituximab-refractory cases. This review aims to explore the pathogenesis, risk factors, and treatment options of PTLD in the alloHSCT setting, finally focusing on adoptive immunotherapy options, namely EBV-specific cytotoxic T-lymphocytes (EBV-CTL) and chimeric antigen receptor T-cells (CAR T). Post-transplant lymphoproliferative disease (PTLD) is a rare and serious complication that occurs as a result of immunosuppression in the setting of allogeneic hematopoietic stem cell transplantation  or solid organ transplantation (SOT) . The estSurvival rates of PTLD have improved with the introduction of the anti-CD20 monoclonal antibody rituximab. However, reports on the potential cure of refractory PTLD are scarce. Today, there is no consensus on how to treat rituximab refractory PTLD, especially in highly aggressive diseases. In the present review, we will focus on PTLD following alloHSCT.The histological classification  includes six types of morphological PTLD: plasmacytic hyperplasia, infectious mononucleosis-like, florid follicular hyperplasia, polymorphic, monomorphic (B-cell or T-/NK-cell types), and classical Hodgkin lymphoma PTLD. From a pathological point of view, monomorphic PTLD is not distinguishable from lymphomas with the same cell of origin in immunocompetent patients .The majority of PTLD arises from B-cells; of note, latent Epstein–Barr virus (EBV) infection is present in 60–80% of the cases. This finding has led to intense research into the oncogenetic implications of EBV in this rare disease . EBV-posHodgkin lymphomas and PTLD of T-lineage (T-PTLD) are the rarest forms of monomorphic PTLD. Due to the rarity of T-PTLD, knowledge about pathogenesis, risk factors, therapy, and prognosis relies predominantly on case reports and small series. T-lymphocytes do not express the EBV receptor CD21; however, some T-PTLD might show aberrant T-cells, which are positive for both CD21 and EBV .The pathophysiology of EBV-positive PTLD has been described. EBV virus is a latent gamma herpes virus with known oncogenic potential. Sero-epidemiology models suggest that 90% of the adult population has been infected, making it the most ubiquitous virus in humans . PrimaryDuring the acute phase of EBV infection, oral mucosa cells and B-cells become infected, leading to a robust immunogenic response with both antibody and cellular upregulation (via specific CD8 and CD4-positive T-cells) ; by mechThe oncogenic potential of EBV, especially in its latent phase, is demonstrated by the association with several malignancies, including endemic Burkitt lymphoma, Hodgkin lymphoma, and HIV-associated lymphomas other than PTLD . MoleculLMP-1 mimics CD40 and constitutively activates important proliferative signaling pathways in the B-cell cycle, such as NF-kB, JAK/STAT, ERK MAPK, IRF, and Wnt ; it has LMP-2A interacts with the B-cell receptor signaling, and its pathway has been linked to anti-apoptosis . UltimatUnder normal circumstances, the over-expression of LMP1 and LMP-2A by B-cells with latent EBV infection promotes a robust CD4 T-cell response that prevents the development of malignancies . HoweverEBV-negative PTLD is less common, often diagnosed later in the post-transplant period, and tends to have a poorer response to immunosuppression withdrawal . The patTwo major features are related to the development of PTLD: (1) EBV serological status of transplant recipient and donor; and (2) depth of T-cell immunosuppression pre- and post-transplant. In particular, EBV serological status is considered the most important risk factor for post-SOT PTLDs  : recipieFor alloHSCT patients, ex vivo or in vivo T-lymphocyte depletion plays a major role. In fact, the use of anti-thymocyte globulin (ATG), commonly employed for matched unrelated donors, strongly affects PTLD development , similarThe role of other immunosuppressant drugs is more uncertain, partly due to different drug schedules and doses. High-intensity immunosuppression in the peri-transplant induction phase appears linked to early PTLDs, while the cumulative immunosuppressant burden in the following years affects late PTLDs . SimilarRecently, reduced intensity conditioning (RIC) regimens have been widely adopted to reduce transplant-related mortality and morbidity, allowing a widening of HSCT indications even to older patients or with a higher comorbidity index. The use of RIC regimens has been proven to be an independent risk factor for EBV-PTLD development, probably due to the more prolonged absence of normally restricting EBV-specific T-lymphocytes . SimilarAdditional risk factors have been identified, such as the presence of the highly oncogenic variant protein LMP-1 in donors, an untreated HCV infection in recipients , and non-Caucasian ethnicity ,30. MoreIt has also been hypothesized that chronic antigen stimulation from viruses other than EBV might play a potential role in EBV-negative PTLDs . In partPTLD occurrence after alloHSCT is primarily associated with the degree of HLA matching, given that more mismatches typically require stronger immunosuppression to prevent GvHD . Thus, tChronic GvHD is a potentially strong risk factor for late-onset PTLDs, both due to immune deregulation from GvHD itself and prolonged immunosuppression . Little In a review of 163 cases, alloHSCT was associated with early-onset T-PTLD, whereas late-onset occurred after immunosuppression with steroids and azathioprine without the administration of calcineurin inhibitors . The majIt is widely assumed that the reduction of the immunosuppression (RIS) is the typical first-line treatment for EBV-positive PTLD, though it may highly increase the risk of GvHD flare in alloHSCT and of rejection in SOT. Moreover, although data are scarce, RIS appears to have limited efficacy, and a specific treatment is necessary in almost the totality of cases.The monoclonal anti-CD20 antibody rituximab is the gold standard of treatment in CD20+ PTLD, polymorphic PTLD, or monomorphic diffuse large B-cell lymphoma-like PTLD resistant to RIS ,38. Howep-value 0.001), as well as its uses any time before transplant   was the histological type in 80% of cases . The maxOverall, several clinical trials are expanding the use of third-party EBV-CTL in the setting of PTLD. A phase III trial is exploring the administration of commercially available off-the-shelf, allogeneic EBV-CTL called Tabelecleucel for EBV+ PTLD patients who failed rituximab or rituximab and chemotherapy . Preliminary results showed an ORR of 50% and a median duration of response not reached in responders . AnotherChimeric antigen receptor T-cell (CAR T-cell) therapy directed against CD19 surface antigen is a novel immunotherapeutic approach that demonstrated promising activity against relapsed and refractory DLBCL ,64,65. ETo date, only one group recently reported successful treatment of PTLD after alloHSCT with donor-derived CAR T-cell therapy . The treFinally, another tumor-associated target is being explored for the treatment of EBV-positive cancers. Preclinical studies showed encouraging results using CAR T-cells directed against surface LMP1 expressed in EBV-positive cancers such as nasopharyngeal carcinoma . An earlNew insights in the pathophysiology of PTLD and in the role of EBV infection have allowed remarkable advances in the treatment and in the improvement of clinical outcomes of this rare, but life-threatening complication of alloHSCT. EBV serological status and immunosuppressive strategies for transplantation remain the most relevant predictive factor for the onset of the disease. However the recent introduction of novel, effective adoptive immunotherapies represent an encouraging option for refractory cases.


Furthermore, the incorporation effect of ZnO:Al (TCO) and CuSCN (HTL) nanolayers was studied for the first time. As a result, the efficiency of the solar cell was maximized from 16.04% to 17.74% by increasing the Jsc and Voc. This work will play an essential role in enhancing the performance of CdTe-based devices with the best performance.A numerical simulation is a valuable tool since it allows the optimization of both time and the cost of experimental processes for time optimization and the cost of experimental processes. In addition, it will enable the interpretation of developed measurements in complex structures, the design and optimization of solar cells, and the prediction of the optimal parameters that contribute to manufacturing a device with the best performance. In this sense, a detailed simulation study was carried out in this work by the Solar Cell Capacitance Simulator (SCAPS). In particular, we evaluate the influence of absorber and buffer thickness, absorber defect density, work function in back contact, R Still, the consumption of the material and the high manufacturing costs of these cells have prompted the search for polycrystalline absorber layers of thin films that can adequately replace silicon. One material of interest is CdTe, which has the highest efficiency in thin-film solar cells, this value being above the one reported for CIGS . CdTe is5/cm) , which e5/cm) . Althoug5/cm) ,5,6,7.3 [2O5 [x [oc [On the other hand, solar cell performance greatly depends on the energy barrier at the absorber/contact interface. Therefore, to reduce this barrier, a back contact with a low resistance and high work function (WF) is necessary. In recent years, many researchers have studied diverse materials employed as back contact layers for CdTe devices ,9,10. So3 , V2O5 [13 [2O5 , NiO [13 [2O5 [x , are use [2O5 [x . These p5 [x [oc . Moreove5 [x [oc ,18.2:F and ZnO:Al) present a good alignment of the conduction band (CB) with the CdS material and a large valence band (VB). Therefore, minimal recombination is expected at the interface, as electrons pass through smoothly and holes are blocked. However, ZnO:Al has a lower refraction index (of around 7–9%) than SnO2:F. Then, this quality could make it more suitable for CdS/CdTe devices because it allows for promoting more significant photon absorption in the CdTe (absorber) layer [On the other side, a transparent electrode (TCO) is used as a front contact to extract the photogenerated electrons to the external charge, usually indium tin oxide (ITO) or fluorine-doped tin oxide (FTO). Nevertheless, both materials possess a high elaboration cost, low electron mobility, and transmittance of around 80% ,20. So, r) layer ,25,26.2:F  [Another essential requirement for a material to be used as front contact is its lattice matching with the buffer layer, so structural parameters should be considered during their coupling. Lattice-matched materials can be grown on top of one another without forming a high density of nonradiative recombination. However, when the growth of lattice-mismatched materials is attempted, many defects result from the relaxation of strain in the crystal structure ,28. In t 3.18 Å) .2 materials. However, several extensions have improved their application capabilities to Si and GaAs solar cells and other materials, such as perovskite [Due to the complexity of solar cells, it is crucial to understand the physics behind these technologies. A helpful tool is a numerical simulation, which allows us to solve the most critical differential equations of solar cells and design and optimize advanced cell structures. SCAPS-1D is a solar cell simulation software conceived at the University of Gent, Belgium. This program was mainly designed for the CdTe and CuInSerovskite ,31. In arovskite ,33.The present investigation yielded an extensive study about the impact of different parameters on the CdTe/CdS device structure by SCAPS-1D. Then, a new configuration proposal (CuSCN/CdTe/CdS/ZnO:Al) was studied for improving the device performance.2:F/CdS/CdTe/Carbon [2:F configuration. This work investigates a CdTe solar cell in superstrate structures to analyze the impact of different parameters on the device’s performance. The reference solar cell configuration in the simulation was as follows: SnOe/Carbon . The devThe SCAPS software solves the fundamental semiconductor equations, the Poisson, the continuity equations for electrons and holes, and the drift equation . The sofTo validate our model, it was necessary to simulate a base solar cell with the typical configuration of CdTe. In this sense, the J-V and EQE curves were calculated by SCAPS and are presented in The film thickness is a critical manufacturing parameter affecting the device’s cost and environmental protection properties. An improvement at thicker absorber layers is observed in the J-V curves of CdTe devices a. The besc results presented in Results show that when the CdTe thickness was increased up to 3000 nm, the Jsc increased, improving the PCE. This is because a larger CdTe layer can absorb more photons of higher wavelengths, increasing the number of charge carriers generated in it. An improvement was observed in almost all the output parameters with the increase in thickness. However, this effect on the FF was not perceived at thicknesses greater than 3000 nm, which could be attributed to a possible increase in Rs. The plotted graph of EQE vs. wavelength for different thicknesses of the CdTe layer is presented in The study of the influence of CdS thickness on the solar cell is essential to improve performance. In this sense, oc. oc from 0.87 V to 0.86 V, Jsc from 24.89 mA/cm2 to 23.22 mA/cm2, FF from 76.18% to 76.09%, and PCE from 16.53% to 15.37%.A linear decrease was observed in almost all the output parameters by the thickness increase a. Howevesc (24.86 mA/cm2 at 10 nm) presented in oc (from 0.41 V to 1.10 V), FF (from 60.6% to 77.52%), and PCE (from 5.99% to 20.12%) is observed with the increase in work function values, while the Jsc remains the same. Therefore, materials with back contact values greater than 5.7 eV are unnecessary.The energy needed to remove an electron of the highest level of the stationary Fermi distribution of a solid is known as the work function. The contacts greatly influence the performance and efficiency of solar cells because these devices are highly dependent on the front and back contact work function . Figure Charge carrier generation and recombination are decisive processes for the performance of CdTe solar cells. The first occurs when sunlight strikes the device, allowing the generation of photo-generated carriers in the absorber layer. The charge carriers are collected and transferred to the external current. However, many of these are lost due to the inadequate quality of the material. Higher defect density, commonly found in low-quality materials, increases the recombination rate, reducing the diffusion length of charge carriers and, thus, their lifetime. To determine the effect of defect density on CdTe solar cells’ performance, it is important to understand the recombination mechanisms  .−3) in the CdTe. These simulations were performed at different defect densities from 1012 to 1017 cm−3  increased as the defect density of the CdTe decreased. As seen in 13 cm−3, which reveals that an efficiency of 17.56% could be achieved only in CdTe films with remarkably low defect densities.As mentioned above, defects in absorber solar cells have a significant impact on efficiency. In this study, numerical simulations were conducted to investigate the influence of bulk defect density (cm017 cm−3 . When an2 (Rs) and from 0 to 3000 Ω cm2 (Rsh) (2) can also reduce the Jsc. On the contrary, the Voc remained the same for values of Rs but increased with higher Rsh. Also, as expected, the FF was mainly affected by the Rs, which directly affected the efficiency. Therefore, we need devices with low or negligible series resistances and very high shunt resistances to obtain high efficiencies.Parasitic resistances have a crucial role in a photovoltaic (PV) system. Series resistance (Rs) is generated by different bulk resistances, such as those of the semiconductor material, metallic material, and contact in the metal–semiconductor . On the m2 (Rsh) . A signi15 to 1 × 1017 cm−3 for CdS (donor densities) and CdTe (acceptor densities), respectively. As can be seen, the main impact was at greater values of acceptor concentration nearby 10T/cm−3, where the utmost efficiency of 17.48% and a Voc of 2.0 V were determined  to replace the common back contact (Cu/Au) [2:F, as mentioned in the introduction section, is commonly used. However, it is necessary to emphasize the characteristics required of a front contact (TCO) for its application in CdTe/CdS solar cells, such as high transparency (>85%), a low resistivity on the order of 104 Ω cm [2:F (superstrate type) which needs a deposit temperature of around 450 °C or ZnO:Al (substrate type) that has the advantage of being deposited at room temperature [2:F due to its greater transparency (cell 2). And the third one, where the CuSCN material works as HTL (cell 3). The J-V curves with different configurations are shown in Using Cu as a p-type dopant at the back contact has been the most productive approach, yielding hole densities of about 10 (Cu/Au) ,54. In a104 Ω cm ,56, goodperature . Therefooc significantly increased by adding the CuSCN layer but slightly decreased in FF. This behavior is because of its high conduction band since the CuSCN operates as an electron reflector. Additionally, the CuSCN and CdTe conductivity (p-type) caused the valence band offset (CuSCN/CdTe) to be negligible. Furthermore, the CuSCN layer became a Cu source for CdTe doping (increasing hole density), which was necessary to improve the Voc in CdTe devices [The typical Cu/Au back contact becomes problematic due to Cu diffusion during device production conducting to degradation and compensation via extra Cu diffusion . For thi devices . Thus, i devices ,54.oc from 0.87 V to 0.94 V. Results by using CuSCN as HTM are in good agreement with some reports where a significant Voc (1 V) was reached [2:F (80%). Nevertheless, there was no noticeable improvement in cell 3 with the CuSCN layer.The output parameters of all CdTe devices are presented in  reached . Figure 2 to analyze their impact on the solar cell performance using the following configurations: CdTe/CdS/SnO2:F (cell 1), CdTe/CdS/ZnO:Al (cell 2), and CuSCN/CdTe/CdS/CdS/ZnO:Al (cell 3). The Rs predominantly result from the available connections or the circuit resistance across the contacts. It is desirable to guarantee a low or negligible Rs to ensure high-efficiency solar cells. However, it is still inconceivable to achieve a perfect FF because of the deficiency of the diode. The Rs were varied from 0 to 10 Ω cmoc remained constant with increasing Rs values. As this value increased from 0 Ω cm2 to 10 Ω cm2, the efficiency decreased from 16.04% to 11.67% (Cell 1), from 16.90% to 11.72% (Cell 2), and from 17.72% to 12.09% (Cell 3). Nevertheless, better performance was shown for the device with the CuSCN layer (compared with the other devices). The effect of adding the CuSCN and ZnO:Al nanolayers in the device design is revealed in In all configurations, the V13 cm−3, a negligible series resistance, a shunt resistance of 3000 Ω cm2, and hole carrier concentration of 1017 cm−3. In addition, the use of ZnO:Al and CuSCN layers was considered. The J-V curve and EQE are presented in As the last step, a new simulation was carried out considering the optimized characteristics obtained in the previous sections, such as thicknesses of 3 µm for CdTe and 25 nm for CdS, a work function of 5.7 eV, a defect density of CdTe of 102:F; cell 2, CdTe/CdS/ZnO:Al; and cell 3, CuSCN/CdTe/CdS/ZnO:Al. First, the influence of different parameters  on cell 1 was studied and explained. Then, the impact of the ZnO:Al and CuSCN was evaluated for cells 2 and 3, respectively. Promising optimized results were achieved with a conversion efficiency from 16.04% (cell 1) to 22.62% (cell 3). Finally, cell 3 demonstrated that incorporating ZnO:Al and CuSCN made it possible to obtain a superior Voc and, consequently, greater efficiency. Furthermore, the third device showed a lower adverse effect against parasitic resistance (Rs). This new proposal will guide the feasible fabrication of higher-efficiency CdTe-based photovoltaic cells.The understanding of CdTe solar cell physics is valuable for achieving greater efficiencies. In this sense, a numerical simulation (SCAPS) of the CdTe device was carried out. The incorporation of the ZnO:Al nanolayer (TCO) and CuSCN nanolayer (HTL) on solar cell performance was compared and studied. The device configurations were: cell 1, CdTe/CdS/SnO


Traffic accidents caused by road icing are a serious global problem, and conventional de-icing methods like spraying chemicals have several limitations, including excessive manpower management, road damage, and environmental pollution. In this study, the carbon nanotubes reinforced de-icing coating for the road system with a self-heating function was developed as part of the development of a new system to prevent accidents caused by road icing. The electrical characteristics of the fabricated coating were analyzed, and the carbon nanotube coating heating performance experiment was conducted to measure the temperature increments by applying a voltage to the coating at a sub-zero temperature using an environmental chamber. In addition, the coating was installed on the road pavement and the applicability was investigated through a heating test in winter. As a result of the experiment, the coating made with the higher carbon nanotube concentration presented higher heating owing to its higher electrical conductivity. In addition, the coating showed sufficient heating performance, although the maximum temperature by Joule heating decreased for the entire coating at sub-zero temperatures. Finally, field tests demonstrated the potential of electrically conductive coatings for de-icing applications. Road systems face their greatest challenges during the winter season in many countries because snowfalls and icing create serious problems in the field of road systems. These challenges involve the mobility of vehicles and the assurance of driver safety. In addition, many road departments are not prepared to immediately remove snow and ice. As a result, snow and ice accumulation on road systems lead to significant financial setbacks. For example, persistent snowfall in northwestern Germany resulted in more than 2000 traffic accidents and a direct economic loss of 100 million Euros in November 2005, and traffic in the northeastern United States was paralyzed by a snowstorm for four days, with a direct economic loss of US $10 billion in January 1996 ,4,5,6. FTo address these issues, many research efforts have been devoted to finding cleaner techniques for safely deicing road systems during winter. Some of the emerging techniques involve the application of embedding electrically heated sheet/grille elements inside the road  and the In our research, we developed a novel coating comprising carbon nanotubes and an epoxy matrix capable of replacing conventional methods of de-icing road systems. We investigated the heating performance of the coating according to atmospheric temperature and its potential applicability through field experiments. We began by examining the electrical properties of the CNT/EP coatings in relation to varying CNT concentrations. Following that, we assessed the heating efficiency of the CNT/EP coating at room temperature, utilizing the Joule effect. To explore the impact of atmospheric temperatures, we measured the heating temperature of the CNT/EP coating under below-freezing conditions. Furthermore, we evaluated the coating’s applicability by observing its performance during winter.3 and a viscosity of 200–450 mPa∙s. The dispersant used acetone with a purity of 99.7% from Samchun Pure Chemical Co., Ltd. .In this study, multi-walled carbon nanotubes were adopted from Nanolab, Inc. . The CNTs have a purity of higher than 85 wt.% , a diameter of 15 nm, and a length of 5–20 µm. Epoxy was sourced from Easy Composites Ltd. , exhibiting a density ranging between 1.12–1.18 g/cmThe CNT/EP coating was fabricated following the procedure shown in 9 Ω. Electrical resistance was assessed based on the current–voltage curves acquired by applying voltages ranging from −10 V to +10 V. The test coating samples were prepared in various CNT concentrations (0.63–5.00 wt.%) with dimensions of 50 mm × 30 mm × 2 mm. High-purity silver paint was applied to both ends of the coating samples to reduce the contact resistance between the coating and the probe point. The electrical conductivity (σ) of the specimens was calculated by σ = L/RA, where L is the length of the coating (m), R is the resistance of the coating (Ω), and A is the area of the coating (m2) [The heating performance due to the Joule effect is based on the electrical conductivity and the applied voltage of the application. Therefore, in our study, we have set the concentration of CNTs and the applied voltage as the main parameters. The resistance of CNT/EP coatings was gauged using a Keithley 2700  for standard resistance, and a Keithley 2450  for high resistance levels exceeding 10ing (m2) ,37. The As shown in A thermal infrared camera  was used to record the heat distribution across the samples during Joule heating, resulting in thermal images. To investigate the effect of atmospheric temperature on the heating performance, the infrared thermal imaging camera and CNT/EP coating were set in the environmental chamber, and the temperature increments of the CNT/EP coating by applied voltage was measured with respect to the environmental temperature (−20 °C to +20 °C) as shown in −8 S/m are insulator, and those with electrical conductivities above 10−8 S/m are conductive. At low concentration, lower than 0.25 wt.% of CNTs, the coatings showed non-conductivity. The electrical conductivity of the coating rapidly increased when the CNT concentration rose to between 0.25 to 1.0 wt.% based on increases in the CNT networks. This sharp increase in the electrical conductivity of CNT/EP coating is because of the formation of a percolation threshold [The addition of CNT in the epoxy metrics can significantly improve the electrical conductivity of the coating because of the superior electrical conductivity and high aspect ratio of the CNT . The elehreshold ,40,41,42Q = I2Rt = V2t/R, where Q is Joule heat, I is current, R is resistance, V is applied voltage and t is operating time). Another important aspect of the Joule heating system is the heating ratio [According to Joule’s law, the CNT/EP coating converts electric energy into thermal energy after loading voltage, which mainly shows rapidly increasing temperature stage of the coating ,44. Figung ratio ,46. FiguP = V2/R. The relationship between the applied voltage and the power is comparable to that between the maximum temperature of the coating and the voltages shown in −3) can be calculated as electric power per unit volumetric [To evaluate the applicability of CNT/EP coatings based on Joule heating, a deeper exploration of thermal efficiency is required, as it signifies the de-icing capability in relation to the energy consumed. lumetric ,49. Figulumetric . The slolumetric ,52,53. Alumetric ,55. ThisMany studies related to Joule heating performance of CNT composites have been conducted ,57,58,590 and t1, respectively. The resistance of EC01 and EC02 was 960 Ω and 1960 Ω at 20 °C, respectively. The resistance of EC02 is about twice that of EC01. The experiment was carried out during an evening in winter 2021. As a result of the heating test, the surface temperature of the coating increased above 10 °C from about −5 °C. After the voltage was applied, it heated up rapidly for 200 s, and the EC01 sample showed a higher temperature than EC02. It should show a similar heating pattern for the same applied voltage, but the EC01 sample showed a higher and more even temperature. This is because the dispersion of the CNTs in EC01 sample was more evenly distributed than that of the EC02 sample which can be confirmed by comparing the heating distribution of the coating as shown in To demonstrate the feasibility of the proposed method, we placed CNT/EP coating on the road pavements as shown in The aim of this study was to examine the relationship between the concentration of CNTs and supply voltage with respect to the heating and electrical performance of CNT/EP coating, and to assess the impact of atmospheric temperature on this relationship. The goal was to provide an alternative solution to the issues caused by chemical de-icing agents in road systems. By dispersing CNTs in an epoxy matrix, we have successfully fabricated highly electrically conductive coatings. Measurement of the variation of electrical conductivity with CNT concentration showed that the percolation threshold occurred when CNT concentration was between 0.5 wt.% and 1.0 wt.%. To evaluate the applicability of CNT/EP coatings based on Joule heating, thermal efficiency was investigated at room temperature, as it signifies the de-icing capability in relation to the energy consumed. Results show that the CNT/EP coating has high thermal efficiency by attaining maximum temperatures with relatively low electric power. As a result of the heating test of the CNT/EP coating according to atmospheric temperature, it was found that a decrease in atmospheric temperature reduced the heating performance of the coating. Nevertheless, it was confirmed that the heating performance of the coating remained above freezing even under sub-zero temperatures. Finally, application tests confirmed that the de-icing coating developed and verified in this study present good heating performance and applicability. As a result of the application test at sub-zero temperature, it was proved that the coating can reach the de-icing temperature and it is possible to respond according to the installation environment. Through these results, it was confirmed that the developed coating could be used not only to prevent ice on the road system as an eco-friendly coating but also may be applied to structures in various fields.


Dear Editors,Acute myeloid leukemia (AML) can be classified into multiple genetic subtypes based on recurrent pathogenic structural variants (SVs), copy number abnormalities (CNAs), and single nucleotide variants (SNVs) that inform prognostication and clinical management , 2. Karyn = 55, 52%), 23 (22%) had AML with myelodysplasia-related changes (AML-MRC), 21 (20%) had relapsed AML and 6 (6%) had therapy related AML (Supplementary Tables NUP98 (11p15.4) rearrangement and 7 had a KMT2A (11q23.3) rearrangement with gene partner MLLT10 (10p12.31) in 3 cases, ELL (19p13.11) in one case, MLLT6 (17q12) in one case and MLLT3 (9p21.3) in 2 cases. Four had t, 3 had inv(3)(q21.3q26.2) or t including a single case with inv(3) with BCR::ABL1, 3 had inv(16)(p13.1q22) or t, and a single case each with either t or KAT6A rearrangement involving 8p11.2 . In 3 other cases, MPseq missed low-level trisomy 8 found in 3–5% of cells by FISH and detected by karyotype  Table . Of the pe Table . Since MNUP98 rearrangement, 2 cases with a TP53 deletion, and one case each with a KMT2A::MLLT10 fusion and an inv(8) resulting in a KAT6A rearrangement, and a 7q deletion. Additionally, karyotype analysis detected 4 cases with a 17p deletion, which presumptively included loss of the TP53 locus, that were not identified by FISH or MPseq. There was 100% concordance for some AML-associated rearrangements with consistent and cytogenetically detectable breakpoints including t, t, inv(16)(p13.1q22), inv(3)(q21.3q26.2), and t , KMT2A rearrangements (75%) and trisomy 8 (81%) , FISH has increased sensitivity compared to MPseq if the abnormality is targetable by the available FISH probe.When evaluating cytogenetic results obtained by each methodology individually, the concordance between MPseq and FISH was 93.3%, between FISH and karyotype was 85.7% and between MPseq and karyotype was 82.9% Fig. . FISH mi21) Fig.  demonstrseq Fig. . Reduced1%) Fig. . FISH anOf the 5 cases with discordant results between MPseq compared to karyotype plus FISH Table , the ELNKMT2C, CUX1, and EZH2, located on 7q and the most frequently gained genes included MYC and/or CCDC26, RAD21, and TRPS1, located on chromosome 8. MECOM was the most frequently rearranged gene in our AML cohort  with focal gains identified by MPseq of 8q24 involving MYC and/or the nearby long noncoding RNA CCDC26. The partial gain of CCDC26 in 7q-55 was also associated with a cryptic ins resulting in an insertion between BCL11B into the CCDC26. We also observed 2 cases with iAMP21 by MPseq (case 5q-65 and 5q-147)  and KRAS. Rare or novel SVs were identified in 20 cases (19.0%) including case KAT6Ar-128, which was found to have a cryptic inv(8) resulting in a KAT6A::SORBS3 fusion. A single case (simple K-132) had a ZMYND11::MBTD1 fusion. SVs disrupting NF1 were found in 2 cases , RUNX1 in 2 cases , non-GATA2 MECOM rearrangements in 6 cases  and an ASXL1 rearrangement resulting in a partial deletion of the gene in one case (5q/7q-87). Finally, MPseq characterized each of 6 NUP98 rearrangements revealing KMD5A partner gene in 3 cases and NSD1 partner in 3 cases. Recurrent focal deletions of AML associated genes not appreciated by karyotyping were common , TP53 and KMT2A are used. Alterations in TP53 are associated with significantly inferior outcomes and treatment responses in AML and biallelic alteration in TP53 AML confers the worse outcomes among CK AMLs [TP53 locus is important for prognostication and may soon impact clinical management.In summary, we identified five cases 4.8%) of discordance between MPseq compared to karyotype with FISH when evaluating recurrent AML-associated genetic abnormalities. Greater discordance was identified when comparing karyotype to FISH (14.3%) or karyotype to MPseq 17.1%) individually. These findings contrast previous studies which have reported limited value of FISH in AML when an adequate karyotype is available [.1% indiv% of discNUP98 rearrangements are associated with poor outcome [NUP98 rearrangements have not yet been incorporated into ELN guidelines. Similarly, KMT2A partial tandem duplications have been reported in 10% of AML and MDS and are associated with poor outcome [MYC amplification, associated with a CK and poor outcome [BCL11B::MYC rearrangements reported as a driver in ambiguous leukemia [ZMYND11::MBTD1 fusion [Although we demonstrate that MPseq does not currently add additional prognostic value above FISH when compared to a comprehensive AML FISH panel, risk stratification guidelines will likely evolve to include novel abnormalities of prognostic significance detected by NGS. For example, growing evidence suggests  outcome , but NUP outcome . Other r outcome , BCL11B:leukemia , ZMYND111 fusion  and iAMP1 fusion .We demonstrate that karyotype analysis when combined with FISH remains a robust laboratory approach in the evaluation of AML. While the use of MPseq did not add significant value above FISH using current ELN guidelines, NGS technologies such as WGS will continue to identify novel high-risk AML subgroups, which will likely enhance future risk stratification guidelines.Supplementary MethodsSupplementary FiguresSupplementary TablesSupplementary Table 4


Yamahai‐shubo, the source of yeast used to produce a Japanese traditional rice wine, Yamahai‐shikomi sake. The bacterial strains were nitrate‐reducing Pseudomonas sp. 61‐02, Leuconostoc mesenteroides LM‐1, Lactiplantibacillus plantarum LP‐2, and Latilactobacillus sakei LS‐4. We examined fermentation factors for Yamahai‐shubo and Yamahai‐shikomi sake samples to compare the suitability of their bacterial combination (16 variations). As a result of principal component analysis, we found that two major groups were formed; one containing strain LP‐2 and the other containing strain LS‐4, and that strains LP‐2 and LS‐4 were important in the Yamahai‐shikomi sake in the presence of strains 61‐02 and LM‐1. Then, we investigated the effects of strains LP‐2 and LS‐4 on the concentration of organic acids  in Yamahai‐shikomi sake. Only in lactic acid, a tendency to decrease with a smaller proportion of LS‐4 strains in Yamahai‐shubo was observed. Subsequently, their effect on the concentration of diacetyl, crucial for aroma, was investigated between the LP‐2 and LS‐4 strains. The sample prepared in the absence of strain LS‐4 exhibited the lowest concentration of diacetyl. This result was supported by the statistical analysis for the sensory scores performed for aroma of each Yamahai‐shikomi sake sample. In conclusion, strain LP‐2 plays a more significant role in improving Yamahai‐shikomi sake quality with strains LM‐1 and 61‐02 rather than strain LS‐4 in Yamahai‐shubo preparation and Yamahai‐shikomi sake brewing.This study investigated the interactions of four bacteria strains isolated from  Leuconostoc mesenteroides LM‐1, Lactiplantibacillus plantarum LP‐2, and Latilactobacillus sakei LS‐4) isolated from Yamahai‐shubo. We examined fermentation factors for Yamahai‐shubo and Yamahai‐shikomi sake samples to compare the suitability of their bacterial combination (16 variations). In conclusion, strain LP‐2 plays a more significant role in improving Yamahai‐shikomi sake quality with strains LM‐1 and 61‐02 rather than strain LS‐4 in Yamahai‐shubo preparation and Yamahai‐shikomi sake brewing.This study investigated the interactions of four bacteria strains (nitrate‐reducing Pseudomonas sp. 61‐02,  It is known to need more time than Sokujo‐shubo because lactic acid is derived from lactobacilli included Yamahai‐shubo, and it increases the risk of putrefaction of nonpreferable microorganisms, causing an off‐flavor , e.g., Leuconostoc mesenteroides, gradually and then preferentially grow, producing only a small amount of lactic acid. Subsequently, bacillus‐form lactic acid bacteria (bacilli‐LAB), e.g., Latilactobacillus sakei  by adding koji and steamed rice with water, it becomes more significant to examine the diversity of bacteria as well as yeasts in the present brewing of Yamahai‐shikomi sake with Yamahai‐shubo to maintain the quality of the sake  in Yamahai‐shubo preparation and showed the requirements of two strains, 61‐02 and LM‐1. In the evaluation process, we performed principal component analysis for samples of Yamahai‐shubo and Yamahai‐shikomi sake. Next, we examined how Lp. plantarum LP‐2 and Ll. sakei LS‐4 contributed to the quality of sake with the coexistence of Pseudomonas sp. strain 61‐02 and Le. mesenteroides LM‐1 in Yamahai‐shikomi sake brewing. Finally, we showed the significance of Lp. plantarum LP‐2 over Ll. sakei LS‐4 and demonstrated that the quality of Yamahai‐shikomi sake can be controlled by LAB through studying various combinations of those strains in Yamahai‐shubo.With this background, 22.1Pseudomonas sp. 61‐02 and three LAB strains ] were used in this study. They were previously isolated from Yamahai‐shubo described before ] and 255 mL of water with the addition of KNO3 (60 mg/L). koji is a type of rice that is made from koji mold  with polished rice made by the industrial koji manufacturing system . Fermentation with koji followed the same method described before ] were prepared as follows. Strain 61‐02 was independently cultured in PN medium  for 3 days at 15°C  for 3 days at 20°C and were added to the initial mixture of Yamahai‐shubo at a cell concentration of 103 cells/mL. Yamahai‐shubo preparation was totally completed in 25 days with the initial mixture of Yamahai‐shubo after adding any of the nitrate‐reducing bacterial strain 61‐02 and/or three LAB bacterial strains  added; see Table Four bacterial strains [nitrate‐reducing Yamahai‐shubo preparation was kept at 8°C until day 4. After day 5, the preparation was warmed up by contact with hot water at 65°C to increase the temperature by one degree per day until it reached 21°C on day 16. After holding the temperature of the mixture at 21°C for another 5 days, the temperature was decreased to 15°C on day 21, to 13°C on day 22, to 12°C on day 23, and was then maintained at 12°C until day 25. The yeast strain KZ‐06 U was separately cultured in koji extract (sake meter −60) for 2 days at 30°C and added between days 10 and 15 of the preparation to reach a cell concentration of 106 cells/mL when the nitrite concentration of the Yamahai‐shubo preparation was less than 1.0 mg/L. The timing of yeast addition in the absence of strain 61‐02 was the same as the timing of yeast addition in the presence of strain 61‐02. The final preparation reached approximately 500 mL in volume and was called “Yamahai‐shubo.” For use at the industrial scale, the process was considered complete at day 25, even if the disappearance of nitrite was delayed and the yeast growth period was shortened.The temperature during The cell numbers of strain 61‐02 and three LAB strains  were determined by the method described previously  were mixed at three different times  to smoothly complete the fermentation  was carried out at 15°C from day 1 to day 2. Both the naka  and the tome  feeding procedures were carried out by adding the components at 10°C. After the tome procedure , sake brewing was monitored by measuring the weight reduction of the fermentation mixture, corresponding to CO2 evolution for 25 days. The temperature of the fermentation mixture was increased by about 0.5°C for another 6 days, kept at 13°C for an additional 7 days, and then promptly lowered to 10°C while the temperature was maintained for an additional 12 days. After sake brewing (25 days), the samples were separated by centrifugation , and the supernatants were called “Yamahai‐shikomi” sake samples . Diacetyl was derivatized by Shinwa DS‐DA  and analyzed using the same gas chromatography system. All data are the averages of two or three independent experiments.Basic fermentation factors for sake meter (SM), ethanol (Alc), total acidity (TA), amino acidity (AA), and Ultraviolet absorption (UV) were determined by the standard method established by National Research Institute of Brewing .The qualities of 2.5https://www.bellcurve.jp/), employing fermentation factors  and sensory scores at the final point (25 days) of Yamahai‐shubo  was performed with Excel Statistical analysis 2008 software for Windows . Changes in three fermentation factors are depicted in Figure Pseudomonas sp. 61‐02, following the method as described in Materials and Methods. Nitrite is known to be essential in Yamahai‐shubo preparation process; it is converted from nitrate, which is derived from PN medium for the preparation of strain 61‐02  and inhibits the growth of wild yeasts that are concomitant in the initial mixture for Yamahai‐shubo. As shown in Figure Yamahai‐shubo preparation and drastically decreased before the second half. However, the coculture of strain LM‐1 only with strain 61‐02 was found to retard the decrease in the concentration of nitrite  in the presence and absence of strain 61‐02 .The four bacterial strains previously found in e Figure . It is ns Figure . Furthers Figure . In addi2 Figure . HoweverYamahai‐shubo preparation in Table Yamahai‐shubo utilizing the advantages of strain LP‐2 without strain LS‐4.Comparing the components at the final point (25 days) of Yamahai‐shikomi sake similarly as Yamahai‐shubo , we performed principal component analysis (PCA) by using fermentation factors for 16 samples of Yamahai‐shubo and Yamahai‐shikomi sake, as shown in Tables To investigate the suitability of the bacterial combination of the nitrate‐reducing Yamahai‐shubo preparations and Yamahai‐shikomi sake samples contained strains 61‐02 and LM‐1  in Yamahai‐shikomi sake and Yamahai‐shubo samples were determined for each sample  was higher than that of LP2 + LS4 (2:1), there was no significant difference between them  isolated from Yamahai‐shubo samples is very important for producing Yamahai‐shikomi sake. In particular, the isolation of different bacteria from Yamahai‐shubo samples and their practical use in sake brewing make it possible to produce Yamahai‐shikomi sake under artificial control in the recent process.The steady supply of Yamahai‐shubo samples contain various kinds of identified and unidentified bacteria, and their bacterial diversities vary depending on the sources from which they are derived. It is well known that the quality of Yamahai‐shikomi sake depends on the diversities of those bacteria. Therefore, it is necessary to grow advantageous bacteria to prepare appropriate Yamahai‐shubo samples and to achieve the desired sake quality.Pseudomonas sp. 61‐02, cocci‐LAB Le. mesenteroides LM‐1, bacilli‐LAB Lp. plantarum LP‐2, and bacilli‐LAB Ll. sakei LS‐4) from the Yamahai‐shubo sample  that produces lactic acid more slowly. We clarified the turnover of those bacteria, along with the amount of nitrite, through this study.We previously isolated four bacterial strains , Yamahai‐shubo was diluted nearly 20 times in Yamahai‐shikomi sake. Thus, the higher acidity of Yamahai‐shubo was supposed to be unable to directly keep the acidity of Yamahai‐shikomi sake. On the other hand, the majority of the acids in Yamahai‐shikomi sake were most likely produced by yeasts coexisting during Yamahai‐shikomi sake brewing under higher acidic environment, resulting in increased acidity. This was consistent with the fact that the viable cell numbers of most bacteria in Yamahai‐shubo became extremely decreased after 25 days  and Sokujo‐shubo affect sake qualities; e.g., yeasts grown in Yamahai‐shubo showed more enhanced tolerance to ethanol than those in Sokujo‐shubo  induction capacity .The significance of LAB as well as yeasts has been recognized in winemaking. In particular, the contribution of 5Yamahai‐shubo produced Yamahai‐shikomi sake with more favorable aromas and tastes than that of another bacilli‐LAB strain, LS‐4. Furthermore, it is likely that strain LP‐2 in Yamahai‐shubo played a significant role in improving sake quality by reducing diacetyl more than strain LS‐4 in Yamahai‐shikomi sake brewing.We found that the addition of the nitrate‐reducing bacterium strain 61‐02, the cocci‐LAB strain LM‐1, and the bacilli‐LAB strain LP‐2 to There is no funding to report for this submission.The authors declare no conflict of interest associated with this manuscript.


Bifidobacterium catenulatum subspecies kashiwanohense (B. kashiwanohense) was originally isolated from infant feces. However, only a few strains have been described, and the characteristics of this subspecies have been poorly investigated. Here, we characterized genotypes and phenotypes of 23 B. kashiwanohense-associated strains, including 12 newly sequenced isolates. Genome-based analysis clarified the phylogenetic relationship between these strains, revealing that only 13 strains are genuine B. kashiwanohense. We defined specific marker sequences and investigated the worldwide prevalence of B. kashiwanohense based on metagenome data. This revealed that not only infants but also adults and weaning children harbor this subspecies in the gut. Most B. kashiwanohense strains utilize long-chain xylans and possess genes for extracellular xylanase (GH10), arabinofuranosidase and xylosidase (GH43), and ABC transporters that contribute to the utilization of xylan-derived oligosaccharides. We also confirmed that B. kashiwanohense strains utilize short- and long-chain human milk oligosaccharides and possess genes for fucosidase (GH95 and GH29) and specific ABC transporter substrate-binding proteins that contribute to the utilization of a wide range of human milk oligosaccharides. Collectively, we found that B. kashiwanohense strains utilize both plant- and milk-derived carbohydrates and identified key genetic factors that allow them to assimilate various carbohydrates.Bifidobacteria are prominent members of the human gut microbiota throughout life. The ability to utilize milk- and plant-derived carbohydrates is important for bifidobacterial colonization of the infant and adult gut. The  Recent studies have demonstrated that their ability to utilize microbiota-accessible carbohydrates is a key factor enabling their stable persistence in the human gut.6–8Bifidobacteria are commonly detected as prominent members of the human gut throughout life. Bifidobacterial colonization has been associated with a reduced risk of obesity, 9–12 Microbiota analysis of infants and adults revealed that the bifidobacterial species colonizing the human gut change with age.14 Specifically, Bifidobacterium breve, Bifidobacterium bifidum, and Bifidobacterium longum subspecies infantis  are generally predominant in infants, whereas Bifidobacterium adolescentis, and B. pseudocatenulatum are prevalent in adults. B. longum subspecies longum  is distributed in both infant and adults.15To date, hundreds of bifidobacterial strains have been isolated, and the characteristics of major bifidobacterial species have been described. Most of these studies involved comparative and functional genome analyses.17 Molecular mechanisms of utilization of these plant-derived carbohydrates have only just started to be elucidated. Based on previous studies, enzymes belonging to glycoside hydrolase (GH) family 43 (GH43) and others  are involved in xylan-related glycan utilization,17 whereas enzymes belonging to GH13 are associated with starch utilization.6 Furthermore, ATP-binding cassette (ABC) transporters, usually encoded in the same operon as these GHs, also play important roles in the uptake of these oligosaccharides.17Recent studies demonstrated that adult-prevalent bifidobacterial species can utilize plant-derived carbohydrates, including xylan-based oligosaccharides , and starch-related carbohydrates .B. pseudocatenulatum possesses more abundant GH43 genes and ABC transporters for xylan-based oligosaccharides utilization than other adult-associated bifidobacteria,17 suggesting that this species assimilates a broader range of xylan-based oligosaccharides than other bifidobacterial species. On the other hand, B. adolescentis possesses numerous GH13 genes18 and accumulates in resistant starch granules in the human gut, suggesting starch preference of this species.19Homologs of the glycosidase and transporter genes are shared among adult-associated bifidobacteria. However, each adult-associated bifidobacterial species exhibits different glycan preferences. 20 HMOs are non-digestible oligosaccharides present in breast milk that include various glycan structures. Fucosyllactose  is the main component of HMO, and genetic factors responsible for its utilization  have been intensively investigated and characterized.21By contrast, the infant-associated species do not utilize plant-derived carbohydrates as well as the adult-prevalent species. However, they readily utilize human milk oligosaccharides (HMO).B. infantis utilizes HMO most efficiently compared with other Bifidobacterium species. The subspecies imports various HMOs via a number of ABC transporters, where an intracellular glycosidase subsequently hydrolyzes the imported oligosaccharides into monosaccharides.23 Consequently, B. infantis assimilates the broadest range of HMO components compared with other bifidobacteria; however, the gene maintenance costs involved with equipping various transporters may be high.24B. bifidum is the second most efficient HMO metabolizer among bifidobacteria. The species hydrolyzes HMOs using extracellular glycosidases, and the resultant mono- and di-saccharides are then taken up by its limited number of transporters.25 The species breaks down most HMOs with lower protein-equipping cost than B. infantis; however, the resultant extracellular saccharides can be utilized by other gut microbes, and B. bifidum does not assimilate HMO efficiently in competition. In contrast, B. breve cannot utilize long-chain HMOs , and only a subset of B. breve strains can utilize FL.26 The FL-utilizing B. breve strains import FL via the well-characterized ABC transporter,20 and the imported FL is subsequently hydrolyzed to lactose and fucose by an intracellular GH95 fucosidase. The utilization system is efficient since B. breve strains can utilize the main HMOs  relying on a limited number of transporters and a glycosidase.Inter- and intra-species variation in the utilization of various HMO components by bifidobacterial species and the underlying molecular mechanisms have been reported. B. kashiwanohense, which we are going to characterize in this study, was isolated from Japanese infant feces in 2011.27 The species was later reclassified as a subspecies of B. catenulatum based on digital DNA – DNA hybridization analysis.28 This subspecies has been rarely detected in humans, and only a few strains have been isolated from infant feces. Some studies have reported that B. kashiwanohense strains can utilize FL.29 However, the utilization of other carbohydrates, and the global and age-dependent prevalence of this subspecies, are not well understood.B. kashiwanohense. We performed comparative genome analyses of 23 B. kashiwanohense-associated strains, including 12 newly sequenced isolates. Based on the phylogenetic analysis, the strains were classified into four groups, and only 13 strains were sensu stricto B. kashiwanohense. Metagenome data-based investigation of worldwide prevalence revealed that while B. kashiwanohense is not often detected in infants and adults, the subspecies is predominant in some weaning children. Most B. kashiwanohense strains can utilize long-chain xylans and possess an extracellular xylanase, as well as ABC transporters and intercellular GHs known to contribute to the utilization of xylan-based oligosaccharides. We also confirmed that B. kashiwanohense can utilize short- and long-chain HMOs and investigated the underlying molecular mechanisms.In the current study, we characterized the genotypes and phenotypes of B. kashiwanohense-associated strains by analyzing their genome sequences ,27 7 strains of Bifidobacterium catenulatum subspecies catenulatum , and 3 strains that have previously been identified as B. kashiwanohense.31We assessed the phylogenetic relationship among 23 equences . Of thesB. kashiwanohense type strain JCM 15,439T and 11 newly sequenced strains. Cluster II included B. catenulatum strains . Strain PV20–2, reported as B. kashiwanohense by Vazquez-Gutierrez et al.,31 was not included in cluster I nor II. The average nucleotide identity (ANI) values of PV20–2 against the strains in cluster I (B. kashiwanohense) and II (B. catenulatum) were approximately 95% , suggesting that PV20–2 should be reclassified as a new subspecies of B. catenulatum (designated as cluster III). The strains N5G01 and N4G05, reported as B. kashiwanohense by Freitas and Hill,30 and 3 putative B. catenulatum strains isolated from Bangladeshi children formed 1 cluster  of 776 core genes of the 23 strains, divided the strains into four major clusters . ClusterB. catenulatum and B. pseudocatenulatum)  of 1989, which is higher than those of the closely related bifidobacterial species  . The pannulatum) . Compariectively .B. kashiwanohense using metagenome data deposited in public databases . We used two complete genome sequences to define the B. kashiwanohense-specific sequence (K-mer) using the bioinformatic tools Kraken238 and Bracken39 . We attempted to detect B. kashiwanohense-specific sequences in 1,193 sets of metagenomic data (data for 289 infants from 6 countries and 904 adults from 14 countries) deposited in the NCBI database  and 11 subjects among 289 infants (3.81%) harbored the bacteria in the gut. We did not observe any differences in the prevalence of this subspecies in different continents. Additionally, we examined the B. kashiwanohense distribution in two metagenome-assembled genomes (MAG) database41. Compared with other Bifidobacteria, fewer B. kashiwanohense MAGs were detected, which supported our result from metagenomic data showing the limited distribution of this subspecies (Supplementary data 4).Next, we examined the worldwide distribution of 14 In the present study, we detected B. kashiwanohense in 3 out of 12 infants during their first 2 years of life  in the subspecies with those of other human-associated bifidobacteria. We visualized the GH gene profile of each strain using a heatmap  and adult-associated species  are separated along the PC1 axis, and B. longum was plotted at the middle of infant- and adult-association species , in comparison with that of strains of bifidobacterial species commonly found in the human gut , all B. kashiwanohense strains, and some B. pseudocatenulatum strains utilize the major HMOs  (B. adolescentis and some B. pseudocatenulatum strains) are unable to utilize these oligosaccharides . Furthermore, we confirmed that most infant-associated species (B. infantis and B. bifidum) did not efficiently utilize plant-derived carbohydrates , whereas B. kashiwanohense and adult-associated bifidobacteria utilized these carbohydrates (green box in We next investigated the growth of uman gut . We confnd 3-FL) , while min silico genome analysis . These transporters and GHs work together to utilize xylan-based oligosaccharides of different sizes and with different side residue modiﬁcations. In the current study, we found that all B. kashiwanohense strains possess homologs of the B. pseudocatenulatum XOS utilization genes involved in XOS uptake and hydrolysis, most of which are encoded in XOS utilization clusters 1 and 2 . The results are shown in Fig. S3. We searched for a gene whose presence corresponded with the LNFP and LNDFH utilization phenotype. No GH genes were associated with the LNFP and LNDFH utilization phenotypes , and no SBP genes were associated with the LNFP utilization phenotype . However, the presence of one SBP gene (ID1343) corresponded with the LNDFH-utilization phenotype .Previous studies have demonstrated the importance of GHs and the ABC transporter substrate-binding protein (SBP) in HMO utilization.22 According to previous studies, there are several SBP subtypes for FL (type I – IV).43 Of note, SBP associated with LNDFH utilization represented a unique subtype (type III) that was only present in B. kashiwanohense strains . In addition, we observed that the LNDFH-non-utilizing strain YIT 13,055 harbors a gene for a FL-SBP that belongs to another subtype (type I) (B. kashiwanohense strains correspond to those in their FL-SBP subtype (B. kashiwanohense (type III) is involved not only in FL but also long-chain HMO utilization (B. kashiwanohense specific FL-SBP (type III) mediates the uptake of long chain HMO, based on the phylogenetic analysis of bifidobacterial SBP.43 Our gene-trait matching approach additionally showed this gene is well conserved among B. kashiwanohense strains.ID1343 is a homolog of SBP that is essential for FL utilization. subtype . The reslization . This isB. kashiwanohense. We found that this subspecies can utilize not only short- and long-chain HMOs but also dietary fiber arabinoxylan and its derived oligosaccharides. By using the gene – trait matching approach, we found the unique metabolic pathways involved in and the key genetic factors for utilizing this wide range of substrates , we assumed that B. kashiwanohense may have a competitive advantage in the complex environment of the human gut. However, our metagenome analysis of B. kashiwanohense distribution suggested that the prevalence of this subspecies is limited in both infants and adults. This suggests that carbohydrate utilization constitutes just one aspect of the competitive advantage in the complex microbial environment, and other factors exist that underpin wide bacterial distribution and predominance in the human gut.Since B. kashiwanohense strains are detectable and predominant in the gut before and after weaning, indicating that B. kashiwanohense may have evolved to adapt to the gut environment during the weaning period. This concept is in good agreement with a recent study,49 showing that a clade of B. longum possessing both HMO- and dietary-fiber-utilizing-genes expanded during the weaning period. It will be interesting to evaluate the association between bifidobacterial carbohydrate utilization expansion during the weaning by using in vitro competitive experiments and/or assessed using future infant cohorts, to support the concept.Our analysis revealed that B. kashiwanohense strains can utilize not only FL but also long-chain HMOs, including LNFP and LNDFH, using the gene – trait matching analysis. These findings are in good agreement with a recent publication by Ojima et al.43 who analyzed the differences in substrate specificity for FL among SBPs. The authors reported that the B. kashiwanohense-specific SBP contributes to the transportation of FL, as well as LNDFH and LNFP I/II, but not LNFP III.43 In the current study, we found little remaining LNFP in the culture supernatant of some B. kashiwanohense strains with specific FL-SBP (type III)  , Fig. S2B. kashiwanohense-associated strains (including 12 new isolates), worldwide (metagenomics) prevalence analysis, and their carbohydrate utilization to characterize the subspecies. The genotype and phenotype analyses revealed that the subspecies can utilize plant-derived carbohydrates and short- and long-chain HMOs. Our detailed characterization of B. kashiwanohense suggests that each bifidobacterial species employs different and unique strategies to colonize the human gut and may contribute to our understanding of the reason for the life-long predominance of bifidobacteria (and the change in species with age).In this study, we conducted comparative genome analysis for 23 2, 5% CO2, and 7% H2, using mGAM broth  containing 0.5 w/vol% glucose and 0.5 w/vol% lactose. Evaluation of Bifidobacterium carbohydrate utilization profiles was performed at 37°C in modified ILS-PIPES  supplemented with the targeted carbohydrates (0.5 w/vol%). Growth curves were evaluated by measuring the OD600 every 30 min using a microplate reader PowerWave 340  in an anaerobic chamber.The strains used in the current study were obtained from the Japan Collection of Microorganisms  and Yakult Culture Collection . The strains were routinely cultured at 37°C in an anaerobic chamber  with 88% N7 Bacteria were grown in a medium containing HMO mixtures for 72 h. According to our previous report,7 the oligosaccharides remaining in the culture supernatant at the end of the experiment were labeled with p-aminobenzoic acid ethyl ester as an ultraviolet light-absorbing compound and quantified using HPLC. The labeled HMO components were separated using a Shimadzu Prominence HPLC system  with an L-column 2 ODS . Modified HMOs were eluted with a 13:87 (vol:vol) mixture of acetonitrile and 100 mM ammonium acetate (pH 4.5) at 40°C and were detected using an SPD-20 A ultraviolet detector (Shimadzu) at 304 nm. The following oligosaccharides were used as controls: 2′-FL (Advanced Protein Technologies), 3-FL , LNFP I , LNDFH I , LNDFH II , LNT , lacto-N-neotetraose , DFL , LNFP II , LNFP III , and N-acetyl glucosamine .The HMO mixture was prepared as described previously.8 A DNA library was prepared using the TrueSeq DNA PCR-Free Library Preparation Kit  and sequenced on MiSeq (Illumina) using the MiSeq Reagent kit V2 (250 bp × 2) (Illumina). The output paired-end reads were assembled into contigs by Unicycler (v0.4.8)50 or A5-miseq (v20160825),51 which were then aligned to the complete genome of B. kashiwanohense JCM 15,439T (accession ID: GCA_001042615.1) using Mauve (2015_02_13).53As described previously, DNA was extracted from bifidobacterial strains using a bead – phenol method.54 For genomic feature comparison among bifidobacterial species (listed in Supplementary data 1), the CDSs number was calculated using Prokka (v1.14.6).55 The number of pan-genome and core-genome genes was evaluated using Roary (v3.13.0)42 using default settings. A phylogenetic tree of B. kashiwanohense-associated strains  was visualized using clinker57 with default settings.The ANIb values between strains  were calculated using pyani (v0.2.9). strains  was cons58 and Bracken (v2.6.0)39 were used to investigate the global distribution of B. kashiwanohense from shotgun metagenomic data . B. kashiwanohense-specific K-mers were defined using B. kashiwanohense JCM 15,439 and APCKJ1. To avoid false positives, the confidence score and the relative abundance cutoff value were carefully adjusted; the confidence score was set to 0.9 and the relative abundance cutoff value to 0.49%, so that the false-positive and false-negative rate for the evaluation dataset14 were 4.9% and 8.2%, respectively. High quality Bifidobacterial MAGs were retrieved from previous reports.41 The MAGs were filtered by contamination (≤5%), completeness (≥90%), number of contigs (≤500), and contig N50 (≥10kb). The B. kashiwanohense MAGs were defined by ANIb value (≥97%) to the B. kashiwanohense type strain  genome.Kraken2 (v2.0.9 beta)n = 350, 59 using dbCAN2.61 The CDSs were considered CAZy genes only if they were annotated using HMMER, DIAMOND, and Hotpep pipeline with default parameters. To normalize the count data of CAZy genes, the “scale” function in R (v4.0.5) was used. To visualize the GH profiles in a heat map format (55, BlastKoala,63 or dbCAN2.Genes in the pan-genome of all bifidobacterial strains n = 350,  were annp format , the “heClick here for additional data file.Click here for additional data file.