Invasive Tissue Research Articles

Postmortem minimally invasive tissue sampling in communities: exploring perceptions of families, funeral workers, religious and community leaders and healthcare providers from Pakistan

BackgroundMinimally invasive tissue sampling (MITS) has increasingly been used to improve the diagnosis of disease and identification of the cause of death, particularly in underserved areas. However, there are multiple barriers to accessing those who die within the community, our study aimed to explore the perceptions and insights of community members and healthcare providers regarding the feasibility of implementing MITS in community settings.MethodsA qualitative exploratory study was conducted. A total of twenty one in-depth interviews were conducted with deceased infants’ parents, elders of the family, religious leaders, community leaders, and funeral workers. Focus group discussions were conducted with health care providers (n = 14) in two peri-urban slum areas of Karachi, Pakistan. The duration of this study was from August to October 2020. Data was analyzed using thematic analysis and was coded and merged into categories forming eight major themes.ResultsIn general, participants viewed minimally invasive tissue sampling (MITS) as beneficial for improving child health, though some had concerns about disrespecting the deceased during sample collection. Misinformation, fear of needles, and medical procedures were major barriers to MITS implementation. To enhance acceptance, community and religious leaders suggested using religious rulings, obtaining parental consent, ensuring confidentiality, and increasing efforts of community engagement. Community healthcare providers, along with funeral workers, recommended providing community members with grief counseling to increase study participation. Besides concerns about sampling interfering with respect for the decease, community members also raised concerns about misinformation. Further, participants provided feedback on the design and appearance of the mobile van used to collect MITS samples from children under the age of five.ConclusionThis study is critical for understanding the challenges associated with implementation of community-based MITS sampling in Pakistan. Integrating the use of a mobile van for sample collection, grief counseling along with community engagement sessions and advocacy will help address community-based misinformation and develop community trust.

A prospective observational study of nurses performing minimally invasive tissue sampling of brain, liver, and lung tissues among deceased neonates and stillbirths in Ethiopia.

With a neonatal mortality rate of 33 per 1,000 live births in 2019, Ethiopia is striving to attain the Sustainable Development Goal target of 12 deaths per 1,000 live births by 2030. A better understanding of the major causes of neonatal mortality is needed to effectively design and implement interventions to achieve this goal. Minimally Invasive Tissue Sampling (MITS), an alternative to conventional autopsy, requires fewer resources and through task-shifting of sample collection from pathologists to nurses, has the potential to support the expansion of pathology-based post-mortem examination and improve mortality data. This paper evaluates the accuracy and adequacy of MITS performed by nurses at a tertiary and general hospital and in the home of the deceased. Nurses in a tertiary and general hospital in Ethiopia were trained in MITS sample collection on neonatal deaths and stillbirths using standardized protocols. MITS sample collection was performed by both pathologists and nurses in the tertiary hospital and by nurses in the general hospital and home-setting. Agreement in the performance of MITS between pathologists and nurses was calculated for samples collected at the tertiary hospital. Samples collected by nurses in the general hospital and home-setting were evaluated for technical adequacy using preestablished criteria. One hundred thirty-nine MITS were done: 125 in hospitals and 14 inside homes. There was a perfect or almost perfect agreement between the pathologists and the nurses in the tertiary hospital using Gwet's agreement interpretation criteria. The adequacy of MITS samples collected by nurses in the general hospital was more than 72% when compared to the preset criteria. The adequacy of the MITS sampling yield ranged from 87% to 91% on liveborn neonatal deaths and 76% for the liver, right and left lungs and 55% for brain tissues in stillbirths. This study demonstrated that task-shifting MITS sample collection to nurses can be achieved with comparable accuracy and adequacy as pathologists. Our study showed that with standardized training and supportive supervision MITS sample collection can be conducted by nurses in a tertiary, general hospital and, at the home of the deceased. Future studies should validate and expand on this work by evaluating task-shifting of MITS sample collection to nurses within community settings and with larger sample sizes.

Development of a Robotic Soft Tissue Collection Mechanism for Disease Detection

This research presents the comprehensive design, development, and testing of an innovative robotic mechanism tailored for the collection of soft tissue samples intended for disease detection applications. The proposed system integrates advanced technologies, such as hydrogels, chambers, and sensors, to enable meticulous and minimally invasive tissue manipulation while concurrently mitigating tissue damage. This paper showcases simulated tissue tests and validates the mechanism on chicken liver tissue. The integration of air sensors, strain gauge sensors, and chambers furnishes real-time feedback on applied forces, tissue interaction dynamics, and alterations in tissue dimensions. The presented work lays the foundation for further advancements in the realm of minimally invasive tissue collection, crucial for precise disease detection.

Performance of a Liquid Biopsy Diagnostic Prediction Model for EBV-Positive Burkitt Lymphoma in Sub-Saharan Africa

Burkitt Lymphoma (BL) is classified into EBV-positive BL (EBL) and EBV-negative BL (sporadic BL), the former being more prevalent in SSA (Al-Khreisat et al 2023). Conventional diagnosis of BL depends on the morphological assessment of invasive tissue biopsy followed by immunohistochemistry (IHC) and fluorescent in situ hybridization (FISH) to reveal the pathognomonic t(8;14), or t(8;22) translocations. This is associated with significant diagnostic delay in regions with limited pathology services (Mawalla et al 2023), often necessitating initiation of treatment based on clinical or histopathological findings without immunohistochemistry (Ogwang et al 2011), with ensuing mismanagement contributing to poor survival outcomes observed in SSA compared to High-Income Countries (HIC) (Sung et al 2021). Here, we sought to explore whether liquid biopsy could be used as an initial diagnostic aid for EBL in settings without easy access to pathology using different EBV DNA parameters and the MYC-Immunoglobulin ( MYC-Ig) gene rearrangement. This prospective study collected whole blood samples prospectively from 221 participants (90 BL and 131 non-BL) enrolled in the Aggressive Infection-Related East Africa Lymphoma (AI-REAL) project (Legason et al 2022) across 5 sites in Tanzania and Uganda. The revised BL diagnostic algorithm (Naresh et al 2021) was used as a gold-standard diagnosis. CfDNA was extracted using the QIAamp Circulating Nucleic Acid Kit (Qiagen, USA), according to the manufacturer's instructions. DNA libraries were prepared with the Thru PLEX Tag-Seq HV kit (Takara Bio Inc. Japan), and hybridization capture was performed using a custom, next-generation sequencing (NGS) panel targeting genes commonly mutated in BL and three EBV genes: EBER1, EBER2, and EBNA2. All sequencing was performed in-country (MiSeq, Illumina, CA, USA) and analyzed in a custom pipeline. Using custom scripts, EBV DNA copies per cell and EBV DNA fragment size ratio were calculated as the proportion of EBV fragments of length 180 - 200 base pairs (bp) divided by the proportion of autosomal fragments within the same size window. The diversity in the fragment size distribution of the EBV and autosomal DNA was calculated and presented as EBV entropy and autosomal entropy. Translocations were called using IgCaller (Nadeu et al 2020) and Genomic Rearrangement Identification Software Suite (GRIDSS) (Cameron et al 2017). Univariate analysis was performed to assess the association between predictor variables (clinical parameters, EBV parameters, and MYC translocation) and a diagnosis of EBL. Ten-fold cross-validation was performed on a liquid biopsy model consisting of all the liquid biopsy variables with the clinical variables included in the model as covariates. Performance metrics for the liquid biopsy model as a diagnostic aid were calculated, and variable importance was determined to assess each variable's relative influence on the model. In our cohort, EBL diagnosis was negatively associated with tumor site (neck) and not significantly associated with tumor site (jaw). A shorter duration of symptoms was associated with a diagnosis of EBL. Presence of MYC translocation t(8;14), higher EBV fragment size ratio, proportion, entropy, and copies per cell for EBER1, EBER2, and EBNA2 were all associated with a diagnosis of EBL.The combined liquid biopsy diagnostic prediction model had an estimated AUC of 90%, with sensitivity between 85%-90%, specificity between 75% - 80%, and positive and negative predictive values of 85% (Figure 1). MYC translocation t(8;14) was the most important diagnostic variable, followed by EBV entropy (Figure 2). This study demonstrates the feasibility of liquid biopsy as a non-invasive diagnostic aid for EBL in settings without easy access to pathology. To strengthen the application of this technology into clinical practice, further analysis is being conducted for weighting and assigning of scores for each predictor variable. This would assist clinicians in making a precise diagnosis, complementing existing diagnostic modalities for EBL, and reducing the rate of misdiagnosis arising from diagnosis based on clinical suspicion and tissue morphology without immunohistochemistry.

The classification of the bladder cancer based on Vision Transformers (ViT)

Bladder cancer is a prevalent malignancy with diverse subtypes, including invasive and non-invasive tissue. Accurate classification of these subtypes is crucial for personalized treatment and prognosis. In this paper, we present a comprehensive study on the classification of bladder cancer into into three classes, two of them are the malignant set as non invasive type and invasive type and one set is the normal bladder mucosa to be used as stander measurement for computer deep learning. We utilized a dataset containing histopathological images of bladder tissue samples, split into a training set (70%), a validation set (15%), and a test set (15%). Four different deep-learning architectures were evaluated for their performance in classifying bladder cancer, EfficientNetB2, InceptionResNetV2, InceptionV3, and ResNet50V2. Additionally, we explored the potential of Vision Transformers with two different configurations, ViT_B32 and ViT_B16, for this classification task. Our experimental results revealed significant variations in the models’ accuracies for classifying bladder cancer. The highest accuracy was achieved using the InceptionResNetV2 model, with an impressive accuracy of 98.73%. Vision Transformers also showed promising results, with ViT_B32 achieving an accuracy of 99.49%, and ViT_B16 achieving an accuracy of 99.23%. EfficientNetB2 and ResNet50V2 also exhibited competitive performances, achieving accuracies of 95.43% and 93%, respectively. In conclusion, our study demonstrates that deep learning models, particularly Vision Transformers (ViT_B32 and ViT_B16), can effectively classify bladder cancer into its three classes with high accuracy. These findings have potential implications for aiding clinical decision-making and improving patient outcomes in the field of oncology.

Comparison of Adjuvant Clindamycin vs Linezolid for Severe Invasive Group A Streptococcus Skin and Soft Tissue Infections.

Linezolid may be an option for severe group A Streptococcus (GAS) infections based on its potent in vitro activity and antitoxin effects, but clinical data supporting its use over clindamycin are limited. This study evaluated treatment outcomes in patients with severe GAS skin and soft tissue infections who received either linezolid or clindamycin. This retrospective single-center cohort study examined patients with GAS isolated from blood and/or tissue cultures with invasive soft tissue infection or necrotizing fasciitis who underwent surgical debridement and received linezolid or clindamycin for at least 48 hours. The primary outcome was percentage change in Sequential Organ Failure Assessment (SOFA) score from baseline through 72 hours of hospitalization. After adjustment for time to first surgical intervention among patients with a baseline SOFA score >0 (n = 23 per group), there was no difference in reduction of SOFA score over the first 72 hours in patients receiving clindamycin vs linezolid. In the entire cohort (n = 26, clindamycin; n = 29, linezolid), there was no difference in inpatient mortality (2% vs 1%) or any secondary outcomes, including duration of vasopressor therapy, intensive care unit length of stay, and antibiotic-associated adverse drug events. There was no difference in reduction of critical illness as measured by SOFA score between baseline and 72 hours among patients treated with clindamycin vs linezolid. Given its more favorable side effect profile, linezolid may be a viable option for the treatment of serious GAS infections and should be further studied.

Community perceptions of postmortem examination and minimally invasive tissue sampling in neonates:a qualitative study in South India

BackgroundPostmortem examination is the gold standard for establishing the cause of death. Minimally Invasive Tissue Sampling (MITS) of organs is a novel approach that can be used as an alternative to postmortem examination. In this study, the community perceptions in two states in South India towards neonatal postmortem and the acceptance towards the MITS technique is studied.MethodsIn-depth interviews were conducted among the participants from Kerala and Karnataka to understand the perception towards postmortem and the acceptance of the novel MITS technique. The interviews were audio recorded, and a thematic analysis was done to identify the overarching themes and codes.ResultsThe knowledge and attitude of participants on conventional and MITS postmortem techniques, reasons for refusal, and the need for raising awareness were identified in the study. Participants favored the MITS techniques as it was less disfiguring and less time-consuming. The major concerns for refusal of conventional postmortem were that the procedure was disfiguring, time-consuming, and caused emotional stress to the parents.ConclusionsParticipants favored the MITS approach over conventional postmortem as it caused less disfigurement and was conducive to the religious practice of burial of the body.

BIOM-25. IDENTIFICATION OF PLASMA-DERIVED PREDICTIVE BIOMARKERS INCLUDING SERPINA3 FOR GLIOMA AND GLIOBLASTOMA

Abstract Advancement in the diagnosis and surveillance of gliomas has been limited posing both diagnostic and therapeutic challenges. Definitive diagnosis, distinction of progression versus pseudoprogression and prognosis assessment still depend on invasive neurosurgical biopsy procedure. Minimally-invasive plasma sampling could minimize the risks associated with invasive tissue sampling. Therefore, we aimed to identify specific plasma protein patterns of low- and high-grade glioma patients, compared to healthy individuals and investigated whether different plasma-derived protein signatures were associated with survival in glioma patients. Plasma samples were from patients with glioma grades I to IV collected during tumor removal and from healthy controls donating blood were processed for Liquid Chromatography Mass Spectrometry (LC/MS) proteomics after depletion of the 14 most abundant plasma proteins, in order to specifically detect low-abundant tumor-derived proteins. Overall, 646 proteins were measured across 104 plasma samples from glioma patients and 57 plasmas from the healthy cohort. PCA showed 2 clusters of samples, one containing low- and high-grade glioma samples and the other containing healthy and some grade IV (GBM) plasma samples. Interestingly, a spatial analysis of tumor location by MRI associated the GBM samples separated from healthy samples with subventricular vicinity. 26 differentially expressed proteins (DEPs) were identified discriminating between glioma and healthy samples and 30 DEPs discriminating GBM and healthy samples. The top most overexpressed proteins in gliomas vs controls were SERPINA3, F13A1, and TKT. Multivariate analysis identified SERPINA3, PPBP and MYH9 as strong discriminators of healthy vs GBM but also of low-grade vs GBM, indicating that these proteins may represent specific plasma biomarkers for GBM. SERPINA3, MYL1 and CLU were associated with poor survival in GBM. In conclusion, we describe sets of plasma derived proteins, in particular SERPINA3, as predictive biomarkers that could be used to assess tumor progression allowing patient-centered treatment options.

The therapeutic effects of gingival mesenchymal stem cells and their exosomes in a chimeric model of rheumatoid arthritis

BackgroundRheumatoid arthritis is a chronic systemic autoimmune disease that involves transformation of the lining of synovial joints into an invasive and destructive tissue. Synovial fibroblasts become transformed, invading and destroying the bone and cartilage of the affected joint(s). Due to the significant role these cells play in the progression of the disease process, developing a therapeutic strategy to target and inhibit their invasive destructive nature could help patients who are afflicted with this debilitating disease. Gingival-derived mesenchymal stem cells are known to possess immunomodulatory properties and have been studied extensively as potential cell-based therapeutics for several autoimmune disorders.MethodsA chimeric human/mouse model of synovitis was created by surgically implanting SCID mice with a piece of human articular cartilage surrounded by RASF. Mice were injected once with either GMSC or GMSCExo at 5–7 days post-implantation. Histology and IHC were used to assess RASF invasion of the cartilage. Flow cytometry was used to understand the homing ability of GMSC in vivo and the incidence of apoptosis of RASF in vitro.ResultsWe demonstrate that both GMSC and GMSCExo are potent inhibitors of the deleterious effects of RASF. Both treatments were effective in inhibiting the invasive destructive properties of RASF as well as the potential for these cells to migrate to secondary locations and attack the cartilage. GMSC home to the site of the implant and induce programmed cell death of the RASF.ConclusionsOur results indicate that both GMSC and GMSCExo can block the pathological effects of RASF in this chimeric model of RA. A single dose of either GMSC or GMSCExo can inhibit the deleterious effects of RASF. These treatments can also block the invasive migration of the RASF, suggesting that they can inhibit the spread of RA to other joints. Because the gingival tissue is harvested with little difficulty, relatively small amounts of tissue are required to expand the cells, the simple in vitro expansion process, and the increasing technological advances in the production of therapeutic exosomes, we believe that GMSCExo are excellent candidates as a potential therapeutic for RA.

A one-pot transcriptional assay method that detects the tumor biomarker FEN1 based on its flap cleavage activity

BackgroundDetection of tumor biomarkers in body fluids is a significant advancement in cancer treatment because it allows diagnosis without invasive tissue biopsies. Nucleases have long been regarded as a potential class of biomarkers that can indicate the occurrence and progression of cancers. Among these, flap endonuclease 1 (FEN1) plays an important role in DNA replication and repair, and also overexpressed in abnormally proliferating cells such as cancer cells. FEN1 is thus considered to be a potential biomarker as well as a target for cancer therapy. ResultsWe developed a novel method for detecting FEN1 based on its specific endonuclease activity which incises bifurcated nucleic acids (flaps), in combination with in vitro transcription. Developed method uses a simple DNA structure (substrate DNA) carrying a short 5′-flap sequence, and a single-stranded sensor DNA encoding the Broccoli light-up aptamer. When the assay mixture was supplied with a FEN1-containing sample, the flap sequence encoding the sense sequence of T7 promoter was cleaved and released from the substrate DNA. Because the sensor DNA was designed to carry the Broccoli RNA aptamer under the antisense sequence of T7 promoter, hybridization of the excised flap onto the sensor DNA initiated the transcription of the Broccoli RNA aptamer, enabling determination of the FEN1 titer based on the fluorescence of transcribed Broccoli aptamer. By using a combination of FEN1-mediated generation of a short oligonucleotide and subsequent oligonucleotide-dependent in vitro transcription, this method could detect FEN1 in biological samples within 1 h. Significance and noveltyDeveloped method enables the detection of FEN1 by a simple one-pot reaction. It can detect sub-nanomolar concentrations of FEN1 within an hour, and has the potential to be used for cancer diagnosis, prognosis, and drug screening. It also enables easy identification of compounds that inhibit FEN1 activity and is thus a versatile platform for screening anti-cancer drugs. We anticipate that the basic principles of this assay can be applied to detect other biomolecules, such as nucleic acids.

Comparison of orthotopic and heterotopic autologous ovarian tissue transplantation outcomes

To compare the outcomes of orthotopic and heterotopic ovarian tissue transplantation (OTT) techniques. Mixed prospective-retrospective cohort study. Academic hospital. A total of 14 recipients of autologous OTT. Of the 14 women, 12 who received orthotopic (n = 6) or heterotopic (n = 6) transplants met the inclusion criteria. All orthotopic transplants and one heterotopic ovarian tissue transplant were performed laparoscopically. Although 5 of the 6 remaining heterotopic transplants were performed subcutaneously under local anesthesia or intravenous sedation, one was performed with robotic assistance. With the exception of one recipient who solely desired restoration of endocrine function, all underwent oocyte retrieval either to cryopreserve oocytes and embryos before the graft function ceased or because they could not otherwise conceive (hysterectomy, radiation damage, and heterotopic transplant). Primary outcome measures were graft function and longevity, and the number of embryos generated per retrieval. The mean age at ovarian tissue harvesting and transplantation was lower in patients with orthotopic vs. heterotopic transplants, although the proportion of transplanted ovarian cortex was lower in heterotopic transplant cases. All grafts restored ovarian endocrine function. Fertilization rates, the number of embryos generated per retrieval, and the mean number of nonarrested embryos were significantly lower in heterotopic OTT. However, time to function and graft longevity were similar between the groups. Although 4 of the 6 women conceived and delivered 7 children among orthotopic ovarian tissue recipients, one recipient had 3 spontaneous live births after heterotopic OTT, presumably because of the induction of function in the remaining menopausal ovary. It appears that orthotopic OTT results in higher gamete and embryo quality. However, the endocrine function restoration rate and longevity are similar between the 2 approaches. When feasible, orthotopic OTT should be preferred for those who intend to conceive, although a less invasive heterotopic OTT can be performed for those who primarily desire ovarian endocrine function.

Concordance between MITS and conventional autopsies for pathological and virological diagnoses

In pandemics or to further study highly contagious infectious diseases, new strategies are needed for the collection of post-mortem tissue samples to identify the pathogen as well as its morphological impact. In this study, an ultrasound-guided minimally invasive tissue sampling (MITS) protocol was developed and validated for post-mortem use. The histological and microbiological qualities of post-mortem specimens were evaluated and compared between MITS and conventional autopsy (CA) in a series of COVID-19 deaths. Thirty-six ultrasound-guided MITS were performed. In five cases more, specimens for histological and virological examination were also obtained and compared during the subsequently performed CA. Summary statistics and qualitative interpretations (positive, negative) were calculated for each organ tissue sample from MITS and CA, and target genes were determined for both human cell count (beta-globin) and virus (SARS-CoV-2 specific E gene). There are no significant differences between MITS and CA with respect to the detectability of viral load in individual organs, which is why MITS can be of utmost importance and an useful alternative, especially during outbreaks of infectious diseases.

Causal effects of genetically predicted endometriosis on breast cancer: a two-sample Mendelian randomization study

This study used a Mendelian randomization (MR) approach to investigate the causal relationship between genetically predicted endometriosis (EMS) and breast cancer risk. A total of 122,977 cases and 105,974 controls were included in the analysis, with gene-level summary data obtained from the Breast Cancer Association Consortium (BCAC). An inverse variance-weighting approach was applied to assess the causal relationship between EMS and breast cancer risk, and weighted median and MR-Egger regression methods were used to evaluate pleiotropy. Results showed a causal relationship between EMS and a decreased risk of overall breast cancer (odds ratio [OR] 0.95; 95% CI 0.90–0.99, p = 0.02). Furthermore, EMS was associated with a lower risk for estrogen receptor (ER)-positive breast cancer in a subgroup analysis based on immunohistochemistry type (OR 0.91; 95% CI 0.86–0.97, p = 0.005). However, there was no causal association between ER-negative breast cancer and survival (OR 1.00; 95% CI 0.94–1.06, p = 0.89). Pleiotropy was not observed. These findings provide evidence of a relationship between EMS and reduced breast cancer risk in invasive breast cancer overall and specific tissue types, and support the results of a previous observational study. Further research is needed to elucidate the mechanisms underlying this association.

Determination of causes of adult deaths using minimally invasive tissue sampling in Gandaki province of Nepal: a multicenter hospital-based study

BackgroundMinimally Invasive Tissue Sampling (MITS) has been successfully used to establish the cause of death in low- and middle-income countries, mostly in stillbirths and neonates. The objective of this study was to determine the causes of death among adults using MITS in the Gandaki province of Nepal and to find out the contribution of MITS to identify the causes of death.MethodsA multicentric hospital-based pilot study was conducted to enroll 100 cases of adult deaths. The specimens of cerebrospinal fluid, blood, brain, lungs, and liver tissue were collected utilizing MITS. These specimens underwent standard histopathological, serological, and microbiological analyses. The findings from MITS, and if available, clinical records and forensic autopsy findings were compiled and the cause of death panel identified the causes of death. The final cause of death allocated to each case was based on the WHO International Medical Certificate of Death.ResultsAmong a total of 100 cases enrolled during the study period, infectious cause attributed to the immediate cause of death in 77 (77%), cardiovascular in 10 (10%), neurological in 8 (8%), malignancy in two (2%), and gastrointestinal and hepatobiliary cause in one (1%) case. The mean age of the cases was 50.8 ± 15.9 years and 76 (76%) were males. MITS established the cause of death in the causal chain of events in 81(81%) cases and identified the cause of death significantly more with infectious than non-infectious causes (p < 0.001).ConclusionsMITS was useful in establishing the cause of death in the majority of adult deaths and the most common cause was infectious disease. Our findings suggest that MITS can be a valuable and alternative tool for mortality surveillance in low-resource settings, where complete diagnostic autopsies are less accepted or less prioritized.

Pan-Immune-Inflammation Index as a Biomarker for Rheumatoid Arthritis Progression and Diagnosis.

Introduction Rheumatoid arthritis (RA) is a chronic autoimmune condition that causes systemic inflammation and affects multiple joints. It is characterized by joint warmth, swelling, pain, and the formation of invasive synovial tissue known as pannus, which contributes to cartilage and bone degradation.Pan-immune-inflammation value (PIV), a marker derived from complete blood count parameters, has shown promise in predicting prognosis in various cancer types and pediatric conditions associated with immune abnormalities. This study aims to explore the relationship between RA, characterized by chronic inflammation and immune system involvement, and PIV, potentially shedding light on novel insights into RA's clinical implications. Methods One hundred four participants, including 64 RA patients (both newly diagnosed and established cases) and 40 healthy controls, were included in the study. Exclusion criteria for RA patients included acute infection, cancer, diabetes, or chronic illness, while control participants were excluded for inflammatory disorders, active infection, diabetes, or malignancy. We assessed disease severity using Disease Activity Score 28 (DAS 28) and obtained complete blood count values, including neutrophil, lymphocyte, platelet, monocyte, and red cell distribution width. C-reactive peptide (CRP) and erythrocyte sedimentation rate were also added.Statistical analyses included correlation assessments, t-tests, Mann-Whitney U tests, and multivariate linear regression. A multiclass receiver operating characteristic analysis determined optimal PIV cut-off values for distinguishing control, remission, and active RA groups, with sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), accuracy, and odds ratios calculated. Results This study comprised a cohort of 104 participants, with a median age of 43.5±17.5. The Remission group was significantly younger than the Control group (p=0.006)but not compared to the Active RA group (p=0.393). CRPlevels were significantly higher in the Active RA group (p<0.001). Neutrophil counts were highest in the Active RA group (p<0.001), as were monocyte counts. Lymphocyte counts were significantly lower in the Active RA group (p<0.001). There were no significant differences in sedimentation rate, hemoglobin, platelet count, and mean platelet volume. PIVwas significantly elevated in the Active RA group (p<0.001) and higher in the Remission group than in the Control group (p=0.001).A PIV value of 353.48 exhibited 71.4% sensitivity, 86.2% specificity, 86.2% PPV, 71.4% NPV, and 78.13% test accuracy for distinguishing active rheumatoid arthritis (p<0.001).A PIV value exceeding 353.48 substantially increased the likelihood of a patient belonging to the active rheumatoid arthritis group, with a 14.62-fold higher probability.Furthermore, the study explored the relationship between clinical and laboratory variables and disease activity in RA patients, finding significant differences inPIV among DAS groups (p=0.025). Conclusions The PIV offers a notable advantage as its constituent parameters are routinely assessed in rheumatoid arthritisand involve cost-effective and straightforward tests. We demonstrated that PIV serves as a valuable marker for distinguishing between remission and active RA when compared to healthy individuals. Additionally, it proved to be an effective tool for assessing disease activity in patients with active rheumatoid arthritis.

Rapid Diagnosis of Bartonella-Induced Hemophagocytic Lymphohistiocytosis Using Next-Generation Sequencing of Plasma

Abstract Hemophagocytic lymphohistiocytosis (HLH) is characterized by uncontrolled activation of macrophages and phagocytosis of hematopoietic cells. Recognition and treatment are often delayed in infectious triggers of HLH because of lack of rapid and reliable diagnostic modalities, as well as the need for invasive tissue sampling. We report a rare case of Bartonella-induced HLH in a kidney transplant patient who presented with fever of unknown origin. The diagnosis was expedited with next-generation sequencing of microbial cell-free DNA. We also describe the successful treatment outcome with antibiotics alone and the promising utility of this newer diagnostic method.

Critical Assessment of Cancer Characterization and Margin Evaluation Techniques in Brain Malignancies: From Fast Biopsy to Intraoperative Flow Cytometry.

Brain malignancies, given their intricate nature and location, present significant challenges in both diagnosis and treatment. This review critically assesses a range of diagnostic and surgical techniques that have emerged as transformative tools in brain malignancy management. Fast biopsy techniques, prioritizing rapid and minimally invasive tissue sampling, have revolutionized initial diagnostic stages. Intraoperative flow cytometry (iFC) offers real-time cellular analysis during surgeries, ensuring optimal tumor resection. The advent of intraoperative MRI (iMRI) has seamlessly integrated imaging into surgical procedures, providing dynamic feedback and preserving critical brain structures. Additionally, 5-aminolevulinic acid (5-ALA) has enhanced surgical precision by inducing fluorescence in tumor cells, aiding in their complete resection. Several other techniques have been developed in recent years, including intraoperative mass spectrometry methodologies. While each technique boasts unique strengths, they also present potential limitations. As technology and research continue to evolve, these methods are set to undergo further refinement. Collaborative global efforts will be pivotal in driving these advancements, promising a future of improved patient outcomes in brain malignancy management.

Treating facial overfilled syndrome with impaired facial expression-Presenting clinical experience with ultrasound imaging.

Facial overfilled syndrome is an adverse event following minimally invasive soft tissue filler injections. It presents in most cases as excess midfacial volume and/or as unnatural smile which is difficult to detect due to the absence of standardized evaluation methods. To showcase how to identify, evaluate, and treat facial overfilled syndrome by utilizing facial ultrasound and simultaneous hyaluronidase injections. Twenty-eight consecutive patients (26 females, 2 males) were enrolled in this study in which facial ultrasound was performed to evaluate the location previously implanted filler material. The position of the oral commissure was objectively measured in relation to bony landmarks, and the severity of lateral canthal lines was assessed by independent and blinded raters. The material was identified in 35.7% inside the subdermal fatty layer, in 28.6% inside the deep supra-periosteal fatty layer, in 10.7% inside the fibrous layer deep to the subdermal fatty layer, whereas in 25.0%, the product was not possible to locate clearly inside one specific layer. On average, 81.6 I.U. [range: 75-150] of hyaluronidase were injected. Lateral canthal line severity was before the treatment 2.28 (1.4) and was after the hyaluronidase treatment 2.02 (1.3) with p = 0.578. The position of the oral commissure increased by 0.60 cm in vertical and by 0.30 cm in horizontal directions (both p < 0.001). Facial overfilled syndrome following aesthetic soft tissue filler injections can present as excess midfacial volume but also as unnatural smile. Targeted hyaluronidase injections into the culprit pockets inside the midfacial soft tissues have shown to re-establish a natural smile, to reduce excess midfacial volume, and to decrease lateral canthal line severity.

Acceptability of minimally invasive autopsy by community members and healthcare workers in Siaya and Kisumu counties, western Kenya, 2017-2018.

Worldwide, nearly six million children under the age of five (<5s) die annually, a substantial proportion of which are due to preventable and treatable diseases. Efforts to reduce child mortality indicators in the most affected regions are often undermined by a lack of accurate cause of death data. To generate timely and more accurate causes of death data for <5s, the Child Health and Mortality Prevention Surveillance (CHAMPS) Network established mortality surveillance in multiple countries using Minimally Invasive Tissue Sampling (MITS) in <5 deaths. Here we present acceptability of MITS by community members and healthcare workers in Siaya and Kisumu counties, western Kenya. From April 2017 to February 2018, we conducted 40 in-depth interviews and five focus group discussions with healthcare workers and community members, before and during CHAMPS implementation. Participants were purposively selected. Field observations to understand traditional death-related practices were also performed. Interviews were transcribed into Nvivo 11.0 for data organization and management. Analysis was guided by the grounded theory approach. Facilitators of acceptability were desire to understand why death occurred, timely performance of MITS procedures, potential for MITS results in improving clinical practice and specific assistance provided to families by the CHAMPS program. However, cultural and religious beliefs highlighted important challenges to acceptability, including CHAMPS teams recruiting after a child's death, rumours and myths, unmet expectations from families, and fear by healthcare workers that some families could use MITS results to sue for negligence. Increasing MITS uptake requires sustained strategies to strengthen the identified facilitators of acceptability and simultaneously address the barriers. MITS acceptance will contribute to better characterization of causes of death and support the development of improved interventions aimed at reducing <5 mortality.

4D Printing of Biocompatible Scaffolds via In Situ Photo-crosslinking from Shape Memory Copolyesters.

The complexity of surgical treatments for large-area soft tissue injuries makes placing large implants into injury sites challenging. Aliphatic polyesters are often used for scaffold preparation in tissue engineering owing to their excellent biodegradability and biocompatibility. Scaffolds with shape-memory effect (SME) can also avoid large-volume trauma during the implantation. However, the complexity and diversity of diseases require more adaptable and precise processing methods. Four-dimensional (4D) printing, a booming smart material additive manufacturing technology, provides a new opportunity for developing shape memory scaffolds. With the aim of personalized or patient-adaptable soft tissues such as blood vessels, we developed a feasible strategy for fabricating scaffolds with fine architectures using 4D printing crosslinkable shape memory linear copolyesters using fused deposition modeling (FDM). To overcome the weak bonding strength of each printed layer during FDM, a catalyst-free photo-crosslinkable functional group derived from biocompatible cinnamic acid was embedded into the linear copolyesters as in situ crosslinking points during FDM printing. Under ultraviolet-assisted irradiation, the resulting 4D scaffold models demonstrated excellent SME, desirable mechanical performance, and good stability in a water environment owing to the chemical bonding between each layer. Moreover, the excellent biocompatibility of the scaffold was evaluated in vitro and in vivo. The developed composite scaffolds could be used for minimally invasive soft tissue repair.