Yaks Are Dependent on Gut Microbiota for Survival in the Environment of the Qinghai Tibet Plateau.

The yak (Poephagus grunniens) has evolved unique adaptations to survive the harsh environment of the Qinghai-Tibetan Plateau, while their gut microorganisms play a crucial role in maintaining the health of the animal. Gut microbes spread through the animal population not only by horizontal transmission but also vertically, which enhances microbial stability and inheritance between generations of the population. Homogenization of gut microbes in different animal species occurs in the same habitat, promoting interspecies coexistence. Using the yak as a model animal, this paper discusses the adaptive strategies under extreme environments, and how the gut microbes of the yak circulate throughout the Tibetan Plateau system, which not only affects other plateau animals such as plateau pikas, but can also have a profound impact on the health of people. By examining the relationships between yaks and their gut microbiota, this review offers new insights into the adaptation of yaks and their ecological niche on the Qinghai-Tibetan plateau.

Identifying Meat from Grazing or Feedlot Yaks Using Visible and Near-infrared Spectroscopy with Chemometrics.

The quality of meat can differ between grazing and feedlot yaks. The present study examined whether spectral fingerprints by visible and near-infrared (Vis-NIR) spectroscopy and chemo-metrics could be employed to identify the meat of grazing and feedlot yaks. Thirty-six 3.5-year-old castrated male yaks (164 ± 8.38 kg) were divided into grazing and feedlot yaks. After 5 months on treatment, liveweight, carcass weight, and dressing percentage were greater in the feedlot than in grazing yaks. The grazing yaks had greater protein content but lesser fat content than feedlot yaks. Principal component analysis (PCA) was able to identify the meat of the two groups to a great extent. Using either partial least squares discriminant analysis (PLS-DA) or the soft independent modeling of class analogies (SIMCA) classification, the meat could be differentiated between the groups. Both the original and processed spectral data had a high discrimination percentage, especially the PLS-DA classification algorithm, with 100% discrimination in the 400-2500 nm band. The spectral preprocessing methods can improve the discrimination percentage, especially for the SIMCA classification. It was concluded that the method can be employed to identify meat from grazing or feedlot yaks. The unerring consistency across different wavelengths and data treatments highlights the model's robustness and the potential use of NIR spectroscopy combined with chemometric techniques for meat classification. PLS-DA's accurate classification model is crucial for the unique evaluation of yak meat in the meat industry, ensuring product traceability and meeting consumer expectations for the authenticity and quality of yak meat raised in different ways.

Patient Cranial Angle and Intrafractional Stability in CyberKnife Robotic Radiosurgery: A Retrospective Analysis.

Purpose: The aim of this study was to investigate whether variations in cranial angles and treatment accuracy during CyberKnife robotic radiosurgery necessitate adjustment of the margins of the planning target volume. Patients and Methods: Data from 66 patients receiving CyberKnife treatment for brain tumors were retrospectively analyzed. Patients were immobilized using a thermoplastic mask and headrest. The cranial angle was measured on planning CT and patients were divided into 2 groups: ≤10° (Group A) and >10° (Group B). Intrafractional motion was recorded using the CyberKnife tracking system over 50 min. Translational and rotational errors were compared between groups, and planning target volume margins were calculated. Results: In Group A, significant translational error differences were found along with the X-axis over time (P < .02). In Group B, significant differences occurred along with the Z-axis (P < .03). No significant rotational or 3-dimensional vector differences were found in either group. Group A had significantly lower Y-axis (P < .045) and roll axis (P < .005) errors compared to Group B. Estimated planning target volume margins in Group A were 0.56 mm (X), 0.46 mm (Y), and 0.47 mm (Z). In Group B, margins were 0.62 mm (X), 0.48 mm (Y), and 0.46 mm (Z). Margins covering 95% of intrafraction motion were 0.49 to 0.50 mm (X, Y, Z) and 0.69 mm (3-dimensional vector) for Group A, and 0.48 to 0.60 mm and 0.79 mm for Group B. With a 1-mm margin, complete coverage was achieved in Group A while 2.1% of vectors in Group B exceeded 1 mm. Conclusion: Adjusting cranial angle to ≤10° during thermoplastic mask molding provided better or similar intrafractional stability compared to >10°.

Prediction of recurrence-related factors for patients with early-stage cervical cancer following radical hysterectomy and adjuvant radiotherapy.

OBJECTIVE: To analyze recurrent factors in patients with clinical early-stage cervical cancer (ESCC) following hysterectomy and adjuvant radiotherapy. METHODS: We collected data from patients with ESCC, staged according to the 2009 Federation International of Gynecology and Obstetrics (FIGO) staging criteria, who underwent hysterectomy followed by adjuvant radiotherapy between 2012 and 2019. These patients were subsequently restaged using the 2018 FIGO criteria. Univariable and multivariable analyses, along with nomogram analyses, were conducted to explore factors associated with recurrence-free survival (RFS). RESULTS: A total of 310 patients met the inclusion criteria, with a median follow-up time of 46 months. Among them, 126 patients with ESCC were restaged to stage III C1 or III C2 after surgery due to lymph node metastasis (LNM) based on the 2018 FIGO staging criteria. Of these, 60 (19.3%) experienced relapse. The 1-, 3-, and 5-year RFS rates were 93.9%, 82.7%, and 79.3%, respectively. Multivariate analysis revealed that the number of positive lymph nodes (LNs), tumor diameter (TD) > 4 cm, and parametrial invasion (PI) were associated with recurrence. The nomogram indicated their predictive value for 3-year and 5-year RFS. Notably, the 5-year recurrence rate (RR) increased by 30.2% in patients with LNM, particularly those with ≥ 3 positive LNs (45.5%). Patients with stage III C2 exhibited a significantly higher RR than those with IIIC1 (56.5% vs. 24.3%, p < 0.001). The 5-year RFS for patients with TD > 4 cm was 65.8%, significantly lower than for those with TD ≤ 4 cm (88.2%). Subgroup analysis revealed higher 5-year RRs in patients with stage III C2 than that in patients with III-C1 (56.5% vs. 24.3%, p < 0.001), demonstrating a significant difference in the RFS survival curve. CONCLUSION: RR in patients with clinical ESCC after hysterectomy followed by adjuvant radiotherapy is correlated with the number of positive LNs, TD > 4 cm, and PI. Emphasis should be placed on the common high-risk factor of LNM association with recurrence after radical hysterectomy in ESCC.

Evaluation of the Position Error of Wearing Surgical Masks During Radiotherapy in Head and Neck Cancer Patients.

Purpose: Wearing a mask during the coronavirus disease 2019 epidemic (COVID-19) is a preventive way to reduce droplet and aerosol transmission. The purpose of this study was to evaluate the position error of wearing a surgical mask during radiotherapy in head and neck cancer patients. Patients and Methods: We collected and analyzed 2351 kV X-ray image records of 81 patients with head and neck cancer who underwent image-guided radiotherapy (IGRT). Patients with/without a surgical mask were divided into the head-neck (HN) mask group and head-neck-shoulder (HNS) mask group. The position error in the X (left-right), Y (superior-inferior), Z (anterior-posterior), 3D (three dimensional) vectors, as well as the pitch and yaw axes were compared between the four groups. Results: We found that patients wearing surgical masks in the HN mask group showed no significant differences in the mean position error of the different types of headrest (p>0.05). In the HNS mask group, only the type C headrest group showed significant differences (P < 0.05). The X axis values were -0.05±0.07 and -0.11± 0.01 cm (P = 0.04), and the pitch axis values were 0.34±0.29° and 0.83±0.08° (P = 0.01). Conclusion: The mean position error of most patients wearing surgical masks was not greater than patients without a surgical mask. Patients wearing while receiving treatment is a low-cost and easy-to-implement prevention method.

Average Daily Gain and Energy and Nitrogen Requirements of 4-Month-Old Female Yak Calves.

There is little information available on milk intake and energy and nitrogen requirements of growing yak calves. This study aimed to fill this important gap, as this information could be beneficial in designing a system to wean yak calves earlier than in natural time. We determined the average daily gain and energy and nitrogen balances and requirements of 4-month-old female yak calves (48.8 ± 2.45 kg, n = 8). The calves were allowed to suck once a day and were fed an ad libitum concentrate: hay diet at a ratio of 60:40. Milk intake averaged 540 ± 26 g/d, yielding 2.28 ± 0.112 MJ/d, which was 13% of the gross energy intake (GEI). The digestible energy intake (DEI):GEI ratio was 0.681, metabolizable energy intake (MEI):DEI was 0.913, and MEI:GEI was 0.621. The average daily gain of the calves was 433 ± 153.1 g/d, which consisted of 78.0 ± 8.99 g protein, 52.7 ± 23.74 g fat, and 302.3 ± 95.1 g water, that is, 18.0% protein, 13.0% fat and 69.8% water. There were 130.7 g of body solids and 9.06 MJ of energy in every kg of body mass gain. Of the MEI, 25.17 kJ were required for 1 g of body mass, 83.40 kJ for 1 g of body solids, and 2.62 kJ for 1 kJ of retained energy (RE), and RE was 36.6% of MEI. The maintenance energy requirement was 5.35 MJ/d, the efficiency of utilization of energy for growth (kg) was 0.72, and the heat increment of feeding for growth was 0.28 (1.55 MJ/d). Digestible nitrogen (N) was 0.685 while retained N (RN) was 0.489 of N intake. The N requirement for maintenance was 11.73 g/d or 0.61 g N/kg0.75 per day, while the biological value (BV) of N was 91.1%. The energy and N requirements for yak calves were relatively low, which could be explained, at least in part, by the high efficiency of utilization of energy and high BV of N when compared to other livestock. These findings could be beneficial in designing early weaning systems for the many Himalayan households depending on yak production for their livelihoods.

Plant-derived Ren's oligopeptide has antiviral effects on influenza virus and SARS-CoV-2.

Influenza virus and SARS-CoV-2 virus are two important viruses that cause respiratory tract diseases. The high-frequency mutation of the two types of viruses leads to failure of the durable immune protection of vaccines, meanwhile it also poses continuous challenges to the development of antiviral drugs. Traditional Chinese medicine contains large number of biologically active compounds, and some of them contain broad-spectrum antiviral ingredients. In this study, we extracted antiviral active ingredients from medicinal and edible plants by biotransformation and enzymatic hydrolysis as a drug, and we named this drug Ren's oligopeptide. Further, we analyzed the antiviral activity of this drug and found that Ren's oligopeptide could inhibit the replication of influenza virus and SARS-CoV-2 virus with high anti-virus activities. In vitro experiments showed that the antiviral activity of the Ren's oligopeptide mainly targets the replication process after virus enters the cell. Therefore, Ren's oligopeptide is a promising drug against influenza and COVID-19.

Global Genome Diversity and Recombination in Mycoplasma pneumoniae.

Genomic changes in Mycoplasma pneumoniae caused by adaptation to environmental or ecologic pressures are poorly understood. We collected M. pneumoniae from children who had confirmed pneumonia in Taiwan during 2017-2020. We used whole-genome sequencing to compare these isolates with a worldwide collection of current and historical clinical strains for characterizing population structures. A phylogenetic tree for 284 strains showed that all sequenced strains consisted of 5 clades: T1-1 (sequence type [ST]1), T1-2 (mainly ST3), T1-3 (ST17), T2-1 (mainly ST2), and T2-2 (mainly ST14). We identified a putative recombination block containing 6 genes (MPN366‒371). Macrolide resistance involving 23S rRNA mutations was detected for each clade. Clonal expansion of macrolide resistance occurred mostly within subtype 1 strains, of which clade T1-2 showed the highest recombination rate and genome diversity. Functional characterization of recombined regions provided clarification of the biologic role of these recombination events in the evolution of M. pneumoniae.

Bacterial factors required for Streptococcus pneumoniae coinfection with influenza A virus.

BACKGROUND: Streptococcus pneumoniae is a common cause of post-influenza secondary bacterial infection, which results in excessive morbidity and mortality. Although 13-valent pneumococcal conjugate vaccine (PCV13) vaccination programs have decreased the incidence of pneumococcal pneumonia, PCV13 failed to prevent serotype 3 pneumococcal disease as effectively as other vaccine serotypes. We aimed to investigate the mechanisms underlying the co-pathogenesis of influenza virus and serotype 3 pneumococci. METHODS: We carried out a genome-wide screening of a serotype 3 S. pneumoniae transposon insertion mutant library in a mouse model of coinfection with influenza A virus (IAV) to identify the bacterial factors required for this synergism. RESULTS: Direct, high-throughput sequencing of transposon insertion sites identified 24 genes required for both coinfection and bacterial infection alone. Targeted deletion of the putative aminotransferase (PA) gene decreased bacterial growth, which was restored by supplementation with methionine. The bacterial burden in a coinfection with the PA gene deletion mutant and IAV in the lung was lower than that in a coinfection with wild-type pneumococcus and IAV, but was significantly higher than that in an infection with the PA gene deletion mutant alone. These data suggest that IAV infection alters host metabolism to benefit pneumococcal fitness and confer higher susceptibility to pneumococcal infection. We further demonstrated that bacterial growth was increased by supplementation with methionine or IAV-infected mouse lung homogenates. CONCLUSIONS: The data indicates that modulation of host metabolism during IAV infection may serve as a potential therapeutic intervention against secondary bacterial infections caused by serotype 3 pneumococci during IAV outbreaks in the future.

Robotics Dexterous Grasping: The Methods Based on Point Cloud and Deep Learning.

Dexterous manipulation, especially dexterous grasping, is a primitive and crucial ability of robots that allows the implementation of performing human-like behaviors. Deploying the ability on robots enables them to assist and substitute human to accomplish more complex tasks in daily life and industrial production. A comprehensive review of the methods based on point cloud and deep learning for robotics dexterous grasping from three perspectives is given in this paper. As a new category schemes of the mainstream methods, the proposed generation-evaluation framework is the core concept of the classification. The other two classifications based on learning modes and applications are also briefly described afterwards. This review aims to afford a guideline for robotics dexterous grasping researchers and developers.

USP11 facilitates colorectal cancer proliferation and metastasis by regulating IGF2BP3 stability.

The abnormal expression of ubiquitin-specific protease 11 (USP11) is thought to be related to tumor development and progression; however, few studies have reported the biological function and clinical importance of USP11 in colorectal cancer (CRC). Therefore, it is necessary to further explore the role of USP11 in CRC. Immunohistochemical staining was used to explore the association between prognosis and USP11 expression in CRC. Cholecystokinin octapeptide (CCK-8), colony formation, transwell, and animal assays were used to study the abilities of proliferation, migration, and invasion in CRC cells. Co-immunoprecipitation assays, Western blotting, ubiquitination assays, and rescue experiments were performed to elucidate the interaction between USP11 and insulin-like growth factor 2 mRNA binding protein 3 (IGF2BP3). We verified that USP11 was overexpressed in CRC tissues and was associated with the depth of tumor invasion and metastasis. USP11 knockdown or overexpression could weaken or reinforce the abilities of proliferation, migration, and invasion in CRC cells in vivo or in vitro. IGF2BP3 was protected by USP11 from degradation via deubiquitination. The rescue experiments revealed that IGF2BP3 overexpression could effectively reverse the decrease in cell proliferation, migration, and invasion caused by USP11 knockdown. Therefore, USP11 might be involved in CRC tumorigenesis and development through a USP11-IGF2BP3 axis pathway, and USP11 overexpression might be a novel indicator for poor prognosis and a potential therapeutic target in CRC patients.

Clonal spread of macrolide-resistant Mycoplasma pneumoniae sequence type-3 and type-17 with recombination on non-P1 adhesin among children in Taiwan.

OBJECTIVES: Mycoplasma pneumoniae is currently the most commonly detected bacterial cause of childhood community-acquired pneumonia in several countries. Of note, clonal expansion of macrolide-resistant ST3 occurred in Japan and South Korea. An alarming surge in macrolide resistance complicates the treatment of pneumonia. We aimed to evaluate the clinical manifestation and clonal relatedness of M. pneumoniae circulating among children in Taiwan. METHODS: We prospectively enrolled 626 children with radiologically confirmed pneumonia between 2017 and 2019. An M. pneumoniae infection was suspected on clinical grounds, and tested by real-time PCR and oropharyngeal swab cultures. We used multilocus sequence typing and whole-genome sequencing to characterize the genetic features of M. pneumoniae. RESULTS: A total of 226 children with M. pneumoniae pneumonia were enrolled. Macrolide resistance was found in 77% (174/226) of patients. Multi-locus sequence typing revealed that ST3 (n = 93) and its single-locus variant ST17 (n = 84) were the predominant clones among macrolide-resistant strains. ST17 presented clinical characteristics comparable to its ancestor ST3. On multivariate analysis, macrolide resistance (OR 3.5; 95% CI 1.4-8.5; p 0.007) was independently associated with fever >72 hours after macrolide treatment. By whole-genome sequencing, prediction analysis of recombination sites revealed one recombination site in ST3 and ST17 compared with M29 (a macrolide-sensitive ST3 strain isolated from China in 2005) containing cytadhesin MgpC-like protein, RepMP4 and RepMP5. ST17 had another recombination site containing an adhesin and RepMP2/3. CONCLUSIONS: In addition to macrolide resistance, ST3 and its ST17 variant might evolve through recombination between repetitive sequences and non-P1 cytadhesins for persistent circulation in Taiwan.

An increase in dietary lipid content from different forms of double-low rapeseed reduces enteric methane emission in Datong yaks on the Qinghai-Tibetan Plateau.

Enteric methane (CH4 ) emission in cattle generally decreases by approximately 1 g/g dry matter intake (DMI) with an increase in dietary lipids of 10 g/kg dry matter (DM). The effect of dietary lipids on CH4 emission in yaks has not been reported and is the subject of this study. Four Datong yaks were used in a 4 × 4 Latin-square design in which the four treatments included restricted intakes of double-low rapeseed differing in form and lipid (ether extract-EE) content: (a) rapeseed meal (EE 32.6 g/kg DM); (b) rapeseed meal and rapeseed cake (EE 45.8 g/kg DM); (c) rapeseed meal and whole cracked rapeseed (EE 54.5 g/kg DM) and (d) rapeseed meal and rapeseed oil (EE 62.7 g/kg DM). The digestibility of feed components did not differ among treatments. The ruminal total volatile fatty acids (p = .082) and acetic acid (p = .062) concentrations tended to be lowest in yaks consuming the diet with highest lipid content. In addition, CH4 production was lowest in this group (p = .004), and declined by 1.75 g/g DMI per 10 g/kg DM reduction in dietary lipid content, a rate substantially faster than in cattle.

Aberrant USP11 expression regulates NF90 to promote proliferation and metastasis in hepatocellular carcinoma.

Growing evidence indicates that deubiquitinase ubiquitin-specific protease 11 (USP11) plays an important role in cellular function by regulating the stability of its substrates. USP11 is dysregulated in many types of cancer and involved in tumor development and progression. We previously showed that USP11 was upregulated in hepatocellular carcinoma (HCC) and promoted HCC cell invasion and metastasis potency. However, the mechanism underlying the role of USP11 in HCC cell metastasis and its function in cell proliferation remain unknown. Here, CCK-8, soft agar assays and nude mouse models showed that USP11 was essential for HCC cells survival and proliferation in vitro and in vivo. Results form mass spectrometry, co-immunoprecipitation, and ubiquitination assays demonstrated that USP11 interacted with nuclear factor 90 (NF90) and promoted its deubiquitination, thereby stabilizing it in HCC cells. Moreover, the effect of USP11 on promoting HCC cells proliferation and metastasis was dependent on NF90, and USP11 expression was positively correlated with NF90 expression in human HCC tissues, as demonstrated via immunohistochemistry. Collectively, the present findings indicated that USP11 binded to and deubiquitinated NF90, thereby stabilizing the protein expression level and promoting HCC cell proliferation and metastasis. NF90 was identified as an important downstream target of USP11. Dysregulated signaling of this novel USP11/NF90 axis might promote HCC proliferation and metastasis, and the axis could be a potential therapeutic target in HCC.

Genomic Insight into the Spread of Meropenem-Resistant Streptococcus pneumoniae Spain23F-ST81, Taiwan.

Incidence of invasive pneumococcal disease caused by antimicrobial-resistant Streptococcus pneumoniae types not included in pneumococcal conjugate vaccines has increased, including a penicillin- and meropenem-resistant serotype 15A-ST63 clone in Japan. During 2013-2017, we collected 206 invasive pneumococcal isolates in Taiwan for penicillin and meropenem susceptibility testing. We found serotypes 15B/C-ST83 and 15A-ST63 were the most prevalent penicillin- and meropenem-resistant clones. A transformation study confirmed that penicillin-binding protein (PBP) 2b was the primary meropenem resistance determinant, and PBP1a was essential for high-level resistance. The rate of serotype 15B/C-ST83 increased during the study. All 15B/C-ST83 isolates showed an ermB macrolide resistance genotype. Prediction analysis of recombination sites revealed 12 recombination regions in 15B/C-ST83 compared with the S. pneumoniae Spain23F-ST81 genome. Pneumococcal clones rapidly recombine to acquire survival advantages and undergo local expansion under the selective pressure exerted by vaccines and antimicrobial drugs. The spread of 15B/C-ST83 is alarming for countries with high antimicrobial pressure.

Adding heat-treated rapeseed to the diet of yak improves growth performance and tenderness and nutritional quality of the meat.

Heat-treated rapeseed was supplemented to indoor fed yaks in winter to test the effect on dry matter intake (DMI), body mass change, and meat quality. Sixteen 3-year-old yak steers (124 ± 15.3 kg) were divided randomly into two groups and were offered either heat-treated rapeseed (HTR) or rapeseed meal (CONT). The yaks were allowed 14 days for adjustment and measurements were made over 120 d. There was no difference in DMI between groups (p = 0.67), but average daily gain tended to be higher (p < 0.056) and feed to gain ratio tended to be lower (p = 0.050) in HTR than in CONT yaks. Meat from HTR yaks was more tender (p = 0.006), had higher intramuscular fat (p = 0.013), and had lower cholesterol content (p = 0.009) than from CONT yaks. In addition, the atherogenic index was lower (0.37 vs. 0.43; p = 0.049), the PUFA:SFA ratio was higher (0.55 vs. 0.37; p = 0.049), and the n-6:n-3 (n-6 PUFA to n-3 PUFA) ratio was lower (2.76 vs. 4.78; p = 0.003) in HTR than in CONT yaks, which all favoured the HTR yaks. Meat from HTR yaks met human health standards of a PUFA:SFA ratio of above 0.4 and n-6:n-3 ratio of less than 4, whereas meat from CONT yaks just missed these standards.

MiR-7-5p-mediated downregulation of PARP1 impacts DNA homologous recombination repair and resistance to doxorubicin in small cell lung cancer.

BACKGROUND: Chemo-resistance is one of the major challenges in the therapy of small cell lung cancer (SCLC). Multiple mechanisms are thought to be involved in chemo-resistance during SCLC treatment, but unfortunately, these mechanisms have not been well elucidated. Herein, we investigated the role of miRNA in the resistance of SCLC cells to doxorubicin (Dox). METHODS: MiRNA microarray analysis revealed that several miRNAs, including miR-7-5p, were specifically decreased in Dox-resistant SCLC cells (H69AR) compared to parental cells (H69). The expression level of miR-7-5p was confirmed by qRT-PCR in Dox-resistant cells (H69AR and H446AR cells) and their parental cells. Bioinformatic analysis indicated that poly ADP-ribose polymerase 1 (PARP1) is a direct target of miR-7-5p. The binding sites of miR-7-5p in the PARP1 3' UTR were verified by luciferase reporter and Western blot assays. To investigate the role of miR-7-5p in the chemo-resistance of SCLC cells to doxorubicin, mimic or inhibitor of miR-7-5p was transfected into SCLC cells, and the effect of miR-7-5p on homologous recombination (HR) repair was analyzed by HR reporter assays. Furthermore, the expression of HR repair factors (Rad51 and BRCA1) induced by doxorubicin was detected by Western blot and immunofluorescent staining in H446AR cells transfected with miR-7-5p mimic. RESULTS: The expression level of miR-7-5p was remarkably reduced (4-fold) in Dox-resistant SCLC cells (H69AR and H446AR cells) compared with that in parental cells (H69 and H446 cells). Poly ADP-ribose polymerase 1 (PARP1) is a direct target of miR-7-5p, and PARP1 expression was downregulated by miR-7-5p. MiR-7-5p impeded Dox-induced HR repair by inhibiting the expression of HR repair factors (Rad51 and BRCA1) that resulted in resensitizing SCLC cells to doxorubicin. CONCLUSIONS: Our findings provide evidence that miR-7-5p targets PARP1 to exert its suppressive effects on HR repair, indicating that the alteration of the expression of miR-7-5p may be a promising strategy for overcoming chemo-resistance in SCLC therapy.

Prophage Excision in Streptococcus pneumoniae Serotype 19A ST320 Promote Colonization: Insight Into Its Evolution From the Ancestral Clone Taiwan 19F-14 (ST236).

Streptococcus pneumoniae 19A ST320, a multidrug-resistant strain with high disease severity that notoriously spread before the use of expanded pneumococcal conjugate vaccines, was derived from a capsular switching event between an international strain Taiwan 19F-14 (ST236) and a serotype 19A strain. However, the molecular mechanisms underlying the adaptive evolution of 19F ST236 to 19A ST320 are unknown. In this study, we compared 19A ST320 to its ancestral clone, 19F ST236, in terms of adherence to respiratory epithelial cells, whole transcriptome, and ability to colonize a young mouse model. Serotype 19A ST320 showed five-fold higher adherence to A549 cells than serotype 19F ST236. High-throughput mRNA sequencing identified a prophage region located between dnaN and ychF in both strains; however, the genes in this region were expressed at significantly higher levels in 19A ST320 than in 19F ST236. Analysis by polymerase chain reaction (PCR) showed that the prophage is able to spontaneously excise from the chromosome and form a circular episome in 19A ST320, but not in 19F ST236. Deletion of the integrase in the prophage of 19A ST320 decreased spontaneous excision and cell adherence, which were restored by complementation. Competition experiments in mice showed that the integrase mutant was six-fold less competitive than the 19A ST320 parent (competitive index [CI]: 0.16; p = 0.02). The 19A ST320 prophage-deleted strain did not change cell adherence capacity, whereas prophage integration strains (integrase mutant and 19F) had decreased expression of the down-stream ychF gene compared to that of 19A ST320. Further deletion of ychF significantly reduced cell adherence. In conclusions, these findings suggest that spontaneous prophage induction confers a competitive advantage to virulent pneumococci.

Generation of Ultrafine Droplets in Femtoliter Scale from a Large Needle with Diameter of 200 Microns.

Ultrafine droplets play important roles in many fields. Here, we prepare ultrafine droplets with volumes in femtoliter scale by applying an electrostatic field between a needle and substrate. The diameter of liquid is reduced significantly to about 1/50 that of the needle tip. By using some solvents consisting of small molecules, ultrafine droplets eject from the needle tip. The volume of the ultrafine droplets depend on the strength of the electrostatic field and properties of the liquid. Ultrafine droplets containing perovskite quantum dots are also ejected on the substrate by using this jetting method. The ultrafine droplets have great potentials in carrying tiny amount of quantum dots and even molecules for various applications.

Using multivariate regression model with least absolute shrinkage and selection operator (LASSO) to predict the incidence of Xerostomia after intensity-modulated radiotherapy for head and neck cancer.

PURPOSE: The aim of this study was to develop a multivariate logistic regression model with least absolute shrinkage and selection operator (LASSO) to make valid predictions about the incidence of moderate-to-severe patient-rated xerostomia among head and neck cancer (HNC) patients treated with IMRT. METHODS AND MATERIALS: Quality of life questionnaire datasets from 206 patients with HNC were analyzed. The European Organization for Research and Treatment of Cancer QLQ-H&N35 and QLQ-C30 questionnaires were used as the endpoint evaluation. The primary endpoint (grade 3(+) xerostomia) was defined as moderate-to-severe xerostomia at 3 (XER3m) and 12 months (XER12m) after the completion of IMRT. Normal tissue complication probability (NTCP) models were developed. The optimal and suboptimal numbers of prognostic factors for a multivariate logistic regression model were determined using the LASSO with bootstrapping technique. Statistical analysis was performed using the scaled Brier score, Nagelkerke R(2), chi-squared test, Omnibus, Hosmer-Lemeshow test, and the AUC. RESULTS: Eight prognostic factors were selected by LASSO for the 3-month time point: Dmean-c, Dmean-i, age, financial status, T stage, AJCC stage, smoking, and education. Nine prognostic factors were selected for the 12-month time point: Dmean-i, education, Dmean-c, smoking, T stage, baseline xerostomia, alcohol abuse, family history, and node classification. In the selection of the suboptimal number of prognostic factors by LASSO, three suboptimal prognostic factors were fine-tuned by Hosmer-Lemeshow test and AUC, i.e., Dmean-c, Dmean-i, and age for the 3-month time point. Five suboptimal prognostic factors were also selected for the 12-month time point, i.e., Dmean-i, education, Dmean-c, smoking, and T stage. The overall performance for both time points of the NTCP model in terms of scaled Brier score, Omnibus, and Nagelkerke R(2) was satisfactory and corresponded well with the expected values. CONCLUSIONS: Multivariate NTCP models with LASSO can be used to predict patient-rated xerostomia after IMRT.