Experimenting with Pig-based Skin Model for Burns. Testing of Mean Literature Findings.

Introduction: Animal models that closely mimic human burn wound healing processes are essential for developing effective burn wound treatments. Pigs are useful animal models for studying burn wound healing. From their extensive literature review, Andrews and Cuttle (2017) reported mean temperature and exposure time values. This study was done to provide initial burn depth for another experiment comparing two burn wound treatments. The secondary goal was to validate a systematic review on porcine burn model standardization. Material and Methods: Six four-week-old Large White x Landrace gilts were housed in a closed structure for 10 days to acclimatize. The procedures were performed under general anesthesia. A round 2.5 cm copper plate welded to an aluminum rod with a wooden handle caused the injuries. The burning device was used to reach a contact temperature of 110 C on the pig's skin. The objective was to create a superficial partial thickness (SPT) burn for 10 seconds (Group 10s) and a deep partial thickness (DPT) burn for 20 seconds (Group 20s) using a plate heated at 110ºC. No stabilizer or pressure controller was used. Wounds were conclusively dressed and harvested 24 hours later. The usual hematoxylin-eosin protocol was used to cut and stain 4-micron sections. Results: A significant difference (p 0.01) was observed in dermis involvement, with a mean of 85.61 % (95% CI= 80.62 to 90.61) for group 10s and 123.71% (95% CI= 114.91 to 132.50) for group 20s. An exposure time of 20 seconds increased dermis depth-related total collagen denaturation by almost 50% compared to 10 seconds. Conclusions: In conclusion, our experiment produced DPT burns in 10 seconds and FT burns in 20 seconds without a pressure application device.

Experimenting with Pig-based Skin Model for Burns. Testing of Mean Literature Findings.

Introduction: Animal models that closely mimic human burn wound healing processes are essential for developing effective burn wound treatments. Pigs are useful animal models for studying burn wound healing. From their extensive literature review, Andrews and Cuttle (2017) reported mean temperature and exposure time values. This study was done to provide initial burn depth for another experiment comparing two burn wound treatments. The secondary goal was to validate a systematic review on porcine burn model standardization. Materials and Methods: Six four-week-old Large White x Landrace gilts were housed in a closed structure for 10 days to acclimatize. The procedures were performed under general anesthesia. A round 2.5 cm copper plate welded to an aluminum rod with a wooden handle caused the injuries. The burning device was used to reach a contact temperature of 110ºC on the pig's skin. The objective was to create a superficial partial thickness (SPT) burn for 10 seconds (Group 10s) and a deep partial thickness (DPT) burn for 20 seconds (Group 20s) using a plate heated at 110ºC. No stabilizer or pressure controller was used. Wounds were conclusively dressed and harvested 24 hours later. The usual hematoxylin-eosin protocol was used to cut and stain 4-micron sections. Results: A significant difference (p 0.01) was observed in dermis involvement, with a mean of 85.61 % (95% CI= 80.62 to 90.61) for group 10s and 123.71% (95% CI= 114.91 to 132.50) for group 20s. An exposure time of 20 seconds increased dermis depth-related total collagen denaturation by almost 50% compared to 10 seconds. Conclusions: In conclusion, our experiment produced DPT burns in 10 seconds and FT burns in 20 seconds without a pressure application device.

Acute Glomerulonephritis Following Systemic Scabies in Two Brothers.

Scabies is a parasitic infestation of the skin with high prevalence in crowded spaces. In some instances, scabies becomes the underlying factor for complicated skin-borne opportunistic pathogens infections in both children and adults. Geographic area and socio-economic factors are determinants of the endemic pattern of this disease. Currently, the treatment of scabies has been under special attention. A combination of oral therapy with Ivermectin and sulfur-based ointments are the gold standard. However, caution is required in patients with kidney impairment. The renal involvement in children with scabies is mainly caused by acute glomerulonephritis. The severity of the nephritic syndrome can lead to other complications. Also, Ivermectin possesses a nephrotoxic effect. Severe hypertension can lead to neurological complications. The aim of our case report is to present two unusual complications in brothers with scabies. We report the cases of two brothers with scabies who presented with severe skin infection that developed acute post infectious glomerulonephritis (APIGN). In addition, one of the brothers presented with posterior reversible encephalopathy syndrome (PRES). The other one developed acute tubule-interstitial acute kidney injury following Ivermectin administration. The evolution of skin lesions was favorable, and kidney function returned to normal in both brothers.

Histologic Evaluation of Early Papilla Healing after Augmentation with Injectable Hyaluronic Acid-A Proof of Concept.

Objectives: This human histological study's purpose was to histologically evaluate papillae's healing after hyaluronic acid (HA) gel augmentation at three healing time points after one injection with hyaDENT BG®. Methods: Fifteen papillae from two patients with stage III, grade B periodontitis have been selected for this study. Every week for three weeks, five papillae were injected once with HA gel, and during the fourth week, the papillae were surgically removed as part of step 3 of the periodontal treatment. The histological analysis was performed on fifteen papillae, with five papillae corresponding to every timepoint of healing (weeks 1, 2, and 3). The primary outcome was considered to be the newly formed collagen fibers. The presence of residual HA, the integrity of epithelium or the presence of erosions/ulcerations, the presence and characteristics of inflammatory infiltrate, the presence of granulomatous reactions, and interstitial edema were considered to be secondary outcomes. Results: From the first to the third week, newly formed connective tissue begins to appear, while the observed HA pools (vesicles) content decreases. The density of inflammatory infiltrate was higher in the first week after injection, decreasing considerably by week 3; however, it was still visible throughout the healing time points. A granulomatous reaction was present in only three samples, while no signs of ulceration or necrosis could be observed; however, epithelial erosions could be observed on some samples after the first week. Conclusions: Papila augmentation with hyaluronic acid promotes new collagen formation from the second week of healing despite some foreign body granulomatous reactions.

Clinical-Pathological Features of Thyroid Neoplasms in Young Patients Diagnosed in a Single Center.

Background and objectives: The aim of this study was to evaluate the clinical-pathological profile in young patients with thyroid cancer. Materials and methods: We realized a retrospective study on patients with thyroid neoplasms who underwent surgery at the "Pius Brinzeu" County Clinical Emergency Hospital in Timisoara, Romania. A comparative analysis of some parameters between two groups, young patients (<45 years) versus patients ≥45 years, was performed. Results: A total of 211 patients met the study inclusion criteria, mostly females (86.26%) with a female/male ratio of 6.81:1. In patients <45 years old (25.64%), papillary thyroid carcinoma was identified in 51.85% of cases; in 53.85% of cases, the tumor was >1 cm; 13.46% had extrathyroidal extension (p = 0.0430); 21.15% capsule invasion (p = 0.1756); 23.08% lympho-vascular invasion (p = 0.0048); and 13.46% of cases locoregional nodal invasion (p = 0.0092). Conclusions: Thyroid cancer in young people was associated with chronic lymphocytic thyroiditis and tumor progression parameters, identifying more cases of extrathyroidal extension, locoregional nodal invasion, lympho-vascular invasion and perineural invasion in young patients compared to older ones. For a better understanding of this pathology and to improve diagnosis and therapeutic management, more studies are needed for these patients.

Obscure Bleeding from a Metastatic Small Bowel Tumor Diagnosed Using Motorized Spiral Enteroscopy: A Case Study and a Literature Review.

Small bowel tumors are relatively rare, representing only around 5% of all gastrointestinal neoplasms, with a progressively increasing incidence. Currently, there are no established guidelines for diagnostic approaches, screening procedures, or management strategies for small bowel tumors. We present here the case of a patient with a rare type of metastatic tumor of the small bowel originating from primary lung adenocarcinoma who presented with abdominal pain, severe iron-deficiency anemia, and melena. The initial investigations, gastroscopy and colonoscopy, failed to identify the bleeding source. The obscure bleeding source and diagnosis were achieved through power motorized spiral enteroscopy (MSE), which allowed the visualization and biopsy of the tumor. Histopathological examination established the presence of a poorly differentiated non-mucinous adenocarcinoma originating from the lung. This case is reported to provide evidence of the efficiency of MSE in the diagnosis of small bowel tumors, with the method providing higher insertion depth in a reduced amount of time.

Feasibility of Conduction System Pacing in Patients with Baseline Bundle Branch Block-A Single-Center Mid-Term Follow-Up Study.

BACKGROUND: The primary prerequisite for a successful conduction system pacing (CSP) procedure is the integrity of the conduction system, which may be impaired if a baseline bundle branch block (BBB) is present. This study aimed to evaluate the feasibility and mid-term performance of permanent CSP in patients with baseline BBB and to compare the results between left bundle branch block (LBBB) and right bundle branch block (RBBB) patterns. MATERIAL AND METHODS: A total of 101 patients with typical BBB and an attempt at CSP were retrospectively reviewed. Procedural characteristics, pacing, sensing parameters, and complications at baseline and after a mid-term follow-up were analyzed. RESULTS: The global procedural success for CSP was 93%. His bundle pacing (HBP) had a significantly lower success rate than left bundle branch area pacing (LBBAP) (50.5% vs. 86%). The paced QRS duration was significantly narrower with HBP. The pacing and sensing thresholds were significantly better with LBBAP. Procedural complications occurred only in the LBBAP group (two acute perforations in the LV cavity and one acute chest pain during lead fixation) without long-term sequelae. The HBP and the LBBAP procedural success rates were higher in the RBBB versus the LBBB group (62.5% vs. 44.9% and 100% vs. 81.5%, respectively). Baseline QRS duration, atrial volumes, and right ventricular diameters were significantly associated with HBP procedural failure. The follow-up pacing and sensing thresholds were similar to the baseline values for all pacing methods and BBB morphology. Only one device-related complication leading to pacing interruption was recorded. CONCLUSION: In patients with bundle branch blocks, CSP is a feasible procedure associated with a high success rate, stable pacing and sensing parameters, and low complication rates over a mid-term follow-up.

Colorectal Carcinomas: Searching for New Histological Parameters Associated with Lymph Node Metastases.

Background and Objectives: Colorectal cancer (CRC) continues to be an essential public health problem. Our study aimed to evaluate the prognostic significance of classic prognostic factors and some less-studied histopathological parameters in CRC. Materials and Methods: We performed a retrospective study on 71 colorectal carcinoma patients who underwent surgery at the "Pius Brînzeu" County Clinical Emergency Hospital in Timișoara, Romania. We analyzed the classic parameters but also tumor budding (TB), poorly differentiated clusters (PDCs) of cells, tumor-infiltrating lymphocytes (TILs), and the configuration of the tumor border on hematoxylin-eosin slides. Results: A high degree of malignancy (p = 0.006), deep invasion of the intestinal wall (p = 0.003), an advanced stage of the disease (p < 0.0001), lymphovascular invasion (p < 0.0001), perineural invasion (p < 0.0001), high-grade TB (p < 0.0001), high-grade PDCs (p < 0.0001), infiltrative tumor border configuration (p < 0.0001) showed a positive correlation with lymph node metastases. Conclusions: The analyzed parameters positively correlate with unfavorable prognostic factors in CRC. We highlight the value of classic prognostic factors along with a series of less-known parameters that are more accessible and easier to evaluate using standard staining techniques and that could predict the risk of relapse or aggressive evolution in patients with CRC.

Invasive Cutaneous Melanoma: Evaluating the Prognostic Significance of Some Parameters Associated with Lymph Node Metastases.

Background and Objectives: This study aimed to assess the clinical-pathological profile of patients with invasive cutaneous melanomas and to identify the parameters with a prognostic role in the lymph nodal spread of this malignant tumor. Materials and Methods: We performed a retrospective study on patients with invasive cutaneous melanomas who underwent surgery in the "Pius Brînzeu" County Clinical Emergency Hospital from Timișoara, Romania, and were evaluated for the status of loco-regional lymph nodes. We selected and analyzed some parameters searching for their relationship with lymph node metastases. Results: We identified 79 patients with invasive cutaneous melanomas (29 men and 50 women, mean age 59.36 years). A percentage of 58.3% of melanomas had Breslow tumor thickness >2 mm; 69.6% of melanomas showed a Clark level IV-V. Tumor ulceration was present in 59.5% of melanomas. A mitotic rate of ≥5 mitoses/mm2 was observed in 48.1% of melanomas. Tumor-infiltrating lymphocytes (TILs), non-brisk, were present in 59.5% of cases and 22.8% of patients had satellite/in-transit metastasis (SINTM). Tumor regression was identified in 44.3% of cases. Lymph nodes metastases were found in 43.1% of patients. Statistical analysis showed that lymph node metastases were more frequent in melanomas with Breslow thickness >2 mm (p = 0.0002), high Clark level (p = 0.0026), mitotic rate >5 mitoses/mm2 (p = 0.0044), ulceration (p = 0.0107), lymphovascular invasion (p = 0.0182), SINTM (p = 0.0302), and non-brisk TILs (p = 0.0302). Conclusions: The Breslow thickness >2 mm, high Clark level, high mitotic rate and ulceration are the most important prognostic factors for lymph nodal spread in cutaneous melanomas. However, some melanomas without these clinical-pathological features can have an unexpected, aggressive evolution, which entails the necessity of close and prolonged clinical follow-up of patients, including those with lesions considered without risk.

Comparison of Type I and Type III Collagen Concentration between Oreochromis mossambicus and Oreochromis niloticus in Relation to Skin Scaffolding.

Background and Objectives: Skin scaffolding can be done using allografts and autografts. As a biological allograft, the skin of Oreochromis niloticus (ON) has been used due to its high type I and III collagen content. Oreochromis mossambicus (OM) is also a member of the Oreochromis family, but not much is known regarding its collagen content. As such, this study aimed to assess and compare the collagen content of the two fish species. Materials and Methods: This is a crossover study comparing the skin collagen contents of the two fish. Young fish were chosen, as they tend to have higher collagen concentrations. The skin samples were sterilized in chlorhexidine and increasing glycerol solutions and analyzed histochemically with Sirius red picrate under polarized light microscopy. Results: 6 young ON and 4 OM specimens were used. Baseline type I collagen was higher for OM, but at maximum sterilization it was higher for ON, with no differences in between Type III collagen was higher for OM across all comparisons with the exception of the last stage of sterilization. Generally, collagen concentrations were higher in highly sterilized samples. Conclusions: OM skin harvested from young fish, with its greater collagen III content may be a better candidate for use as a biological skin scaffold in the treatment of burn wounds, compared to ON.

Influence of Section Thickness on the Accuracy and Specificity of Histometric Parameters Using Confocal Laser Scanning Microscopy in a Canine Model of Experimental Peri-Implantitis-A Proof of Concept.

OBJECTIVES: Tissue breakdown was assessed by confocal laser scanning microscopy (CLSM) using autofluorescence around implants with ligatures, on a dog hemimandible. Influence of section thickness on the accuracy of histometrical observations was also evaluated, in comparison with thin sections in light microscopy. MATERIAL AND METHODS: Three months after tooth extraction, implants were placed. Two months after abutment placement, ligatures were placed with no plaque control. 11 months post-implantation, the animal was sacrificed. Undecalcified thin (30 µm) sections were cut, stained and evaluated by light microscopy to be used as a reference. Additional sections were performed, so that another pair of unstained thick sections resulted (250-300 µm). Tissue loss was assessed using histomorphometric parameters under CLSM and was compared to the light microscopy reference ones. RESULTS: Morphometry confirmed tissue loss more pronounced on the "thick" and quick sections, when compared to the time-consuming and technique-sensitive "thin" ones. CONCLUSIONS: Within the limits of the present study, the adequacy of histometrical observations under CLSM reveal commensurable information about soft-tissue-bone-implant details, when compared to traditional light microscopy histological protocols. The CLSM investigation may seem demanding, yet the richness of data acquired may justify this approach, provided seatbacks caused by improper manipulation of "thick" sections are avoided.

Relationship between Novel Elastography Techniques and Renal Fibrosis-Preliminary Experience in Patients with Chronic Glomerulonephritis.

INTRODUCTION: A renal biopsy represents the gold standard in the diagnosis, prognosis, and management of patients with glomerulonephritis. So far, non-invasive elastographic techniques have not confirmed their utility in replacing a biopsy; however, the new and improved software from Hologic Supersonic Mach 30 is a promising method for assessing the renal tissue's stiffness and viscosity. We investigated whether this elastography technique could reveal renal tissue fibrosis in patients with chronic glomerulonephritis. MATERIALS AND METHODS: Two-dimensional-shear wave elastography (SWE) PLUS and viscosity plane-wave ultrasound (Vi PLUS) assessments were performed in 40 patients with chronic glomerulopathies before being referred for a renal biopsy. For each kidney, the mean values of five stiffness and viscosity measures were compared with the demographic, biological, and histopathological parameters of the patients. RESULTS: In total, 26 men and 14 women with a mean age of 52.35 ± 15.54 years, a mean estimated glomerular filtration rate (eGFR) of 53.8 ± 35.49 mL/min/1.73m2, and a mean proteinuria of 6.39 ± 7.42 g/24 h were included after providing their informed consent. Out of 40 kidney biopsies, 2 were uninterpretable with inappropriate material and were divided into four subgroups based on their fibrosis percentage. Even though these elastography techniques were unable to differentiate between separate fibrosis stages, when predicting between the fibrosis and no-fibrosis group, we found a cut-off value of <20.77 kPa with the area under the curve (AUC) of 0.860, a p < 0.001 with 88.89% sensitivity, and a 75% specificity for the 2D SWE PLUS measures and a cut-off value of <2.8 Pa.s with an AUC of 0.792, a p < 0.001 with 94% sensitivity, and a 60% specificity for the Vi PLUS measures. We also found a cut-off value of <19.75 kPa for the 2D SWE PLUS measures (with an AUC of 0.789, p = 0.0001 with 100% sensitivity, and a 74.29% specificity) and a cut-off value of <1.28 Pa.s for the Vi PLUS measures (with an AUC 0.829, p = 0.0019 with 60% sensitivity, and a 94.29% specificity) differentiating between patients with over 40% fibrosis and those with under 40%. We also discovered a positive correlation between the glomerular filtration rate (eGFR) and 2D-SWE PLUS values (r = 0.7065, p < 0.0001) and Vi PLUS values (r = 0.3637, p < 0.0211). C reactive protein (CRP) correlates with the Vi PLUS measures (r = -0.3695, p = 0.0189) but not with the 2D SWE PLUS measures (r = -0.2431, p = 0.1306). CONCLUSION: Our findings indicate that this novel elastography method can distinguish between individuals with different stages of renal fibrosis, correlate with the renal function and inflammation, and are easy to use and reproducible, but further research is needed for them to be employed routinely in clinical practice.

Histomorphometrical and CBCT Evaluation of Tissue Loss Progression Induced by Consecutive, Alternate Ligatures in Experimental Peri-Implantitis in a Dog Model: A Pilot Study.

OBJECTIVES: Soft and hard tissue breakdown was histologically and radiologically assessed around implants with alternate, consecutively placed ligatures on the same edentulous dog hemimandible. The influence of ligatured implants (LI) on adjacent non-ligatured implants (NLI, as a possible naturally induced peri-implantitis) was also evaluated. MATERIAL AND METHODS: Three months after tooth extraction, five dental implants were placed in the dog hemimandible. Two months after abutment placement, ligatures were placed subsequently two months apart on alternate implants, while both intermediate implants were left without ligatures. Ligatures were kept in place during the entire experiment, and no plaque control measures were taken. Eleven months post-implantation, the animal was sacrificed. Undecalcified ground sections were cut, stained with Masson Goldner and MOVAT Pentachrome and evaluated by light microscopy. Soft and hard tissue loss was assessed using histomorphometric and CBCT parameters. RESULTS: All NLI presented deep false peri-implant pockets on the oral aspect and pronounced vertical bone resorption on the buccal aspect. After 2, 4 and 6 months, during the breakdown period, more than 30% of the bone was lost in LI in all directions, while, despite immediate vicinity, NLI displayed less destruction. Intense inflammation, typical for induced peri-implantitis, was present, with similar intensity in LI as NLI, but in different parts of the lesions. Morphometry confirmed intense soft tissue inflammation, more bone resorption and higher amounts of infiltrated connective tissue in LI when compared with NLI. CONCLUSION: Within the limits of the present pilot study, the adequacy of the experimental dog model based on ligature-induced peri-implantitis was able to be successfully challenged by non-ligature models of spontaneously occurring peri-implant inflammation, while meeting the requirements for experimental designs with a very small numbers of animals. The influence of implants with severe peri-implantitis on adjacent implants resulted in less than expected tissue loss in the latter accession numbers.

Poorly differentiated clusters and tumor budding are important prognostic factors in colorectal carcinomas.

The aim of our study was to assess the prognostic value of the two new grading systems based on the quantification of tumor budding - TB (GBd) and poorly differentiated clusters - PDCs (PDCs-G) in colorectal carcinomas (CRC). We performed a retrospective study on 71 CRC patients who underwent surgery at the Emergency County Hospital, Timișoara. CRC cases were classified based on haematoxylin-eosin slides, using the conventional grading system, GBd and PDCs-G, respectively. We used two-tier and three-tier grading schemes for each system. Subsequently,  we evaluated  associations with other prognostic factors in CRC. Based on the three-tier GBd (GBd-3t)  most cases (34/69, 49.27%) were classified as G3Bd-3t, while based on the conventional grading system, the majority of the cases (55/69, 79.71%) were considered G2. On the other hand, based on the three-tier PDCs-G system (PDCs-G-3t), most cases (31/69, 44.93%) were PDCs-G2-3t. We also noted a more significant association of GBd-3t with other prognostic parameters analyzed, as compared to the conventional grading system. Nodal status, tumor stage, and lymphovascular invasion were strongly correlated with GBd-3t (p=0.0001). Furthermore, we noted that PDCs-G-3t correlated more significantly than the conventional grading system with nodal status (p<0.0001), tumor stage (p=0.0003), lymphovascular invasion (p<0.0001), perineural invasion (p=0.005) and the tumor border configuration (p<0.0001). High GBd and PDCs-G grades correlate directly with other negative prognostic factors in CRC.Thus, these new parameters/classification methods could be used as additional tools for risk stratification in patients with CRC.

Three-Dimensional Virtual Pathology Specimens: Decrease in Student Performance upon Switching to Digital Models.

Several alternatives to formalin-stored physical specimens have been described in medical literature, but only a few studies have addressed the issue of learning outcomes when these materials were employed. The aim of this study was to conduct a prospective controlled study to assess student performance in learning anatomic pathology when adding three-dimensional (3D) virtual models as adjunct teaching materials in the study of macroscopic lesions. Third-year medical students (n = 501) enrolled at the Victor Babes University of Medicine and Pharmacy in Timisoara, Romania, were recruited to participate. Student performance was assessed through questionnaires. Students performed worse with new method, with poorer results in terms of overall (mean 77.6% ±SD 11.8% vs. 83.6% ±10.5) and individual question scores (percentage of questions with maximum score 34.6% ±25.6 vs. 47.7 ± 24.6). This decreased performance was generalizable, as it was observed across all language divisions and was independent of the teaching assistant involved in the process. In an open-ended feedback evaluation of the new 3D specimens, most students agreed that the new method was better, bringing arguments both for and against these models. Although subjectively the students found the novel teaching materials to be more helpful, their learning performance decreased. A wider implementation as well as exposure to the technique and use of virtual specimens in medical teaching could improve the students' performance outcome by accommodating the needs for novel teaching materials for digital natives.

Comparing PRP and bone marrow aspirate effects on cartilage defects associated with partial meniscectomy: a confocal microscopy study on animal model.

AIM: The aim of our study was to assess the therapeutic effects of platelet-rich plasma (PRP) and bone marrow aspirate concentrate (BMAC) in an animal knee lesion complex associating a large osteochondral defect and meniscal defect resulted from partial meniscectomy, a clinical situation that occurs quite often in orthopedic practice. MATERIALS AND METHODS: Twenty-one male rabbits were included in the study, and all underwent initial surgery on the right knee to create the osteochondral defect on the internal femoral condyle, and remove the anterior horn of the internal meniscus, simulating a clinical situation. Rabbits were separated in three study groups: control, PRP group, in which three PRP injections were administered, and BMAC group, in which one single BMAC injection was administered. At the end of the six months follow-up period, knees were harvested and further analyzed using confocal microscopy and three-dimensional (3D) reconstruction of the articular surface. RESULTS: Therapeutic groups had better results concerning articular surface remodeling and joint degeneration indicators in comparison to trauma group. CONCLUSIONS: Our results suggest that using post-operative regenerative therapies does improve final results concerning surface contact remodeling that was investigated using confocal microscopy and should be considered a valid treatment adjuvant in managing patients with this type of lesion complex, as it improves global joint outcome.

Long Non-Coding RNA Expression in Laser Micro-Dissected Luminal A and Triple Negative Breast Cancer Tissue Samples-A Pilot Study.

Background and Objectives: Breast cancer (BC) remains one of the major causes of cancer death in women worldwide. The difficulties in assessing the deep molecular mechanisms involved in this pathology arise from its high complexity and diverse tissue subtypes. Long non-coding RNAs (lncRNAs) were shown to have great tissue specificity, being differentially expressed within the BC tissue subtypes. Materials and Methods: Herein, we performed lncRNA profiling by PCR array in triple negative breast cancer (TNBC) and luminal A tissue samples from 18 BC patients (nine TNBC and nine luminal A), followed by individual validation in BC tissue and cell lines. Tissue samples were previously archived in formalin-fixed paraffin-embedded (FFPE) samples, and the areas of interest were dissected using laser capture microdissection (LCM) technology. Results: Two lncRNAs (OTX2-AS1 and SOX2OT) were differentially expressed in the profiling analysis (fold change of 205.22 and 0.02, respectively, p < 0.05 in both cases); however, they did not reach statistical significance in the individual validation measurement (p > 0.05) when analyzed with specific individual assays. In addition, GAS5 and NEAT1 lncRNAs were individually assessed as they were previously described in the literature as being associated with BC. GAS5 was significantly downregulated in both TNBC tissues and cell lines compared to luminal A samples, while NEAT1 was significantly downregulated only in TNBC cells vs. luminal A. Conclusions: Therefore, we identified GAS5 lncRNA as having a differential expression in TNBC tissues and cells compared to luminal A, with possible implications in the molecular mechanisms of the TNBC subtype. This proof of principle study also suggests that LCM could be a useful technique for limiting the sample heterogeneity for lncRNA gene expression analysis in BC FFPE tissues. Future studies of larger cohort sizes are needed in order to assess the biomarker potential of lncRNA GAS5 in BC.

Vitamin D alleviates oxidative stress in adipose tissue and mesenteric vessels from obese patients with subclinical inflammation.

Obesity is an age-independent, lifestyle-triggered, pandemic disease associated with both endothelial and visceral adipose tissue (VAT) dysfunction leading to cardiometabolic complications mediated via increased oxidative stress and persistent chronic inflammation. The purpose of the present study was to assess the oxidative stress in VAT and vascular samples and the effect of in vitro administration of vitamin D. VAT and mesenteric artery branches were harvested during abdominal surgery performed on patients referred for general surgery (n = 30) that were randomized into two subgroups: nonobese and obese. Serum levels of C-reactive protein (CRP) and vitamin D were measured. Tissue samples were treated or not with the active form of vitamin D: 1,25(OH)2D3 (100 nmol/L, 12 h). The main findings are that in obese patients, (i) a low vitamin D status was associated with increased inflammatory markers and reactive oxygen species generation in VAT and vascular samples and (ii) in vitro incubation with vitamin D alleviated oxidative stress in VAT and vascular preparations and also improved the vascular function. We report here that the serum level of vitamin D is inversely correlated with the magnitude of oxidative stress in the adipose tissue. Ex vivo treatment with active vitamin D mitigated obesity-related oxidative stress.

Monoamine oxidase is a source of oxidative stress in obese patients with chronic inflammation 1.

Obesity is an important preventable risk factor for morbidity and mortality from cardiometabolic disease. Oxidative stress (including in visceral adipose tissue) and chronic low-grade inflammation are the major underlying pathomechanisms. Monoamine oxidase (MAO) has recently emerged as an important source of cardiovascular oxidative stress. The present study was conducted to evaluate the role of MAO as contributor to reactive oxygen species (ROS) production in white adipose tissue and vessels harvested from patients undergoing elective abdominal surgery. To this aim, visceral adipose tissue and mesenteric artery branches were isolated from obese patients with chronic inflammation and used for organ bath, ROS production, quantitative real-time PCR, and immunohistology studies. The human visceral adipose tissue and mesenteric artery branches contain mainly the MAO-A isoform, as shown by the quantitative real-time PCR and immunohistology experiments. A significant upregulation of MAO-A, the impairment in vascular reactivity, and increase in ROS production were found in obese vs. non-obese patients. Incubation of the adipose tissue samples and vascular rings with the MAO-A inhibitor (clorgyline, 30 min) improved vascular reactivity and decreased ROS generation. In conclusion, MAO-A is the predominant isoform in human abdominal adipose and vascular tissues, is overexpressed in the setting of inflammation, and contributes to the endothelial dysfunction.

Poorly differentiated clusters: prognostic significance in colorectal carcinomas immunohistochemistry images.

Colorectal cancer (CRC) remains a major public health burden worldwide, despite increased knowledge on its pathogenesis and advances in therapy. We aimed to evaluate a new histological grading system based on poorly differentiated clusters (PDCs) counting - the PDCs grade (PDCs-G), and its clinicopathological and prognostic significance, compared to the World Health Organisation (WHO) grading system (WHO grade). We reviewed 71 surgical resection specimens for CRC from the Emergency County Hospital "Pius Brînzeu" Timisoara. The cases were graded using the WHO grade and the PDCs-G, with further analysis of their association with the other recognised prognostic parameters. Using the WHO grade, 9% of the analysed cases were G1, 80% G2, 11% G3, and none of the tumours was graded G4, while in the PDCs-G 16% were G1, 45% G2, and 39% G3. In multivariate analysis PDCs-G was significantly associated with the American Joint Committee on Cancer stage of the disease (AJCC stage) (p = 0.0003), depth of invasion (pT) (p = 0.0084), nodal status (LNM) (p < 0.0001), lymphovascular invasion (LVI) (p < 0.0001), perineural invasion (PNI) (p < 0.0052), and tumour border configuration (p < 0.0001). The novel grading system based on PDCs counting is an additional histological tool in the evaluation of CRC and a promising new prognostic factor for these patients.