The Multi-Faceted Nature of Renalase for Mitochondrial Dysfunction Improvement in Cardiac Disease.

The cellular mechanisms and signaling network that guide the cardiac disease pathophysiology are inextricably intertwined, which explains the current scarcity of effective therapy and to date remains the greatest challenge in state-of-the-art cardiovascular medicine. Accordingly, a novel concept has emerged in which cardiomyocytes are the centerpiece of therapeutic targeting, with dysregulated mitochondria as a critical point of intervention. Mitochondrial dysfunction pluralism seeks a multi-faceted molecule, such as renalase, to simultaneously combat the pathophysiologic heterogeneity of mitochondria-induced cardiomyocyte injury. This review provides some original perspectives and, for the first time, discusses the functionality spectrum of renalase for mitochondrial dysfunction improvement within cardiac disease, including its ability to preserve mitochondrial integrity and dynamics by suppressing mitochondrial ΔΨm collapse; overall ATP content amelioration; a rise of mtDNA copy numbers; upregulation of mitochondrial genes involved in oxidative phosphorylation and cellular vitality promotion; mitochondrial fission inhibition; NAD+ supplementation; sirtuin upregulation; and anti-oxidant, anti-apoptotic, and anti-inflammatory traits. If verified that renalase, due to its multi-faceted nature, behaves like the "guardian of mitochondria" by thwarting pernicious mitochondrial dysfunction effects and exerting therapeutic potential to target mitochondrial abnormalities in failing hearts, it may provide large-scale benefits for cardiac disease patients, regardless of the underlying causes.

The Importance of Immunohistochemical Heterogeneous Expression of MMR Protein in Patients with Colorectal Cancer in Stage II and III of the Disease.

Background and objectives: In patients with colorectal cancer (CRC), heterogeneous expression of Mismatch repair (MMR) proteins can manifest itself in several different forms and is not such a rare phenomenon. Therefore, it is very important to recognize the nuclear expression of MMR proteins of different MMR status in order to avoid false positive or false negative results. The aim of this study was to determine the frequency and distribution of heterogeneous expression of MMR proteins in patients with stages II and III of the disease as well as its association with clinical, demographic and pathological characteristics of CRC in relation to proficient and deficient expression of MMR proteins. Material and Methods: The study included 104 cases of colorectal cancer obtained from surgical colectomy material in stages II and III of the disease. Results: From a total of 104 patients with colorectal cancer, immunohistochemical analysis of the expression of all four MMR proteins showed that heterogeneous expression of MMR proteins (as well as deficient immunoreactivity of tumor cells) was present in 12 cases, while proficient expression of MMR proteins was detected in 80 tumors. Conclusions: Our study showed that the only independent predictors of the loss of MMR protein expression were younger patient age and right-sided anatomical location of the tumor. The study also established the existence of heterogeneous expression of MMR proteins in a non-negligible percentage of CRCs (11.5%), where heterogeneous nuclear expression of MMR proteins was described in several different forms.

Distributions of Platelet-Derived Growth Factor Receptor-α Positive Cells and Interstitial Cells of Cajal in the Colon of Rats with Diabetes Mellitus Type 2.

Background and Objectives: Diabetic gastroenteropathy (DG) is a common complication of diabetes mellitus type 2. Interstitial cells are non-neural cells of mesenchymal origin inserted between nerve elements and smooth muscle cells, necessary for normal function and peristaltic contractions in the gastrointestinal (GI) tract. There are at least two types of interstitial cells within the GI muscle layer-interstitial cells of Cajal (ICC) and interstitial platelet-derived growth factor receptor α-positive cells (IPC). The mechanism of diabetic gastroenteropathy is unclear, and interstitial cells disorders caused by metabolic changes in diabetes mellitus (DM) could explain the symptoms of DG (slow intestinal transit, constipation, fecal incontinence). The aim of this study was to identify PDGFRα and c-kit immunoreactive cells in the colon of rats with streptozotocin-nicotinamide-induced diabetes mellitus type 2, as well as to determine their distribution in relation to smooth muscle cells and enteric nerve structures. Materials and Methods: Male Wistar rats were used, and diabetes type 2 was induced by an intraperitoneal injection of streptozotocin, immediately after intraperitoneal application of nicotinamide. The colon specimens were exposed to PDGFRα and anti-c-kit antibodies to investigate interstitial cells; enteric neurons and smooth muscle cells were immunohistochemically labeled with NF-M and desmin antibodies. Results: Significant loss of the intramuscular ICC, myenteric ICC, and loss of their connection in intramuscular linear arrays and around the ganglion of the myenteric plexus were observed with no changes in nerve fiber distribution in the colon of rats with diabetes mellitus type 2. IPC were rarely present within the colon muscle layer with densely distributed PDGFRα+ cells in the colon mucosa and submucosa of both experimental groups. In summary, a decrease in intramuscular ICC, discontinuities and breakdown of contacts between myenteric ICC without changes in IPC and nerve fibers distribution were observed in the colon of streptozotocin/nicotinamide-induced diabetes type 2 rats.

Renalase Challenges the Oxidative Stress and Fibroproliferative Response in COVID-19.

The hallmark of the coronavirus disease 2019 (COVID-19) pathophysiology was reported to be an inappropriate and uncontrolled immune response, evidenced by activated macrophages, and a robust surge of proinflammatory cytokines, followed by the release of reactive oxygen species, that synergistically result in acute respiratory distress syndrome, fibroproliferative lung response, and possibly even death. For these reasons, all identified risk factors and pathophysiological processes of COVID-19, which are feasible for the prevention and treatment, should be addressed in a timely manner. Accordingly, the evolving anti-inflammatory and antifibrotic therapy for severe COVID-19 and hindering post-COVID-19 fibrosis development should be comprehensively investigated. Experimental evidence indicates that renalase, a novel amino-oxidase, derived from the kidneys, exhibits remarkable organ protection, robustly addressing the most powerful pathways of cell trauma: inflammation and oxidative stress, necrosis, and apoptosis. As demonstrated, systemic renalase administration also significantly alleviates experimentally induced organ fibrosis and prevents adverse remodeling. The recognition that renalase exerts cytoprotection via sirtuins activation, by raising their NAD+ levels, provides a "proof of principle" for renalase being a biologically impressive molecule that favors cell protection and survival and maybe involved in the pathogenesis of COVID-19. This premise supports the rationale that renalase's timely supplementation may prove valuable for pathologic conditions, such as cytokine storm and related acute respiratory distress syndrome. Therefore, the aim for this review is to acknowledge the scientific rationale for renalase employment in the experimental model of COVID-19, targeting the acute phase mechanisms and halting fibrosis progression, based on its proposed molecular pathways. Novel therapies for COVID-19 seek to exploit renalase's multiple and distinctive cytoprotective mechanisms; therefore, this review should be acknowledged as the thorough groundwork for subsequent research of renalase's employment in the experimental models of COVID-19.

Vitamins, microelements and the immune system: current standpoint in the fight against coronavirus disease 2019.

Coronavirus disease 2019 (COVID-19) is an acute respiratory disease associated with severe systemic inflammation. The optimal status of vitamins and microelements is considered crucial for the proper functioning of the immune system and necessary for successful recovery. Most patients with respiratory distress in COVID-19 are vitamin and microelement deficient, with vitamin D and Se deficiency being the most common. Anyway, various micronutrient supplements are widely and arbitrarily used for prevention or in the treatment of COVID-19. We aimed to summarise current knowledge about molecular and physiological mechanisms of vitamins (D, A, C, B6, B9 and B12) and microelements (Se, Zn, Cu and Fe) involved in the immune system regulation in consideration with COVID-19 pathogenesis, as well as recent findings related to their usage and effects in the prevention and treatment of COVID-19. In the early course of the pandemic, several, mainly observational, studies reported an association of some micronutrients, such as vitamin C, D and Zn, with severity reduction and survival improvement. Still, emerging randomised controlled trials showed no effect of vitamin D on hospitalisation length and no effect of vitamin C and Zn on symptom reduction. Up to date, there is evidence neither for nor against the use of micronutrients in the treatment of COVID-19. The doses that exceed the recommended for the general population and age group should not be used, except in clinical trials. Benefits of supplementation are primarily expected in populations prone to micronutrient deficiencies, who are, as well, at a higher risk of worse outcomes in COVID-19.

Interstitial Cells of Cajal and Neural Structures in the Human Fetal Appendix.

BACKGROUND/AIMS: The interstitial cells of Cajal (ICC) are located within and around the digestive tract's muscle layers. They function as intestinal muscle pacemakers and aid in the modification of enteric neurotransmission. The appendix's unique position requires an appropriate contraction pattern of its muscular wall to adequately evacuate its contents. We investigated the development and distribution of nervous structures and ICC in the human fetal appendix. METHODS: Specimens were exposed to anti-c-kit (CD117) antibodies to investigate ICC differentiation. Enteric plexuses were examined using anti-neuron-specific enolase, and the differentiation of smooth muscle cells was studied with anti-desmin antibodies. RESULTS: During weeks 13-14, numerous myenteric plexus ganglia form an almost uninterrupted sequence throughout the body and apex of the appendix. Fewer ganglia were present at the submucosal border of the circular muscle layer and within this layer. A large number of ganglia appear within the circular and longitudinal muscle layers in a later fetal period. The first ICC subtypes noted were of the myenteric plexus and the submucous plexus. In the later fetal period, the number of intramuscular ICC markedly rises, and this subtype becomes predominant. CONCLUSIONS: The ICC and nervous structure distribution in the human fetal appendix are significantly different from all other parts of the small and large intestine. The organization of ICC and the enteric nervous system provides the basis for the specific contraction pattern of the muscular wall of the appendix.

Oral supplementation with melatonin reduces oxidative damage and concentrations of inducible nitric oxide synthase, VEGF and matrix metalloproteinase 9 in the retina of rats with streptozotocin/nicotinamide induced pre-diabetes.

Hyperglycemia mediated oxidative stress and pro-angiogenic molecules such as vascular endothelial growth factor (VEGF) and matrix metalloproteinase 9 (MMP9) are considered important for diabetic retinopathy onset and progression. Melatonin is a pineal hormone that regulates circadian and seasonal rhythms and most likely is involved in regulating glucose metabolism. We aimed to evaluate the potential benefit of melatonin supplementation to the pre-diabetic retina by assessing melatonin effects on lipid peroxidation (thiobarbituric acid reactive substances, TBARS), protein oxidation (advanced oxidation protein products, AOPP) and concentrations of inducible nitric oxide synthase (iNOS), VEGF and MMP9 in the retina of rats with pre-diabetes. Pre-diabetes was induced by streptozotocin (45 mg/kg, i.p.) following nicotinamide injection (110 mg/kg, i.p.). Beside mild hyperglycemia, lower serum insulin, increased fructosamine and lower HDL cholesterol, the present study demonstrated decreased serum melatonin in pre-diabetic rats, as well as, increased concentration of retinal TBARS, AOPP, iNOS, VEGF, and MMP9. Oral supplementation with melatonin (85 µg/animal/day) caused melatonin and HDL cholesterol levels to rise in treated rats and reduced levels of fasting serum glucose and fructosamine. It also affected serum insulin and quantitative insulin sensitivity check index (QUICKI) in treated groups but had no significant effect on non-fasting glucose. Finally, supplementation with melatonin reduced concentrations of TBARS, AOPP, iNOS, VEGF, and MMP9 in significant level, thereby exerting an overall positive effect on oxidative stress and pro-angiogenic signaling in the pre-diabetic retina. Thus, oral melatonin might be considered in an early treatment or in the prevention of retinal changes associated with pre-diabetes.

Development of interstitial cells of Cajal in the human digestive tract as the result of reciprocal induction of mesenchymal and neural crest cells.

Neural crest cells (NCC) can migrate into different parts of the body and express their strong inductive potential. In addition, they are multipotent and are able to differentiate into various cell types with diverse functions. In the primitive gut, NCC induce differentiation of muscular structures and interstitial cells of Cajal (ICC), and they themselves differentiate into the elements of the enteric nervous system (ENS), neurons and glial cells. ICC develop by way of mesenchymal cell differentiation in the outer parts of the primitive gut wall around the myenteric plexus (MP) ganglia, with the exception of colon, where they appear simultaneously also at the submucosal border of the circular muscular layer around the submucosal plexus (SMP) ganglia. However, in a complex process of reciprocal induction of NCC and local mesenchyma, c-kit positive precursors are the first to differentiate, representing probably the common precursors of ICC and smooth muscle cells (SMC). C-kit positive precursors could represent a key impact factor regarding the final differentiation of NCC into neurons and glial cells with neurons subsequently excreting stem cell factor (SCF) and other signalling molecules. Under the impact of SCF, a portion of c-kit positive precursors lying immediately around the ganglia differentiate into ICC, while the rest differentiate into SMC.

Expression of CD34 and CD146 vascular markers contributes to the immunological function of the human palatine tonsil.

The fundamental function of the palatine tonsil is the immune response to airborne and foodborne pathogenic agents. Small blood vessels have an important role in the provision of a special microenvironment in which the immune response occurs. In this study, we investigated the expression of vascular markers CD34 and CD146 and basal lamina marker - type IV collagen - in the small blood vessels of the human palatine tonsil in the context of their role in the immunological function of the tonsil. The tonsils were collected after tonsillectomy from ten patients with chronic tonsillitis, aged 18-28 years. Five-µm-thick paraffin sections were routinely stained with haematoxylin and eosin, while the studied markers (CD34, CD146 and type IV collagen) were detected immunohistochemically using LSAB2/HRP method. CD34 was expressed equally in the capillaries within and below the crypt epithelium, in lymphoid follicles and in high endothelial venules localized para- and interfollicularly. CD146 molecule was expressed on the luminal surface of endothelial cells in the capillaries of the crypt epithelium, while its expression in high endothelial venules was seen on the luminal and lateral surfaces of the cuboidal endothelial cells. In contrast to the basal lamina of intraepithelial capillaries, where collagen IV-immunopositivity is mostly seen as a continuing line, the basal lamina of high endothelial venules was seen as a two- or three-layered structure beneath the cuboidal endothelial cells. The specifics of expression of CD34, CD146, and type IV collagen confirm the morphofunctional specialization of endothelium in crypt epithelium capillaries, and also in endothelium of high endothelial venules, which is directly associated with the role of these vessels in the immune function of the tonsil.

The effect of bilberries on diabetes-related alterations of interstitial cells of Cajal in the lower oesophageal sphincter in rats.

Diabetic gastroenteropathy involves not only the parasympathetic and sympathetic autonomic nerves, but also enteric neurons, smooth muscle cells and interstitial cells of Cajal (ICC). ICC are the cells of mesenchymal origin that occur within and around the muscle layers in the gastrointestinal tract. The objective of the present study was to investigate the alterations of ICC in the lower oesophageal sphincter (LOS) of streptozotocin-nicotinamide non-insulin-dependent diabetes rats. Moreover, we investigated possible ICC in rats with the same type of diabetes, treated with bilberry fruit extract, bearing in mind that its hypoglycemic effect had been already proven. Male Wistar rats (10 weeks old) were used, and diabetes was induced by an intraperitoneal injection of streptozotocin, immediately after intraperitoneal application of nicotinamide. The specimens were exposed to anti-c-kit antibodies to investigate the distribution of ICC, and the smooth muscle cells were immunohistochemically labelled using anti-desmin antibodies. Intramuscular ICC were very abundant in the LOS of rats. They were spindle-shaped, with two long processes connecting them into long linear sequences. In the LOS of diabetic rats, intramuscular ICC were rarely present and linear cell-cell connections between these cells were completely missing. In groups treated with bilberry, the number and distribution of ICC were exactly the same as in the above described rats with induced diabetes. In summary, a decrease of intramuscular ICC, discontinuities and breakdown of contacts between ICC were observed in streptozotocin-nicotinamide induced diabetes rats and in groups treated with bilberry. Bilberry fruit extract was shown to have hypoglycemic activity, but without any protective effects on ICC in the LOS of diabetic rats.

Tensor fascia lata flap is a workhorse for defects after inguinal lymph node block dissection.

Introduction: Enlarged inguinal lymph nodes very often present a site of metastatic disease. Inguinal lymph node block dissection is a demanding procedure, which usually requires at least one of reconstructive modalities. Among different reconstruction options we selected the tensor fascia lata (TFL) musculocutaneous flap. Objective: The paper aims at presenting a series of inguinal block dissections, followed by immediate reconstruction, using the TFL flap, and evaluation of tumor type, flap dimension, complication rate and the duration of hospital stay. Methods: We present a consecutive case series of 25 conducted block dissections. The defects were reconstructed using TFL flap, because of the extent and site of the tissue defects, reliability of the flap, and potentially primarily infected exulcerated tumors. Results: The reconstruction was successful in all cases, the incidence of surgical complications was 16%, no further complications, such as lymphedema or gait disturbances, were noted. Primary skin tumors were predominant (13 cases), followed by genitalia tumors (four cases). The male sex was more frequently affected (14 vs. 11 cases). Conclusion: Having in mind that TFL presents as a flap of adjustable size, length, shape, and volume, with negligible donor site morbidity, and after comparing of our results to those of other authors, we advise broader use of TFL flap. As a reliable flap, not too difficult to harvest, with a low complication rate, it must be taken into consideration regarding the benefits for the patient, and, on the other hand, the surgery cost and duration.

Primary reconstruction of neck defect after excision of metastatic melanoma of unknown primary site with regional pectoral myocutaneous flap.

Introduction: Metastatic melanoma of unknown primary (MMUP) is already a well described oncologic phenomenon in the literature, whereas tissue defects' reconstructions on the neck region always present a challenge for the reconstructive surgeon. Two cases of giant metastatic, skin infiltrative neck tumor masses are presented. In both cases MMUP was diagnosed. Both intraoperative tissue defects were reconstructed using pectoralis major (PM) regional flap. Outline of cases: The first patient was admitted with giant tumor mass on the right side of the neck. The fast growing mass appeared two months prior to the admission. Thorough examination showed no signs of primary tumor. Removal surgery was performed and the defect was reconstructed using the PM musculocutaneous flap. The second patient was admitted with large tumor mass on the left side of the neck. Thorough examination displayed no signs of any primary tumor. After the excision, the tumor mass and subsequent neck dissection, reconstruction followed, using the pedicled PM muscle flap and partial thickness skin transplants. There were no major complications in either case. The histopathological examinations presented metastatic melanoma diagnoses. Conclusion: Clinical outcome of MMUP described in literature is rather variable. Different studies have shown that prognosis in patients with MMUP is better than that in patients with diagnosed primary melanoma with metastatic disease. Therefore, the best initial course of action in those cases would be surgery, according to oncological principles, if possible. Neck defects' reconstructions should fulfill both functional and esthetic demands. Due to the reliability and low cost of the procedure, PM regional flap presents a very good and trustworthy reconstruction modality.

Ultrastructure of the human palatine tonsil and its functional significance.

The human palatine tonsils represent a mucosa-associated lymphoid tissue with a significant function in mucosal protection against alimentary and airborne pathogens. The ultrastructure of different morphological compartments in the human palatine tonsil was studied in eighteen tonsils obtained from the patients who had undergone elective tonsillectomy due to chronic tonsillitis. The tonsillar specimens were analyzed by scanning and transmission electron microscopy. The results showed the presence of tight junctions between superficial epithelial cells of the oropharyngeal tonsillar surface. The crypt epithelium is a sponge-like structure infiltrated by non-epithelial cells, mostly lymphocytes, and is characterized by the presence of small pores - microcrypts occupied by large microvillus cells and/or lymphocytes. Antigen-presenting Langerhans cells with typical intracytoplasmic Birbeck granules were also found in the crypt epithelium. The lymphoid follicles are composed of lymphocytes and two types of non-lymphoid follicular cells: small fibroblast-like cells and large cells, morphologically consistent with antigen-bearing follicular dendritic cells or macrophages. The interfollicular areas consisted of a dense network of reticular cells and reticular fibers; many lymphocytes were interspersed between the reticular fibers. In addition to arterioles and high endothelial venules in the interfollicular lymphoid tissue, some fenestrated capillaries were seen intraepithelially and subepithelially. The complex ultrastructure of the human palatine tonsil provides a microenvironment necessary for antigen uptake, antigen processing and immune response.

Reconstruction of full thickness abdominal wall defect following tumor resection: a case report.

INTRODUCTION: Reconstruction of a full thickness abdominal wall defect is a demanding procedure for general and also for plastic surgeons, requiring vigorous planning and reconstruction of three layers. CASE OUTLINE: We present a case of a 70-year-old patient with a huge abdominal wall tumor with 40 years evolution. Surgery was performed under general anesthesia. Full thickness abdominal defect appeared after the tumor resection. Reconstruction followed in the same act. The defect was reconstructed using a combination of techniques, including omental flap, fascia lata graft, local skin flaps and skin grafts. After surgery no major complications were noted, only a partial skin flap loss, which was repaired using partial thickness skin grafts. The final result was described by the patient as very good, without hernia formation. CONCLUSION: Omenthoplasty, abdominal wall reconstruction in combination with free fascia lata graft and skin grafts can be one of good options for the reconstruction of full thickness abdominal wall defects.

Second look procedure for large burn defect by banana peel pericranial flap based on one artery.

INTRODUCTION: Scalp and calvarial defects may result from trauma, thermal or electrical burns, resection of benign or malignant tumors, infections or radionecrosis. Reconstruction of large scalp defects is a demanding procedure. The reconstructive "ladder" are applicable to scalp and calvarial defects reconstruction. CASE OUTLINE: A 68-year-old female was admitted to our clinic due to the nine-day old scalp burn wound, incurred under unclear circumstances. Third degree burn wound affected the left frontal-parietal, temporal and part of the occipital region with carbonification of the whole left ear lobe.The treatment was carried out in two stages. Radical full thickness necrectomy of the scalp was performed, the defect margins were curetted to the active bleeding, and the ear lobe was amputated.The defect sized 23 x 15 cm was reconstructed using the"banana peel"transposition galea-cutaneous flap from the remainder of the scalp, which was based only on the right occipital artery.Two months after the surgery the appearance was satisfactory, and all wounds were healed. CONCLUSION: Designing of large-scale flaps is very hazardous, especially in elderly people. Scalp reconstruction based on one artery has to be planned in detail and performed when the possibility of complication is reduced to minimum. Our case report underlines possible reconstruction as delayed procedure even with the exposed bone (second look procedure), as well as the reconstruction of half scalp with the local flap based on one pericranial artery.

Appearance of interstitial cells of Cajal in the human midgut.

Several subtypes of the interstitial cells of Cajal (ICC) form networks that play a role in gastrointestinal motor control. ICC express c-kit and depend on signaling via Kit receptors for development and phenotype maintenance. At 7-8 weeks of development, c-kit-immunoreactive (c-kit-IR) cells are present in the human oesophagus, stomach and proximal duodenum wall. In the remaining small and large bowel, c-kit-IR cells appear later. The object of the present study is to determine the timing of the appearance of c-kit-IR ICC in the parts of the digestive tube originating from the midgut (distal duodenum, jejunum, ileum and proximal colon). Specimens were obtained from eight human embryos and 11 fetuses at 7-12 weeks of gestational age. The specimens were exposed to anti-c-kit antibodies to investigate ICC differentiation. The differentiation of enteric neurons and smooth muscle cells was immunohistochemically examined by using anti-PGP9,5 and anti-desmin antibodies, respectively. In the distal duodenum, jejunum and ileum, c-kit-IR cells emerged at week 9 at the level of the myenteric plexus in the form of a thin row of cells encircling the inception of the ganglia. These cells were multipolar or spindle-shaped with two long processes and corresponded to the ICC of the myenteric plexus. In the proximal colon, c-kit-IR cells emerged at week 9-10 in the form of two parallel belts of cells extending at the submucosal plexus and the myenteric plexus levels. We conclude that ICC develop following two different patterns in the human midgut.