Molecular mechanisms of carfilzomib-induced cardiotoxicity in mice and the emerging cardioprotective role of metformin.

Carfilzomib (Cfz), an irreversible proteasome inhibitor licensed for relapsed/refractory myeloma, is associated with cardiotoxicity in humans. We sought to establish the optimal protocol of Cfz-induced cardiac dysfunction, to investigate the underlying molecular-signaling and, based on the findings, to evaluate the cardioprotective potency of metformin (Met). Mice were randomized into protocols 1 and 2 (control and Cfz for 1 and 2 consecutive days, respectively); protocols 3 and 4 (control and alternate doses of Cfz for 6 and 14 days, respectively); protocols 5A and 5B (control and Cfz, intermittent doses on days 0, 1 [5A] and 0, 1, 7, and 8 [5B] for 13 days); protocols 6A and 6B (pharmacological intervention; control, Cfz, Cfz+Met and Met for 2 and 6 days, respectively); and protocol 7 (bortezomib). Cfz was administered at 8 mg/kg (IP) and Met at 140 mg/kg (per os). Cfz resulted in significant reduction of proteasomal activity in heart and peripheral blood mononuclear cells in all protocols except protocols 5A and 5B. Echocardiography demonstrated that Cfz led to a significant fractional shortening (FS) depression in protocols 2 and 3, a borderline dysfunction in protocols 1 and 4, and had no detrimental effect on protocols 5A and 5B. Molecular analysis revealed that Cfz inhibited AMPKα/mTORC1 pathways derived from increased PP2A activity in protocol 2, whereas it additionally inhibited phosphatidylinositol 3-kinase/Akt/endothelial nitric oxide synthase pathway in protocol 3. Coadministration of Met prevented Cfz-induced FS reduction and restored AMPKα phosphorylation and autophagic signaling. Conclusively, Cfz decreased left ventricular function through increased PP2A activity and inhibition of AMPKα and its downstream autophagic targets, whereas Met represents a novel promising intervention against Cfz-induced cardiotoxicity.

Canagliflozin attenuates the progression of atherosclerosis and inflammation process in APOE knockout mice.

BACKGROUND: Sodium glucose co-transporter2 inhibitors reduce the incidence of cardiovascular events in patients with type 2 diabetes mellitus based on the results of recent cardiovascular outcome studies. Herein, we investigated the effects of long-term treatment with canagliflozin on biochemical and immunohistochemical markers related to atherosclerosis and atherosclerosis development in the aorta of apolipoprotein E knockout (Apo-E(-/-)) mice. METHODS: At the age of 5 weeks, mice were switched from normal to a high-fat diet. After 5 weeks, Apo-E(-/-) mice were divided into control-group (6 mice) treated with 0.5% hydroxypropyl methylcellulose and Cana-group (7 mice) treated with canagliflozin (10 mg/kg per day) per os. After 5 weeks of intervention, animals were sacrificed, and heart and aorta were removed. Sections stained with hematoxylin-eosin (H&E) were used for histomorphometry whereas Masson's stained tissues were used to quantify the collagen content. Immunohistochemistry to assess MCP-1, CD68, a-smooth muscle actin, MMP-2, MMP-9, TIMP-1 and TIMP-2 expression was carried out and q-PCR experiments were performed to quantify mRNA expression. RESULTS: Canagliflozin-group mice had lower total-cholesterol, triglycerides and glucose levels (P < 0.01), while heart rate was significantly lower (P < 0.05). Histomorphometry revealed that one in seven Cana-group mice versus four in six control mice developed atheromatosis, while aortic root plaque was significantly less, and collagen was 1.6 times more intense in canagliflozin-group suggesting increased plaque stability. Immunohistochemistry revealed that MCP-1 was significantly less expressed (P < 0.05) in the aortic root of canagliflozin-group while reduced expression of a-actin and CD68 was not reaching significance (P = 0.15). VCAM-1 and MCP-1 mRNA levels were lower (P = 0.02 and P = 0.07, respectively), while TIMP-1/MMP-2 ratio expression was higher in canagliflozin-group approaching statistical significance (P = 0.07). CONCLUSIONS: Canagliflozin attenuates the progression of atherosclerosis, reducing (1) hyperlipidemia and hyperglycemia, and (2) inflammatory process, by lowering the expression of inflammatory molecules such as MCP-1 and VCAM-1. Moreover, canagliflozin was found to increase the atherosclerotic plaque stability via increasing TIMP-1/MMP-2 ratio expression.

Mammary gland epithelial changes in thyroidectomized female rats.

PURPOSE: Numerous clinical studies have addressed the relationship of hypothyroidism and breast cancer with conflicting results. In the present experimental study we sought to determine whether absolute hypothyroidism established for a long period of time leads to epithelial alterations of the mammary gland. METHODS: Thirty five female Wistar rats were allocated to be subjected to either thyroidectomy (N=20) or not (N=15). The rats were kept alive for a period of 3 months in a weather controlled environment. Serum T3, T4, follicular stimulating hormone (FSH) and estradiol levels were measured at baseline and 10 days after thyroidectomy. Mammary glands were obtained at the end of the experiment and reviewed by an expert pathologist. RESULTS: Both serum FSH and estradiol levels were lower 10 days after thyroidectomy; however, only FSH values were significantly lower in the thyroidectomized animals. Pathological analysis revealed significantly increased atrophy and periductal fibrosis of the mammary gland among thyroidectomized animals. CONCLUSION: This is the first in vivo experimental study that reveals an association between the thyroid and mammary glands. Future studies should address the proteomic relationship that connects them.

Transarterial embolization with sorafenib in animal livers: a pharmacokinetics study.

PURPOSE: To assess the safety and feasibility of the targeted delivery of the antiangiogenic drug sorafenib to the liver using transarterial chemoembolization methodology as a novel approach to hepatocellular carcinoma (HCC) therapy. MATERIALS AND METHODS: Seven healthy New Zealand white rabbits were used in the study. After placement of a catheter in the common hepatic artery, six rabbits were treated with chemoembolization of sorafenib in iodized oil (Lipiodol) (sorafenib dose 0.1 mg/kg), and one rabbit received Lipiodol only. Liquid chromatography tandem mass spectrometry was used to measure the concentration of sorafenib in the peripheral blood and liver tissue 24 hours and 72 hours after treatment. Histochemical staining of the liver sections and biochemical measurements were performed. RESULTS: The administration of sorafenib in Lipiodol emulsions by transarterial chemoembolization resulted in sorafenib concentrations of 794 ng/g ± 240 and 64 ng/g ± 15 in the liver tissue 24 hours and 72 hours after treatment. The average liver-to-serum ratios 24 hours and 72 hours after treatment were approximately 14 and 22. The histochemical staining of the liver tissue sections and aspartate aminotransferase, alanine aminotransferase, γ-glutamyltransferase and total bilirubin concentrations indicated no significant liver damage. CONCLUSIONS: Transarterial chemoembolization with sorafenib in Lipiodol is an effective methodology for the localized delivery of this drug to the liver and has possible practical implications in therapeutic interventions for the treatment of hepatocellular carcinoma.

Comparative Analysis of Three Workload Measurement Methodologies in Surgical Pathology: Conclusions and Implications on Public Health Care and Costing of Pathology Services.

OBJECTIVES: To carry out a comparative analysis between 3 different workload measurement systems in surgical pathology: the Resource-Based Relative Value Scale (RBRVS), the Level 4 Equivalent (L4E), and the Automatable Activity-Based Approach to Complexity Unit Scoring (AABACUS). The RBRVS is one of the most widely used systems in terms of attempting to measure workload, whereas it has been proposed as a means of costing (and thus setting reimbursement rates) of surgical pathology services in Greece, despite being widely criticized for its inaccurate design. METHODS: Surgical pathology workload for 1 representative month at Evaggelismos General Hospital was assessed using both the RBRVS and the 2 newer methods. RESULTS: Pearson correlation showed a high level of correlation (0.902, P < .01) between the L4E and AABACUS but less so between either of those and the RBRVS (0.712 and 0.626, respectively; P < .01). The highest level of discrepancy was observed in the subspecialties of genitourinary, breast, dermatopathology, and gastrointestinal pathology. In addition, total and average working hours as calculated by the RBRVS were significantly lower compared with the other 2 systems. CONCLUSIONS: The RBRVS tends to underestimate actual workload as a result of its inability to take specific workload parameters into account, such as slide count or the need for intradepartmental consultation.

Case report: skeletal muscle metastasis from follicular thyroid carcinoma presenting as synovial sarcoma.

Differentiated thyroid carcinomas tend to remain localized and usually are of slow progression with excellent long-term survival. The major sites of distant metastases are cervical lymph nodes, lungs and bones and the minor sites include the brain, liver, pericardium, skin, kidney, pleura and muscle. Skeletal muscle metastases from differentiated thyroid carcinoma, are exceedingly rare. In this report, a 42-year-old woman with follicular thyroid cancer that had had a total thyroidectomy and radioiodine ablation nine years ago was presented with a painful right thigh mass and negative PET/CT scan. The patient had also lung metastases during the follow-up period which were treated with surgery, chemotherapy and radiation therapy. An MRI scan of the right thigh showed a deep-seated lobulated mass with cystic regions, bleeding elements and strong heterogeneous post contrast administration enhancement. Due to the similarities in clinical manifestations and imaging features between soft tissue tumors and skeletal muscle metastases, the case was initially misdiagnosed in favor of synovial sarcoma. Histopathological, immunohistochemistry and molecular analysis of the soft tissue mass confirmed to be a thyroid metastasis and, as a result, a final diagnosis of skeletal muscle metastasis was provided. Even though the probability of a skeletal muscle metastasis from thyroid cancer approaches zero, this study aims to raise the awareness to the medical community that these events do in fact occur in the clinical setting and should be considered in the differential diagnosis of patients with thyroid carcinomas.

Treatment of an unresectable cardiac angiosarcoma using paclitaxel and image guided radiotherapy: Case report and literature review.

Primary cardiac angiosarcomas (PCAs) are rare tumors that are typically found in the right atrium between the third and the fifth decade of life. While surgical removal of the tumor combined with adjuvant chemotherapy and/or radiotherapy is the treatment of choice, most of the patients present with unresectable tumors and metastatic disease carrying a dismal prognosis with a median survival of less than 1 year. Doxorubicin and ifosfamide based chemotherapy combined with radiotherapy is currently employed in these patients, but no standardized treatment algorithms exist. In this report, we present the management of a patient with an unresectable PCA treated using weekly paclitaxel (120 mg) combined with radiotherapy (60 Gy in 30 fractions) delivered by a helical TomoTherapy system. Follow-up imaging studies showed a remarkable tumor regression which allowed for surgical excision of the tumor 10 months post treatment. Histopathological examination of the resected mass showed no viable tumor cell. On the latest follow-up study, 12 months post treatment, no sign of disease progression (local or distant) was found, and the patient is in good clinical condition.

Understanding the Role of Radio-Sensitizing Nanoparticles in Enhancing Pathologic Response in Soft Tissue Sarcomas.

High-atomic-number (Z) nanoparticles produce a cascade of low-energy secondary electrons and characteristic X-rays when ionized by X-ray irradiation. These secondary particles deposit their energy in the vicinity of the nanoparticles and, provided that the latter are selectively accumulated within tumor cells, this results in increased DNA damage and tumor cell deaths. This study reviews the utilization of high-Z nanoparticles in the treatment of soft tissue sarcomas (STS). Both in vitro and in vivo experiments demonstrated that the dose is enhanced by approximately 1.2 when polyethelyne glycol (PEG)-modified gold nanoparticles, and from 1.4 to 1.8 when hafnium oxide nanoparticles (NBTXR3, Nanobiotix SA, France) are introduced into tumor cells and activated by X-ray beams. In a phase 2/3 clinical trial investigating the therapeutic benefit of using nanoparticles in preoperative external beam radiotherapy for locally advanced STS, the proportion of patients with a pathological complete response in their resected tumor was doubled when NBTXR3 nanoparticles were used. Additionally, a higher percentage of patients with complete tumor resection was observed in the NBTXR3 plus radiotherapy group. Similar toxicity profiles were found for both the NBTXR3 plus radiotherapy and the radiotherapy alone patient groups. The incorporation of radio-sensitizing nanoparticles in the preoperative radiotherapy of STS could enhance treatment outcomes.

"Starting from the Image": A Tele-pathology Pre-graduate Course Aimed at Motivating Medical Students.

In the tele-course entitled "Starting from the image", medical students are confronted with practical tasks in relevant professional contexts. Initially, a macroscopic or microscopic image of a patient case is presented to learners who then receive relevant information on the patient's history, clinical findings, and other laboratory tests. A pathologist actively discusses the pathological findings; then, a clinician explains their implications for the patient's individualized treatment and prognosis. In this way, pathology's interaction with other medical specialties is highlighted. Students declared that through these simulated professional practice experiences, they strengthened their decision-making skills. Educators should consider upgrading from information-based teaching to practice-focused instruction.

Delivery of Porphyrins Through Self-Assembling Peptide Hydrogels for Accelerated Healing of Experimental Skin Defects In Vivo.

INTRODUCTION: The care and healing of skin defects resulting from different causes has been the object of research to achieve rapid and complete skin regeneration. Hydrogels have been used for their ability to maintain hydration during wound healing, absorb wound exudate, and cover the underlying tissue without adherence while being transparent. In this study, we evaluated the efficacy of a hydrogel (H) with encapsulated porphyrin (H+P) on a rat model of surgically-induced skin defects. METHODS: Four round 6 mm diameter skin defects were performed under general anesthesia on the dorsal area of 24 three-month-old "Young" and 24 twelve-month-old "Mature" male rats. Each age group was separated into the Control, H, and H+P groups, n=8 each, where no therapy, H, or H+P was respectively applied daily for 20 days. Digital photographs and skin biopsies were taken on the third, seventh, 10th, and 20th postoperative days and evaluated by planimetry, histology, and immunohistochemistry. RESULTS: Planimetry results demonstrated significantly decreased perimeter, diameter, and area measurements (p<0.005) of group H+P compared to Control and H groups on days 10 and 20 in the young rats, while in the mature rats, the significant differences were evident earlier (perimeter third day p<0.05; diameter and area seventh day p<0.05 and p<0.005, respectively vs. H). Granulation and scar tissue formation were also reduced in the H+P groups although they were not statistically significant. CONCLUSIONS: The application of H+P on the skin defects benefited the healing process in both young and mature animal groups, as evidenced by the statistically significant findings of planimetry. The beneficial healing process was more pronounced in the mature animals, both in the level of statistical significance as well as regarding time (evident already on the third day of healing), probably due to porphyrin assisting the reduced healing rate, which is observed in organisms of advanced age.

Adipose-Derived Stem Cells and Tacrolimus Improve Nerve Regeneration in a Rat Sciatic Nerve Defect Model.

This study compared the effect of undifferentiated adipose-derived stem cells (ADSCs) vs tacrolimus (FK506) in peripheral nerve regeneration in a rat sciatic nerve complete transection model. Forty Wistar rats were equally distributed in four groups. In the SHAM surgery group, the sciatic nerve was exposed and no further intervention was done. In the conduit-alone group (the SLN group), a 10-mm nerve gap was created and bridged with a fibrin conduit filled in with normal saline. In the FK506 group, the fibrin conduit was injected with soluble FK506. In the ADSC group, the conduit was impregnated with undifferentiated ADSCs. Nerve regeneration was assessed by means of walking track analysis, electromyography, and neurohistomorphometry. Clinically and microscopically, nerve regeneration was achieved in all groups at 12 weeks. Walking track analysis confirmed functional recovery in the FK506 and ADSC groups, but there was no difference between them. Recovery in function was also achieved in the SLN group, but it was inferior (P<.05). Electromyography demonstrated superior nerve regeneration in the FK506 and ADSC groups compared with the SLN group (P<.05), with no difference between the FK506 and ADSC groups. Similarly, histology showed no difference between the FK506 and ADSC groups, although both outperformed the SLN group (P<.05). No complications were observed. Successful peripheral nerve regeneration can be accomplished after a 10-mm nerve defect treated with nerve conduits. Superior nerve regeneration may be expected when the conduits are loaded with undifferentiated ADSCs or FK506, with similar outcomes for ADSCs and FK506. [Orthopedics. 2023;46(6):e353-e361.].

Epinephrine, vasopressin, and nitroglycerin improve neurologic outcome in porcine asphyxial cardiac arrest.

AIM: The aim of the present study was to assess whether the combination of epinephrine, vasopressin, and nitroglycerin would improve initial resuscitation success, 24-hour survival, and neurologic outcome compared with epinephrine alone in a swine model of asphyxial cardiac arrest (CA). MATERIALS AND METHODS: This prospective randomized experimental study was conducted at a laboratory research department. Twenty male Landrace/Large-White pigs 12 to 15 weeks of age were investigated. Asphyxial CA was induced by occlusion of the endotracheal tube. Pigs remained untreated for 4 minutes before attempting resuscitation by unclamping the endotracheal tube, mechanical ventilation, chest compressions, and epinephrine (group E) or a combination of epinephrine with vasopressin and nitroglycerin (group EVN) administered intravenously. In case of restoration of spontaneous circulation, the animals were supported for 30 minutes and then observed for 24 hours. RESULTS: Coronary perfusion pressure and mean arterial pressure were significantly increased during cardiopulmonary resuscitation in group EVN. In both groups, restoration of spontaneous circulation and survival rates were comparable (P value, nonsignificant). At 24 hours after CA, neurologic deficit score was significantly better in animals treated with the combination pharmacotherapy (P < .001). Brain histologic damage score was also higher in group EVN compared with group E (P < .001). Total histologic damage score and neurologic deficit score showed a statistical significant correlation (P < .001). CONCLUSION: In this porcine model of asphyxial CA, the addition of nitroglycerin to vasopressin and epinephrine maintained elevated coronary perfusion pressure during asphyxia CA and resulted in significantly better neurologic and histopathologic outcome in comparison with epinephrine alone.

Myxoid spindle cell sarcoma with ETV6-NTRK3 fusion.

Myxoid spindle cell sarcoma is a rare sarcoma with a demanding histopathologic diagnosis due to the absence of pathognomic immunohistochemistry markers. Genetics include complex karyotypic alterations without characteristic molecular abnormalities for this entity. NTRK alterations are rare findings with great clinical importance since they can be therapeutically targeted with two NTRK inhibitors. Herein we present a case of an adult unclassified myxoid spindle cell sarcoma with ETV6/NTRK3 fusion gene, which is a molecular finding characteristic for infantile fibrosarcoma.

Asynchronous E-learning after synchronous E-learning in the pathology course. When is the proper time for this transition?

BACKGROUND: Recordings of live streaming e-lessons of pathology at medical school of National and Kapodistrian University of Athens, Greece are uploaded to the e-class portfolio of each student enrolled in the course. We measured the number of views each video received and noticed that this number exceeded the number of enrolled students. Our main aim was to investigate the correlation between the upload of an educational video and the views it got so as to determine when the proper time is for professors to e-share or upload an educational video for the students. MATERIALS AND METHODS: We measured the number of views of the recorded e-lessons when all videos were uploaded, with a frequency of 15 days. We used analysis of variances statistical analysis to find the significance of the amount of time each video had been uploaded on the virtual platform of the course. We also applied t-tests to assess the significance of alteration of the number of views related to the amount of time until the examinations. RESULTS: Time was a statistically significant factor in the impact of an educational video. The two-factor analysis without interaction measured P ≃ 0.001, proving the strong correlation between time and the increase of views. As the examination date was approaching, there was a statistically significant increase in the number of views of the videotaped e-lessons. Almost 50% of the views of each of the videos took place in the two-week examination period of the course. CONCLUSIONS: The educational videos that contained the core learning concepts of the pathology course should be uploaded first. The complex learning points of the pathology course must be available at the beginning of the semester. Additionally, recordings of videos covering the complex learning points of the course should be uploaded as an additional tool of asynchronous e-learning for the students who choose to watch their former e-lessons to prepare for the examinations.

Participation and Interactivity in Synchronous E-Learning Pathology Course During the COVID-19 Pandemic.

PURPOSE OF THE ARTICLE: Due to the COVID-19 outbreak, educational institutions had to utilize online platform solutions to deliver their curriculum. We conducted this study to explore participation and interactivity in a synchronous e-learning non-mandatory participation course in pathology at a medical school in Greece. The knowledge acquired is expected to be instrumental in the development of educational practices. MATERIALS AND METHODS: The data for this study were gathered through the recorded video archives of the synchronous e-lessons. We observed online participation at seven time points during each of the assessed e-lessons. Moreover, we identified and categorized the professor's/students' interactivity patterns according to content. RESULTS: The maximum number of students participating in the first e-lesson was N = 196. We recorded a reduction of N = 91 students, approximately 46%, in maximum student participants from the second observed e-lesson, and an additional decrease of N = 28 students, approximately 27%, from the third observation. Participation numbers continued to lessen. Even though there was a statistically significant difference in the mean percentage of students participating between the seven time points of each e-lesson, the difference in the mean percentage of students' online participation between the seven e-lessons assessed was not statistically significant. This indicates a consistent e-audience. Evidence of interactivity was summarized in a table, and each professor-students interaction was classified according to its content. We found that the professor posed questions to his students every 2-5 minutes during every synchronous e-lesson and e-tutorial observed, and students wrote 3-6 answers in chat in response to each question. Students asked more questions as more synchronous e-learning classes took place, with limited exceptions. CONCLUSION: From our perspective, our observations set the basis for further research to enhance our understanding of the aspects of the e-learning environment towards the formulation of policies for higher-quality education. PLAIN TEXT: Our pathology department places high value on the quality of education that the medical students receive. Due to the COVID-19 pandemic, our department had to deploy e-learning modalities for curriculum delivery. Thus, we conducted this research to evaluate a pathology e-learning class in terms of students' participation and the interactivity dynamics between them and the professor. We used statistics to measure participation during each e-lesson and identified recurring patterns of interactivity. We avoided imposing our predetermined interpretations of the data in this study so as to present an accurate depiction of the aspects of the e-learning environment. We were very pleased to identify a steady e-audience despite the drop-out rate from one e-lesson to the next, as well as strong, increasing interactivity patterns between the students and the professor, as students posed more and more questions from one e-lesson to the next. We are looking forward to future studies that address the e-learning procedure's challenges and provide evidence of its effectiveness and quality.

The Modulating Effect of Ursodeoxycholic Acid on Liver Tissue Cyclooxygenase-2 Expression Following Extended Hepatectomy.

INTRODUCTION: Hepatic regeneration is a complex process involving a multitude of well-timed molecular operations. Ursodeoxycholic acid (UDCA) is postulated to exert a protective effect against oxidative stress and enzymatic degradation of the extracellular matrix, in turn potentiating the regenerative response. The aim of the present animal study is to evaluate the impact of UDCA administration in liver tissue expression of cyclooxygenase-2 (COX-2) in a setting of acute liver failure achieved by 80% hepatectomy. MATERIALS AND METHODS: Twenty-four adult male Sprague-Dawley rats were randomly assigned to an experimental (UDCA) and a control group. Animals in the UDCA received oral pretreatment with UDCA for 14 days via feeding tube, while animals in the control group received saline. All animals underwent resection of approximately 80% of the liver parenchyma. Tissue and blood sample collection were performed 48 hours postoperatively. RESULTS: The postoperative mitotic index and Ki-67 levels were found to be elevated in the UDCA group (43±11.4 and 13.7±24.7 versus 31±16.7 and 7.6±5.7), albeit without any statistical significance. Pretreatment with UDCA significantly decreased COX-2 expression levels (p=0.28) as well as serum tumor necrosis factor α (TNFα) levels (37.3±10.9 pg/mL versus 75.4±14.4 pg/mL, p=0.004). COX-2 expression score was observed to be weakly correlated to Ki-67 levels in both groups. Although COX-2 expression score was not correlated with serum TNFα levels in the control group, animals pretreated with UDCA exhibited moderate correlation (r=0.45). CONCLUSION: Preoperative administration of UDCA exerts a suppressive effect on tissue expression of COX-2 following 80% hepatectomy and enforces a positive correlation between COX-2 and serum TNFα levels, suggesting that UDCA preconditions liver tissue to display an enhanced regenerative response to circulating cytokines, most notably TNFα. The weak association of COX-2 with Ki-67 expression levels suggests that COX-2 may be of secondary importance during the early phases of liver regeneration.

Hydroxyethyl starch 6% (130/0.4) ameliorates acute lung injury in swine hemorrhagic shock.

BACKGROUND: Traumatic hemorrhage induces acute lung injury. The aim of this study was to assess whether lactated Ringer's solution or 6% hydroxyethyl starch 130/0.4 would have different effects on acute lung injury following hemorrhagic shock. METHODS: Twenty healthy pigs (19 ± 2 kg) were subjected to hemorrhage and were randomly allocated to two groups: Group A (10 pigs) who received lactated Ringer's solution and Group B (10 pigs) who received hydroxyethyl starch 130/0.4. Hemodynamic response and serum lactate were measured at predetermined phases. Four hours after fluid resuscitation animals were euthanized. Lungs were harvested, and tissue samples were collected. Focal thickening of the alveolar membranes, vascular congestion, number of activated neutrophils, alveolar edema, interstitial neutrophil infiltration, intraalveolar infiltration, and alveolar hemorrhage were assessed. Each feature was given a score from 0 to 3 (0 = absence, 3 = severe). The wet/dry ratio was also calculated, and with the use of Evans blue dye extravasation method, capillary permeability was assessed. RESULTS: The total histology score of Group A differed significantly from that of Group B, being significantly lower in Group B animals P = 0.048. The wet/dry weight ratio was significantly higher in the lactated Ringer's group (median [range]) (Group A, 5.1 [0.5]; Group B, 4.9 [0.3]; P = 0.009). The Evans blue dye extravasation method was utilized to study the lung capillary permeability. The animals in Group B showed a marked reduction in microvascular capillary permeability compared with the animals in Group A (Group A, 58.5 [21] mg/g; Group B, 51.5 [14] mg/g; P = 0.017). CONCLUSIONS: Our study indicates that resuscitation after hemorrhagic shock with hydroxyethyl starch 130/0.4 led to less lung edema and less microvascular permeability in this swine model.

Functional modulation of gut microbiota in diabetic rats following dietary intervention with pistachio nuts (Pistacia vera L.).

BACKGROUND: Gut microbiota holds a key-role in numerous biological functions and has emerged as a driving force for the development of diabetes. Diet contributes to gut microbiota diversity and functionality providing a tool for the prevention and management of the disease. The study aimed to investigate the effect of a dietary intervention with pistachio nuts, a rich source of monounsaturated fatty acids, dietary fibers and phytochemicals on gut microbiota composition in the rat model of Type 1 Diabetes. METHODS: Male Wistar rats were randomly assigned into four groups: healthy animals which received control diet (CD) or pistachio diet (PD), and diabetic animals which received control diet (DCD) or pistachio diet (DPD) for 4 weeks. Plasma biochemical parameters were determined and histological examination of liver and pancreas was performed at the end of the dietary intervention. Adherent intestinal microbiota populations in jejunum, ileum, caecum and colon were analyzed. Fecal microbiota populations at the beginning and the end of the study were determined by microbiological analysis and 16S rRNA sequencing. RESULTS: Diabetic animals of both groups exhibited high plasma glucose and low insulin concentrations, as well as characteristic pancreatic lesions. Pistachio supplementation significantly increased lactobacilli and bifidobacteria populations in jejunum, ileum and caecum (p < 0.05) and normalized microbial flora in all examined intestinal regions of diabetic animals. After 4 weeks of supplementation, populations of bifidobacteria and lactobacilli were increased in feces of both healthy and diabetic animals, while enterococci levels were decreased (p < 0.05). Next Generation Sequencing of fecal samples revealed increased and decreased counts of Firmicutes and Bacteroidetes, respectively, in healthy animals that received the pistachio diet. Actinobacteria OTUs were higher in diabetic animals and increased over time in the pistachio treated groups, along with increased abundance of Bifidobacterium. Lactobacillus, Turicibacter and Romboutsia populations were elevated in healthy animals administered the pistachio nuts. Of note, relative abundance of Bacteroides was higher in healthy than in diabetic rats (p < 0.05). CONCLUSION: Dietary pistachio restored normal flora and enhanced the presence of beneficial microbes in the rat model of streptozotocin-induced diabetes.

The Effects of Ursodeoxycholic Acid Pretreatment in an Experimental Setting of Extended Hepatectomy: A Feasibility Study.

Introduction Liver regeneration is an exceptionally complex process, orchestrated by a multitude of growth factors and cytokines. Tumor necrosis factor-alpha (TNF-a) and interleukin-6 (Il-6) have a pivotal role in the initiation of the regenerative response. Ursodeoxycholic acid (UDCA) exhibits a liver protective effect that enhances liver growth after injury. The aim of the present study is to evaluate the effect of UDCA in the circulating levels of TNF-a and Il-6 in rats undergoing extended 80% hepatectomy. Materials and methods Twenty-two male Sprague Dawley rats were randomly assigned in an experimental (UDCA group) and a control group. Mice in the UDCA-group received oral pretreatment of UDCA for two weeks preoperatively at a dosage of 25 mg/kg/day. An 80% hepatic resection was performed in both groups by resecting the middle, inferior right, and left lateral liver lobes. The experiment ended 48 hours postoperatively. Results UDCA pretreatment significantly depressed circulating levels of both TNF-a and Il-6 after the conclusion of the experiment as compared to the control group (p=0.001 and p=0.01, respectively). Furthermore, TNF-a levels were significantly reduced before the institution of liver injury (p=0.02). Mice in the UDCA-group exhibited better liver growth as demonstrated by significantly increased Ki-67 and mitotic rate (p=0.04 and p=0.02, respectively). Finally, the liver regeneration rate (LRR) was significantly elevated in the experimental group (UDCA group, 54.5% vs control group, 35.8%; p=0.002) signifying enhanced liver growth kinetics. Conclusion UDCA reduces the expression of TNF-a and Il-6 during the priming phase of liver regeneration. An 80% hepatectomy model of acute liver failure exhibited enhanced liver regeneration in the experimental group, plausibly due to the immunomodulatory effects of UDCA.

Cranioectodermal dysplasia: a probable ciliopathy.

Cranioectodermal dysplasia (CED), also known as Sensenbrenner syndrome, is a rare autosomal recessive genetic disorder characterized by typical craniofacial, skeletal and ectodermal defects, and tubulointerstitial nephritis leading to early end-stage renal failure. We report on a new familial case of a 9-year-old patient and two fetuses of 23 and 19 weeks of gestation respectively. Hypohidrosis was an additional ectodermal finding is the patient with CED. Postmortem findings in the two fetuses included acromesomelic shortening, craniofacial characteristics with absence of craniosynostosis, small kidneys with tubular and glomerular microscopic cysts, persistent ductal plate with portal fibrosis in the liver, small adrenals and roughly unremarkable histopathology of the physeal growth plate. Posterior fossa anomalies were additional findings in this patient and included an enlarged cisterna magna and a posterior fossa cyst. The above findings, in association with renal cysts, persistent ductal plate and portal fibrosis, introduce CED, a nonlethal genetic skeletal disorder of yet unknown molecular origin, as a possible member of the expanding group of ciliopathies.