High comorbidity burden and low incidence of adverse events in primary shoulder arthroplasty in tertiary care at a university hospital.

Background: Reducing perioperative risk is in the focus of primary arthroplasty caregivers. Primary shoulder arthroplasty patients are considered among those with the lowest perioperative risk. Nonetheless, in tertiary care centres and university settings patients with significant comorbidities are being treated. It remains unclear whether the overall comorbidity burden is truly higher and if this results in an increased frequency of adverse events (AE). As a result, we conducted a study to assess the comorbidity burden and the frequency of major perioperative adverse events and predictive factors following primary shoulder arthroplasty in the university setting. Methods: A retrospective cohort analysis was conducted on patients undergoing primary shoulder arthroplasty for a non-trauma, non-tumour indication from January 1st, 2014 through December 31st, 2018. Administrative data were recorded to assess comorbidity burden and revision surgery within the first postoperative year. Major adverse events were routinely recorded on a weekly basis by the treating physicians. Descriptive and comparative statistical analyses were performed. The cohort was compared against a large North American sample. Results: Of 386 patients who underwent 400 primary shoulder arthroplasties 14 (3.5 %) experienced adverse events. While AE were distributed equally among anatomical and reverse shoulder arthroplasties, no adverse event was recorded in the 34 hemiarthroplasty patients. The cohort showed an increased comorbidity burden in international comparison. Peptic ulcer disease was significantly associated with AE, while mild liver disease experienced a trend towards AE. Conclusion: We found an increased comorbidity burden and a low rate of AE for primary shoulder arthroplasty in a tertiary care and university setting. The distinct role of peptic ulcer disease in this cohort and the trend in mild liver disease merit further investigation in larger samples. The findings underscore the importance of perioperative risk assessment and management. Level of evidence: III, retrospective cohort study.

T peripheral helper (Tph) cells, a marker of immune activation in cancer and autoimmune disorders.

T peripheral helper (Tph) cells are a newly discovered subtype of CD4+ T cells that have emerged as the counterpart of T follicular helper (Tfh) cells in the peripheral tissues. These two cell types share some common characteristics, such as high levels of PD1 and CXCL13 expression, but differ in the expression of transcription factors and chemokine receptors. Tph cells have been studied in relation to B cells' effector functions, including cytokines production and antibody-mediated immune responses. However, their role in the inflammatory-mediated development of malignancies remains poorly understood. Tph cells were initially identified in the synovium of rheumatoid arthritis patients and have since been found to be expanded in several autoimmune diseases. They have been linked to a worse prognosis in autoimmune conditions, but intriguingly, their presence has been correlated with better outcomes in certain types of cancer. The functions of Tph cells are still being investigated, but recent data suggests their involvement in the assembly of tertiary lymphoid structures (TLS). Furthermore, their interaction with B cells, which have been mainly described as possessing a memory phenotype, promotes their development. In this review, we explore the role of Tph cells in peripheral immune responses during cancer and autoimmune disorders.

Implementation of back to living well, a community-based program for the tertiary prevention of low back pain: a study protocol.

BACKGROUND: The current literature supports the effectiveness of exercise, education, and self-management interventions for the long-term management of persistent low back pain. However, there is significant uncertainty about the implementation of interventions related to barriers, facilitators, and patient's preferences. This study will evaluate the Back to Living Well program implementation from a participant and organizational perspective. More specifically we address the following objectives: 1) identify program barriers and facilitators from participants' perspectives, 2) identify factors related to program, personal and contextual factors that contribute to negative and positive outcomes, and outcome trajectories, 3) identify factors influencing participants' selection of an in-person or e-health program, and 4) evaluate program specific barriers and facilitators from the organization and care delivery perspectives. METHODS: This study will utilize a mixed-method convergent design including a longitudinal cohort strand and a longitudinal qualitative interview strand. The RE-AIM framework will be used to assess program implementation. Participants (n = 90, 1:1: in person or virtual) who choose to register in the program as well as staff (n = 10 to 15) involved in the delivery of the program will be invited to participate. Participants will participate in a 12-week physical activity, education, and self-management program. Implementation outcomes will be measured at 3-, 6-, 12-months, and six months after the end of the follow-ups. Interview scripts and directed content analysis will be constructed based on the Theoretical Domains Framework and the Neuromatrix Model of Pain, Theoretical Domains Framework. Staff interviews will be constructed and analyzed using the Consolidated Framework for Implementation Research. Participants will also complete pain, disability, quality of life and psychological questionnaires, wear an activity tracker at all time points, and complete weekly pain and activity limitation questions using a mobile application. DISCUSSION: The study results will provide evidence to inform potential future implementation of the program. An effective, appropriately targeted, and well implemented exercise program for the long-term management (i.e., tertiary prevention) of LBP could minimize the burden of the condition on patients, the health care system and society. TRIAL REGISTRATION: ClinicalTrials.gov NCT05929846. This (Registration Date: July 3 2023) study has been approved by the Hamilton Integrated Research Ethics Board Project ID#15,354.

Tertiary Lymphoid Structures in Brain Metastases of Lung Cancer: Prognostic Significance and Correlation With Clinical Outcomes.

BACKGROUND/AIM: The prognosis of patients with brain metastases (BMs) originating from lung cancer remains poor, despite advancements in treatment strategies. The role of tertiary lymphoid structures (TLSs) within the tumor immune microenvironment of BMs has not been extensively explored. PATIENTS AND METHODS: This study utilized patient-derived clinical samples from 17 patients with histologically confirmed BMs of lung cancer, undergoing surgical resection. Immunohistochemistry was employed to analyze the presence and characteristics of TLS and tumor-infiltrating lymphocytes (TILs) within BM tissues, correlating these with clinical outcomes. RESULTS: TLSs, albeit in their immature form, were identified within BM tissues, distinguishing them from their mature counterparts in primary lung cancer tissues. A significant correlation between TLS density (but not TIL density) and improved postoperative survival was observed, underscoring the potential of TLS density as an independent prognostic marker. Furthermore, TLS density did not correlate with the Graded Prognostic Assessment (GPA) index, suggesting its unique prognostic value beyond conventional predictors. CONCLUSION: Our findings reveal the presence of TLSs in lung cancer-derived BMs and highlight their prognostic significance, independent of the GPA index. The identification of TLS within the unique central nervous system tumor microenvironment offers new insights into the immune landscape of BMs and suggests potential avenues for immunotherapeutic interventions targeting these structures to improve patient outcomes.

Characteristics and clinical significance of tertiary lymphoid structures in OSCC.

OBJECTIVES: This study aimed to investigate the characteristics of tertiary lymphoid structures (TLSs) in oral squamous cell carcinoma (OSCC) and their association with clinical and pathological features. MATERIALS AND METHODS: 12 TLS-related chemokines in TCGA database were analyzed to investigate the TLSs in OSCC. The density, maturity, and location of TLSs in a large cohort of 189 OSCC patients (114 of which had clinical and prognostic information) were assessed. And the significance between TLSs and clinicopathologic characteristics was analyzed. RESULTS: Bioinformatics and analysis showed that TLSs were associated with better clinical outcomes in OSCC. Histological staining and analysis showed that the overall survival rate of the high-density group (71/112, 63.4%) was significantly higher (p < 0.0001) than that of the low-density group (41/112, 36.6%), and the high-density group had fewer lymph node metastases (50.0%/68.3%, p = 0.021). And TLSs were divided into 4 types according to the maturity and location. Different types of TLSs are associated with prognosis (OS, p < 0.0001), clinical features (T stage, p = 0.028; degree of differentiation, p = 0.043), and precancerous lesion types (OSF, p = 0.049) of OSCC patients. CONCLUSION: TLSs were closely associated with better OSCC prognosis, and a more systematic classification may better guide the formulation of further treatment options.

Factors associated with chronotype, job burnout, and perceived stress among nurses in Chinese tertiary hospitals: A multicenter cross-sectional study.

Shift work is a recognized work pattern for nurses worldwide. The disruption of shift workers' biological clocks usually leads to sleep disorders and affects their awareness at work. Eveningness and occupational stress might be effective in causing burnout syndrome. Therefore, this study aimed to evaluate the chronotype, job burnout and perceived stress among Chinese tertiary hospital nurses, and understand the predictors of circadian rhythm in this group. Between July and September 2020, 23 hospitals were randomly selected from 113 tertiary hospitals in Hunan Province. Twenty-five percent of the nurses working in each hospital were targeted for selection. 28.1% and 17.6% of nurses reported eveningness type and morningness type, respectively. The scores for emotional exhaustion, depersonalization, and perceived stress of eveningness nurses were higher than those of morningness counterparts. Eveningness nurses also reported a lower sense of personal accomplishment. Risk factors of eveningness included being under 30 years old, never exercising, having the stressors of late-night shifts and career development, higher levels of emotional exhaustion, sleep latency, sleep duration, and hypnotic use. Shifts may be unavoidable for nurses, nevertheless, understanding the predictors and related factors of chronotype for nurses is necessary for nursing educators and managers to develop a reasonable shift system and appropriate measures to assist nurses in adjusting their work.

Predictors, prevalence and outcome of hypertensive disorders in pregnancy in Nigerian tertiary health facilities: A secondary analysis of the Maternal and Perinatal Database for Quality, Equity and Dignity Programme.

OBJECTIVE: Determine prevalence, risk factors and outcomes of hypertensive disorders in pregnancy (HDP). DESIGN: Cross-sectional analysis of data captured in the Maternal and Perinatal Database for Quality, Equity and Dignity (MPD-4-QED) between September 2019 and August 2020. SETTING: Fifty-four referral level facilities in Nigeria. POPULATION: Women whose pregnancy ended (irrespective of the location or duration of pregnancy) or who were admitted within 42 days of delivery. METHODS: Descriptive statistics and multilevel mixed-effects logistic regression models. MAIN OUTCOME MEASURES: Prevalence of HDP, sociodemographic and clinical factors associated with HDP and perinatal outcomes. RESULTS: Among the 71 758 women 6.4% had HDP and gestational hypertension accounted for 49.8%. Preeclampsia and eclampsia were observed in 9.5% and 7.0% of all pregnancies, respectively. The predictors of HDP were age over 35 years (OR1.96, 95% CI 1.82-2.12; p < 0.001), lack of formal educational (OR 1.18, 95% CI 1.06-1.32; p = 0.002), primary level of education (OR 1.20, 95% CI 1.03-1.4; p < 0.002), nulliparity (OR 1.21, 95% CI 1.12-1.31; p < 0.001), grand-multiparity (OR 1.36, 95%CI 1.21-1.52; p < 0.001), previous caesarean section (OR 1.26, 95%CI 1.15-1.38; p < 0.001) and previous miscarriage (OR 1.22, 95% CI 1.13-1.31; p < 0.001). Overall 3.7% of the patients with HDP died, with eclampsia having the highest case fatality rate of 27.9%. Stillbirth occurred in 11.9% of pregnancies with hypertensive disorders. CONCLUSIONS: Hypertensive disorders in pregnancy are not uncommon in Nigeria. They are associated with adverse outcomes with over one-quarter of women with eclampsia dying. The main predictors include older age, poor education, extremes of parity and previous CS or miscarriage. Maternal and perinatal outcomes are poor with about a quarter developing complications and about 1 in 10 having stillbirths.

Trends in childhood intussusception in a Nigerian tertiary hospital.

Background: Early presentation, high rate of successful non-operative treatment, low morbidity and mortality in childhood intussusception is common in High and Upper Middle-Income Countries but not in many Lower middle- and Low-income countries. Aim: To assess the trends in the profile, treatment modalities and outcomes of intussusception in our hospital. Materials and methods: Retrospective study over a 12-year period divided into two 6-year periods. Data entry/analysis was done using SPSS and various indices were compared between these two periods. Two-tailed t-test for two independent means was used to compare means while two-tailed Fisher exact tests were used to compare categorical variables. Results were presented as tables, means, ranges, percentages and a p-value less than 0.05 was deemed statistically significant. Results: There was a significant increase in the proportion of successful non-operative treatment (18.6% vs 34%, p=0.03), reduction in the incidence of operative manual reduction (27.1% vs 12.8%; p=0.026), reduction in operative treatment (78.5% vs 63.9%, p=0.034), increased utilization of pre-intervention ultrasound (75% vs96.7%, p<0.0001) and reduction in hospital stay duration (10.47 ±7.95days vs 7.24±4.86 days; p=0.004). Conclusions: Contribution of successful non-operative treatment to the overall treatment of intussusception significantly increased while that of operative manual reduction significantly reduced and bowel resection showed no change. Preoperative utilization of ultrasonography significantly increased while mean duration of admission reduced significantly, but late presentation, morbidity and mortality rates had no significant changes.

Automated immunoassay of serum NY-ESO-1 and XAGE1 antibodies for predicting clinical benefit with immune checkpoint inhibitor (ICI) in advanced non-small cell lung cancer.

BACKGROUND: NY-ESO-1 and XAGE1 cancer/testis antigens elicit humoral and cellular immune responses in NSCLC patients. We aimed to predict clinical benefit with ICI monotherapy, using an automated immunoassay of NY-ESO-1/XAGE1 antibodies (Abs). METHODS: This study enrolled 99 NSCLC patients who received nivolumab after chemotherapy, including 21 patients harboring EGFR, ALK, or KRAS alterations. The cutoff value (10 units/mL) of NY-ESO-1 and XAGE1 Ab was determined based on Ab levels in non-malignant controls, and NY-ESO-1/XAGE1 Abs in NSCLC were measured before nivolumab. Differences in PFS and OS between the Ab-positive and Ab-negative groups were retrospectively analyzed using Cox regression analysis after applying inverse probability of treatment weighting (IPTW). RESULTS: NY-ESO-1/XAGE1 Abs were positive in 28 NSCLC, who responded more highly to nivolumab than the Ab-negatives (response rate 50.0% vs. 15.5 %, p < 0.0007). The IPTW-adjusted positives and negatives for NY-ESO-1/XAGE1 Abs were 24.5 and 70.2, respectively. The Ab-positives showed longer IPTW-adjusted PFS (HR = 0.59, 95 % CI: 0.39-0.90, p = 0.014) and IPTW-adjusted OS (HR = 0.51, 95 % CI: 0.32-0.81, p = 0.004) than the Ab-negatives. Among NSCLC harboring driver genes, the Ab-positives (n = 10) showed longer PFS (HR = 0.34, 95 % CI: 0.13-0.89, p = 0.029) and OS (HR = 0.27, 95 % CI: 0.098-0.75, p = 0.012) than the Ab-negatives (n = 11). CONCLUSION: Our immunoassay of NY-ESO-1/XAGE1 Abs is probably useful for predicting the clinical benefit with nivolumab in NSCLC, including those harboring driver genes. These results suggest that our immunoassay may be useful in ICI monotherapy for NSCLC.

Dietary perception and knowledge among cardiac patients visiting tertiary care hospital outpatient department (OPD).

Objective: To appraise the dietary perception and knowledge of cardiac patients visiting Outpatient department of a tertiary care hospital. Methods: A cross sectional study of 466 patients attending cardiac outdoor clinic at a tertiary care cardiac hospital in Rawalpindi, Pakistan were selected through convenient sampling from January to April 2021. Patients included in the study were above 18 years of age and had a cardiac disease. A structured questionnaire in English and Urdu was drafted with questions related to patient demographics, dietary perception and knowledge. Results: Among 466 patients, 60% (n=280) were males and 40% (n=186) were females whereas 14% (n=66) of the patients were uneducated and 15% (n=68) had completed postgraduate education. In this study 23% (n=106) of the participants used internet to search nutrition related information and 72 % (n=339) of the patients consider their lifestyle to be healthy. Majority (n=261, 56%) of the patients had no idea regarding sodium consumption. 46% (n=215) of the patients had no knowledge regarding the effects of bakery items on cardiac health. Conclusion: Dietary Knowledge plays a predominant role in the management of cardiac disease. The study concluded that cardiac patients had inadequate knowledge regarding dietary intake in cardiac disease and has high prevalence of dietary myths and misconceptions. Strategic plan including nutritional awareness program and intensive counseling sessions should be designed to increase dietary knowledge of cardiac patients.

Resilience and social support as protective factors against suicidal ideation among tertiary students during COVID-19: a cross-sectional study.

BACKGROUND: Suicidal ideation is a depression symptom which represents a key (cognitive) component of suicidality and plays an important role in suicide risk detection, intervention, and prevention. Despite existing research showing the importance of certain factors of depression symptoms and suicidal ideation, less is known about the interaction between the various risk and protective factors. The aim of the study was to examine whether living conditions characteristics and personal circumstances during the COVID-19 pandemic predicted the presence of depression symptoms and suicidal ideation among tertiary students and whether resilience and social support can mitigate the detrimental effects of difficult life circumstances. METHOD: A large online cross-sectional study was conducted in March 2021 among 4,645 Slovenian tertiary students. Hierarchical multiple regression and hierarchical logistic regression methods were used to assess and compare the effect of life circumstances variables, as opposed to resilience and social support, on depression symptoms and suicidal ideation. RESULTS: Female gender, single relationship status, living alone, a higher degree of household conflict, having a history of mental illness and chronic disease diagnosis were significant predictors of depression scores. All but gender were also predictors of suicidal ideation. Household conflict and a history of mental illness were the factors showing the strongest effect in both cases. On the other hand, social support and, in particular, resilience proved to be strong protective factors against depression symptoms and suicidal ideation. After accounting for one's resilience and social support, the explained variance in depression scores was more than doubled, while the harmful effect of household conflict and history of mental illness significantly decreased. CONCLUSIONS: The findings stress the importance of one's resilience and social support and explain why some people manage to maintain mental well-being despite finding themselves in difficult life circumstances, which was the case for many tertiary students during the COVID-19 pandemic. These insights may inform preventive efforts against developing suicidal ideation and may be used as support for the design and implementation of interventions for improving resilience and social support from childhood onward.

Caesarean section rates in a tertiary teaching hospital in northern Uganda: a retrospective analysis using the robson ten group classification system.

BACKGROUND: The Robson Ten Groups Classification System (RTGCS) is increasingly used to assess, monitor, and compare caesarean section (CS) rates within and between healthcare facilities. We evaluated the major contributing groups to the CS rate at Gulu Regional Referral Hospital (GRRH) in Northern Uganda using the RTGCS. METHODS: We conducted a retrospective analysis of all deliveries from June 2019 through July 2020 at GRRH, Gulu city, Uganda. We reviewed files of mothers and collected data on sociodemographic and obstetric variables. The outcome variables were Robson Ten Groups (1-10) based on parity, gestational age, foetal presentation, number of foetuses, the onset of labour, parity and lie, and history of CS. RESULTS: We reviewed medical records of 3,183 deliveries, with a mean age of 24.6 ± 5.7 years. The overall CS rate was 13.4% (n = 427). Most participants were in RTGCS groups 3 (43.3%, n = 185) and 1 (29.2%, n = 88). The most common indication for CS was prolonged labour (41.0%, n = 175), followed by foetal distress (19.9%, n = 85) and contracted pelvis (13.6%, n = 58). CONCLUSION: Our study showed that GRRH patients had a low-risk obstetric population dominated by mothers in groups 3 and 1, which could explain the low overall CS rate of 13.4%. However, the rates of CS among low-risk populations are alarmingly high, and this is likely to cause an increase in CS rates in the future. We recommend group-specific interventions through CS auditing to lower group-specific CS rates.

Persisting Hypercalcemia and Hyperparathyroidism after Kidney Transplantation Have a Negative Impact on Graft and Patient Survival.

Hyperparathyroidism (HPT) with hypercalcemia, often deemed irreversible and detrimental to graft survival post-kidney transplantation (KT), prompts pre-transplant parathyroidectomy in hypercalcemic patients. In this retrospective analysis of 1212 kidney transplant recipients (KTRs) between 2006 and 2019, the incidence and effect of persistent HPT and hypercalcemia on graft and patient survival, and risk factors for persistence were analyzed until 60 months of follow up (FU). At KT, 5.7% (n = 69) had no HPT, 32.7% (n = 396) had HPT without hypercalcemia and 37.0% (n = 448) had HPT with hypercalcemia. At 2 years FU, 26.4% (n = 320) of patients had no HPT and 6% (n = 73) had HPT with hypercalcemia. Dialysis and dialysis duration were linked to HPT development, while dialysis, KT waiting time and donor type correlated with persisting hypercalcemia after KT. KTRs with normalized PTH and recovered hypercalcemia had improved death-censored graft survival (p < 0.001) and overall patient survival (p < 0.001). HPT with hypercalcemia is frequent at time of KT with normalization of PTH and calcium in a substantial proportion of patients after a KT. These findings question the routine pre-KT parathyroidectomy for suspected parathyroid autonomy. Persisting HPT, especially with hypercalcemia, adversely affects graft and patient survival, suggesting the need for more aggressive treatment of HPT, especially in cases of persisting hypercalcemia.

Profile of gastroduodenal perforation patients admitted in a rural tertiary care hospital: An observational cross-sectional study.

Background: Gastro-duodenal perforation is a common surgical emergency that remains a formidable health burden worldwide with significant morbidity and mortality. Ulcer disease remains the most common cause of gastro-duodenal perforation. Diagnosing the presence of H. pylori can help eradicate the infection from the community at large and thereby reduce the chances of gastro-duodenal perforation. Aims: To assess the clinical presentation of gastro-duodenal perforation patients and to evaluate the detection of Helicobacter pylori infection by available investigations. Materials and Methods: A descriptive observational study was conducted among 80 patients presenting with clinical features suggestive of gastro-duodenal perforation and confirmed by clinical, radiological basis and operative findings admitted at a rural tertiary care hospital during 2019-2020. Detailed history was taken from the patient/party, clinically examined, and blood/tissue samples were investigated. The patients were managed with standard treatment modality in the studied institute. Data were collected, compiled, and entered MS Excel and analyzed using appropriate software. Descriptive analysis was done in the form of proportion for categorical variables, mean or median for continuous variables. Result: Cases of gastro-duodenal perforations were more among middle to later age of life, mostly affecting married male patients hailed from rural area and unskilled workers. History of intake of spicy food, prolonged starvation, history of NSAID use were common among them. Majority of the patients had history of pain abdomen in the past suggesting of PUD and history of taking variety group of acid reducing agents. Most of them presented with epigastric pain, vomiting, abdominal distension along with other signs of peritonitis. Obliteration of liver dullness and free gas under right dome of diaphragm was also noted in large proportion among them. Majority of cases were found positive for H. pylori on Histology (85%), followed by rapid urease test (RUT) (80%) and a positivity of 72.5% and 68.8% on serum IgG and IgA antibody respectively. Rapid Urease Test was more sensitive as well as specific in diagnosing of H. pylori than antibody detection test. Conclusion: Early detection of H. pylori infection and treatment with potent anti H. pylori therapy postoperatively has been found to be adequate.

Nurse academics' self-reported insights into the teaching of undergraduate nursing numeracy: A national cross-sectional study.

AIMS: The aim of this research was to explore the teaching of undergraduate nursing numeracy in tertiary education settings in Australia. Specifically, it explored and identified: (1) the most common basic numeracy concepts taught, (2) additional training and resources to support numeracy teaching, (3) who is best placed to teach numeracy and (4) the preferred methods of teaching medication calculations. BACKGROUND: Nurse academics are required to teach nursing numeracy to undergraduate nursing students who enter university unprepared to accurately calculate medication dosages. It is important that students understand numeracy concepts as this is then applied to contextualised clinical applications. Nurse academics teach basic numeracy; however, the literature reveals that nurse academics do not consider themselves mathematics teachers and that experts in this area are better suited to teaching this skill. There are a dearth of studies about the nurse academics who conduct the teaching and this study seeks to fill that gap by exploring firsthand the nurse academics' self-reported insights into the teaching of undergraduate nursing numeracy. DESIGN: Cross-sectional survey. The setting was Australian universities that deliver an accredited undergraduate nursing degree. METHODS: Purposive sampling was used to recruit Australian nurse academics (n = 170), sessional or permanent who currently teach all aspects of nursing numeracy and medication calculations to undergraduate nursing students. Data were collected between Nov 2023 - Feb 2024 using an online survey platform and analysed using a descriptive content analysis. RESULTS: Nurse academics taught basic arithmetic most commonly (92 %), yet most (90 %) had not received professional development or additional training in how to teach these concepts. To assist with numeracy teaching, resources were requested (47 %) as were the need for mathematics learning support staff (82 %). The formula method was most commonly taught (91 %), however, most participants (94 %) were willing to learn and to teach other methods of calculating medications. CONCLUSIONS: This research suggests an evidenced-based education framework be created to support and guide nurse academics when teaching all aspects of nursing numeracy and medication calculations. Building the teaching capacity of nurse academics in this vital area will enhance student competence and contribute to patient safety.

Identification and Validation of a Novel Tertiary Lymphoid Structures-Related Prognostic Gene Signature in Hepatocellular Carcinoma.

Background: Hepatocellular carcinoma (HCC) is one of the most common malignant tumors originating from the digestive system. Tertiary lymphoid structures (TLS), non-lymphoid tissues outside of the lymphoid organs, are closely connected to chronic inflammation and tumorigenesis. However, the detailed relationship between TLS and HCC prognosis remained unclear. In this study, we aimed to construct a TLS-related gene signature for predicting the prognosis of HCC patients. Methods: The Cancer Genome Atlas (TCGA) clinical data from 369 HCC tissues and 50 normal liver tissues were utilized to examine the differential expression of TLS-related genes. Based on least absolute shrinkage and selection operator (LASSO) Cox regression analysis, the prognostic model was constructed using the TCGA cohort and validated in the GSE14520 cohort and International Cancer Genome Consortium (ICGC) cohort. The Kaplan-Meier (KM) and receiver operating characteristic (ROC) curves were employed to validate the predictive ability of the prognostic model. Furthermore, Cox regression analysis was applied to identify whether the TLS score could be employed as an independent prognosis factor. A nomogram was developed to predict the survival probability of HCC patients. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were performed for TLS-related genes. Genetic mutation analysis, the CIBERSORT algorithm, and single-sample gene set enrichment analysis (ssGSEA) were used to assess the tumor mutation landscape and immune infiltration. Finally, the role of the TLS score in HCC therapy was investigated. Results: Six genes were included in the construction of our prognostic model (CETP, DNASE1L3, PLAC8, SKAP1, C7, and VNN2), and we validated its accuracy. Survival analysis showed that patients in the high-TLS score group had a significantly better overall survival than those in the low-TLS score group. Univariate, multivariate Cox regression analysis and the establishment of a nomogram indicated that the TLS score could independently function as a potential prognostic marker. A significant association between TLS score and immunity was revealed by an analysis of gene alterations and immune cell infiltration. In addition, two subtypes of the TLS score could accurately predict the effectiveness of sorafenib, transcatheter arterial chemoembolization (TACE), and immunotherapy in HCC patients. Conclusion: In this research, we conducted and validated a prognostic model associated with TLS that may be helpful for predicting clinical outcomes and treatment responsiveness for HCC patients.

Prediction and assessment of deleterious and disease causing nonsynonymous single nucleotide polymorphisms (nsSNPs) in human FOXP4 gene: An in - silico study.

In humans, FOXP gene family is involved in embryonic development and cancer progression. The FOXP4 (Forkhead box protein P4) gene belongs to this FOXP gene family. FOXP4 gene plays a crucial role in oncogenesis. Single nucleotide polymorphisms are biological markers and common determinants of human diseases. Mutations can largely affect the function of the corresponding protein. Therefore, the molecular mechanism of nsSNPs in the FOXP4 gene needs to be elucidated. Initially, the SNPs of the FOXP4 gene were extracted from the dbSNP database and a total of 23124 SNPs was found, where 555 nonsynonymous, 20525 intronic, 1114 noncoding transcript, 334 synonymous were obtained and the rest were unspecified. Then, a series of bioinformatics tools (SIFT, PolyPhen2, SNAP2, PhD SNP, PANTHER, I-Mutant2.0, MUpro, GOR IV, ConSurf, NetSurfP 2.0, HOPE, DynaMut2, GeneMANIA, STRING and Schrodinger) were used to explore the effect of nsSNPs on FOXP4 protein function and structural stability. First, 555 nsSNPs were analyzed using SIFT, of which 57 were found as deleterious. Following, PolyPhen2, SNAP2, PhD SNP and PANTHER analyses, 10 nsSNPs (rs372762294, rs141899153, rs142575732, rs376938850, rs367607523, rs112517943, rs140387832, rs373949416, rs373949416 and rs376160648) were common and observed as deleterious, damaging and diseases associated. Following that, using I-Mutant2.0 and MUpro servers, 7 nsSNPs were found to be the most unstable. GOR IV predicted that these seven nsSNPs affect protein structure by altering the protein contents of alpha helixes, extended strands, and random coils. Following DynaMut2, 5 nsSNPs showed a decrease in the ΔΔG value compared with the wild-type and were found to be responsible for destabilizing the corresponding protein. GeneMANIA and STRING network revealed interaction of FOXP4 with other genes. Finally, molecular dynamics simulation analysis revealed consistent fluctuation in RMSD and RMSF values, Rg and hydrogen bonds in the mutant proteins compared with WT, which might alter the functional and structural stability of the corresponding protein. As a result, the aforementioned integrated comprehensive bioinformatic analyses provide insight into how various nsSNPs of the FOXP4 gene change the structural and functional properties of the corresponding protein, potentially proceeding with the pathophysiology of human diseases.

Sustained innate interferon is an essential inducer of tertiary lymphoid structures.

Tertiary lymphoid structures (TLS) resemble follicles of secondary lymphoid organs and develop in nonlymphoid tissues during inflammation and cancer. Which cell types and signals drive the development of TLS is largely unknown. To investigate early events of TLS development in the lungs, we repeatedly instilled p(I:C) plus ovalbumin (Ova) intranasally. This induced TLS ranging from lymphocytic aggregates to organized and functional structures containing germinal centers. We found that TLS development is independent of FAP+ fibroblasts, alveolar macrophages, or CCL19 but crucially depends on type I interferon (IFN-I). Mechanistically, IFN-I initiates two synergistic pathways that culminate in the development of TLS. On the one hand, IFN-I induces lymphotoxin (LT)α in lymphoid cells, which stimulate stromal cells to produce the B-cell-attracting chemokine CXCL13 through LTßR-signaling. On the other hand, IFN-I is sensed by stromal cells that produce the T-cell-attracting chemokines CXCL9, CXCL10 as well as CCL19 and CCL21 independently of LTßR. Consequently, B-cell aggregates develop within a week, whereas follicular dendritic cells and germinal centers appear after 3 weeks. Thus, sustained production of IFN-I together with an antigen is essential for the induction of functional TLS in the lungs.

The impact of tertiary lymphoid structures on tumor prognosis and the immune microenvironment in non-small cell lung cancer.

Non-small cell lung cancer (NSCLC) is a common malignancy whose prognosis and treatment outcome are influenced by many factors. Some studies have found that tertiary lymphoid structures (TLSs) in cancer may contribute to prognosis and the prediction of immunotherapy efficacy However, the combined role of TLSs in NSCLC remains unclear. We accessed The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases to obtain mRNA sequencing data and clinical information as the TCGA cohort, and used our own sample of 53 advanced NSCLC as a study cohort. The samples were divided into TLS+ and TLS- groups by pathological tissue sections. Patients of the TLS+ group had a better OS (p = 0.022), PFS (p = 0.042), and DSS (p = 0.004) in the TCGA cohort, and the results were confirmed by the study cohort (PFS, p = 0.012). Furthermore, our result showed that the count and size of TLSs are closely associated with the efficacy of immunotherapy. In addition, the TLS+ group was associated with better immune status and lower tumor mutation load. In the tumor microenvironment (TME), the expression levels of CD4+ T cells and CD8+ T cells of different phenotypes were associated with TLSs. Overall, TLSs are a strong predictor of survival and immunotherapeutic efficacy in advanced NSCLC, and T cell-rich TLSs suggest a more ordered and active immune response site, which aids in the decision-making and application of immunotherapy in the clinic.