Neutrophilic granulocyte-derived B-cell activating factor supports B cells in skin lesions in hidradenitis suppurativa.

BACKGROUND: Hidradenitis suppurativa (HS) is a chronic inflammatory disease characterized by painful inflamed nodules, abscesses, and pus-draining tunnels appearing in axillary, inguinal, and perianal skin areas. HS lesions contain various types of immigrated immune cells. OBJECTIVE: This study aimed to characterize mediators that support lesional B/plasma cell persistence in HS. METHODS: Skin samples from several cohorts of HS patients and control cohorts were assessed by mRNA sequencing, quantitative PCR on reverse-transcribed RNA, flow cytometry, and immunohistofluorescence. Blood plasma and cultured skin biopsy samples, keratinocytes, dermal fibroblasts, neutrophilic granulocytes (neutrophils), monocytes, and B cells were analyzed. Complex systems biology approaches were used to evaluate bulk and single-cell RNA sequencing data. RESULTS: Proportions of B/plasma cells, neutrophils, CD8+ T cells, and M0 and M1 macrophages were elevated in HS lesions compared to skin of healthy and perilesional intertriginous areas. There was an association between B/plasma cells, neutrophils, and B-cell activating factor (BAFF, aka TNFSF13B). BAFF was abundant in HS lesions, particularly in nodules and abscesses. Among the cell types present in HS lesions, myeloid cells were the main BAFF producers. Mechanistically, granulocyte colony-stimulating factor in the presence of bacterial products was the major stimulus for neutrophils' BAFF secretion. Lesional upregulation of BAFF receptors was attributed to B cells (TNFRSF13C/BAFFR and TNFRSF13B/TACI) and plasma cells (TNFRSF17/BCMA). Characterization of the lesional BAFF pathway revealed molecules involved in migration/adhesion (eg, CXCR4, CD37, CD53, SELL), proliferation/survival (eg, BST2), activation (eg, KLF2, PRKCB), and reactive oxygen species production (eg, NCF1, CYBC1) of B/plasma cells. CONCLUSION: Neutrophil-derived BAFF supports B/plasma cell persistence and function in HS lesions.

High-dimensional single cell mass cytometry analysis of the murine hematopoietic system reveals signatures induced by ageing and physiological pathogen challenges.

BACKGROUND: Immune ageing is a result of repetitive microbial challenges along with cell intrinsic or systemic changes occurring during ageing. Mice under 'specific-pathogen-free' (SPF) conditions are frequently used to assess immune ageing in long-term experiments. However, physiological pathogenic challenges are reduced in SPF mice. The question arises to what extent murine experiments performed under SPF conditions are suited to analyze immune ageing in mice and serve as models for human immune ageing. Our previous comparisons of same aged mice with different microbial exposures, unambiguously identified distinct clusters of immune cells characteristic for numerous previous pathogen encounters in particular in pet shop mice. RESULTS: We here performed single cell mass cytometry assessing splenic as secondary and bone marrow as primary lymphoid organ-derived leukocytes isolated from young versus aged SPF mice in order to delineate alterations of the murine hematopoietic system induced during ageing. We then compared immune clusters from young and aged SPF mice to pet shop mice in order to delineate alterations of the murine hematopoietic system induced by physiological pathogenic challenges and those caused by cell intrinsic or systemic changes during ageing. Notably, distinct immune signatures were similarly altered in both pet shop and aged SPF mice in comparison to young SPF mice, including increased frequencies of memory T lymphocytes, effector-cytokine producing T cells, plasma cells and mature NK cells. However, elevated frequencies of CD4+ T cells, total NK cells, granulocytes, pDCs, cDCs and decreased frequencies of naïve B cells were specifically identified only in pet shop mice. In aged SPF mice specifically the frequencies of splenic IgM+ plasma cells, CD8+ T cells and CD4+ CD25+ Treg were increased as compared to pet shop mice and young mice. CONCLUSIONS: Our study dissects firstly how ageing impacts both innate and adaptive immune cells in primary and secondary lymphoid organs. Secondly, it partly distinguishes murine intrinsic immune ageing alterations from those induced by physiological pathogen challenges highlighting the importance of designing mouse models for their use in preclinical research including vaccines and immunotherapies.

Correction to: High-dimensional single cell mass cytometry analysis of the murine hematopoietic system reveals signatures induced by ageing and physiological pathogen challenges.

An amendment to this paper has been published and can be accessed via the original article.

Wild immunology assessed by multidimensional mass cytometry.

A great part of our knowledge on mammalian immunology has been established in laboratory settings. The use of inbred mouse strains enabled controlled studies of immune cell and molecule functions in defined settings. These studies were usually performed in specific-pathogen free (SPF) environments providing standardized conditions. In contrast, mammalians including humans living in their natural habitat are continuously facing pathogen encounters throughout their life. The influences of environmental conditions on the signatures of the immune system and on experimental outcomes are yet not well defined. Thus, the transferability of results obtained in current experimental systems to the physiological human situation has always been a matter of debate. Studies elucidating the diversity of "wild immunology" imprintings in detail and comparing it with those of "clean" lab mice are sparse. Here, we applied multidimensional mass cytometry to dissect phenotypic and functional differences between distinct groups of laboratory and pet shop mice as a source for "wild mice". For this purpose, we developed a 31-antibody panel for murine leukocyte subsets identification and a 35-antibody panel assessing various cytokines. Established murine leukocyte populations were easily identified and diverse immune signatures indicative of numerous pathogen encounters were classified particularly in pet shop mice and to a lesser extent in quarantine and non-SPF mice as compared to SPF mice. In addition, unsupervised analysis identified distinct clusters that associated strongly with the degree of pathogenic priming, including increased frequencies of activated NK cells and antigen-experienced B- and T-cell subsets. Our study unravels the complexity of immune signatures altered under physiological pathogen challenges and highlights the importance of carefully adapting laboratory settings for immunological studies in mice, including drug and therapy testing. © 2016 International Society for Advancement of Cytometry.

Telerehabilitation in Physiotherapy Science: A Scoping Review.

The advancement of computer science technologies and telemedical devices has led to an increase in the use of telerehabilitation (TR) as a therapeutic intervention. In our days, TR interventions can be considered as alternative solutions to face-to-face therapy. The primary aim of this study is to evaluate whether TR can be effective in physiotherapy. This can be adjudicated by investigating the use of the TR applications, their cost effect, and the level of effectiveness each one of them can provide. Randomized controlled trials that were published between 2003 and 2023 in the English language and used TR as the intervention were collected from online databases (MEDLINE, Physiotherapy Evidence Database (PEDro), and Cochrane) to be reviewed. Twenty of them met the criteria and were included in the study. Studies meeting the inclusion criteria were categorized by the body system investigated. Out of the 20 studies that met the inclusive criteria, five are related to the musculoskeletal system, six are related to the nervous system, two are related to proprioception and balance, one is related to the respiratory system, one is related to the cardiovascular system, two are related to pelvic floor control, and three are related to autoimmune disorders. Studies have shown that implementing TR has resulted in significant improvements in terms of functionality, muscle strength, endurance, and self-improvement. Proprioception, autoimmune diseases, and cardiovascular health have shown the most improvement. The most commonly used tools for implementing TR are gamified virtual reality (VR) and digital apps. However, there are some disadvantages, such as the lack of personal contact and the cost involved. We found that TR has the potential to positively impact various health disorders, making it a suitable form of therapy for people who can't receive in-person treatment. Nonetheless, it cannot replace traditional physiotherapy, nor does it hold the same value as it.

Prediction of Near-Term Breast Cancer Occurrence using Subtraction of Temporally Sequential Digital Mammograms.

Breast cancer remains one of the leading cancers for women worldwide. Fortunately, with the introduction of mammography, the mortality rate has significantly decreased. However, earlier breast cancer prediction could effectively increase the survival rates, improve patient outcomes, and avoid unnecessary biopsies. For that purpose, prediction of breast cancer, using subtraction of temporally sequential digital mammograms and machine learning, is proposed. A new dataset was collected with 192 images from 32 patients (three screening rounds, with two views of each breast). This dataset included precise annotation of each individual malignant mass, present in the most recent mammogram, with the two priors being radiologically evaluated as normal. The most recent mammogram was considered as the "future" screening round and provided the location of the mass as the ground truth for the training. The two previous mammograms, the "current" and the "prior", were processed and a new, difference image was formed for the prediction. Ninety-six features were extracted and five feature selection algorithms were combined to identify the most important features. Ten classifiers were tested in leave-one-patient-out and k-fold-patient cross-validation (k = 4 and 8). Ensemble Voting achieved the highest performance in the prediction of the development of breast mass in the next screening round, with 85.7% sensitivity, 83.7% specificity, 83.7% accuracy and 0.85 AUC. The proposed methodology could lead to a new mammography-based model that could predict the short-term risk for developing a malignancy, thus providing an earlier diagnosis.

Epidemiological trends in cutaneous lymphomas in Greece.

Primary cutaneous lymphomas (PCLs) are a heterogenous group of non-Hodgkin lymphomas arising in the skin from T- or B-lymphocytes, for which there is limited epidemiological data available. To describe the disease characteristics and estimate annual incidence rates (IRs) and temporal trends of PCLs and their subtypes in Attica, Greece. A retrospective analysis of all PCL patients, diagnosed in Attica's main haemopathology referral centre from 2009 to 2021, was conducted. In total, 1,189 patients were included; 725 males and 464 females (males__females=1.56). The median age at diagnosis was 62 years. The annual IR was 2.2 new cases per 100,000 individuals. Most patients (n=979, 82.3%) were diagnosed with cutaneous T-cell lymphoma (CTCL) with a crude IR of 1.8 new cases per 100,000 person-years. Mycosis fungoides (MF) was the most common subtype (n=817, 68.7%), followed by lymphomatoid papulosis (LyP) (n=59, 5.0%). The crude IR for MF was 1.5 new cases per 100,000 person-years. Cutaneous B-cell lymphomas (CBCLs) accounted for 17.6% (n=210) of all PCLs (IR: 0.4 new cases per 100,000 person-years). PCL, CTCL and MF incidence rates increased from 2009 to 2019, followed by a decrease in 2020-2021. The incidence rate of CBCL increased steadily during the study period. The annual IRs of PCL in Greece were higher than those reported in other studies from Europe, America and Asia. The increase in IRs from 2009 to 2019 may reflect physicians' improved diagnostic efficiency. The COVID-19 pandemic may be the reason for the decline in PCL, CTCL and MF diagnoses from 2020 to 2021.

String-of-pearls locking plate and cerclage wire stabilization of periprosthetic femoral fractures after total hip replacement in six dogs.

OBJECTIVE: To report use of, and outcome after, string-of-pearls (SOP™) plate and multiple cerclage wire fixation for treatment of periprosthetic femoral fractures (PFF) associated with total hip replacement (THR) in dogs. STUDY DESIGN: Case series. ANIMALS: Dogs (n=6) with PFF after THR. METHODS: Clinical records (2005-2010) and radiographic evaluations of dogs that had PFF associated with THR, treated with a SOP™ plate and cerclage wires were retrospectively reviewed. Clinical and radiographic postoperative assessments were performed 4, 12, and 24 weeks postoperatively. Telephone follow-up was performed >12 months postoperatively. RESULTS: Three fractures occurred intraoperatively and 3 occurred postoperatively. One SOP™ failed at 2 weeks necessitating revision using 2 parallel SOP™ implants. One dog was euthanatized because of quadriceps muscle tie-down at 6 weeks. Other dogs were free of lameness with full range of motion of the stifle and hip joints at final clinical examination, and positive outcomes were maintained at >12 month telephone questionnaire. There was no evidence of implant failure and positive evidence of fracture healing at final radiographic follow-up. CONCLUSIONS: Stabilization resulted in bone healing in 5 dogs; failure in 1 dog may be attributable to technical error. Optimal technical guidelines for use of the SOP™ in this circumstance are unknown, particularly where fracture configuration varies.

Total hip replacement after failed femoral head and neck excision in two dogs and two cats.

OBJECTIVE: To document outcome in 2 dogs and 2 cats after conversion of femoral head and neck excision (FHNE) to total hip replacement (THR). STUDY DESIGN: Case series. ANIMALS: Dogs (n = 2) and 2 cats. METHODS: For 1 dog and 2 cats, THR was performed using cemented acetabular and femoral components. Noncemented acetabular and cemented femoral components were used in 1 dog. RESULTS: All animals presented with severe hip pain and chronic lameness after unsuccessful FHNE. Potential causes of suboptimal outcome after FHNE were incomplete resection of the femoral neck (n = 3) and fibrous adhesions involving the sciatic nerve (n = 2). Post-FHNE remodeling of the proximal femur and acetabulum necessitated unconventional modifications of surgical technique. In all 4 cases, final clinical outcome and radiographic reassessment were satisfactory. Aseptic loosening of the acetabular bone-cement interface necessitating surgical revision was the only complication noted in 1 cat. CONCLUSIONS: Despite severe preoperative pain and chronic functional impairment in all cases, conversion of FHNE to THR produced marked clinical improvement including return to unrestricted exercise within 12 weeks of surgery.

Automatic Breast Mass Segmentation and Classification Using Subtraction of Temporally Sequential Digital Mammograms.

OBJECTIVE: Cancer remains a major cause of morbidity and mortality globally, with 1 in 5 of all new cancers arising in the breast. The introduction of mammography for the radiological diagnosis of breast abnormalities, significantly decreased their mortality rates. Accurate detection and classification of breast masses in mammograms is especially challenging for various reasons, including low contrast and the normal variations of breast tissue density. Various Computer-Aided Diagnosis (CAD) systems are being developed to assist radiologists with the accurate classification of breast abnormalities. METHODS: In this study, subtraction of temporally sequential digital mammograms and machine learning are proposed for the automatic segmentation and classification of masses. The performance of the algorithm was evaluated on a dataset created especially for the purposes of this study, with 320 images from 80 patients (two time points and two views of each breast) with precisely annotated mass locations by two radiologists. RESULTS: Ninety-six features were extracted and ten classifiers were tested in a leave-one-patient-out and k-fold cross-validation process. Using Neural Networks, the detection of masses was 99.9% accurate. The classification accuracy of the masses as benign or suspicious increased from 92.6%, using the state-of-the-art temporal analysis, to 98%, using the proposed methodology. The improvement was statistically significant (p-value < 0.05). CONCLUSION: These results demonstrate the effectiveness of the subtraction of temporally consecutive mammograms for the diagnosis of breast masses. Clinical and Translational Impact Statement: The proposed algorithm has the potential to substantially contribute to the development of automated breast cancer Computer-Aided Diagnosis systems with significant impact on patient prognosis.

A Review of Computer-Aided Breast Cancer Diagnosis Using Sequential Mammograms.

Radiologists assess the results of mammography, the key screening tool for the detection of breast cancer, to determine the presence of malignancy. They, routinely, compare recent and prior mammographic views to identify changes between the screenings. In case a new lesion appears in a mammogram, or a region is changing rapidly, it is more likely to be suspicious, compared to a lesion that remains unchanged and it is usually benign. However, visual evaluation of mammograms is challenging even for expert radiologists. For this reason, various Computer-Aided Diagnosis (CAD) algorithms are being developed to assist in the diagnosis of abnormal breast findings using mammograms. Most of the current CAD systems do so using only the most recent mammogram. This paper provides a review of the development of methods to emulate the radiological approach and perform automatic segmentation and/or classification of breast abnormalities using sequential mammogram pairs. It begins with demonstrating the importance of utilizing prior views in mammography, through the review of studies where the performance of expert and less-trained radiologists was compared. Following, image registration techniques and their application to mammography are presented. Subsequently, studies that implemented temporal analysis or subtraction of temporally sequential mammograms are summarized. Finally, a description of the open access mammography datasets is provided. This comprehensive review can serve as a thorough introduction to the use of prior information in breast cancer CAD systems but also provides indicative directions to guide future applications.

Identification and Classification of Benign and Malignant Masses based on Subtraction of Temporally Sequential Digital Mammograms.

Breast cancer remains the leading cause of cancer deaths and the second highest cause of death, in general, among women worldwide. Fortunately, over the last few decades, with the introduction of mammography, the mortality rate of breast cancer has significantly decreased. However, accurate classification of breast masses in mammograms is especially challenging. Various Computer-Aided Diagnosis (CAD) systems are being developed to assist radiologists with the accurate classification of breast abnormalities. In this study, classification of benign and malignant masses, based on the subtraction of temporally sequential digital mammograms and machine learning, is proposed. The performance of the algorithm was evaluated on a dataset created for the purposes of this study. In total, 196 images from 49 patients, with precisely annotated mass locations and biopsy confirmed malignant cases, were included. Ninety-six features were extracted and five feature selection algorithms were employed to identify the most important features. Ten classifiers were tested using leave-one-patient-out and 7-fold cross-validation. Neural Networks, achieved the highest classification performance with 90.85% accuracy and 0.91 AUC, an improvement compared to the state-of-the-art. These results demonstrate the effectiveness of the subtraction of temporally consecutive mammograms for the classification of breast masses as benign or malignant.

HIV-related Merkel cell carcinoma: A report of three cases from the UK.

Merkel cell carcinoma (MCC) of the skin is a rare, aggressive and often fatal neuroendocrine skin cancer. The incidence of MCC has significantly increased in the last decades. Factors that have been associated with the development of MCC include infection with Merkel Cell polyomavirus (MCPyV), ultraviolet exposure, hematologic malignancies and immunosuppression.We present three cases of patients living with HIV who were diagnosed with MCC. HIV cases associated with MCC have been rarely reported and to our knowledge, not yet before in the UK.

Digital subtraction of temporally sequential mammograms for improved detection and classification of microcalcifications.

BACKGROUND: Our aim was to demonstrate that automated detection and classification of breast microcalcifications, according to Breast Imaging Reporting and Data System (BI-RADS) categorisation, can be improved with the subtraction of sequential mammograms as opposed to using the most recent image only. METHODS: One hundred pairs of mammograms were retrospectively collected from two temporally sequential rounds. Fifty percent of the images included no (BI-RADS 1) or benign (BI-RADS 2) microcalcifications. The remaining exhibited suspicious findings (BI-RADS 4-5) in the recent image. Mammograms cannot be directly subtracted, due to tissue changes over time and breast deformation during mammography. To overcome this challenge, optimised preprocessing, image registration, and postprocessing procedures were developed. Machine learning techniques were employed to eliminate false positives (normal tissue misclassified as microcalcifications) and to classify the true microcalcifications as BI-RADS benign or suspicious. Ninety-six features were extracted and nine classifiers were evaluated with and without temporal subtraction. The performance was assessed by measuring sensitivity, specificity, accuracy, and area under the curve (AUC) at receiver operator characteristics analysis. RESULTS: Using temporal subtraction, the contrast ratio improved ~ 57 times compared to the most recent mammograms, enhancing the detection of the radiologic changes. Classifying as BI-RADS benign versus suspicious microcalcifications, resulted in 90.3% accuracy and 0.87 AUC, compared to 82.7% and 0.81 using just the most recent mammogram (p = 0.003). CONCLUSION: Compared to using the most recent mammogram alone, temporal subtraction is more effective in the microcalcifications detection and classification and may play a role in automated diagnosis systems.

Dynamic changes of CTCs in patients with metastatic HR(+)/HER2(-) breast cancer receiving salvage treatment with everolimus/exemestane.

PURPOSE: Detection of CTCs represents a poor prognostic factor in patients with early and metastatic breast cancer (mBC) and treatment with everolimus-exemestane (E/E) is an established effective treatment in hormone receptor-positive/HER2-negative mBC patients. The effect of E/E on CTCs in mBC patients was prospectively investigated. METHODS: CTCs from 50 pre-treated patients with mBC receiving E/E were analyzed using the CellSearch (CS) platform and triple immunofluorescence (IF) staining for cytokeratin, M30 and Ki67 expression to assess their proliferative and apoptotic status. RESULTS: CTCs (by CS) were detected in 64% of patients before treatment and E/E administration resulted in their decreased prevalence [(n = 18; 36%, p = 0.004) and (n = 7; 19.4%, p = 0.019) post-1st and post-3rd treatment cycle, respectively] whereas it was significantly increased at disease progression (PD: 61%) compared to post-1st and post-3rd cycle (p = 0.049 and p = 0.021, respectively). Ki67-positive CTCs were detected in 60%, 60%, 17% and 50% of patients before treatment, post-1st, post-3rd cycle and at PD, respectively, while the opposite was observed for M30-positive CTCs (0% at baseline, 10% after the 1st cycle, 50% after the 3rd cycle and 0% at PD). The detection of even ≥ 1 CTC/5 ml after one cycle was associated with decreased PFS (3.3 vs 9.0 months, p = 0.025) whereas the detection of even ≥ 2 CTCs at PD was associated with decreased OS (32.4 vs 19.5 months; p = 0.009). CONCLUSIONS: The combination of E/E resulted in early elimination of proliferating CTCs in mBC patients and this effect was associated with a favorable clinical outcome.

Guidelines for the Execution of True Spherical Osteotomies Using a Modified Dome Blade Design.

OBJECTIVES: The purpose of this study was to explore the trigonometric principles of the spherical osteotomy, establish guidelines for its application and test the guidelines on bone models using a new blade design. We propose a new rule of osteotomies incorporating the outlined geometric principles, and applicable to the use of spherical cuts in veterinary orthopaedic surgery. MATERIALS AND METHODS: The trigonometric principles for the execution of neutral, closing and opening spherical osteotomies were explored in silico. A modification of the existing commercially available dome blade was designed and manufactured such that it facilitated the performance of spherical osteotomy with a minimized blade radius. A pilot study was performed whereby the modified dome blade was used to create spherical osteotomy in canine radial bone models. The surfaces of the osteotomy models were laser-scanned using a three-dimensional (3D) scanner; the resultant scans were imported into and analysed using a commercial 3D analysis software. The accuracy of osteotomy execution was measured as the distance between the targeted centre of osteotomy and the actual centre of osteotomy as found on the 3D scans. RESULTS: By utilizing the geometric principles of spherical osteotomy, an accurate osteotomy position was achieved. The centre of the spherical cut performed on bone models was confirmed to be within 5% tolerance of the location as planned in silico demonstrating the accurate and relevant clinical application of geometric principles. CLINICAL SIGNIFICANCE: The trigonometric guidelines for the execution of spherical osteotomy can be applied in a pre-clinical environment with accuracy. The new guidelines combined with the proposed new rule for spherical osteotomy utilizing the new blade design are translatable into clinical application, permitting the surgeon to accurately plan osteotomy application while mitigating the significant loss of bone-to-bone contact during correction of torsional deformities inherent in the principles of dome osteotomy use.

SLAMF7 and IL-6R define distinct cytotoxic versus helper memory CD8+ T cells.

The prevailing 'division of labor' concept in cellular immunity is that CD8+ T cells primarily utilize cytotoxic functions to kill target cells, while CD4+ T cells exert helper/inducer functions. Multiple subsets of CD4+ memory T cells have been characterized by distinct chemokine receptor expression. Here, we demonstrate that analogous CD8+ memory T-cell subsets exist, characterized by identical chemokine receptor expression signatures and controlled by similar generic programs. Among them, Tc2, Tc17 and Tc22 cells, in contrast to Tc1 and Tc17 + 1 cells, express IL-6R but not SLAMF7, completely lack cytotoxicity and instead display helper functions including CD40L expression. CD8+ helper T cells exhibit a unique TCR repertoire, express genes related to skin resident memory T cells (TRM) and are altered in the inflammatory skin disease psoriasis. Our findings reveal that the conventional view of CD4+ and CD8+ T cell capabilities and functions in human health and disease needs to be revised.

The H-Y Antigen in Embryonic Stem Cells Causes Rejection in Syngeneic Female Recipients.

Pluripotent stem cells are promising candidates for cell-based regenerative therapies. To avoid rejection of transplanted cells, several approaches are being pursued to reduce immunogenicity of the cells or modulate the recipient's immune response. These include gene editing to reduce the antigenicity of cell products, immunosuppression of the host, or using major histocompatibility complex-matched cells from cell banks. In this context, we have investigated the antigenicity of H-Y antigens, a class of minor histocompatibility antigens encoded by the Y chromosome, to assess whether the gender of the donor affects the cell's antigenicity. In a murine transplant model, we show that the H-Y antigen in undifferentiated embryonic stem cells (ESCs), as well as ESC-derived endothelial cells, provokes T- and B cell responses in female recipients.