A Comprehensive Exploration of Therapeutic Strategies in Inflammatory Bowel Diseases: Insights from Human and Animal Studies.

Inflammatory bowel disease (IBD) is a collective term for a group of chronic inflammatory enteropathies which are characterized by intestinal inflammation and persistent or frequent gastrointestinal signs. This disease affects more than 3.5 million humans worldwide and presents some similarities between animal species, in particular, dogs and cats. Although the underlying mechanism that triggers the disease is not yet well understood, the evidence suggests a multifactorial etiology implicating genetic causes, environmental factors, microbiota imbalance, and mucosa immune defects, both in humans and in dogs and cats. Conventional immunomodulatory drug therapies, such as glucocorticoids or immunosuppressants, are related with numerous adverse effects that limit its long-term use, creating the need to develop new therapeutic strategies. Mesenchymal stromal cells (MSCs) emerge as a promising alternative that attenuates intestinal inflammation by modulating inflammatory cytokines in inflamed tissues, and also due to their pro-angiogenic, anti-apoptotic, anti-fibrotic, regenerative, anti-tumor, and anti-microbial potential. However, this therapeutic approach may have important limitations regarding the lack of studies, namely in veterinary medicine, lack of standardized protocols, and high economic cost. This review summarizes the main differences and similarities between human, canine, and feline IBD, as well as the potential treatment and future prospects of MSCs.

Dual production of human mesenchymal stromal cells and derived extracellular vesicles in a dissolvable microcarrier-based stirred culture system.

BACKGROUND & AIMS: Cell therapies based on mesenchymal stromal cells (MSCs) have gained an increasing therapeutic interest in the context of multiple disorders. Nonetheless, this field still faces important challenges, particularly concerning suitable manufacturing platforms. Here, we aimed at establishing a scalable culture system to expand umbilical cord-derived Wharton's jelly MSC (MSC(WJ)) and their derived extracellular vesicles (EVs) by using dissolvable microcarriers combined with xeno(geneic)-free culture medium. METHODS: MSC(WJ) isolated from three donors were cultured at a starting density of 1 × 106 cells per spinner flask, i.e., 2.8 × 103 cells per cm2 of dissolvable microcarrier surface area. After a 6-day expansion period of MSC(WJ), extracellular vesicles (EVs) were produced for 24 h. RESULTS: Taking advantage of an intermittent agitation regimen, we observed high adhesion rates to the microcarriers (over 90% at 24 h) and achieved 15.8 ± 0.7-fold expansion after 6 days of culture. Notably, dissolution of the microcarriers was achieved through a pectinase-based solution to recover the cell product, reducing the hurdles of downstream processing. MSC identity was validated by detecting the characteristic MSC immunophenotype and by multilineage differentiation assays. Considering the growing interest in MSC-derived EVs, which are known to be mediators of the therapeutic features of MSC, this platform also was evaluated for EV production. Upon a 24-h period of conditioning, secreted EVs were isolated by ultrafiltration followed by anion-exchange chromatography and exhibited the typical cup-shaped morphology, small size distribution (162.6 ± 30.2 nm) and expressed EV markers (CD63, CD9 and syntenin-1). CONCLUSIONS: Taken together, we established a time-effective and robust scalable platform that complies with clinical-grade standards for the dual production of MSC(WJ) and their derived EV.

[Recurrent Bacteremia after Cyanoacrylate Sclerotherapy of Ectopic Duodenal Varices]. / Bacteriemias Recorrentes após Escleroterapia de Varizes Ectópicas Duodenais com Cianoacrilato.

Sclerotherapy with N-butyl-2-cyanoacrylate is the first-line endoscopic approach for gastric and ectopic variceal bleeding, but it can be associated with local or systemic complications. Episodes of transient bacteremia after the procedure are frequent, but documented cases of recurrent bacteremia are rare. The authors report a 47-year-old female patient with liver cirrhosis who underwent duodenal sclerotherapy with cyanoacrylate after upper gastrointestinal bleeding. Subsequently, she developed five episodes of bacteremia with unknown origin. A definitive diagnosis of recurrent bacteremia due to cyanoacrylate was only possible after an exhaustive study to exclude other infectious foci. This case highlights a rare complication in an unusual topography (ectopic varices) and with a high number of episodes of bacteremia. A multidisciplinary management was paramount due to the patient's high surgical and anesthetic risk, comorbidities, and surgical aggressiveness.

A escleroterapia com N-butil-2-cianoacrilato é a abordagem endoscópica de primeira linha nas hemorragias por varizes gástricas ou ectópicas, podendo estar associada a complicações locais ou sistémicas. São frequentes situações de bacteriemia transitória após o procedimento, mas são raros os casos documentados de bacteriemias recorrentes. O caso relata uma doente de 47 anos com cirrose hepática submetida a esclerose de varizes duodenais com cianoacrilato após hemorragia digestiva alta. Posteriormente, desenvolveu cinco episódios de bacteriémia sem foco evidente. O diagnóstico definitivo de bacteriemias recorrentes com ponto de partida no cianoacrilato só foi possível após estudo exaustivo para excluir outros focos infeciosos. Este caso evidencia uma complicação rara, numa topografia também incomum (varizes ectópicas) e com um elevado número de episódios de bacteriémia. Destaca-se a importância de uma abordagem multidisciplinar para gerir esta doente, tendo em conta as comorbilidades, o elevado risco anestésico e a agressividade cirúrgica para extração do foco infecioso.

Bioprocessing Considerations towards the Manufacturing of Therapeutic Skeletal and Smooth Muscle Cells.

Tissue engineering approaches within the muscle context represent a promising emerging field to address the current therapeutic challenges related with multiple pathological conditions affecting the muscle compartments, either skeletal muscle or smooth muscle, responsible for involuntary and voluntary contraction, respectively. In this review, several features and parameters involved in the bioprocessing of muscle cells are addressed. The cell isolation process is depicted, depending on the type of tissue (smooth or skeletal muscle), followed by the description of the challenges involving the use of adult donor tissue and the strategies to overcome the hurdles of reaching relevant cell numbers towards a clinical application. Specifically, the use of stem/progenitor cells is highlighted as a source for smooth and skeletal muscle cells towards the development of a cellular product able to maintain the target cell's identity and functionality. Moreover, taking into account the need for a robust and cost-effective bioprocess for cell manufacturing, the combination of muscle cells with biomaterials and the need for scale-up envisioning clinical applications are also approached.

Wear of Titanium Implant Platforms with Different Abutment Connections and Abutment Materials: A Pilot Study.

The most commonly used material in dental implants and their abutments is titanium. Zirconia is a more aesthetic alternative to titanium abutments; however, it is much harder. There are concerns that zirconia could damage the surface of the implant over time, especially in less stable connections. The aim was to evaluate the wear of implants with different platforms connected to titanium and zirconia abutments. A total of six implants were evaluated, two of each connection type: external hexagon, tri-channel, and conical connections (n = 2). Half of the implants were connected to zirconia abutments, and the other half to titanium abutments (n = 3). The implants were then cyclically loaded. The implant platforms were evaluated by digital superimposing micro CT files and calculating the area of the loss surface (wear). In all the implants, a statistically significant loss of the surface area (p = 0.028) was observed when comparing the area before and after cyclic loading. The average lost surface area was 0.38 mm2 with titanium abutments and 0.41 mm2 with zirconia abutments. The average lost surface area was 0.41 mm2 with the external hexagon, 0.38 mm2 with the tri-channel, and 0.40 mm2 with the conical connection. In conclusion, the cyclic loads induced implant wear. However, neither the type of abutment (p = 0.700) nor the connection (p = 0.718) influenced the amount of surface area lost.

Peritoneal Dialysis Exit-Site Care Protocols in Portugal and Its Association with Catheter-Related Infections.

INTRODUCTION: Exit-site infection (ESi) prevention is a key factor in lowering the risk of peritonitis. This study aimed to evaluate the associations between exit-site (ES) care protocols and the annual incidence rates of ESi and peritonitis in Portugal. METHODS: We performed a national survey using two questionnaires: one about the incidence of catheter-related infections and the other characterizing patients' education and ES care protocols. RESULTS: In 2017 and 2018, 14 Portuguese units followed 764 and 689 patients. ESi incidence rate was 0.41 episodes/year, and the peritonitis incidence rate was 0.37. All units monitor catheter-related infections on a yearly basis, use antibiotic prophylaxis at the time of catheter placement, and treat nasal carriage of S. aureus, although with different approaches. Screening for nasal carriage of S. aureus is performed by 12 units, and daily topical antibiotic cream is recommended by 6 out of 14 of the units. We did not find statistical differences in ESi/peritonitis, comparing these practices. The rate of ESis was lower with nonocclusive dressing immediately after catheter insertion, bathing without ES dressing, with the use of colostomy bags in beach baths and was higher with the use of bath sponge. The peritonitis rate was lower with bathing without ES dressing and if shaving of the external cuff was performed in the presence of chronic ESi. CONCLUSIONS: We found potential proceedings associated with ESi and peritonitis. A regular national audit of peritoneal dialysis units is an important tool for clarifying the best procedures for reduction of catheter-related infections.

It is not always "essential" - regarding a hypertensive urgency.

Hypertension is, in a minority of cases, secondary to an identifiable cause. In this context, the aetiology of the blood pressure elevation is essential since it may be treatable. We present a case of a young woman with hypertension secondary to fibromuscular dysplasia (FMD) of the renal artery in which the endovascular treatment was crucial for its management.

3D Cancer Models: Depicting Cellular Crosstalk within the Tumour Microenvironment.

The tumour microenvironment plays a critical role in tumour progression and drug resistance processes. Non-malignant cell players, such as fibroblasts, endothelial cells, immune cells and others, interact with each other and with the tumour cells, shaping the disease. Though the role of each cell type and cell communication mechanisms have been progressively studied, the complexity of this cellular network and its role in disease mechanism and therapeutic response are still being unveiled. Animal models have been mainly used, as they can represent systemic interactions and conditions, though they face recognized limitations in translational potential due to interspecies differences. In vitro 3D cancer models can surpass these limitations, by incorporating human cells, including patient-derived ones, and allowing a range of experimental designs with precise control of each tumour microenvironment element. We summarize the role of each tumour microenvironment component and review studies proposing 3D co-culture strategies of tumour cells and non-malignant cell components. Moreover, we discuss the potential of these modelling approaches to uncover potential therapeutic targets in the tumour microenvironment and assess therapeutic efficacy, current bottlenecks and perspectives.

Patient-Derived Explants of Colorectal Cancer: Histopathological and Molecular Analysis of Long-Term Cultures.

Colorectal cancer (CRC) is one of the most common cancers worldwide. Although short-term cultures of tumour sections and xenotransplants have been used to determine drug efficacy, the results frequently fail to confer clinically useful information. Biomarker discovery has changed the paradigm for advanced CRC, though the presence of a biomarker does not necessarily translate into therapeutic success. To improve clinical outcomes, translational models predictive of drug response are needed. We describe a simple method for the fast establishment of CRC patient-derived explant (CRC-PDE) cultures from different carcinogenesis pathways, employing agitation-based platforms. A total of 26 CRC-PDE were established and a subset was evaluated for viability (n = 23), morphology and genetic key alterations (n = 21). CRC-PDE retained partial tumor glandular architecture and microenvironment features were partially lost over 4 weeks of culture. Key proteins (p53 and Mismatch repair) and oncogenic driver mutations of the original tumours were sustained throughout the culture. Drug challenge (n = 5) revealed differential drug response from distinct CRC-PDE cases. These findings suggest an adequate representation of the original tumour and highlight the importance of detailed model characterisation. The preservation of key aspects of the CRC microenvironment and genetics supports CRC-PDE potential applicability in pre- and co-clinical settings, as long as temporal dynamics are considered.

Application of LDH assay for therapeutic efficacy evaluation of ex vivo tumor models.

The current standard preclinical oncology models are not able to fully recapitulate therapeutic targets and clinically relevant disease biology, evidenced by the 90% attrition rate of new therapies in clinical trials. Three-dimensional (3D) culture systems have the potential to enhance the relevance of preclinical models. However, the limitations of currently available cellular assays to accurately evaluate therapeutic efficacy in these models are hindering their widespread adoption. We assessed the compatibility of the lactate dehydrogenase (LDH) assay in 3D spheroid cultures against other commercially available readout methods. We developed a standardized protocol to apply the LDH assay to ex vivo cultures, considering the impact of culture growth dynamics. We show that accounting for growth rates and background release levels of LDH are sufficient to make the LDH assay a suitable methodology for longitudinal monitoring and endpoint assessment of therapeutic efficacy in both cell line-derived xenografts (xenospheres) and patient-derived explant cultures. This method has the added value of being non-destructive and not dependent on reagent penetration or manipulation of the parent material. The establishment of reliable readout methods for complex 3D culture systems will further the utility of these tumor models in preclinical and co-clinical drug development studies.

Heterotypic Tumor Spheroids in Agitation-Based Cultures: A Scaffold-Free Cell Model That Sustains Long-Term Survival of Endothelial Cells.

Endothelial cells (ECs) are an important component of the tumor microenvironment, playing key roles in tumor development and progression that span from angiogenesis to immune regulation and drug resistance. Heterotypic tumor spheroids are one of the most widely used in vitro tumor microenvironment models, presenting improved recapitulation of tumor microenvironments compared to 2D cultures, in a simple and low-cost setup. Heterotypic tumor spheroid models incorporating endothelial cells have been proposed but present multiple limitations, such as the short culture duration typically obtained, the use of animal-derived matrices, and poor reproducibility; the diversity of culture conditions employed hinders comparison between studies and standardization of relevant culture parameters. Herein, we developed long-term cultures of triple heterotypic spheroids composed of the HCC1954 tumor cell line, human fibroblasts, and ECs. We explored culture parameters potentially relevant for EC maintenance, such as tumor cell line, seeding cell number, cell ratio, and agitation vs. static culture. In HCC1954-based spheroids, we observed maintenance of viable EC for up to 1 month of culture in agitation, with retention of the identity markers CD31 and von Willebrand factor. At the optimized tumor cell:fibroblast:EC ratio of 1:3:10, HCC1954-based spheroids had a higher EC area/total spheroid area at 1 month of culture than the other cell ratios tested. EC maintenance was tumor cell line-dependent, and in HCC1954-based spheroids it was also dependent on the presence of fibroblasts and agitation. Moreover, vascular endothelial growth factor (VEGF) supplementation was not required for maintenance of EC, as the factor was endogenously produced. ECs co-localized with fibroblasts, which accumulated preferentially in the core of the spheroids and secreted EC-relevant extracellular matrix proteins, such as collagen I and IV. This simple model setup does not rely on artificial or animal-derived scaffolds and can serve as a useful tool to explore the culture parameters influencing heterotypic spheroids, contributing to model standardization, as well as to explore molecular cross talk of ECs within the tumor microenvironment, and potentially its effects on drug response.

Diffuse Calcinosis from Total Thyroidectomy and Secondary Hypocalcaemia.

Hypoparathyroidism is a rare endocrine disorder and its main cause is cervical surgery such as thyroidectomy. The incidence of hypoparathyroidism after total thyroidectomy varies, and is reported to be between 0.3 and 6.3%. In terms of brain imaging, hypoparathyroidism can cause calcification mainly of the basal ganglia, with other areas rarely affected. Concerning extracerebral calcification, few studies have investigated the prevalence of visceral calcifications. We describe a case of a woman with secondary hypocalcaemia who presented with a transient ischaemic attack (TIA) and the investigation revealed an extensive calcinosis of the brain and the vascular structures responsible for the event. LEARNING POINTS: Iatrogenic hypoparathyroidism with hypocalcaemia is an important complication of total thyroidectomy. Hypocalcaemia manifestations can vary from asymptomatic to life-threatening conditions.Hypoparathyroidism (with hypocalcaemia) can cause cerebral calcification mainly of basal ganglia, with other areas of the brain rarely affected.Extracerebral calcifications are under-studied.

An Atypical Presentation of Diffuse Large B-Cell Lymphoma.

Diffuse large B-cell lymphoma (DLBCL) is the most common type of non-Hodgkin's lymphoma (NHL) and it can metastasize to extranodal sites. The involvement of the adrenal glands is rare. In this report, we discuss the case of a 65-year-old man with complaints of asthenia, anorexia, hypersudoresis, and a weight loss of 10 kg in the month prior to his presentation. Suprarenal insufficiency and bilateral masses with heterogeneous contrast uptake in the adrenal glands were documented on a thoracoabdominopelvic CT. Infectious causes and functioning tumors were excluded. After an exhaustive study, DLBCL was diagnosed. Through this case report, the authors intend to sound the alert on the existence of a rare presentation of DLBCL.

Patient-derived ovarian cancer explants: preserved viability and histopathological features in long-term agitation-based cultures.

Ovarian carcinoma (OvC) remains a major therapeutic challenge due to its propensity to develop resistance after an initial response to chemotherapy. Interactions of tumour cells with the surrounding microenvironment play a role in tumour survival, invasion capacity and drug resistance. Cancer models that retain tissue architecture and tumour microenvironment components are therefore essential to understand drug response and resistance mechanisms. Herein, our goal was to develop a long-term OvC patient-derived explant (OvC-PDE) culture strategy in which architecture and cell type heterogeneity of the original tumour would be retained. Samples from 25 patients with distinct OvC types and one with a benign tumour, were cultured for 30 days in agitation-based culture systems with 100% success rate. OvC-PDE cultures retained the original tumour architecture and main cellular components: epithelial cells, fibroblasts and immune cells. Epithelial cells kept their original levels of proliferation and apoptosis. Moreover, the major extracellular components, such as collagen-I and -IV, were retained in explants. OvC-PDE cultures were exposed to standard-of-care chemotherapeutics agents for 2 weeks, attesting the ability of the platform for drug assays employing cyclic drug exposure regimens. We established an OvC-PDE dynamic culture in which tumour architecture and cell type heterogeneity were preserved for the different OvC types, replicating features of the original tumour and compatible with long-term drug exposure for drug efficacy and resistance studies.

Symptomatic Pneumocephalus as a Complication of Lumbar Epidural Anaesthesia.

INTRODUCTION: Lumbar epidural anaesthesia is a commonly used technique for analgesia during labour. One of the rare complications of this technique is pneumocephalus. CASE DESCRIPTION: We report the case of a 35-year-old female admitted to the Emergency Department with severe headache associated with fast head movements. Five days previously she had a eutocic delivery and lumbar epidural anaesthesia was performed. A brain computed tomography (CT) scan showed pneumocephalus and she was admitted to the hospital ward. A brain CT scan on the fourth day of hospitalization showed resolution of ventricular pneumocephalus. DISCUSSION: The most frequently occurring symptom with pneumocephalus is headache associated with fast brain motion resulting from air injection and meningeal irritation. When there is clinical suspicion of pneumocephalus, a brain CT scan should be performed for the diagnosis. LEARNING POINTS: Pneumocephalus is the presence of air in the intracranial cavity and its development after spinal or epidural anaesthesia is extremely infrequent.Headache that occurs in the setting of lumbar epidural anaesthesia should not be labelled as post-dural puncture headache.The suspicion of pneumocephalus, based on the characteristics of the headache, should be maintained to obtain an emergent brain CT scan.

Total digital workflow in the fabrication of a partial removable dental prostheses: A case report.

This report describes a clinical and laboratory protocol used in the fabrication of a removable partial dental prosthesis with a digital workflow in a 73-year-old patient. The metal framework was produced with a selective laser melting technique. For quality assurance and discrepancy analysis, the framework was superimposed and compared to the respective digital design file, and the printed model was also compared to the digital impression and rendered as a 3D colour map. Differences were detected in the framework on the clasp of tooth 17 (upper right second molar) and on the printed model on the interproximal surfaces of the abutments, particularly on tooth 17 and palatine area. The use of this digital workflow allowed for the achievement of an removable partial dental prosthesis with a good fit and occlusion with minimal adjustments, with the reduction of both clinical and laboratory time. Further studies are needed to gain a better understanding of these techniques.

Sphere-plane methodology to evaluate the wear of titanium of dental implants: a research proposal.

OBJECTIVE: Titanium is the most commonly used material to manufacture dental implants and abutments. Recently, zirconia abutments have been manufactured with better aesthetic properties. However, zirconia abutments are harder than titanium implants; therefore, they could wear the implant surface. Therefore, this article aims to describe a sphere-plane system that can be used to assess the wear that different abutment materials cause in the titanium of dental implants when submitted to cyclic loading. This method can be used to simulate the oral cavity, where the abutment (sphere) applies loads onto the implant (titanium plane). The spheres were made of different materials (titanium and zirconia), and the specimens were loaded for 4,000,000 cycles. The scar size and area on titanium planes were measured with stereoscopic images and analysed through profilometry. RESULTS: The wear of titanium planes was similar when tested against zirconia or titanium spheres. The sphere-plane system is a method that can be used to evaluate and quantify the wear of the titanium of dental implants, and compared with methods that use real implants, this system is simpler and less expensive. This method could facilitate further research to evaluate the wear of titanium against different materials and under different testing conditions.

Family rituals in pediatric epilepsy: Links to parental competence and adaptation.

This cross-sectional study examined the associations between family ritual meaning and 3 indicators of parental adaptation (anxious and depressive symptoms and quality of life) via 2 dimensions of parental competence-satisfaction and efficacy-in parents of children with epilepsy. Two hundred Portuguese parents of children diagnosed with epilepsy for at least 6 months completed self-report measures assessing the main study variables. Our results showed that when parents reported stronger family ritual meaning, they also reported higher levels of parental satisfaction and efficacy, which were in turn were associated with lower psychological distress (anxious and depressive symptoms) and better quality of life of parents. This pattern of results was significant regardless of the severity of children's epilepsy, age group, and family socioeconomic level. In the context of pediatric epilepsy, empirical evidence was found for the role of family ritual meaning in being directly and indirectly associated with parents' adaptation outcomes by higher levels of parental satisfaction and efficacy. Focusing intervention targets on what families naturally do and recognize as their own ritual meaning may be a favorable route to address those at risk of psychological distress and lower quality of life. (PsycINFO Database Record

The psychological costs of comparisons: Parents' social comparison moderates the links between family management of epilepsy and children's outcomes.

Parents play a key role in how children deal with epilepsy. When diagnosed with health conditions, people seek comparison information from fellow patients and families, and this information has consequences for how they evaluate their situation. This study examined the moderating role of parents' social comparison orientation in the associations between family management (parental perceptions of family life difficulties and child's daily life) and adaptation outcomes of children with epilepsy (HRQoL and perceived stigma). Participants included 201 dyads of children with epilepsy and either their mother or father. The results showed that when parents perceived higher difficulties managing their child's epilepsy and/or reported that their child was more affected by this condition, children reported higher perceived stigma and worse HRQoL only when parents had a higher social comparison orientation. Our results are innovative in showing that when parents have a higher social comparison orientation, their children may be at increased risk of poorer outcomes.