Intense Inflammatory Reaction Research Articles

Immunological aspects of COVID-19 in children

The authors present a review of the literature on the peculiarities of immunity in children, which may explain the differences in the symptoms of COVID-19 in childhood. In children, COVID-19 tends to occur more easily than in adults. However, as in the years of the pandemic, and in the post-pandemic period, there were quite a few cases of complicated course with the need for hospitalization, and sometimes with fatal outcomes. Infants under the age of 1 year account for the highest percentage of hospitalizations among children. Currently, the analysis of the results of comparing the clinical manifestations of the disease and laboratory data is continuing. Basically, the features of COVID-19 in children are explained by the immaturity of the immune system and the atrial fibrillation of the respiratory tract. It is known that in childhood there are fewer angiotensin converting enzyme type 2 receptors (ACE 2, ACE 2), with which the coronavirus enters the human body. The disease caused by SARS-CoV-2 in children is often accompanied by gastrointestinal manifestations. A small percentage of sick children may develop an intense inflammatory reaction called “multisystem inflammatory syndrome”, also known as MIS-C. In this condition, high titers of antibodies to RBD SARS-CoV-2 and autoantibodies to free interleukin (IL) -1Ra are detected. These data should be taken into account when clarifying the prevention and treatment regimens aimed at preventing complications in the pediatric population.

Mesenchymal stem cell-based adjunctive therapy for Pseudomonas aeruginosa-induced keratitis: A proof-of-concept in-vitro study

PurposePseudomonas aeruginosa-induced keratitis is one of the most severe and challenging forms of corneal infection, owing to its associated intense inflammatory reactions leading to corneal necrosis and dense corneal scar with loss of vision. Since mesenchymal stem cells (MSCs) are reported to possess antimicrobial and immunomodulatory properties, they can be tested as an adjuvant treatment along with the antibiotics which are the current standard of care. This study aims to investigate the anti-bacterial and immunomodulatory roles of human bone marrow MSC-derived conditioned medium (MSC-CM) in P. aeruginosa-infected human corneal epithelial cells (HCECs) in vitro. MethodsThe effect of MSC-CM on the growth of clinical isolates of P. aeruginosa was evaluated by colony-forming unit assay. The expression of inflammatory cytokines (IL-6 and TNF-α) and an antimicrobial peptide (Lipocalin 2) in lipopolysaccharide-treated MSCs and HCECs was analyzed through ELISA. Corneal epithelial repair following infection with P. aeruginosa was studied through scratch assay. ResultsCompared to control (P. aeruginosa (5*105) incubated in DMEM (1 ml) at 37 °C for 16 h), MSC-CM significantly: i) inhibits the growth of P. aeruginosa (159*109 vs. 104*109 CFU/ml), ii) accelerates corneal epithelial repair following infection with P. aeruginosa (9% vs. 24% closure of the wounded area after 12 h of infection), and iii) downregulates the lipopolysaccharide-induced expression of IL-6, TNF-α and Lipocalin 2 in HCECs. A combination of MSC-CM with an antibiotic, Ciprofloxacin moderately regulated the expression of IL-6, TNF-α, and Lipocalin 2. ConclusionMSC-CM holds promise as an adjunctive therapeutic approach for P. aeruginosa-induced corneal epithelial damage.

Pilonidal Sinus at The Appendectomy Incision

Pilonidal sinus is a disease that progresses with inflammation due to hairs that penetrate the epidermis layer of the skin. This case report presented the diagnosis and treatment process of the incisional pilonidal sinus case, which occurred at an appendectomy incision. A 47-year-old male patient was admitted with purulent discharge and erythema for about three days in the appendectomy incision. The patient underwent appendectomy with a right paramedian incision approximately eight years ago, and recurrent abscesses developed in the incision scar in the postoperative period. The skin and subcutaneous tissues, including the abscess sac, were surgically excised. In the excision material taken from the patient, there was an intense inflammatory reaction with hair covering the entire dermis and erosion and ulceration of the epidermis.

A critical role for host-derived cystathionine-β-synthase in Staphylococcus aureus-induced udder infection

Cystathionine-β-synthase (CBS) catalyzes the first step of the transsulfuration pathway. The role of host-derived CBS in Staphylococcus aureus (S. aureus)-induced udder infection remains elusive. Herein, we report that S. aureus infection enhances the expression of CBS in mammary epithelial cells in vitro and in vivo. A negative correlation is present between the expression of CBS and inflammation after employing a pharmacological inhibitor/agonist of CBS. In addition, CBS achieves a fine balance between eliciting sufficient protective innate immunity and preventing excessive damage to cells and tissues preserving the integrity of the blood-milk barrier (BMB). CBS/H2S reduces bacterial load by promoting the generation of antibacterial substances (ROS, RNS) and inhibiting apoptosis, as opposed to relying solely on intense inflammatory reactions. Conversely, H2S donor alleviate inflammation via S-sulfhydrating HuR. Finally, CBS/H2S promotes the expression of Abcb1b, which in turn strengthens the integrity of the BMB. The study described herein demonstrates the importance of CBS in regulating the mammary immune response to S. aureus. Increased CBS in udder tissue modulates excessive inflammation, which suggests a novel target for drug development in the battle against S. aureus and other infections.

Eosinophil Granule Proteins Involvement in Acute Appendicitis-An Allergic Disease?

Several pieces of evidence point to an allergic component as a trigger of acute appendicitis. As the Th2 immune response is characterized by eosinophil mobilization to the target organ and release of their cationic granule proteins, it is reasonable to investigate if the degranulation of eosinophils could be associated with the local injury. The primary aim of this study is to evaluate the participation of eosinophils granules proteins in acute appendicitis, both at local and systemic levels and the secondary aim is to evaluate the diagnostic accuracy of eosinophils granules proteins for the detection of acute appendicitis, as well as for distinguishing between complicated and uncomplicated acute appendicitis. Eosinophil-derived neurotoxin (EDN), eosinophil cationic protein (ECP) and eosinophil peroxidase (EP) are the most well-known eosinophil granule proteins. From August 2021 to April 2022, we present a prospective single-center study to evaluate the EDN, ECP, and EP concentrations simultaneously in appendicular lavage fluid (ALF) and the serum of 22 patients with acute phlegmonous appendicitis (APA), 24 with acute gangrenous appendicitis (AGA), and 14 normal controls. Concerning EDN, no differences were found between groups. ECP concentrations in ALF and serum were significantly higher in the histologically confirmed acute appendicitis compared to the control groups (p < 0.0001 and p < 0.0001, respectively). In ALF, no differences were found between ECP levels in APA: 38.85 ng/mL (IQR 26.50-51.77) and AGA 51.55 ng/mL (IQR 39.55-70.09) groups (p = 0.176). In the serum, no difference was found between ECP levels at APA: 39 ng/mL (IQR 21.30-56.90) and AGA: 51.30 ng/mL (IQR 20.25-62.59) (p = 0.100). For EP, the concentrations in ALF (p < 0.001) and serum (p < 0.001) were both higher in acute appendicitis compared to the control. In ALF, no difference was found between APA: 240.28 ng/mL (IQR 191.2-341.3) and AGA: 302.5 (IQR 227.7-535.85) (p = 0.236). In the serum, no differences were found between APA: 158.4 ng/mL (IQR 111.09-222.1) and AGA: 235.27 (IQR 192.33-262.51) (p = 0.179). Globally, the ALF concentrations were higher than serum concentrations, reflecting an intense inflammatory local reaction in AA. The optimal ECP cut-off for discriminating between acute appendicitis and the controls was >11.41 ng/mL, with a sensitivity of 93.5%, but with a specificity for identifying appendicitis of 21.4%, good discriminative power (AUC = 0.880). For EP, the optimal cut-off was >93.20 ng/mL, with a sensitivity of 87%, but with a specificity of 14.3% (AUC = 0.901), excellent discriminative power. For the diagnosis of perforated AA, the discriminative power of ECP and EP serum concentrations are weak (AUC = 0.562 and AUC = 0.664, respectively). Concerning the presence of peritonitis, the discriminative power of ECP and EP serum concentrations is acceptable, respectively: AUC = 0.724 and AUC = 0.735. Serum levels of EDN (p = 0.119), ECP (p = 0.586) and EP (p = 0.08) in complicated appendicitis were similar to uncomplicated appendicitis. Serum concentrations of ECP and EP can be added to decision-making AA diagnosis. A Th2-type immune response is present in AA. These data bring forward the role of an allergic reaction in the pathogenesis of acute appendicitis.

Physicochemical and histological analysis of an experimental endodontic repair material containing 45S5 bioactive glass.

This study aimed to evaluate the maximum compressive strength, the modulus of elasticity, pH variation, ionic release, radiopacity and biological response of an experimental endodontic repair cement based on 45S5 Bioglass®. An in vitro and in vivo study with an experimental endodontic repair cement containing 45S5 bioactive glass was conducted. There were three endodontic repair cement groups: 45S5 bioactive glass-based (BioG), zinc oxide-based (ZnO), and mineral trioxide aggregate (MTA). In vitro tests were used to evaluate their physicochemical properties: compressive strength, modulus of elasticity, radiopacity, pH variation, and the ionic release of Ca+ and PO4. An animal model was used to evaluate the bone tissue response to endodontic repair cement. Statistical analysis included the unpaired t-test, one-way ANOVA and Tukey's test. BioG showed the lowest compressive strength and ZnO showed the highest radiopacity among the groups, respectively (p < 0.05). There were no significant differences in the modulus of elasticity among the groups. BioG and MTA maintained an alkaline pH during the 7days of evaluation, both at pH 4 and in a pH 7 buffered solutions. PO4 was elevated in BioG, peaking at 7days (p < 0.05). Histological analysis showed less intense inflammatory reactions and new bone formation in MTA. BioG showed inflammatory reactions that decreased over time. These findings suggest that the BioG experimental cement had good physicochemical characteristics and biocompatibility required for bioactive endodontic repair cement.

Osseointegration evaluation of an experimental bone graft material based on hydroxyapatite, reinforced with titanium-based particles.

Bone graft materials are more and more frequently used in dentistry for improving the periodontal support and for creating a bone support favorable for the insertion of dental implants. The experimental study carried out on laboratory animals aimed to evaluate the biocompatibility and the manner of integration of an experimental bone augmentation material, based on hydroxyapatite (HAp), reinforced with titanium-based particles by comparison with a commercial synthetic graft material already existing on the profile market, also based on HAp. We noticed a common pattern of evolution, although there were differences related to the speed of new bone tissue formation and implicitly the morphological elements captured at the two moments of time. In the presence of both synthetic materials, ossification also begins from the center of the cavity at distance from the margins of the bone defect, with a common pattern with an appearance with the presence of osteon-like structures. The experimental material generally determined a more intense initial inflammatory reaction, followed by the generation of a repair bone tissue with a denser appearance but with a less uniform structure and a greater number of residual particles.

Cutaneous protothecosis in a dog successfully treated with oral itraconazole in pulse dosing

BackgroundProtothecosis is a rare infectious disease caused by unicellular, achlorophyllous, microalgae of the genus Prototheca, ubiquitously distributed in nature. The algae are emerging pathogens, whose incidence is increasing in both human and animal populations and serious systemic infections related to this pathogen have been increasingly described in humans in recent years. After mastitis in dairy cows, canine protothecosis is the second most prevalent form of the protothecal disease in animals. Here, we report the first case of chronic cutaneous protothecosis due to P. wickerhamii in a dog in Brazil, successfully treated with a long-term therapy with itraconazole in pulse.Case presentationUpon clinical examination, exudative nasolabial plaque, ulcered, and painful lesions in central and digital pads and lymphadenitis were observed in a 2-year-old mixed-breed dog, with a 4-month history of cutaneous lesions and contact with sewage water. Histopathological examination revealed intense inflammatory reaction, with numerous spherical to oval, encapsulated structures stained with Periodic Acid Schiff, compatible with Prototheca morphology. Tissue culture on Sabouraud agar revealed yeast-like, greyish-white colonies after 48 h of incubation. The isolate was subjected to mass spectrometry profiling and PCR-sequencing of the mitochondrial cytochrome b (CYTB) gene marker, leading to identification of the pathogen as P. wickerhamii. The dog was initially treated with oral itraconazole at a dosage of 10 mg/kg once daily. After six months, the lesions resolved completely, yet recurred shortly after cessation of therapy. The dog was then treated with terbinafine at a dose of 30 mg/kg, once daily for 3 months, with no success. The resolution of clinical signs, with no recurrence over a 36-months follow-up period, was achieved after 3 months of treatment with itraconazole (20 mg/kg) in pulse intermittently on two consecutive days a week.ConclusionsThis report highlights the refractoriness of skin infections by Prototheca wickerhamii with therapies proposed in the literature and suggests a new treatment option with oral itraconazole in pulse dosing for long-term disease control successfully performed in a dog with skin lesions.

Brown Spider Venom Phospholipase-D Activity upon Different Lipid Substrates.

Brown spider envenomation results in dermonecrosis, characterized by an intense inflammatory reaction. The principal toxins of brown spider venoms are phospholipase-D isoforms, which interact with different cellular membrane components, degrade phospholipids, and generate bioactive mediators leading to harmful effects. The Loxosceles intermedia phospholipase D, LiRecDT1, possesses a loop that modulates the accessibility to the active site and plays a crucial role in substrate. In vitro and in silico analyses were performed to determine aspects of this enzyme's substrate preference. Sphingomyelin d18:1/6:0 was the preferred substrate of LiRecDT1 compared to other Sphingomyelins. Lysophosphatidylcholine 16:0/0:0 was preferred among other lysophosphatidylcholines, but much less than Sphingomyelin d18:1/6:0. In contrast, phosphatidylcholine d18:1/16:0 was not cleaved. Thus, the number of carbon atoms in the substrate plays a vital role in determining the optimal activity of this phospholipase-D. The presence of an amide group at C2 plays a key role in recognition and activity. In silico analyses indicated that a subsite containing the aromatic residues Y228 and W230 appears essential for choline recognition by cation-π interactions. These findings may help to explain why different cells, with different phospholipid fatty acid compositions exhibit distinct susceptibilities to brown spider venoms.

Differential invasiveness & expression of antimicrobial peptides in Shigella serotypes.

The study of Shigella pathogenesis at present is severely hampered by the lack of a relevant animal model that replicates human bacillary dysentery. Different Shigella serogroups cause varying severity of clinical illness. Ex vivo colonization of Shigella flexneri, S. dysenteriae and S. sonnei were characterized in human paediatric colonic pinch biopsies in the in vitro organ culture (IVOC) model to study the invasiveness of Shigella by gentamicin protection assay (GPA). Furthermore, the expression of antimicrobial peptides (AMPs) in response to different serotypes of Shigella was also studied in IVOC model. IVOC explants were inoculated with 109 colony forming units of different serotypes of Shigella and recovery of bacteria studied. Histopathological analysis was carried out to study inflammatory immune responses. GPA was done to elucidate the invasiveness of different serotypes of Shigella. Secretions of AMPs were measured by enzyme-linked immunosorbent assay (ELISA). Western blotting was performed to check the expression of AMPs and nuclear factor kappa B in IVOC explants. After 24 h post-infection, the colon biopsies showed intense inflammatory reaction. In both IVOC and GPA, S. dysenteriae 1 was the most invasive as compared to S. flexneri and S. sonnei. S. sonnei was the least invasive. ELISA demonstrated that S. sonnei dampened the HBD (human β-defensin)-2 responses whereas there was augmentation by S. dysenteriae and there was a modest but non-significant increase by S. flexneri. A modest increase in HBD-3 by S. sonnei and S. flexneri was observed but was not found to be significant. However, western blotting data showed upregulation of all AMPs by all serotypes. Western blotting is more sensitive than ELISA. In the present study, differences in invasiveness and AMP production induced by different serotypes of Shigella were found. Human intestinal IVOC represents a model system to investigate early interaction between pathogenic bacteria and the human gut.

2221. Use of PCR and MLST analysis to establish Treponema pallidum as the cause of a painful oral lesion

Abstract Background A syphilitic chancre is described classically as a single, indurated, and painless ulcer at the site of Treponema pallidum inoculation. Methods We present a 39-year-old heterosexual, monogamous, HIV-negative woman with a painful tongue lesion of one month’s duration. To exclude a malignancy, a biopsy was performed. Results Histopathology showed intense inflammatory reaction, and T. pallidum immunocytochemistry demonstrated a high density of spirochetes in the subepithelial connective tissue. Subsequent testing revealed a reactive anti-T. pallidum IgG EIA and an RPR titer of 1:8. PCR amplification and multi-locus sequence typing (MLST) confirmed that the visualized spirochetes were T. pallidum. This particular MLST strain pattern has been identified twice before and named ST-109; however, no clinical information is available. Diagnosis and appropriate treatment were delayed since syphilis was not suspected initially. When the results became available, the patient was treated with 2.4 million units of benzathine penicillin given IM with complete resolution of the lesion. The male partner, who reported no symptoms, had a positive T. pallidum EIA with an RPR titer of 1:64 and was also treated with penicillin. Within four months, a woman from an adjacent town presented to urgent care with three perineal severely painful lesions, testing negative for HIV, Herpes Simplex Virus (HSV) by PCR of an ulcer swab, and chlamydia and gonorrhea on a urine sample by a NAAT. The anti-T. pallidum EIA was reactive, and the RPR titer was 1:4. The lesions resolved with IM penicillin treatment. Conclusion The observations reported here indicate that a subset of early syphilis cases may present with painful lesions. Despite the commonly held belief that syphilitic chancres are painless, several case series have reported tender lesions, single or multiple, without evidence of concurrent HSV infection to explain the pain. This is the first PCR and MLST sequencing analysis of a T. pallidum strain within a painful chancre. Given the rise in the number of cases of syphilis in the USA, it is essential that clinicians be aware of this atypical presentation. It remains to be determined whether such painful lesions are only associated with certain T. pallidum genotypes. Disclosures All Authors: No reported disclosures.

Role of osteopontin in the process of pulpal healing following tooth replantation in mice.

IntroductionThe role of osteopontin (OPN) following severe injury remains to be elucidated, especially its relationship with type I collagen (encoded by the Col1a1 gene) secretion by newly-differentiated odontoblast-like cells (OBLCs). In this study, we examined the role of OPN in the process of reparative dentin formation with a focus on reinnervation and revascularization after tooth replantation in Opn knockout (KO) and wild-type (WT) mice.MethodsMaxillary first molars of 2- and 3-week-old-Opn KO and WT mice (Opn KO 2W, Opn KO 3W, WT 2W, and WT 3W groups) were replanted, followed by fixation 3–56 days after operation. Following micro-computed tomography analysis, the decalcified samples were processed for immunohistochemistry for Ki67, Nestin, PGP 9.5, and CD31 and in situ hybridization for Col1a1.ResultsAn intense inflammatory reaction occurred to disrupt pulpal healing in the replanted teeth of the Opn KO 3W group, whereas dental pulp achieved healing in the Opn KO 2W and WT groups. The tertiary dentin in the Opn KO 3W group was significantly decreased in area compared with the Opn KO 2W and WT groups, with a significantly low percentage of Nestin-positive, newly-differentiated OBLCs during postoperative days 7–14. In the Opn KO 3W group, the blood vessels were significantly decreased in area and pulp healing was disturbed with a failure of pulpal revascularization and reinnervation.ConclusionsOPN is necessary for proper reinnervation and revascularization to deposit reparative dentin following severe injury within the dental pulp of erupted teeth with advanced root development.

P.80 Respiratory chain dysfunction in dermatomyositis is associated with mitochondrial DNA depletion

Patients with dermatomyositis suffer from reduced aerobic metabolism contributing to impaired muscle function, which has been linked to cytochrome c oxidase (COX) deficiency in muscle tissue. This mitochondrial respiratory chain dysfunction is typically seen in perifascicular regions, which also show the most intense inflammatory reaction along with capillary loss and muscle fiber atrophy. The objective of this study was to investigate the pathobiology of the oxidative phosphorylation deficiency in dermatomyositis. Muscle biopsy specimens with perifascicular COX deficiency from five juveniles and seven adults with early untreated dermatomyositis were investigated. Immunohistochemical analyses of subunits in the respiratory chain were performed including complex I (subunit NDUFB8), complex II (succinate dehydrogenase, subunit SDHB) and complex IV (COX, subunit MTCO1). The results showed a profound deficiency of complex I and IV in the perifascicular regions with enzyme histochemical COX deficiency, whereas succinate dehydrogenase activity and complex II were preserved. In situ hybridization of mitochondrial RNA showed depletion of mitochondrial DNA (mtDNA) transcripts in the perifascicular regions. Analysis of mtDNA by next generation deep sequencing and quantitative PCR in affected muscle regions showed an overall reduction of mtDNA copy number particularly in the perifascicular regions. In conclusion, the respiratory chain dysfunction in dermatomyositis muscle is associated with mtDNA depletion causing deficiency of complex I and IV which are partially encoded by mtDNA, whereas complex II which is entirely nuclear gene encoded is preserved. The depletion of mtDNA indicates a perturbed replication of mtDNA partially explaining the muscle pathology and the disturbed aerobic metabolism.

Clinical and histological comparative outcomes after injections of poly-L-lactic acid and calcium hydroxyapatite in arms: A split side study.

Although much has been published on the use of poly-L-lactic acid (PLLA) and calcium hydroxyapatite (CaHA) for off-face indications, questions remain regarding their exact mechanisms of action in subcutaneous tissue and their comparative efficacy. To present the clinical and histological results of the PLLA and CaHA injections into the opposing arms of the same patients. Five women received superficial subcutaneous injections of PLLA into the left arm and CaHA into the right arm. After three sessions, the clinical and histological outcomes were analyzed. After the first session, three patients showed improvement in the right arm (CaHA), but at the end of study, two patients showed better results in the left arm (PLLA). Histologically, moderate to intense lymphocytic and giant cell infiltrate, as well as collagen and elastic fiber neoformation, were observed equally near the particles of both products. Dermis had no inflammatory or fiber alterations. In this study, there were no clinical differences between these two fillers. Despite current thinking and previous histological studies, we found both products produced moderate to intense inflammatory reaction, as well as collagenic/elastogenic fiber neoformation, only in the subcutaneous tissue of the immediate vicinity and surrounding the individual filler particles.

ORAL LEUKOPLAKIA IN A SMOKING PATIENT USING INHALED FORMOTEROL AND BUDESONIDE

A 67-year-old non-White female patient was referred to our service for evaluation of a white oral lesion. The patient reported being a heavy smoker for 56 years and a 15-year history of asthma controlled with inhaled formoterol fumarate and budesonide for 15 years. Extraoral examination showed no relevant alterations. Intraoral examination revealed a nondetachable white verrucous plaque on the right side of the soft palate close to the limit with the hard palate. The lesion measured approximately 1.5 × 1.0 cm and had regular borders and well-defined limits. The differential diagnosis included oral leukoplakia and verrucous carcinoma. A biopsy was performed under local anesthesia. Histologic examination revealed epithelium with hyperkeratosis, acanthosis, and an intense inflammatory reaction in the lamina propria, suggesting oral leukoplakia. The lesion was excised and the clinical and microscopic analyses confirmed the diagnosis of oral leukoplakia. No evidence of recurrence was observed at 19-month follow-up.

In Vivo Evaluation of PCL Vascular Grafts Implanted in Rat Abdominal Aorta.

Electrospun tissue-engineered grafts made of biodegradable materials have become a perspective search field in terms of vascular replacement, and more research is required to describe their in vivo transformation. This study aimed to give a detailed observation of hemodynamic and structural properties of electrospun, monolayered poly-ε-caprolactone (PCL) grafts in an in vivo experiment using a rat aorta replacement model at 10, 30, 60 and 90 implantation days. It was shown using ultrasound diagnostic and X-ray tomography that PCL grafts maintain patency throughout the entire follow-up period, without stenosis or thrombosis. Vascular compliance, assessed by the resistance index (RI), remains at the stable level from the 10th to the 90th day. A histological study using hematoxylin-eosin (H&E), von Kossa and Russell–Movat pentachrome staining demonstrated the dynamics of tissue response to the implant. By the 10th day, an endothelial monolayer was forming on the graft luminal surface, followed by the gradual growth and compaction of the neointima up to the 90th day. The intense inflammatory cellular reaction observed on the 10th day in the thickness of the scaffold was changed by the fibroblast and myofibroblast penetration by the 30th day. The cellularity maximum was reached on the 60th day, but by the 90th day the cellularity significantly (p = 0.02) decreased. From the 60th day, in some samples, the calcium phosphate depositions were revealed at the scaffold-neointima interface. Scanning electron microscopy showed that the scaffolds retained their fibrillar structure up to the 90th day. Thus, we have shown that the advantages of PCL scaffolds are excellent endothelialization and good surgical outcome. The disadvantages include their slow biodegradation, ineffective cellularization, and risks for mineralization and intimal hyperplasia.

Respiratory chain dysfunction in perifascicular muscle fibres in patients with dermatomyositis is associated with mitochondrial DNA depletion.

Patients with dermatomyositis (DM) suffer from reduced aerobic metabolism contributing to impaired muscle function, which has been linked to cytochrome c oxidase (COX) deficiency in muscle tissue. This mitochondrial respiratory chain dysfunction is typically seen in perifascicular regions, which also show the most intense inflammatory reaction along with capillary loss and muscle fibre atrophy. The objective of this study was to investigate the pathobiology of the oxidative phosphorylation deficiency in DM. Muscle biopsy specimens with perifascicular COX deficiency from five juveniles and seven adults with DM were investigated. We combined immunohistochemical analyses of subunits in the respiratory chain including complex I (subunit NDUFB8), complex II (succinate dehydrogenase, subunit SDHB) and complex IV (COX, subunit MTCO1) with in situ hybridisation, next generation deep sequencing and quantitative polymerase chain reaction (PCR). There was a profound deficiency of complexes I and IV in the perifascicular regions with enzyme histochemical COX deficiency, whereas succinate dehydrogenase activity and complex II were preserved. In situ hybridisation of mitochondrial RNA showed depletion of mitochondrial DNA (mtDNA) transcripts in the perifascicular regions. Analysis of mtDNA by next generation deep sequencing and quantitative PCR in affected muscle regions showed an overall reduction of mtDNA copy number particularly in the perifascicular regions. The respiratory chain dysfunction in DM muscle is associated with mtDNA depletion causing deficiency of complexes I and IV, which are partially encoded by mtDNA, whereas complex II, which is entirely encoded by nuclear DNA, is preserved. The depletion of mtDNA indicates a perturbed replication of mtDNA explaining the muscle pathology and the disturbed aerobic metabolism.

ИСПОЛЬЗОВАНИЕ ПЛАЗМЫ, ОБОГАЩЕННОЙ ТРОМБОЦИТАМИ, ПРИ ЛЕЧЕНИИ ОЖОГОВ В ЭКСПЕРИМЕНТЕ

Objective. To evaluate the effectiveness of platelet-rich plasma injections in the treatment of wounds in experimental animals with burns. Materials and methods. The experiment was carried out on 30 sexually mature male Wistar rats, which, after simulation of infected burn wound with boiling water and excision of necrotic tissue to pinpoint bleeding, hemostasis, on the second day, were divided into the main and control groups, depending on the chosen strategy of subsequent treatment. Wound defect of the animals in the control group (15 rats) was treated and bandages soaked in 0.02% decamethoxin solution were applied. Platelet-rich plasma was additionally injected into the wound area of the animals in the main group (15 rats) on the 1&lt;sup&gt;st&lt;/sup&gt;, 3&lt;sup&gt;rd&lt;/sup&gt;, 5&lt;sup&gt;th&lt;/sup&gt; day after necrectomy. The remaining plasma was applied to the wound surface followed by closure with a polyvinyl chloride film and gauze bandage. Wound dressings were performed daily. The study involved a histological examination of the wound site and surrounding tissues the collection of which was carried out on the 1&lt;sup&gt;st&lt;/sup&gt;, 3&lt;sup&gt;rd&lt;/sup&gt;, 7&lt;sup&gt;th&lt;/sup&gt;, 14&lt;sup&gt;th&lt;/sup&gt; day after the removal of necrotic tissues. Results. In animals of the main group, the treatment of which involved the use of platelet-rich plasma, on the 3rd day after necrectomy in the wound area a decrease in the depth of injury that extended only of to subcutaneous tissue, a decrease in the activity of the inflammatory reaction in the tissues, followed by the proliferation of new epidermis and almost complete healing of wound defect up to 14 days were determined. For comparison, pathological changes in the tissues of the injured area of the animals in the control group were more pronounced with signs of muscle injury. In addition, during the entire observation period in this group of animals, an intense inflammatory reaction with low proliferative activity persisted. Conclusion. The effectiveness of platelet-rich plasma injections as an element of complex local treatment of burn wounds in rats has been experimentally confirmed. What this paper adds The expediency and effectiveness of platelet-rich plasma in the treatment of burns was histologically substantiated in the work on a specific experimental model of rats. In particular, its ability to positively influence the paranecrotic zone, reduce the intensity of inflammatory reaction in tissues, actively restore skin appendages, and stimulate the processes of proliferation and epithelization was established.

Adenosquamous inflammatory mammary carcinoma in bitch: Can CA 15-3, CRP and LDH improve prognosis?

Inflammatory mammary carcinoma (IMC) is characterized by the presence of any subtype of aggressive carcinoma in association with an intense inflammatory reaction, in addition to the presence of tumor emboli. Considering IMC as a type of cancer with chronic inflammation, the complex relationship between inflammation, and alterations in cellular metabolism have highlighted the rationale assessment of cancer antigen 15-3 (CA 15-3), c-reactive protein (CRP), and lactate dehydrogenase (LDH). The present report had the objective to describe a rare case of adenosquamous carcinoma in mammary gland of a 10-year-old bitch, 7 kg, sexually intact, mixed-breed and how CA 15-3, CRP and LHD helped to determinate the clinical presentation of inflammatory carcinoma and the prognosis. Complete clinical staging, cell blood count and renal, and hepatic biochemistry was performed and did not reveal any significant abnormalities. CA 15-3 (9.46 IU/mL), CRP (5.3 mg/L), and LDH (287 U/L) were all above the references values reported by the literature. Although adenosquamous carcinoma of the mammary gland with inflammatory presentation in dogs is rare, the association of serum biomarkers could improve the patient's prognosis evaluation.

Lentigo Maligna on a Difficult Location of the Face Treated with 5% Imiquimod: A Case Report of 2-Year Follow-Up.

Topical imiquimod is an off-label alternative treatment for lentigo maligna used when surgery cannot be performed. Severe inflammatory response induced by this drug may generate many complaints and force patients to discontinue use. We present a case in which interval treatment with 5% topical imiquimod was implemented for severe inflammatory response. An 82-year-old Caucasian woman presented with a large, irregularly pigmented lesion on her left cheek within the scar of a previously excised melanocytic lesion diagnosed as junctional nevus. Based on dermatoscopical examination confirmed by histopathological description, lentigo maligna was diagnosed. Since the lesion was large and covered the lower left eyelid and due to the risk of disfigurement, the patient refused surgical excision. Therefore, treatment with imiquimod 5% once daily, five times per week, was offered. After 5 weeks, the treatment was stopped because of intense inflammatory reaction and ulceration. On the follow-up visit after 3 months, videodermatoscopical examination revealed changes in the pigmentation of the lentigo maligna including the presence of residual gray dots and fading of the previous dark brown and black colors. The inflammatory response had almost resolved. The second course of treatment with imiquimod five times a week was implemented again for 5 weeks, and after a 2-month interval the third course with the same regimen was started. Total clearance of the lesion was achieved, which was confirmed by videodermatoscopical examination. During the 2-year follow-up, no relapse was observed based on dermatoscopical examination. We propose interval treatment with topical imiquimod 5% lentigo maligna for severe inflammatory reactions in patients with contraindications to surgery. This could help patients overcome this typical response effect and decrease their cessation of treatment.