Polymorphonuclear Infiltration Research Articles

Synovial tissue features associated with poor prognosis in inflammatory arthritis

BackgroundInflammatory arthritis encompasses a group of immune-mediated diseases characterized by chronic joint inflammation. Despite having pathogenic mechanisms in common, the prognosis of rheumatoid arthritis (RA), psoriatic arthritis (PsA), and undifferentiated arthritis (UA) could be different regarding progression to chronic, to erosive, or to self-limited disease. Our aim was to evaluate the potential association of synovial tissue (ST) inflammatory cell infiltrate, the presence of ectopic lymphoid neogenesis (LN +) structures, and poor prognosis factors (PPF) in patients with RA, PsA, and UA.MethodsWe conducted a retrospective study including patients with active arthritis (RA, PsA, UA) who had ST obtained by rheumatological arthroscopy or ultrasound-guided biopsy. Clinical, demographic, and immunohistochemical data of the synovium was evaluated. Patients with biological therapy at the time of synovial biopsy were excluded. PPF in patients with RA and UA were defined by the presence of anti-cyclic citrullinated peptide antibodies and/or rheumatoid factor, development of bone erosions, or requirement of biological therapy during the follow-up. PPF in patients with PsA were defined as the presence of high levels of acute-phase reactants (ESR/CRP), dactylitis or nail involvement at the time of biopsy, development of bone erosion, or requirement of biological therapy during the follow-up.ResultsA total of 88 patients were included: 26 RA, 33 PsA, and 29 UA. All patients were followed up for 5 years after the biopsy. Fourteen (53.84%) RA patients had PPF, and 17 (65.38%) had LN + . LN + was associated with PPF (p 0.038) and biologic therapy initiation (p 0.018). A total of 14 (43.75%) PsA patients had PPF. CD15 infiltrate (410.68 [SD 477.63] cells/mm2) was associated with PPF (p 0.008) in PsA patients. Sixteen (55.17%) patients with UA had PPF, and 13 (44.82%) had LN + . In this group, synovial CD68 + macrophages cells density was negatively correlated with DAS28-CRP (r = − 0.346, p 0.042).ConclusionsThe presence of LN + and higher CD15 + polymorphonuclear cells infiltrate was associated with PPF in RA and PsA, respectively. No associations were found for UA. These findings suggest a great heterogeneity of the ST features and its pathogenic implications in the subtypes of inflammatory arthritis.

Reinforcement of colon anastomosis healing with leukocyte platelet-rich fibrin in rabbit model

AimThis study investigated the regenerative efficacy of leukocyte platelet-rich fibrin (L-PRF) on colon anastomotic healing in rabbits. Main methodsThirty-six healthy male white New Zealand rabbits were subjected to complete transactions of the ascending colon. The rabbits were equally divided into two groups: the control group, where the transected colon ends were anastomosed by a simple interrupted suture pattern, and the L-PRF-treated group, in which L-PRF was wrapped entirely around the anastomotic line. The postoperative acute pain scale was assessed using the Bristol Rabbit Pain Scale before surgery and at each four-hour interval post-operatively. After euthanizing the rabbits, the adhesion degree score, anastomotic bursting pressure, and stenosis degree of the anastomotic colon were assessed, and histopathological examination at the 7th, 14th, and 28th days postoperatively. Key findingsRabbits in both groups showed a significant increase in pain scores compared to baseline. Postoperatively, the L-PRF group exhibited significantly lower pain scores, adhesion scores, and stenosis degrees than the control group. However, the anastomotic bursting pressure was significantly higher in the L-PRF group. Re-epithelialization, polymorphonuclear neutrophil infiltration, granulation tissue formation, and collagen deposition scores were improved considerably in the L-PRF group compared to the control group. Immunostaining of growth factor expression was significantly lower in the control than in the L-PRF group. SignificanceThe L-PRF can augment collagen deposition, re-epithelialize the mucosa, promote angiogenesis, reduce adhesions, and diminish the stenosis degree scores. Therefore, it can be considered a promising aid in healing bowel anastomoses.

Chitosan thread in the healing of the cecal wall of rabbits (Oryctolagus cuniculus) submitted to cecorrhaphy.

The development and evaluation of synthesis materials are crucial to reducing the morbidity and magnitude of post-enterorrhaphy surgical complications. Despite the possibility of production, chitosan thread has not yet been used in enterorrhaphy, and its effects on intestinal healing have not been evaluated. Therefore, this study aimed to evaluate the effects of chitosan thread on the intestinal wall repair of rabbits submitted to cecorrhaphy. For this, 42 rabbits were allocated into two groups with 21 animals. One group was submitted to cecorrhaphy with chitosan suture thread (CG) and the other with poliglecaprone suture thread (PG). The occurrence of postoperative complications, the intensity of edema, cellular response, formation of granulation tissue, as well as the deposition and maturation of collagen fibers, and the intensity of vascular endothelial growth factor (VEGF-α) expression, were evaluated during the intestinal wall repair process. The evaluations occurred on the 5th, 15th, and 25th postoperative (PO) days. The animals did not develop peritonitis, but adherence was observed in six animals from CG and seven from PG, with no difference between groups. The polymorphonuclear infiltrate showed higher intensity and higher amount of type III collagen fibers in CG on the 15th PO day. In contrast, a lower amount of type I collagen fibers was observed in CG samples on the 25th PO day. Therefore, the chitosan thread used for cecorrhaphy in rabbits results in minimal postoperative complications, presents biocompatibility, and bioactively assists the tissue repair process of the cecal wall, inducing minimal tissue reaction, stimulating the deposition of type III collagen fibers in the proliferative phase, with sustained VEGF-α expression, but with reduced deposition of type I fibers, indicating a delay in collagen maturation.

0.01% Hypochlorous Acid Treats Aspergillus fumigatus Keratitis in Rats by Reducing Fungal Load and Inhibiting the Inflammatory Response.

To investigate the antifungal and anti-inflammatory effects of 0.01% hypochlorous acid (HCLO) on rats with Aspergillus fumigatus keratitis. The time-kill assay and broth microdilution procedures were used in vitro to demonstrate that 0.01% HCLO was fungicidal and fungistatic. The severity of the disease was evaluated in vivo using a clinical score and slit-lamp photographs. Fungal load, polymorphonuclear neutrophil infiltration, and the production of related proteins were determined using colony plate counting, in vivo confocal microscopy, periodic acid-Schiff staining, fungal fluorescence staining, immunofluorescence staining, myeloperoxidase assay, and Western blotting. In vitro, 0.01% HCLO can destroy A. fumigatus spores in 1minute. The optical density of the 0.01% HCLO group was significantly lower than that of the phosphate-buffered saline control group (P < 0.01), and no visible mycelium was observed using a fluorescence microscope. 0.01% HCLO reduced the severity of A. fumigatus keratitis in rats by decreasing the clinical score, fungal loading (periodic acid-Schiff, plate count, and fungal fluorescence staining), and inhibiting neutrophil infiltration and activity (immunofluorescence staining and myeloperoxidase). Furthermore, the Western blot analysis revealed that 0.01% HCO decreased protein expression levels of tumor necrosis factor-α and IL-1β. According to our findings, 0.01% HCLO can kill A. fumigatus spores in vitro. It has antifungal and anti-inflammatory effects on A. fumigatus keratitis in rats. It also inhibited A. fumigatus growth; decreased neutrophil infiltration, tumor necrosis factor-α, and IL-1β expression; and provided a potential treatment for fungal keratitis. This study provides a potential treatment for fungal keratitis in the clinic.

Beneficiary Effects of Colchicine on Inflammation and Fibrosis in a Mouse Model of Kidney Injury

Introduction: Low-grade inflammation is seen in many chronic illnesses, including chronic kidney disease (CKD). We have recently reported on beneficiary effects of anti-inflammatory treatment in the interleukin (IL-) 1 pathway on anemia as well as CKD extent in a mouse model. Colchicine has been shown to have beneficiary effects in several inflammatory conditions through various mechanisms, including inhibition of tubulin polymerization as well as caspase-1-mediated IL-1 activation. Methods: Kidney injury (KI) was induced by administering an adenine diet to 8-week-old C57BL/6J mice treated with colchicine (Col) (30 µg/kg) or saline injections for 3 weeks, generating 4 groups: C, Ccol, KI, and KIcol. Results: KI animals had an increase in inflammation indices in the blood (neutrophils), liver, and kidneys (uromodulin, IL-6, pSTAT3). Increased kidney tubulin polymerization and caspase-1 in KI, as well as kidney Mid88 and IRAK4 (downstream of IL-1), were inhibited in KIcol. Kidney macrophage and polymorphonuclear infiltration (positive for F4/80 and MPO, respectively), the percentage of fibrotic area, and TGFβ mRNA levels were lower in KIcol versus KI. Conclusions: Colchicine inhibited tubulin polymerization and caspase-1 activation and attenuated kidney inflammation and fibrosis in a mouse model of adenine-induced KI. Given its reported safety profile for long-term anti-inflammatory therapy without increasing infection tendency, it may serve as novel therapeutic approach in CKD.

Endometrial inflammatory response to insemination in jennies

In the jenny introduction of semen into the uterus triggers an endometrial inflammatory response characterized by polymorphonuclear (PMN) infiltration. The presence of cryoprotective agents in frozen-thawed semen may increase the inflammatory reaction. The objective of this study was to evaluate the effects of two cryoprotective agents in the induction of endometrial inflammatory response in jennies inseminated with extended semen or sham-inseminated with extender only or with deionized water. Eight Creole donkeys were inseminated or sham-inseminated in successive cycles with each one of the following treatments: DW: deionized water; B: a commercial cryopreservation extender containing glycerol (BotuCrio®, Botupharma); LDL: a cryopreservation extender that contains low density lipoproteins instead of glycerol (UNAM-extender); FS: fresh semen in a extender which contains no cryoprotective agents (INRA96, France); S-B: Semen extended in Botucrio; and S-LDL: Semen extended in UNAM-extender. The order in which jennies received each treatment was randomized. In each case, four 0.5 ml straws were used for deep insemination in the uterine horn ipsilateral to ovulation. In the case of treatments that included semen, each straw contained 200 million sperm cells. After each treatment the jennies were administered 5 mg of PGF2α at day 5 post-ovulation to induce return to estrus in order to administer the next treatment, until each jenny had received allthe treatments. Endometrial cytology samples were obtained before each treatment (time 0) and at 8, 12, 24 and 48-h after treatment. Smears were fixed and stained with Diff-Quik. Five fields were randomly chosen for microscopic observation. The total number of PMN and the total number of cells of any type were counted. The proportions of PMN in the smears were compared using the "generalized linear models" procedure of the SPSS program. Predictors were treatment, post-treatment time, treatment x time interaction. The jenny was used as a block. For multiple comparisons, the Bonferroni method was used. Extenders with cryoprotective agents and no semen resulted in higher proportion (p˂ 0.05) of PMN than the same extenders containing semen (B 43% vs S-B 26.9 % LDL 36% vs S-LDL 25.3%). The duration of significantly elevated PMN was also higher when extenders contained no semen. The extender with no cryoprotective agent (INRA96) also resulted in a higher proportion of PMN (39.6 %) than S-B and S-LDL. DW resulted in a proportion of PMN (44.8%) similar to those of the extenders containing no semen, and higher than those of the extenders containing semen. The results suggest that the presence of semen modulates and reduces the endometrial inflammatory response of jennies. The cryoprotective agents by themselves do not appear to exacerbate the inflammatory response, as a similar response was produced by an extender containing no cryoprotective agent and even by DW.

Does the uterine inflammatory response differ between mares and donkey jennies inseminated with frozen donkey semen?

Frozen donkey semen produces poor fertility rates in jennies (0-11%) while showing normal fertility in mares (33-53%). This may be related to interspecific differences in the interaction of the uterus with frozen semen. This study aimed to evaluate and compare the inflammatory process in the endometrium of donkey jennies and mares inseminated with cryopreserved donkey semen. A total of 36 estrous cycles from 12 Pega donkey jennies and 36 estrous cycles from 12 mares (Breton and Mangalarga), aged between 3 and 14 years old were used. Once ovulation was detected, the animals were inseminated deeply in the horn ipsilateral to ovulation with thawed donkey semen containing 1 billion sperm. The following parameters were evaluated in both species at ovulation/before insemination (time 0h) and after insemination (times 6h, 12h, 24h, 48h, 72h, 96h and 120h): uterine vascularization, endometrial cytology, serum progesterone levels and pregnancy rate. Normally distributed data were analyzed using a mixed-model ANOVA and a Wilcoxon test for non-normally distributed data, using R software. Donkey jennies showed a higher percentage of colored pixels in the uterine body region at T6 compared to mares. Also, the percentage of inflammatory cells was higher in the donkey jenniescompared to the equine cytology at T12 (52.75 versus 37.44) and T48 (16.00 versus 3.98). The inflammatory peak was reached at T6 in both species (55.77 versus 57.31) but remained elevated until T12 only in donkeys. Although a complete resolution of the inflammation occurred within 48 hours after AI in both species, the percentage of inflammatory cells remained higher in jennies than in mares. An eosinophilic uterine infiltrate was present in donkeys, while it was not observed in mares. In addition, donkey jennies showed a significant increase in monocytes at T12, compared to the other times. Also, jennies showed a higher mean P4 level (13 ng/ml), compared to the mares (6 ng/ml), both species had progesterone concentrations compatible with the maintenance of pregnancy. The pregnancy rate 14 days after AI was higher in mares than in donkeys, reaching a rate of 54.55% (6/11) versus 0% (0/12), respectively. In conclusion, the inflammatory uterine response in the presence of frozen donkey semen differs between mares and jennies. Jennies have a later inflammatory resolution, marked by the presence of intense polymorphonuclear and eosinophilic infiltrates. These differences may contribute to the lower fertility rates in this species.

Hypertrophic pachymeningitis due to IgG4-related disease (RD-IgG4). A case report

IntroductionHypertrophic pachymeningitis (HP) is a clinico-radiological entity characterized by a thickening of the dura mater that may be focal or diffuse and manifested by a variety of neurological syndromes. Aetiologically, it is classified as infectious, neoplastic, autoimmune, and idiopathic. Many of these formerly idiopathic cases have been shown to fall into the spectrum of IgG4-related disease. ObjectiveTo describe the case of a patient attended for neurological involvement due to hypertrophic pachymeningitis with initial diagnosis of inflammatory myofibroblastic tumour and final diagnosis of IgG4-related disease. CaseA 25-year-old woman with neurological symptoms of 3 years’ evolution characterized initially by right hypoacusis, evolving with headache and diplopia. Magnetic resonance imaging (MRI) of the encephalon showed pachymeningeal thickening with involvement of vasculo-nervous structures in the tip of the cerebellum, cavernous sinus, ragged foramen, and optic chiasm. The patient presented for consultation with the result of an incisional biopsy that reported a proliferative lesion combining fibrous elements of fascicular or swirling arrangement with collagenized streaks with dense, lymphoplasmacytic infiltrate and some macrophages, with negative staining for ALK 1, with a diagnosis of inflammatory myofibroblastic tumour.Due to suspicion of IgG4-related disease (IgG4-RD) the biopsy was sent for review and pertinent complementary studies were requested. Biopsy reviewNon storiform fibrosis, predominantly lymphoplasmacytic infiltrate, histiocytes, and polymorphonuclear infiltrate in sectors, without granulomas or atypia. Staining for germs negative. Immunohistochemistry with 50–60 IgG4+/HPF cells and range of 15%–20%, CD68+ in histiocytes, CD1a−, S100−.The patient presented deterioration of visual acuity due to ophthalmic nerve involvement, so glucocorticoid treatment was started in pulses and rituximab with regression of symptoms and imaging improvement of the lesions. ConclusionHP is a clinical imaging syndrome with variable symptoms and aetiologies that poses a diagnostic challenge.In this case the initial diagnosis was inflammatory myofibroblastic tumour, which is a neoplasm of variable behaviour, locally aggressive, and can metastasize; it is one of the main differential diagnoses of IgG4-related disease because they share anatomopathological features, including storiform fibrosis.IgG4-RD is an immune-mediated condition that can have single or multiple involvement. Its diagnosis is complex when it presents with single organ involvement or in non-typical organs (CNS, meninges) in which data are scarce, as in the case of our patient with single organ involvement of the CNS.Although there are classification criteria to guide non-specialists in the diagnosis, the sum of the clinical picture, imaging, laboratory, pathological anatomy, and immunohistochemistry will always be evaluated together for a definitive diagnosis.

Different cellular and immunohistochemical abdominal wall cicatrization parameters evaluation in comparison with sublay, onlay, and ipom technique in an experimental rat model.

Incisional hernia (IH) occurs when there is a partial or complete solution of continuity of a fascia previously incised. Systematic reviews demonstrate that surgical treatment of IHs with the use of meshes are approximately 16%. Meta-analyses have demonstrated the superiority of mesh placement using sublay technique, but without a pathophysiological explanation. Thus, we aim to evaluate the different techniques of mesh positioning in an experimental model. Fifty rats were distributed into five groups; control; simulation (SM)-submitted to laparotomy only; onlay-the mesh was positioned in onlay fashion; retromuscular (SL)-the mesh was positioned in a sublay fashion; intraperitoneal (IPOM)-positioning of the mesh adjacent to the transversalis fascia, inside the cavity. After 60days, adhesions, tensiometry, histology, and immunohistochemistry were addressed. The IPOM group had the most adhesions, together with the SL group, with significantly relevant results. The SL group had higher values of tensiometric evaluation, while the IPOM group had the lowest mean in the tensiometry evaluation, being even lower than the SM group. Regarding histological and immunohistochemical findings, the SL group had a higher pixel number count compared to the groups, with statistical significance, in addition to higher expression of polymorphonuclear infiltrate and CD68 markers. The mesh positioning in sublay compartment is associated with the development of more pronounce minimum tensile force required for detaching the surrounding abdominal wall tissues it was incorporated. The intensity of these findings correlates to the different histological and immunohistochemical profiles observed following each repair, since SL group was characterized by a higher proportion of collagen, inflammatory, and reparative elements. Characterizing these pro-healing elements and its counterparts will allow the development of new therapeutic tools which could be added to the still far-from-ideal current therapeutic options for IH treatment.

Protective Effects of Interleukin-1 Blockade on Group B Streptococcus-Induced Chorioamnionitis and Subsequent Neurobehavioral Impairments of the Offspring.

Group B Streptococcus (GBS) is one of the most common bacteria isolated in human chorioamnionitis. Placental infection due to GBS is a major risk factor for fetal organ injuries, preterm birth, perinatal morbidity and mortality, and life-long multiorgan morbidities. Preclinical and clinical studies have shown that GBS-induced infection drives polymorphonuclear (PMN) cell infiltration within the placenta, the hallmark of human chorioamnionitis. In preclinical and clinical studies, the upregulation of interleukin(IL)-1β in the placenta and maternal/fetal blood was associated with a high risk of neurodevelopmental impairments in the progeny. We hypothesized that targeted IL-1 blockade administered to the dam alleviates GBS-induced chorioamnionitis and the downstream fetal inflammatory response syndrome (FIRS). IL-1 receptor antagonist (IL-1Ra) improved the gestational weight gain of GBS-infected dams and did not worsen the infectious manifestations. IL-1Ra reduced the IL-1β titer in the maternal sera of GBS-infected dams. IL-1Ra decreased the levels of IL-1β, IL-6, chemokine (C-X-C motif) ligand 1 (CXCL1), and polymorphonuclear (PMN) infiltration in GBS-infected placenta. IL-1Ra treatment reduced the IL-1β titer in the fetal sera of GBS-exposed fetuses. IL-1 blockade also alleviated GBS-induced FIRS and subsequent neurobehavioral impairments of the offspring without worsening the outcome of GBS infection. Altogether, these results showed that IL-1 plays a key role in the physiopathology of live GBS-induced chorioamnionitis and consequent neurobehavioral impairments.

Targeting IL-1β as an immunopreventive and therapeutic modality for K-ras-mutant lung cancer.

K-ras–mutant lung adenocarcinoma (KM-LUAD) is associated with abysmal prognosis and is tightly linked to tumor-promoting inflammation. A human mAb, canakinumab, targeting the proinflammatory cytokine IL-1β, significantly decreased the risk of lung cancer in the Canakinumab Anti-inflammatory Thrombosis Outcomes Study. Interestingly, we found high levels of IL-1β in the lungs of mice with K-rasG12D–mutant tumors (CC-LR mice). Here, we blocked IL-1β using an anti–IL-1β mAb in cohorts of 6- or 14-week-old CC-LR mice to explore its preventive and therapeutic effect, respectively. IL-1β blockade significantly reduced lung tumor burden, which was associated with reprogramming of the lung microenvironment toward an antitumor phenotype characterized by increased infiltration of cytotoxic CD8+ T cells (with high IFN-γ and granzyme B expression but low programmed cell death 1 [PD-1] expression) while suppressing neutrophils and polymorphonuclear (PMN) myeloid-derived suppressor cells. When querying the Cancer Genome Atlas data set, we found positive correlations between IL1B expression and infiltration of immunosuppressive PMNs and expression of their chemoattractant, CXCL1, and PDCD1 expressions in patients with KM-LUAD. Our data provide evidence that IL-1β blockade may be a preventive strategy for high-risk individuals and an alternative therapeutic approach in combination with currently available treatments for KM-LUAD.

Term Infant with Bilateral Parenchymal Brain Hemorrhages.

Term Infant with Bilateral Parenchymal Brain Hemorrhages.

Safety profile of intravenous administration of live Pichia pastoris cells in mice.

Pichia pastoris has been widely used to produce antigenic proteins aimed to integrate subunit vaccines. Moreover, increasing interest in large-scale vaccine production at the lowest cost is rapidly focusing in the development of yeast surface display (YSD) systems for delivery of antigens. In this scenario, the safety of live yeast administration must be warranted, however, such information is very scarce. Here, we assess the intravenous administration (i.v.) of live P. pastoris cells in order to trace dissemination in BALB/c mice and to evaluate the immune response raised against the yeast compared to the well-defined pathogen Candida albicans. Our results demonstrate dissemination of P. pastoris to the heart, kidney, and spleen, but it is quickly eliminated during the first 48 h postinfection (hpi), with persistence in the liver along with mild mononuclear (MN) and polymorphonuclear (PMN) infiltrate, which was resolved at 144 hpi. In vivo delayed-type hypersensitivity test (DTH) or in vitro antigenic stimulation of mice splenocytes demonstrate that transient infection of P. pastoris did not induce a cell-mediated immune response nor increase the level of circulating IgG or IgM. These results demonstrate the innocuous profile of P. pastoris and support its use as a safe delivery system for vaccine development.

Некротичний ентероколіт у передчасно народжених дітей, які померли: причини, фактори ризику летальності, гістологічні зміни в стінці кишки

Despite advances in the diagnosis and treatment of necrotizing enterocolitis (NEC), the associated morbidity and mortality rates remain high. Purpose - to establish risk factors for mortality of necrotizing enterocolitis in preterm born infants, as well as to analyze histological changes of the intestinal lining. Materials and methods. The course of NEC in 21 preterm neonates who died of this disease (group 1, n=21) over a period of 3 years was analyzed. To establish risk factors for mortality rate health indicators of children in group 1 were compared with the course of NEC in children who survived with similar stages of the disease (group 2, n=43). The following research methods were used: general clinical, laboratory, instrumental, histological and statistical. Results. Our data show that the main causes of severe stages of NEC in preterm infants is infection, often in combination with severe asphyxia. The identified risk factors for mortality allowed to establish that the risk of death for children with NEC was associated with: male sex (OR=4.675; χ2=7.679; p=0.006) - increases the risk for mortality by 4 time; inflammatory changes in the placenta (OR=6.139; χ2=10.501; p=0.002) - increases the risk by 6 times; red blood cell transfusion in children (OR=8.262; χ2=8.557; p=0.004) - increases the risk by 8 times; thrombocytopenia (OR=4.320; χ2=4.866; p=0.028) - increases the risk by 4 time; the developmen of multiple organ system failure (OR=12.364; χ2=17.578; p&lt;0.001) and DIC syndrome (OR=10.725; χ2=14.592; p&lt;0.001) - increases the risk by 12 and 11 times, respectively; the positive symptoms - oedema of the anterior abdominal wall (OR=14.025; χ2=19.258; p&lt;0.001) and vasodilation of the anterior abdominal l wall (OR=5.333; χ2=5.444; p=0.02) - increases the risk by 14 and 5 times, respectively; the intestinal pneumatosis on abdominal when x-ray detected (OR=6.840; χ2=6.867; p=0.009) and the peritoneal effusion detected by abdominal ultrasound (OR=8.750; χ2=14.448; p&lt;0.001) - increases the risk of mortality by 7 and 9 times, respectively. During histological examination of the intestinal wall with NEC lymphohistiocytic infiltration of submucosa indicates perinatal hypoxia and its crucial role in the thanatogenesis of the disease, while polymorphonuclear segmental neutrophil infiltration is associated with perinatal infection. In 15 children (71.4%) changes of both types were noted, which indicates mixed etiology of intestinal lesions. Conclusions. Study results confirmed that necrotizing enterocolitis is a serious disease of newborns with a high mortality rate. The severe forms of NEC occur against the background of infection in combination with hypoxia. The obtained risk factors for the mortality rate of NEC allow to improve the prognosis of the course of this disease, will provide an opportunity to identify children who need increased attention of doctors to the treatment and further management of these patients with the use of preventive technologies that can prevent catastrophic consequences. The presence of congenital intestinal defects in combination with premature birth contribute to the development and aggravate the course of NEC, up to the development of stage III and a negative prognosis of the disease. The research was carried out in accordance with the principles of the Helsinki Declaration. The study protocol was approved by the Local ethics committee of all participating institutions. The informed consent of the parents of patient was obtained for conducting the studies. No conflict of interests was declared by the authors. Key words: necrotizing enterocolitis, preterm neonates, risk factors for mortality, histological changes.

Treatment of experimentally induced partial-thickness burns in rats with different silver-impregnated dressings.

To evaluate the morphometric, macroscopic and microscopic aspects of experimentally induced partial-thickness burns in rats treated with different silver-based dressings. Wistar rats were used, divided into six treatments: saline (NaCl 0.9%); silver sulfadiazine 1%; Silvercel; Mepilex Ag; Aquacel Ag and Acticoat. The animals were monitored daily and euthanized at 7, 14 and 30 days after injury induction (DAI). At 7 DAI, necrosis/crust was greater in control, silver sulfadiazine and Mepilex Ag treatments, granulation tissue was induced by Aquacel Ag, polymorphonuclear infiltrate (PMN) infiltration was intensified by Mepilex Ag; mononuclear infiltrate (MN) infiltration and angiogenesis were increased by Silvercel. At 14 DAI, hemorrhage was decreased by Silvercel and Mepilex Ag, PMN infiltration increased by Acticoat. At 30 DAI, angiogenesis was greater in the Acticoat treatment and fibroblasts were increased by Acticoat and Mepilex Ag. Collagen was induced at 14 DAI by silver sulfadiazine and Aquacel Ag and, at 30 DAI, by silver sulfadiazine and Silvercel treatments. Silvercel and Acticoat presented better results than the other products. However, all the dressings were better than the control at some point during the process, and may contribute to the healing of partial thickness burns. Silvercel and Aquacel Ag treatments induced better cosmetic outcomes regarding wound closure and scarring.

Tissue healing changes on wounds in rats after treatment with Hancornia speciosa latex in cream-gel formulation

Hancornia speciosa latex has shown pharmacological potential in wound healing processes due to its angiogenic, osteogenic, and anti-inflammatory activities. The aims of this study were to carry out a cream-gel formulation with 5, 10 and 25% of H. speciosa serum latex and to evaluate its potential to stimulate the skin regeneration in rats' wounds. One hundred and twenty rats were divided into five groups: neutral control with saline (G1), cream-gel based on H. speciosa latex serum at 5% m/v (G2), cream-gel at 15% m/v (G3), cream-gel at 25% m/v (G4), and cream-gel (G5). The animals were euthanized at three, seven, 14 and 21 days after the injury induction, and some parameters were analyzed: wound contraction, necrosis, fibrin, polymorphonuclear and mononuclear infiltrates, fibroblast, angiogenesis, hemorrhage, and collagen. The therapeutic treatment with cream-gel at 15 and 25% is beneficial in the inflammatory phase of healing processes since it increased the angiogenesis and proliferation of mononuclear infiltrations in wounds. Regarding wound contraction, the treatment with cream-gel (5 and 15%) induced a higher rate of contraction in the proliferative phase. The 15% cream-gel formulation stimulated a greater production of collagen in the injured tissues. H. speciosa cream-gel is a low-cost herbal medicine which can aid in tissue repair.

Co-relation between clinical parameters and histological parameters with outcome in severe alcoholic hepatitis patients

Background and Aim: Alcoholic hepatitis (AH) is characterized by recent onset of jaundice, with or without other signs of liver decompensation (ascites and/or encephalopathy), in patients with chronic alcohol abuse. Severe AH is defined by a modified Madey’s discriminant function of 32 or higher and has poor prognosis. Alcoholic hepatitis histologic score is a new approach which includes four histologic features (bridging fibrosis or cirrhosis, hepatocellular or canalicular/ductular bilirubinostasis, severe polymorphonuclear infiltration and megamitochonia) that allows patient classification into three groups with different risks of death within 90 days.In the present study,we aimed to correlate histologic and clinical parameters with outcomes in severe alcoholic hepatitis patients, which in turn will predict the prognosis of the patients.

Clinical and pathomorphological analysis of deaths from COVID-19 in 2020

The aim of the work – to conduct clinical and pathomorphological analysis of deaths from COVID-19 in 2020. Materials and methods. We analyzed 41 case histories and results of pathological-anatomical examination of patients who were died of COVID-19 during 2020. Results. The lethal outcome of COVID-19 disease was recorded at day 22 (16; 27) of the disease. Among the dead, there is a high percentage of men (73.2 %), early old age and middle old age patients (75.6 %) with comorbid pathology (92.7 %). Early lung damage with COVID-19 in the deceased was determined by pronounced interstitial and interstitial-alveolar edema, the presence of erythrocyte stasis in the pulmonary microvessels, blood clots and hypoperfusion leukocyte stasis, as well as the presence of erythrocytes in the alveoli. Bilateral polysegmental subtotal viral pneumonia in 90.2 % of dead patients was characterized by significant edema and thickening of the alveolar walls with their moderate infiltration by lymphocytes, focal peribronchial and perivascular inflammatory polymorphonuclear infiltration, multiple and small exfoliated alveolar epithelium (87.8 %), as well as metaplasia of a few alveolocytes preserved on the luminal surface of the alveoli (82.9 %). Every tenth person who died of COVID-19 had signs of secondary bacterial microflora. In 85.4 % of patients who died on day 22–27 of the disease focal or sublobar pneumofibrosis was diagnosed. In those who died due to COVID-19, multiorgan failure was characterized by focal necrosis of the renal tubular epithelium (73.2 %), focal lymphocytic-leukocyte infiltration (12.2 %) and renal microvascular thrombosis (17.1 %), focal centro-lobular necrosis (90.2 %) and focal lymphocytic-leukocyte infiltration of lobes (7.3 %) of the liver. Thrombotic complications were confirmed in 22.0 % of deceased patients: ischemic cerebral infarction, transmural myocardial infarction, pulmonary embolism, deep vein thrombosis of the lower extremities under the pathology. These thrombotic complications were not diagnosed during life in all patients. The majority of deaths due to COVID-19 had morphological signs of chronic cardiovascular pathology. Ischemic heart disease and hypertension during the life of patients were not diagnosed in all cases. Conclusions. Early lung damage in COVID-19 in the deceased was determined by pronounced interstitial-alveolar edema, blood clots and leukocyte stasis in microvessels, less often – the presence of “hyaline membranes”. In 90.2 % of the dead patients bilateral polysegmental subtotal pneumonia with edema and lymphocytic infiltration of the pulmonary interstitium, inflammatory peribronchial and perivascular focal polymorphonuclear infiltrates, foci of atelectasis and dyscryphaseses was found. In 9.7 % of patients bilateral subtotal viral-bacterial fibrinous-purulent bronchopneumonia developed. In those who died on the 22nd–27th day of the disease focal pneumofibrosis was determined. Pathomorphologically, thrombotic complications, which were not diagnosed in all patients during their lifetime, were confirmed in 22.0 % of deceased patients. Most deaths from COVID-19 had morphological signs of chronic cardiovascular disease.

256 Effects of Transition Period Nutrition on Uterine Health

Abstract The transition from gestation to lactation, also known as the transition period, is a critical time for dairy cows. This phase is typically defined as 3 wk before parturition through 3 wk after parturition. Peak disease incidence (shortly after parturition) corresponds with the time of greatest negative energy balance (NEB), the peak in blood concentrations of nonesterified fatty acids, and the greatest acceleration of milk yield. Decreased fertility in the face of increasing milk production may be attributable to greater severity of postpartal NEB resulting from inadequate transition management or increased rates of disease. The depth and duration of NEB is highly related to dry matter intake. Formulation and delivery of appropriate diets that limit total energy intake to requirements but also provide proper intakes of all other nutrients (including the most limiting amino acids Met and Lys) before calving can help lessen the extent of NEB after calving. Cows that received controlled-energy diets during the last 3 wk prepartum had shorter days to pregnancy than cows that consumed high-energy diets in this time period. Additionally, a fully acidified prepartum diet formulated with a higher Ca concentration improved reproductive performance and uterine immune function in the postpartum period through the decrease of days to first ovulation, a tendency to decrease service per conception rate, improved glandular morphology, a tendency to increase polymorphonuclear neutrophil infiltration, and a tendency to increase the tight-junction protein occludin. Supplementation of rumen-protected methionine during the transition period resulted in improved uterine immune function through improved glandular morphology, increased neutrophil infiltration number after calving, and discovery of neutrophil extracellular trap formation in bovine endometrial tissue. In conclusion, nutritional strategies during the transition impact uterine health and fertility in dairy cows.

Structural organization of periodontium and morphological changes in its components in modeled periodontitis and its correction in rats

Introduction. The aim of our research was to study peculiar features of morphological changes in the components of the periodontium in the modeled acute periodontitis and its correction by stem cells.&#x0D; Materials and Methods. The results of histological study of periodontal tissues of the animals with acute experimental periodontitis, which was corrected using human mesenchymal stem cells as well as rat muscle stem cells, were analysed. Depending on the treatment, the animals were divided into 4 groups: the 1st group – the animals without pathology (intact); the 2nd – the animals with modeled acute periodontitis; the 3rd – the animals with periodontitis corrected by human mesenchymal stem cells, the 4th – the animals with periodontitis corrected using rat muscle stem cells.&#x0D; Results and Discussion. According to the results of the research it was established that correction by human mesenchymal stem cells and rats muscular stem cells had positive effect on a course of the modeled pathological process. The morphological changes characteristic of periodontitis gradually reduced, i.e. the normal blood supply to the tissues restored, the arteries were less full-blooded, the tone of their walls decreased. Consequently, the swelling of the tissues gradually decreased becoming of original size. The polymorphonuclear infiltration fields decreased and gradually disappeared. Osteoblasts promoted restoration of the bone structure of the jaw alveolar process. The size of the periodontal pocket decreased, and in some areas it fully closed as a result of complete adhesion of the mucous membrane of the gums to the tooth, which rose on its surface upwards.&#x0D; Conclusions. In the experimental modeling of acute periodontitis the destructive changes in the epithelium and connective tissue stroma, impaired blood flow, inflammatory response in the gum tissue were present. The correction by means of human mesenchymal stem cells as well as rat muscle stem cells had positive effect on the modeled pathological process: the morphological changes characteristic of periodontitis gradually reduced, normal blood supply to tissues restored, polymorphonuclear infiltration fields gradually decreased and disappeared. The bone structure of the jaw alveolar process restored owing to osteoblasts. The size of the periodontal pocket decreased; in some cases, they closed completely as a result of complete adhesion of the mucous membrane of the gums to the tooth, which rose on its surface upwards.