bluuebunny/crossref_metadata_2025 huggingface dataset:
Contact Mechanism of Rail Grinding with Open-Structured Abrasive Belt Based on Pressure Grinding Plate
Authors: Wu Zhiwei , Fan Wengang , Qian Chang , Hou Guangyou
Published: March 2023

The current research of abrasive belt grinding rail mainly focuses on the contact mechanism and structural design. Compared with the closed structure abrasive belt grinding, open-structured abrasive belt grinding has excellent performance in dynamic stability, consistency of grinding quality, extension of grinding mileage and improvement of working efficiency. However, in the contact structure design, the open-structured abrasive belt grinding rail using a profiling pressure grinding plate and the closed structure abrasive belt using the contact wheel are different, and the contact mechanisms of the two are different. In this paper, based on the conformal contact and Hertz theory, the contact mechanism of the pressure grinding plate, abrasive belt and rail is analyzed. Through finite element simulation and static pressure experiment, the contact behavior of pressure grinding plate, abrasive belt and rail under single concentrated force, uniform force and multiple concentrated force was studied, and the distribution characteristics of contact stress on rail surface were observed. The results show that under the same external load, there are three contact areas under the three loading modes. The outer contour of the middle contact area is rectangular, and the inner contour is elliptical. In the contact area at both ends, the stress is extremely small under a single concentrated force, the internal stress is drop-shaped under a uniform force, and the internal stress under multiple concentration forces is elliptical. Compared with the three, the maximum stress is the smallest and the stress distribution is more uniform under multiple concentrated forces. Therefore, the multiple concentrated forces is the best grinding pressure loading mode. The research provides support for the application of rail grinding with open-structured abrasive belt based on pressure grinding plate, such as contact mechanism and grinding pressure mode selection.
Development of glycan-targeted nanoparticles as a novel therapeutic opportunity for gastric cancer treatment
Authors: dos Santos Sofia Nascimento , Junior Dino Seigo Gushiken , Pereira Jhonatas Pedrosa Marim , Iadocicco Natália Miranda , Silva André Henrique , do Nascimento Tatielle , Dias Luís Alberto Pereira , de Oliveira Silva Flávia Rodrigues , Ricci-Junior Eduardo , Santos-Oliveira Ralph , Bernardes Emerson Soares
Published: March 2023

Chemotherapy resistance remains a major cause of therapeutic failure in gastric cancer. The combination of genetic material such as interference RNAs (iRNAs) to silence cancer-associated genes with chemotherapeutics has become a novel approach for cancer treatment. However, finding the right target genes and developing non-toxic, highly selective nanocarrier systems remains a challenge. Here we developed a novel sialyl-Tn-targeted polylactic acid—didodecyldimethylammonium bromide nanoparticle (PLA-DDAB) nanoparticles (NPs) loaded with dsRNA targeting ST6GalNac-I and/or galectin-3 genes. Using single photon emission computed tomography (SPECT), we have demonstrated that 99mtechnetium radiolabeled sialyl-Tn-targeted nanoparticles can reach the tumor site and downregulate ST6GalNAc-I and galectin-3 RNA expression levels when injected intravenously. Furthermore, using an in vivo gastric tumor model, these nanoparticles increased the effectiveness of 5-FU in reducing tumor growth. Our findings indicate that cancer-associated glycan-targeted NPs loaded with dsRNA targeting ST6GalNAc-I and/or galectin-3 in combination with standard chemotherapy, have the potential to become a novel therapeutic tool for gastric cancer.
Comparative chloroplast genomics of 34 species in subtribe Swertiinae (Gentianaceae) with implications for its phylogeny
Authors: Yang Lucun , Deng Shengxue , Zhu Yongqing , Da Qilin
Published: March 2023

Background

Subtribe Swertiinae, a medicinally significant and highly speciose Subtribe of family Gentianaceae. Despite previous extensive studies based on both morphology and molecular data, intergeneric and infrageneric relationships within subtribe Swertiinae remain controversial.

Methods

Here, we employed four newly generated Swertia chloroplast genomes with thirty other published genomes to elucidate their genomic characteristics.

Results

The 34 chloroplast genomes were small and ranged in size from 149,036 to 154,365 bp, each comprising two inverted repeat regions (size range 25,069–26,126 bp) that separated large single-copy (80,432–84,153 bp) and small single-copy (17,887–18,47 bp) regions, and all the chloroplast genomes showed similar gene orders, contents, and structures. These chloroplast genomes contained 129–134 genes each, including 84–89 protein-coding genes, 37 tRNAs, and 8 rRNAs. The chloroplast genomes of subtribe Swertiinae appeared to have lost some genes, such as rpl33, rpl2 and ycf15 genes. Comparative analyses revealed that two mutation hotspot regions (accD-psaI and ycf1) could serve as effective molecular markers for further phylogenetic analyses and species identification in subtribe Swertiinae. Positive selection analyses showed that two genes (ccsA and psbB) had high Ka/Ks ratios, indicating that chloroplast genes may have undergone positive selection in their evolutionary history. Phylogenetic analysis showed that the 34 subtribe Swertiinae species formed a monophyletic clade, with Veratrilla, Gentianopsis and Pterygocalyx located at the base of the phylogenetic tree. Some genera of this subtribe, however, were not monophyletic, including Swertia, Gentianopsis, Lomatogonium, Halenia, Veratrilla and Gentianopsis. In addition, our molecular phylogeny was consistent with taxonomic classification of subtribe Swertiinae in the Roate group and Tubular group. The results of molecular dating showed that the divergence between subtrib Gentianinae and subtrib Swertiinae was estimated to occur in 33.68 Ma. Roate group and Tubular group in subtribe Swertiinae approximately diverged in 25.17 Ma.

Conclusion

Overall, our study highlighted the taxonomic utility of chloroplast genomes in subtribe Swertiinae, and the genetic markers identified here will facilitate future studies on the evolution, conservation, population genetics, and phylogeography of subtribe Swertiinae species.
The construction of patient quality management model in oral post anesthesia care unit in China: a grounded theory approach
Authors: Zhang Qi , Tang Heshu , Yao Zhiqing , Han Wei
Published: March 2023

Background

Nowadays, people have paid more and more attention to the quality of physical and mental health recovery after oral surgery anesthesia. As a remarkable feature of patient quality management, it can effectively reduce the risk of postoperative complications and pain in Post Anesthesia Care Unit (PACU). However, the patient management model in oral PACU remains unknown, especially in China. The purpose of this study is to explore the management elements of patient quality management in the oral PACU and to construct the management model.

Methods

Strauss and Corbin’s grounded theory method was used to explore the experiences of three anesthesiologists, six anesthesia nurses and three administrators working in oral PACU. Twelve semi-structured interviews were conducted using face-to-face in a tertiary stomatological hospital from March to June, 2022. The interviews were transcribed and thematically analysed according to QSR NVivo 12.0 qualitative analysis tool.

Results

Three themes and ten subthemes were identified through an active analysis process, including three of the core team members: stomatological anesthesiologists, stomatological anesthesia nurses and administrators, three of the main functions: education and training, patient care and quality control and four of the team operation processes: analysis, plan, do, check.

Conclusion

The patient quality management model of the oral PACU is helpful for the professional identity and career development of stomatological anesthesia staff in China, which can accelerate the professional development of oral anesthesia nursing quality. According to the model, the patient’s pain and fear will decrease, meanwhile, safety and comfort will increase. It can make contributions to the theoretical research and clinical practice in the future.
APC mutation correlated with poor response of immunotherapy in colon cancer
Authors: Li Bing , Zhang Guoliang , Xu Xuejie
Published: March 2023

Objective

APC (adenomatous polyposis coli) gene mutation is a central initialization in colon cancer tumorigenesis. However, the connection between APC gene mutation and immunotherapy efficacy for colon cancer remains unknown. This study aimed to explore the impact of APC mutation on immunotherapy efficacy for colon cancer.

Methods

Colon cancer data from The Cancer Genome Atlas (TCGA) and Memorial Sloan Kettering Cancer Center (MSKCC) were used for the combined analysis. Survival analysis was performed to evaluate the association between APC mutation and immunotherapy efficacy in colon cancer patients. The expressions of immune check point molecules, tumor mutation burden (TMB), CpG methylation level, tumor purity (TP), microsatellite instability (MSI) status and tumor-infiltrating lymphocyte (TIL) in the two APC status were compared to evaluate the associations between APC mutation and immunotherapy efficacy indicators. Gene set enrichment analysis (GSEA) was performed to identify signaling pathways related to APC mutation.

Results

APC was the most frequently mutated gene in colon cancer. The survival analysis demonstrated that APC mutation was correlated with a worse immunotherapy outcome. APC mutation was associated with lower TMB, lower expression of immune check point molecules (PD-1/PD-L1/PD-L2), higher TP, lower MSI-High proportion and less CD8 + T cells and follicular helper T cells infiltration. GSEA indicated that APC mutation up-regulated mismatch repair pathway, which may play a negative role in evoking an antitumor immune response.

Conclusion

APC mutation is associated with worse immunotherapy outcome and inhibition of antitumor immunity. It can be used as a negative biomarker to predict immunotherapy response.
Isolated massive histiocytes renal interstitial infiltration: a case report of an unexpected cause of acute kidney injury in a kidney transplant recipient
Authors: Martins Luis E. M. , Moyses-Neto Miguel , Costa Roberto S. , Traina Fabiola , Romao Elen A.
Published: March 2023

Background

Acute kidney injury is a frequent cause of hospital readmission in kidney transplant recipients (KTR), usually associated with infections and graft rejection. Herein, we report a case of an unusual cause of acute kidney injury in a KTR (massive histiocytes renal interstitial infiltration).

Case presentation

A 40-year-old woman was submitted to a second kidney transplant. One year after surgery, she presented asthenia, myalgia, and fever, haemoglobin 6.1 g/dL; neutrophils: 1.3 × 109/µL; platelets: 143 × 109/µL; blood creatinine 11.8 mg/dL, requiring dialysis. A kidney biopsy revealed diffuse histiocytic infiltration, which was assumed due to dysregulated immunological activation triggered by infections. The patient had multiple infections, including cytomegalovirus infection (CMV), aspergillosis, bacteraemia, and urinary tract infections, which could trigger the immune response. Haemophagocytic lymphohistiocytosis (HLH) was ruled out. The present case highlights the occurrence of isolated massive renal interstitial infiltration of histiocytes that does not meet the criteria for HLH or other related pathologies.

Conclusions

Renal histiocyte activation and infiltration may have been initiated by an immunological mechanism similar to what occurs in HLH and infectious processes. The present case highlights the occurrence of isolated massive renal interstitial infiltration of histiocytes that does not meet the criteria for HLH or other related pathologies.
Mobile health to promote physical activity in people post stroke or transient ischemic attack – study protocol for a feasibility randomised controlled trial
Authors: Thurston Charlotte , Bezuidenhout Lucian , Humphries Sophia , Johansson Sverker , von Koch Lena , Häger Charlotte K. , Holmlund Lisa , Sundberg Carl Johan , Garcia-Ptacek Sara , Kwak Lydia , Nilsson Michael , English Coralie , Conradsson David Moulaee
Published: March 2023

Background

Physical activity is essential to improve health and reduce the risk of recurrence of stroke or transient ischemic attack (TIA). Still, people post stroke or TIA are often physically inactive and the availability of physical activity promotion services are often limited. This study builds on an existing Australian telehealth-delivered programme (i-REBOUND– Let’s get moving) which provides support for home-based physical activity for people post stroke or TIA. The aim of this study is to test the feasibility, acceptability, and preliminary effects of a mobile Health (mHealth) version of the i-REBOUND programme for the promotion of physical activity in people post stroke or TIA living in Sweden.

Methods

One hundred and twenty participants with stroke or TIA will be recruited via advertisement. A parallel-group feasibility randomised controlled trial design with a 1:1 allocation ratio to 1) i-REBOUND programme receiving physical exercise and support for sustained engagement in physical activity through behavioural change techniques, or 2) behavioural change techniques for physical activity. Both interventions will proceed for six months and be delivered digitally through a mobile app. The feasibility outcomes (i.e., reach, adherence, safety and fidelity) will be monitored throughout the study. Acceptability will be assessed using the Telehealth Usability Questionnaire and further explored through qualitative interviews with a subset of both study participants and the physiotherapists delivering the intervention. Clinical outcomes on preliminary effects of the intervention will include blood pressure, engagement in physical activity, self-perceived exercise self-efficacy, fatigue, depression, anxiety, stress and health-related quality of life and will be measured at baseline and at 3, 6 and 12 months after the baseline assessments.

Discussion

We hypothesise that the mHealth delivery of the i-REBOUND programme will be feasible and acceptable in people post stroke/TIA living in rural and urban regions of Sweden. The results of this feasibility trial will inform the development of full-scale and appropriately powered trial to test the effects and costs of mHealth delivered physical activity for people after stroke or TIA.

Trial registration

ClinicalTrials.gov Identifier: NCT05111951. Registered November 8, 2021.
Deprescribing psychotropic medicines for behaviours that challenge in people with intellectual disabilities: a systematic review
Authors: Adams Danielle , Hastings Richard P. , Maidment Ian , Shah Chetan , Langdon Peter E.
Published: March 2023

Background

Clear evidence of overprescribing of psychotropic medicines to manage behaviours that challenges in people with intellectual disabilities has led to national programmes within the U.K. such as NHS England’s STOMP to address this. The focus of the intervention in our review was deprescribing of psychotropic medicines in children and adults with intellectual disabilities. Mental health symptomatology and quality of life were main outcomes.

Methods

We reviewed the evidence using databases Medline, Embase, PsycINFO, Web of Science, CINAHL and Open Grey with an initial cut-off date of 22nd August 2020 and an update on 14th March 2022. The first reviewer (DA) extracted data using a bespoke form and appraised study quality using CASP and Murad tools. The second reviewer (CS) independently assessed a random 20% of papers.

Results

Database searching identified 8675 records with 54 studies included in the final analysis.

The narrative synthesis suggests that psychotropic medicines can sometimes be deprescribed. Positive and negative consequences were reported. Positive effects on behaviour, mental and physical health were associated with an interdisciplinary model.

Conclusions

This is the first systematic review of the effects of deprescribing psychotropic medicines in people with intellectual disabilities which is not limited to antipsychotics. Main risks of bias were underpowered studies, poor recruitment processes, not accounting for other concurrent interventions and short follow up periods. Further research is needed to understand how to address the negative effects of deprescribing interventions.

Trial registration

The protocol was registered with PROSPERO (registration number CRD42019158079)
The association between a priori dietary patterns and psychological disorders in military personnel
Authors: Parastouei Karim , Rostami Hosein , Chambari Mahla
Published: March 2023

Background

Studies have identified a high prevalence of poor mental health, including depression, anxiety, and stress in military occupations. A low quality diet is one of the potential factors related to mental disorders. This study aimed to investigate the association of a priori dietary patterns, including dietary approach to stop hypertension (DASH), the Mediterranean diet (MD), dietary inflammatory index (DII), and healthy eating index-2015 (HEI-2015) with the odds of depression, anxiety, and stress in military staff.

Methods

This cross-sectional study was performed on a total of 400 military staff, aged 38.67 ± 5.22 (ranging from 30 to 60) years, recruited from Iranian military centers. The dietary intake of participants and adherence to the DASH, MD, DII, and HEI-2015 was measured using a 168-item food frequency questionnaire (FFQ). Mental health was evaluated with the use of the Depression, Anxiety, and Stress Scale − 21 (DASS-21).

Results

The prevalence of depression, anxiety, and stress was 64.5%, 63.2%, and 61.3%, respectively. Individuals with the highest adherence to HEI-2015, compared to those with the lowest adherence, were significantly at lower odds of anxiety (OR = 0.51, 95%CI: 0.27–0.96, p = 0.03), while higher adherence to DII was related to a 2.74-fold increased odds of anxiety (OR = 2.74, 95%CI: 1.06–7.04, p = 0.03). Compared to those with the lowest adherence (quartile 1), quartile 2 of the HEI-2015 was associated with decreased odds of stress (p = 0.04). No association was identified between dietary patterns and depression.

Conclusion

Greater adherence to HEI-2015 and lower adherence to DII are associated with lower odds of anxiety in military staff.
Association between impairment and self-rated health: a brazilian population study considering type, origin, and degree of limitation
Authors: Anderle Paula , Ziegelmann Patrícia Klarmann , de Goulart Bárbara Niegia Garcia
Published: March 2023

Background

Perceived health is a well-known, low-cost measure in public health, and has been used in several studies on individuals with impairment. Although many studies have related impairment to self-rated health (SRH), few have considered the origin and degree of limitation of the impairment. This study examined whether physical, hearing, or visual impairments—when analyzed according to origin (congenital or acquired) and degree of limitation (with or without)—are associated with the SRH status.

Methods

This cross-sectional study used data of 43,681 adult individuals from the Brazilian National Health Survey (NHS, 2013). The outcome SRH was dichotomized into poor (including the regular, poor, and very poor responses) or good (including the good and very good responses). Crude and adjusted (for socio-demographic characteristics and chronic diseases history) prevalence ratios (PR) estimates were evaluated using Poisson regression models with the robust variance estimator.

Results

Poor SRH prevalence was estimated at 31.8% (95%CI:31.0–33.0) among the non-impaired population, 65.6% (95%CI:60.6–70.0) among individuals with physical impairment, 50.3% (95%CI:45.0–56.0) for people with hearing impairment, and 55.3% (95%CI:51.8–59.0) for the visually impaired. Individuals with congenital physical impairment—with or without limitations—presented the strongest association with the poorest SRH status. Participants with non-limiting, congenital hearing impairment showed a protective factor to poor SRH (PR = 0.40 95%CI: 0.38–0.52). Individuals with acquired visual impairment with limitations demonstrated the strongest association with poor SRH (PR = 1.48 95%CI:1.47–1.49). Among the impaired population, middle-aged participants showed a stronger association with poor SRH than older adult participants.

Conclusions

Impairment is associated with poor SRH status, especially among people with physical impairment. The origin and degree of limitation of each type of impairment differently impacts SRH among the impaired population.
Staged replacement of both hips and both knees in patients with rheumatoid arthritis
Authors: Cao Jian , Wang Wenqi , Feng Wei , Xu Hong , Wang Duan , Zhou Zongke
Published: March 2023

Background

Patients with rheumatoid arthritis (RA) undergoing bilateral total hip arthroplasty (THA) and total knee arthroplasty (TKA) are an uncommon population, and their outcomes are also difficult to predict. The purpose of this study was to evaluate whether both bilateral cementless THA and cemented posterior-stabilized TKA (PS-TKA) can provide reliable outcomes for RA patients.

Methods

We retrospectively reviewed 30 RA patients (60 hips and 60 knees) who underwent both elective bilateral cementless THA and cemented PS-TKA, with a minimum follow-up of 2 years. Clinical, patient-reported, and radiographic data were retrospectively analyzed.

Results

The mean follow-up was 84 months (range, 24–156). By the last follow-up, the post-operative range of motion, Harris Hip Score, Knee Society Score (KSS) clinical, KSS functional, Western Ontario and McMaster Universities Index of Osteoarthritis (WOMAC) hip, and WOMAC knee scores were significantly improved compared to the preoperative values. All patients achieved the ability to walk. In addition, overall satisfaction scores on a 100-point scale were 92.5 after THA and 89.6 after TKA. Only one patient underwent revision surgery due to knee joint instability, and all replaced hips and knees were radiographically stable by the assessment of the radiolucent line. The proportion of implants that did not suffer loosening or require revision surgery was 99.2% during the 84-month follow-up, based on Kaplan-Meier analysis.

Conclusions

Our study suggests that bilateral cementless THA and cemented PS-TKA provides reliable mid-long-term clinical, patient-reported, and radiographic outcomes in RA patients, with high survivorship and patient satisfaction.
Nurses’ knowledge, attitudes, and practice with regards to nutritional management of diabetes mellitus
Authors: Farzaei Mahsa , Shahbazi Shahla , Gilani Neda , Ostadrahimi Alireza , Gholizadeh Leila
Published: March 2023

Background

The prevalence of diabetes is increasing rapidly worldwide. Nurses work collaboratively with multidisciplinary teams to improve diabetes management. Yet, little is known about nurses’ role in nutritional management of diabetes. This study aimed to evaluate nurses’ knowledge, attitudes, and practice (KAP) toward nutritional management of diabetes.

Methods

This cross-sectional study was conducted with 160 nurses, who were recruited between July 4 and July 18, 2021 from two referral tertiary teaching hospitals in Iran. A validated paper-based self-reported questionnaire was used to assess nurses’ KAP. Data were analyzed using descriptive statistics and multiple linear regression analysis.

Results

The mean knowledge score of nurses about nutritional management of diabetes was 12.16 ± 2.83, and 61.2% showing a moderate knowledge level on nutritional management of diabetes. The mean attitudes score was 60.68 ± 6.11, with 86.92% of participants demonstrating positive attitudes. The mean practice score of study participants was 44.74 ± 7.81, with 51.9% having a moderate level of practice. Higher knowledge scores were observed among male nurses (B = -7.55, p = 0.009) and those with blended learning as a preferred learning method (B = 7.28, p = 0.029). Having an opportunity to provide education to patients with diabetes during shifts affected nurses’ attitudes positively (B = -7.59, p = 0.017). Practice scores were higher among nurses who perceived themselves competent in the nutritional management of diabetes (B = -18.05, p = 0.008).

Conclusion

Nurses’ knowledge and practice in the nutritional management of diabetes should be increased to help improve the quality of the dietary care and patient education they provide these patients. Further studies are needed to confirm the results of this study both in Iran and internationally.
Healthcare provider perceptions on the implementation of the universal test-and-treat policy in South Africa: a qualitative inquiry
Authors: Nicol Edward , Mehlomakulu Vuyelwa , Jama Ngcwalisa Amanda , Hlongwa Mbuzeleni , Basera Wisdom , Pass Desiree , Bradshaw Debbie
Published: March 2023

Background

South Africa had an estimated 7.5 million people living with HIV (PLHIV), accounting for approximately 20% of the 38.4 million PLHIV globally in 2021. In 2015, the World Health Organization recommended the universal test and treat (UTT) intervention which was implemented in South Africa in September 2016. Evidence shows that UTT implementation faces challenges in terms of human resources capacity or infrastructure. We aim to explore healthcare providers (HCPs)’ perspectives on the implementation of the UTT strategy in uThukela District Municipality in KwaZulu-Natal province.

Methods

A qualitative study was conducted with one hundred and sixty-one (161) healthcare providers (HCPs) within 18 healthcare facilities in three subdistricts, comprising of Managers, Nurses, and Lay workers. HCPs were interviewed using an open ended-survey questions to explore their perceptions providing HIV care under the UTT strategy. All interviews were thematically analysed using both inductive and deductive approaches.

Results

Of the 161 participants (142 female and 19 male), 158 (98%) worked at the facility level, of which 82 (51%) were nurses, and 20 (12.5%) were managers (facility managers and PHC manager/supervisors). Despite a general acceptance of the UTT policy implementation, HCPs expressed challenges such as increased patient defaulter rates, increased work overload, caused by the increased number of service users, and physiological and psychological impacts. The surge in the workload under conditions of inadequate systems’ capacity and human resources, gave rise to a greater burden on HCPs in this study. However, increased life expectancy, good quality of life, and immediate treatment initiation were identified as perceived positive outcomes of UTT on service users. Perceived influence of UTT on the health system included, increased number of patients initiated, decreased burden on the system, meeting the 90-90-90 targets, and financial aspects.

Conclusion

Health system strengthening such as providing more systems’ capacity for expected increase in workload, proper training and retraining of HCPs with new policies in the management of patient readiness for lifelong ART journey, and ensuring availability of medicines, may reduce strain on HCPs, thus improving the delivery of the comprehensive UTT services to PLHIV.
Engineering Escherichia coli for efficient assembly of heme proteins
Authors: Ge Jianzhong , Wang Xiaolu , Bai Yingguo , Wang Yaru , Wang Yuan , Tu Tao , Qin Xing , Su Xiaoyun , Luo Huiying , Yao Bin , Huang Huoqing , Zhang Jie
Published: March 2023

Background

Heme proteins, such as hemoglobin, horseradish peroxidase and cytochrome P450 (CYP) enzyme, are highly versatile and have widespread applications in the fields of food, healthcare, medical and biological analysis. As a cofactor, heme availability plays a pivotal role in proper folding and function of heme proteins. However, the functional production of heme proteins is usually challenging mainly due to the insufficient supply of intracellular heme.

Results

Here, a versatile high-heme-producing Escherichia coli chassis was constructed for the efficient production of various high-value heme proteins. Initially, a heme-producing Komagataella phaffii strain was developed by reinforcing the C4 pathway-based heme synthetic route. Nevertheless, the analytical results revealed that most of the red compounds generated by the engineered K. phaffii strain were intermediates of heme synthesis which were unable to activate heme proteins. Subsequently, E. coli strain was selected as the host to develop heme-producing chassis. To fine-tune the C5 pathway-based heme synthetic route in E. coli, fifty-two recombinant strains harboring different combinations of heme synthesis genes were constructed. A high-heme-producing mutant Ec-M13 was obtained with negligible accumulation of intermediates. Then, the functional expression of three types of heme proteins including one dye-decolorizing peroxidase (Dyp), six oxygen-transport proteins (hemoglobin, myoglobin and leghemoglobin) and three CYP153A subfamily CYP enzymes was evaluated in Ec-M13. As expected, the assembly efficiencies of heme-bound Dyp and oxygen-transport proteins expressed in Ec-M13 were increased by 42.3–107.0% compared to those expressed in wild-type strain. The activities of Dyp and CYP enzymes were also significantly improved when expressed in Ec-M13. Finally, the whole-cell biocatalysts harboring three CYP enzymes were employed for nonanedioic acid production. High supply of intracellular heme could enhance the nonanedioic acid production by 1.8- to 6.5-fold.

Conclusion

High intracellular heme production was achieved in engineered E. coli without significant accumulation of heme synthesis intermediates. Functional expression of Dyp, hemoglobin, myoglobin, leghemoglobin and CYP enzymes was confirmed. Enhanced assembly efficiencies and activities of these heme proteins were observed. This work provides valuable guidance for constructing high-heme-producing cell factories. The developed mutant Ec-M13 could be employed as a versatile platform for the functional production of difficult-to-express heme proteins.
Hyperglycemia induces PFKFB3 overexpression and promotes malignant phenotype of breast cancer through RAS/MAPK activation
Authors: Cheng Xiao , Jia Xiupeng , Wang Chunnian , Zhou Shangyan , Chen Jiayi , Chen Lei , Chen Jinping
Published: March 2023

Background

Breast cancer is the most common tumor in women worldwide. Diabetes mellitus is a global chronic metabolic disease with increasing incidence. Diabetes mellitus has been reported to positively regulate the development of many tumors. However, the specific mechanism of hyperglycemic environment regulating breast cancer remains unclear. PFKFB3 (6-phosphofructose-2-kinase/fructose-2, 6-bisphosphatase 3) is a key regulatory factor of the glycolysis process in diabetes mellitus, as well as a promoter of breast cancer. So, we want to explore the potential link between PFKFB3 and the poor prognosis of breast cancer patients with hyperglycemia in this study.

Methods

Cell culture was utilized to construct different-glucose breast cancer cell lines. Immunohistochemistry was adopted to analyze the protein level of PFKFB3 in benign breast tissues, invasive ductal carcinoma with diabetes and invasive ductal carcinoma without diabetes. The Kaplan–Meier plotter database and GEO database (GSE61304) was adopted to analyze the survival of breast cancer patients with different PFKFB3 expression. Western blot was adopted to analyze the protein level of PFKFB3, epithelial–mesenchymal transition (EMT)-related protein and extracellular regulated protein kinases (ERK) in breast cancer cells. Gene Set Cancer Analysis (GSCA) was utilized to investigate the potential downstream signaling pathways of PFKFB3. TargetScan and OncomiR were utilized to explore the potential mechanism of PFKFB3 overexpression by hyperglycemia. Transfections (including siRNAs and miRNA transfection premiers) was utilized to restrain or mimic the expression of the corresponding RNA. Cell functional assays (including cell counting, MTT, colony formation, wound-healing, and cell migration assays) were utilized to explore the proliferation and migration of breast cancer cells.

Results

In this study, we demonstrated that the expression of PFKFB3 in breast cancer complicated with hyperglycemia was higher than that in breast cancer with euglycemia through cell experiment in vitro and histological experiment. PFKFB3 overexpression decreased the survival period of breast cancer patients and was correlated with a number of clinicopathological parameters of breast cancer complicated with diabetes. PFKFB3 promoted the proliferation and migration of breast cancer in a hyperglycemic environment and might be regulated by miR-26. In addition, PFKFB3 stimulated epithelial-mesenchymal transition of breast cancer in a hyperglycemic environment. In terms of downstream mechanism exploration, we predicted and verified the cancer-promoting effect of PFKFB3 in breast cancer complicated with hyperglycemia through RAS/MAPK pathway.

Conclusions

In conclusion, PFKFB3 could be overexpressed by hyperglycemia and might be a potential therapeutic target for breast cancer complicated with diabetes.
The CLDN5 gene at the blood-brain barrier in health and disease
Authors: Hashimoto Yosuke , Greene Chris , Munnich Arnold , Campbell Matthew
Published: March 2023

The CLDN5 gene encodes claudin-5 (CLDN-5) that is expressed in endothelial cells and forms tight junctions which limit the passive diffusions of ions and solutes. The blood–brain barrier (BBB), composed of brain microvascular endothelial cells and associated pericytes and end-feet of astrocytes, is a physical and biological barrier to maintain the brain microenvironment. The expression of CLDN-5 is tightly regulated in the BBB by other junctional proteins in endothelial cells and by supports from pericytes and astrocytes. The most recent literature clearly shows a compromised BBB with a decline in CLDN-5 expression increasing the risks of developing neuropsychiatric disorders, epilepsy, brain calcification and dementia. The purpose of this review is to summarize the known diseases associated with CLDN-5 expression and function. In the first part of this review, we highlight the recent understanding of how other junctional proteins as well as pericytes and astrocytes maintain CLDN-5 expression in brain endothelial cells. We detail some drugs that can enhance these supports and are being developed or currently in use to treat diseases associated with CLDN-5 decline. We then summarise mutagenesis-based studies which have facilitated a better understanding of the physiological role of the CLDN-5 protein at the BBB and have demonstrated the functional consequences of a recently identified pathogenic CLDN-5 missense mutation from patients with alternating hemiplegia of childhood. This mutation is the first gain-of-function mutation identified in the CLDN gene family with all others representing loss-of-function mutations resulting in mis-localization of CLDN protein and/or attenuated barrier function. Finally, we summarize recent reports about the dosage-dependent effect of CLDN-5 expression on the development of neurological diseases in mice and discuss what cellular supports for CLDN-5 regulation are compromised in the BBB in human diseases.
A diagnostic support system based on pain drawings: binary and k-disease classification of EDS, GBS, FSHD, PROMM, and a control group with Pain2D
Authors: Emmert D. , Szczypien N. , Bender Tim T. A. , Grigull L. , Gass A. , Link C. , Klawonn F. , Conrad R. , Mücke M. , Sellin J.
Published: March 2023

Background and objective

The diagnosis of rare diseases (RDs) is often challenging due to their rarity, variability and the high number of individual RDs, resulting in a delay in diagnosis with adverse effects for patients and healthcare systems. The development of computer assisted diagnostic decision support systems could help to improve these problems by supporting differential diagnosis and by prompting physicians to initiate the right diagnostic tests. Towards this end, we developed, trained and tested a machine learning model implemented as part of the software called Pain2D to classify four rare diseases (EDS, GBS, FSHD and PROMM), as well as a control group of unspecific chronic pain, from pen-and-paper pain drawings filled in by patients.

Methods

Pain drawings (PDs) were collected from patients suffering from one of the four RDs, or from unspecific chronic pain. The latter PDs were used as an outgroup in order to test how Pain2D handles more common pain causes. A total of 262 (59 EDS, 29 GBS, 35 FSHD, 89 PROMM, 50 unspecific chronic pain) PDs were collected and used to generate disease specific pain profiles. PDs were then classified by Pain2D in a leave-one-out-cross-validation approach.

Results

Pain2D was able to classify the four rare diseases with an accuracy of 61–77% with its binary classifier. EDS, GBS and FSHD were classified correctly by the Pain2D k-disease classifier with sensitivities between 63 and 86% and specificities between 81 and 89%. For PROMM, the k-disease classifier achieved a sensitivity of 51% and specificity of 90%.

Conclusions

Pain2D is a scalable, open-source tool that could potentially be trained for all diseases presenting with pain.
APOBEC3-mediated mutagenesis in cancer: causes, clinical significance and therapeutic potential
Authors: Butler Kelly , Banday A. Rouf
Published: March 2023

Apolipoprotein B mRNA-editing enzyme, catalytic polypeptides (APOBECs) are cytosine deaminases involved in innate and adaptive immunity. However, some APOBEC family members can also deaminate host genomes to generate oncogenic mutations. The resulting mutations, primarily signatures 2 and 13, occur in many tumor types and are among the most common mutational signatures in cancer. This review summarizes the current evidence implicating APOBEC3s as major mutators and outlines the exogenous and endogenous triggers of APOBEC3 expression and mutational activity. The review also discusses how APOBEC3-mediated mutagenesis impacts tumor evolution through both mutagenic and non-mutagenic pathways, including by inducing driver mutations and modulating the tumor immune microenvironment. Moving from molecular biology to clinical outcomes, the review concludes by summarizing the divergent prognostic significance of APOBEC3s across cancer types and their therapeutic potential in the current and future clinical landscapes.
Targeting IGF1R signaling enhances the sensitivity of cisplatin by inhibiting proline and arginine metabolism in oesophageal squamous cell carcinoma under hypoxia
Authors: Fang Kang , Sun Mingchuang , Leng Zhuyun , Chu Yuan , Zhao Ziying , Li Zhaoxing , Zhang Yunwei , Xu Aiping , Zhang Zehua , Zhang Li , Chen Tao , Xu Meidong
Published: March 2023

Background

Cisplatin (DDP)-based chemotherapy is commonly adopted as the first-line treatment for patients with oesophageal squamous cell carcinoma (OSCC), but the high rate of drug resistance limits its clinical application and the underlying mechanisms at play remain unclear. The aims of this study were to elucidate the role of abnormal signal transmission and metabolism in the chemoresistance of OSCC under hypoxia and to identify targeted drugs that enhance the sensitivity of DDP chemotherapy.

Methods

Upregulated genes in OSCC were determined by RNA sequencing (RNA-seq), the Cancer Genome Atlas (TCGA) database, immunohistochemistry (IHC), real-time quantitative PCR (RT-qPCR), and western blotting (WB). The clinicopathological significance of insulin-like growth factor-I receptor (IGF1R), argininosuccinate synthetase 1 (ASS1), and pyrroline-5-carboxylate reductase 1 (PYCR1) in OSCC was analysed using tissue micriarray (TMA). Metabolic abnormalities were determined by untargeted metabolomics analysis. The DDP-resistance role of IGF1R, ASS1, and PYCR1 in OSCC was investigated in vitro and in vivo.

Results

Generally, tumour cells exist in a hypoxic microenvironment. By genomic profiling, we determined that IGF1R, as a receptor tyrosine kinase (RTK), was upregulated in OSCC under low-oxygen conditions. Clinically, enhanced IGF1R expression was associated with higher tumour stages and a poorer prognosis in OSCC patients, and its inhibitor, linsitinib, showed synergistic effects with DDP therapy in vivo and in vitro. Since oxygen-deprivation frequently lead to metabolic reprogramming, we further learned via metabolomics analysis that abnormal IGF1R pathways promoted the expression of metabolic enzymes ASS1 and PYCR1 by the transcriptional activity of c-MYC. In detail, enhanced expression of ASS1 promotes arginine metabolism for biological anabolism, whereas PYCR1 activates proline metabolism for redox balance, which maintains the proliferation ability of OSCC cells during DDP treatment under hypoxic conditions.

Conclusion

Enhanced expression of ASS1 and PYCR1 via IGF1R pathways rewired arginine and proline metabolism, promoting DDP resistance in OSCC under hypoxia. Linsitinib targeting IGF1R signaling may lead to promising combination therapy options for OSCC patients with DDP resistance.
HES1-mediated down-regulation of miR-138 sustains NOTCH1 activation and promotes proliferation and invasion in renal cell carcinoma
Authors: Liu Shuangjie , Dou Lei , Miao Miao , Man Xiaojun , Wei Baojun , Jiang Zhaowei , Ouyang Yongze , Ozaki Toshinori , Yu Meng , Zhu Yuyan
Published: March 2023

Background

Although the aberrant activation of NOTCH1 pathway causes a malignant progression of renal cell carcinoma (RCC), the precise molecular mechanisms behind the potential action of pro-oncogenic NOTCH1/HES1 axis remain elusive. Here, we examined the role of tumor suppressive miR-138–2 in the regulation of NOTCH1-HES1-mediated promotion of RCC.

Methods

This study employed bioinformatics, xenotransplant mouse models, ChIP assay, luciferase reporter assay, functional experiments, real-time PCR and Western blot analysis to explore the mechanisms of miR-138–2 in the regulation of NOTCH1-HES1-mediated promotion of RCC, and further explored miR-138–2-containing combination treatment strategies.

Results

There existed a positive correlation between down-regulation of miR-138 and the aberrant augmentation of NOTCH1/HES1 regulatory axis. Mechanistically, HES1 directly bound to miR-138–2 promoter region and thereby attenuated the transcription of miR-138-5p as well as miR-138–2-3p. Further analysis revealed that miR-138-5p as well as miR-138–2-3p synergistically impairs pro-oncogenic NOTCH1 pathway through the direct targeting of APH1A, MAML1 and NOTCH1.

Conclusions

Collectively, our current study strongly suggests that miR-138–2 acts as a novel epigenetic regulator of pro-oncogenic NOTCH1 pathway, and that the potential feedback regulatory loop composed of HES1, miR-138–2 and NOTCH1 contributes to the malignant development of RCC. From the clinical point of view, this feedback regulatory loop might be a promising therapeutic target to treat the patients with RCC.
Exercise in adults admitted to hospital with diabetes‐related foot ulcers: a pilot study of feasibility and safety
Authors: Aitken Emily , Hiew Jonathan , Hamilton Emma J , Manning Laurens , Ritter Jens Carsten , Raby Edward , Gittings Paul M
Published: March 2023

Background

Diabetes‐related foot ulcers result in significant mortality, morbidity and economic costs. Pressure offloading is important for ulcer healing, but patients with diabetes‐related foot ulcers are presented with a dilemma, because whilst they are often advised to minimise standing and walking, there are also clear guidelines which encourage regular, sustained exercise for patients with diabetes. To overcome these apparently conflicting recommendations, we explored the feasibility, acceptability and safety of a tailored exercise program for adults admitted to hospital with diabetes‐related foot ulcers.

Methods

Patients with diabetes‐related foot ulcers were recruited from an inpatient hospital setting. Baseline demographics and ulcer characteristics were collected, and participants undertook a supervised exercise training session comprising aerobic and resistance exercises followed by prescription of a home exercise programme. Exercises were tailored to ulcer location, which complied with podiatric recommendations for pressure offloading. Feasibility and safety were assessed via recruitment rate, retention rate, adherence to inpatient and outpatient follow up, adherence to home exercise completion, and recording of adverse events.

Results

Twenty participants were recruited to the study. The retention rate (95%), adherence to inpatient and outpatient follow up (75%) and adherence to home exercise (50.0%) were all acceptable. No adverse events occurred.

Conclusions

Targeted exercise appears safe to be undertaken by patients with diabetes‐related foot ulcers during and after an acute hospital admission. Recruitment in this cohort may prove challenging, but adherence, retention and satisfaction with participation in exercise were high.

Trial registration

The trial is registered in the Australian New Zealand Clinical Trials Registry (ACTRN12622001370796).
Host-mediated RNA editing in viruses
Authors: Zhu Tongtong , Niu Guangyi , Zhang Yuansheng , Chen Ming , Li Chuan-Yun , Hao Lili , Zhang Zhang
Published: March 2023

Viruses rely on hosts for life and reproduction, cause a variety of symptoms from common cold to AIDS to COVID-19 and provoke public health threats claiming millions of lives around the globe. RNA editing, as a crucial co-/post-transcriptional modification inducing nucleotide alterations on both endogenous and exogenous RNA sequences, exerts significant influences on virus replication, protein synthesis, infectivity and toxicity. Hitherto, a number of host-mediated RNA editing sites have been identified in diverse viruses, yet lacking a full picture of RNA editing-associated mechanisms and effects in different classes of viruses. Here we synthesize the current knowledge of host-mediated RNA editing in a variety of viruses by considering two enzyme families, viz., ADARs and APOBECs, thereby presenting a landscape of diverse editing mechanisms and effects between viruses and hosts. In the ongoing pandemic, our study promises to provide potentially valuable insights for better understanding host-mediated RNA editing on ever-reported and newly-emerging viruses.