Novel Immunohistochemical and Morphological Approaches in a Retrospective Study of Post-Mortem Myocarditis.

Background and Objectives: This study presents a retrospective analysis of 26 autopsy cases from a single centre, primarily focusing on forensic cases, with a majority of male individuals. Materials and Methods: We systematically analysed autopsy reports and cardiac tissue slides using haematoxylin-eosin stain and immunohistochemistry for CD3, CD163, and IL-6. The histological assessment evaluated key variables such as inflammation severity, necrosis, and background changes using a standardised grading system. Quantitative analysis of immunohistochemical markers was performed, calculating the percentage of positively stained cells within the inflammatory infiltrate. Results: The average age was 51.6 years, slightly skewed towards older males. The fatalities varied widely, with sudden death and drug abuse being the most common conditions linked to myocarditis findings on histological examination. A strong correlation was found between the severity of inflammation (measured by size within a myocardium section) and the scoring system based on the number of inflammatory foci per section (p ≤ 0.001). Most cases showed mild to minimal fibrosis, with some exhibiting moderate to severe fibrosis, arteriosclerosis, and myocyte hypertrophy. The presence of protein CD3 in the inflammatory infiltrate revealed a moderate inverse correlation between the CD3 values and the severity of inflammation and necrosis, and a strong inverse correlation with neutrophil levels. CD3 levels were higher in sudden death cases and lower in cases with numerous inflammatory foci, highlighting the discreet nature of lymphocytic myocarditis. Macrophage presence, assessed using CD163, showed a moderate inverse correlation with neutrophil levels and significant differences between sudden death and non-sudden death cases. Macrophage-rich inflammation was observed in cases with pneumonia/bronchopneumonia-associated lesions. IL-6 expression showed a moderate direct correlation with inflammation severity (p = 0.028), severity of necrosis (p = 0.005), and the number of inflammatory foci per section (p = 0.047). A moderate inverse correlation was found between CD3 and IL-6 expression (p = 0.005). Conclusions: These findings highlight the need for a unique immunohistochemical approach in forensic cases of myocarditis, differing from guidelines for endomyocardial biopsies due to diverse inflammatory cells. The study suggests exploring inflammatory chemokines within myocarditis foci for their significance in clinical scenarios. Specifically, IL-6, a crucial pro-inflammatory interleukin, correlated significantly with the severity of inflammation and necrosis (p < 0.05). This study provides novel and valuable insights into the histopathological and immunological markers of myocarditis in autopsy cases.

Vitamin D Status among 2-18-Year-Old Romanian Pediatric Patients: A Single-Center Study.

An adequate vitamin D level is essential for optimal bone mass formation during growth. The present study aimed to assess (i) the sex-specific, age-specific, and potential seasonal (spring, summer, winter) influences on the pediatric circulating levels of 25-hydroxyvitamin D (25(OH)D); (ii) determine the frequency of pediatric patients with vitamin D deficiency (VDD) or insufficiency (VDI); and (iii) quantify the association between age category, sex, and season types and susceptibility to VDD and VDI, respectively. Laboratory data were collected on serum 25(OH)D levels in children aged between 2 and 18 years (n = 1674) who underwent blood sampling following admission to a university pediatric hospital in Cluj-Napoca (Romania) between January and June 2023. VDD (<20 ng/mL) was observed in 27% of pediatric patients. Among toddlers and preschoolers (2-5 years), VDD was 11%, while it was 33% among school-aged children (6-11 years) and 39% among adolescents (12-18 years). We found a significant difference in the frequencies of vitamin D status between females and males (p = 0.006). Also, we found significant associations of vitamin D status with age categories (p < 0.0001) and seasonal variations (p = 0.03). After adjusting for season of blood collection, the multinomial logistic regression model showed that children aged 6-11 years old (adjusted OR = 7, 95% CI: (4.9, 9.4)), children aged 12-18 years old (adjusted OR = 14, 95% CI: (9.3, 19.6)), and females (adjusted OR = 1.43, 95% CI: (1.10, 1.86)) were significantly associated with higher odds of VDD. In conclusion, the study revealed a significant difference in the frequency of VDD and VDI among pediatric patients older than six years, with a significant difference according to sex and season, being more pronounced among girls and during the winter and spring seasons.

Neutrophilic Myocarditis: Insights from a Forensic Centre's Retrospective Study.

BACKGROUND: Neutrophilic myocarditis often stems from bacterial or fungal infections, and it is typically detectable through blood cultures or analyses of the primary infection site. However, research specifically addressing the morphological features of acute myocarditis in complex sepsis cases is scarce, with existing studies primarily dating back to the pre-antibiotic era. METHODS: This study constitutes a retrospective and descriptive analysis encompassing 22 forensic cases. We collected data from forensic reports emphasising clinical details, disease history, gross observations, and histopathological findings. RESULTS: The results show that using positive-air-pressure ventilation could be related to cardiac inflammation (45.45%, 10/22). Despite large-spectrum antibiotic therapy, the blood samples were positive for Staphylococcus aureus (MRSA strain), Klebsiella pneumoniae (ESBL strain), Acinetobacter baumannii, and Pseudomonas aeruginosa. Colonies developed in the myocardium of 36% of the patients (8/22), where 4 of them had septic emboli. Fungal myocarditis accompanied bacterial infections (2/8) and were unsuspected clinically. Background changes, such as interstitial fibrosis and arteriosclerosis, were associated with a greater degree of inflammation and septic embolism. CONCLUSION: Neutrophilic myocarditis in patients with emerging sepsis is linked to fatal virulent infections, where bacteria and/or fungi contaminate and impair the myocardium syncytium. Prolonged hospitalisation and positive-air-pressure ventilation may be a risk factor for this condition and needs further research.

Biocomposite Material Based on Lactococcus lactis sp. Immobilized in Natural Polymer Matrix for Pharmaceutical Removal from Aqueous Media.

Ecosystems are negatively impacted by pharmaceutical-contaminated water in different ways. In this work, a new biosorbent obtained by immobilizing Lactococcus lactis in a calcium alginate matrix was developed for the removal of pharmaceuticals from aqueous solutions. Ethacridine lactate (EL) was selected as the target drug. Lactococcus Lactis biomass was chosen for the biosorbent synthesis for two reasons: (i) the microbial biomass used in the food industry allows the development of a low-cost biosorbent from available and renewable materials, and (ii) there is no literature mentioning the use of Lactococcus Lactis biomass immobilized in natural polymers as a biosorbent for the removal of pharmaceuticals. The characterization of the synthesized biosorbent named 5% LLA was performed by scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR) analysis. Additionally, particle size and the point of zero charge were established. Batch biosorption investigations showed that using 5% LLA at an initial pH of 3.0 and a biosorbent dose of 2 g/L resulted in up to 80% EL removal efficiency for all EL initial concentrations (20-60 mg/L). Four equilibrium isotherms, given in the order of Redlich-Peterson > Freundlich > Hill > Temkin, are particularly relevant for describing the experimental data for EL biosorption on the 5% LLA biosorbent using correlation coefficient values. Kinetic parameters were determined using kinetic models such as pseudo-first-order, pseudo-second-order, Elovich, Avrami and Weber-Morris. The pseudo-second-order kinetics model provides the greatest fit among the evaluated equations, with correlation coefficients greater than 0.99. According to the study's findings, the developed biocomposite is a potentially useful material for the removal of pharmaceuticals from aqueous matrices.

Natural Polymers as Carriers for Encapsulation of Volatile Oils: Applications and Perspectives in Food Products.

The technique of encapsulating different materials into matrices that can both protect and release their contents under specific circumstances is known as encapsulation. It serves the primary function of shielding delicate components from outside influences, including heat, light, and humidity. This can be accomplished by a variety of procedures that, depending on the method and materials selected, result in the creation of particles with various structures. The materials used for encapsulation in food applications must be of high quality, acceptable for human consumption, and stable during processing and storage. The most suitable natural polymers for food applications are carbohydrates, proteins, or mixtures thereof. Volatile oils are end products of plant metabolism, accumulated and stored in various plant organs, cells, or secretory tissues. These are natural and are characterized by the scent of the aromatic plants they come from. Because of their antibacterial and antioxidant qualities, they are being utilized more and more in the food and pharmaceutical industries. Since volatile oils are highly sensitive to environmental changes, they must be stored under specific conditions after being extracted from a variety of plant sources. A promising method for increasing the applicability of volatile oils is their encapsulation into colloidal particles by natural polymers such as carbohydrates and proteins. Encapsulation hides the unfavorable taste of nutrients while shielding delicate dietary ingredients from the effects of heat, moisture, oxygen, and pH. This technique results in improved stability for volatile oils that are often sensitive to environmental factors and offers the possibility of using them in an aqueous system even if they are insoluble in water. This paper aims to provide an overview of the current advances in volatile oil encapsulation technologies and presents a variety of natural polymers used in the food industry for encapsulation. Also, a distinct section is created to highlight the current advances in dairy products enriched with encapsulated volatile oils.

The Importance of Neonatal Screening for Galactosemia.

Galactosemia is an inborn metabolic disorder caused by a deficient activity in one of the enzymes involved in the metabolism of galactose. The first description of galactosemia in newborns dates from 1908, ever since complex research has been performed on cell and animal models to gain more insights into the molecular and clinical bases of this challenging disease. In galactosemia, the newborn appears to be born in proper health, having a window of opportunity before developing major morbidities that may even be fatal following ingestion of milk that contains galactose. Galactosemia cannot be cured, but its negative consequences on health can be avoided by establishing precocious diagnosis and treatment. All the foods that contain galactose should be eliminated from the diet when there is a suspicion of galactosemia. The neonatal screening for galactosemia can urge early diagnosis and intervention, preventing complications. All galactosemia types may be detected during the screening of newborns for this disorder. The major target is, however, galactose-1-phosphate uridyltransferase (GALT) deficiency galactosemia, which is diagnosed by applying a combination of total galactose and GALT enzyme analysis as well as, in certain programs, mutation screening. Most critically, infants who exhibit symptoms suggestive of galactosemia should undergo in-depth testing for this condition even when the newborn screening shows normal results. The decision to enroll global screening for galactosemia among the specific population still faces many challenges. In this context, the present narrative review provides an updated overview of the incidence, clinical manifestations, diagnosis, therapy, and prognosis of galactosemia, questioning under the dome of these aspects related to the disease the value of its neonatal monitoring.

Use of Yoghurt Enhanced with Volatile Plant Oils Encapsulated in Sodium Alginate to Increase the Human Body's Immunity in the Present Fight Against Stress.

(1) Background: The COVID-19 pandemic and the imposition of strict but necessary measures to prevent the spread of the new coronavirus have been, and still are, major stress factors for adults, children, and adolescents. Stress harms human health as it creates free radicals in the human body. According to various recent studies, volatile oils from various aromatic plants have a high content of antioxidants and antimicrobial compounds. An external supply of antioxidants is required to destroy these free radicals. The main purpose of this paper is to create a yoghurt with high antioxidant capacity, using only raw materials from Romania; (2) Methods: The bioactive components used to enrich the cow milk yoghurt were extracted as volatile oils out of four aromatic plants: basil, mint, lavender and fennel. Initially, the compounds were extracted to determine the antioxidant capacity, and subsequently, the antioxidant activity of the yoghurt was determined. The 2,2-diphenyl-1-picrylhy-drazyl (DPPH) method was used to determine the antioxidant activity; (3) Results: The results show that cow milk yoghurt enhanced with volatile oils of basil, lavender, mint and fennel, encapsulated in sodium alginate has an antioxidant and antimicrobial effect as a staple food with multiple effects in increasing the body's immunity. The antioxidant activity proved to be considerably higher than the control sample. The highest antioxidant activity was obtained on the first day of the analysis, decreasing onwards to measurements taken on days 10 and 20. The cow milk yoghurt enriched with volatile basil oil obtained the best results; (4) Conclusions: The paper shows that yoghurts with a high antioxidant capacity were obtained, using only raw materials from Romania. A healthy diet, compliance with safety conditions and finding appropriate and safe methods to increase the body's immunity is a good alternative to a major transition through harder times, such as pandemics. The creation of food products that include natural antioxidant compounds combines both the current great possibility of developing food production in Romania and the prevention and reduction of the effects caused by pandemic stress in the human body.

Ganglioneuroblastoma in children.

INTRODUCTION: Neuroblastoma ranks third among pediatric malignancies. CASE REPORT: The case of a 3-year-old child is presented, who suddenly had frequent, unproductive, emetic cough; fever; and weight loss. Lung X-ray showed an opacity situated in the posterior superior mediastinum. Thoracic ultrasound revealed a slightly inhomogeneous, hypoechoic mass located in the posterior superior mediastinum. Computed tomography evidenced a tumor mass with homogeneous appearance in the costo-vertebral groove. Histological examination confirmed the diagnosis of ganglioneuroblastoma. CONCLUSION: Although history and clinical examination provided few elements, diagnosis was made based on imaging and histopathological examination.

The Romanian version of the Juvenile Arthritis Multidimensional Assessment Report (JAMAR).

The Juvenile Arthritis Multidimensional Assessment Report (JAMAR) is a new parent/patient-reported outcome measure that enables a thorough assessment of the disease status in children with juvenile idiopathic arthritis (JIA). We report the results of the cross-cultural adaptation and validation of the parent and patient versions of the JAMAR in the Romanian language. The reading comprehension of the questionnaire was tested in 15 JIA parents and patients. Each participating centre was asked to collect demographic, clinical data and the JAMAR in 100 consecutive JIA patients or all consecutive patients seen in a 6-month period and to administer the JAMAR to 100 healthy children and their parents. The statistical validation phase explored descriptive statistics and the psychometric issues of the JAMAR: the three Likert assumptions, floor/ceiling effects, internal consistency, Cronbach's alpha, interscale correlations, test-retest reliability, and construct validity (convergent and discriminant validity). A total of 310 JIA patients (11.9% systemic, 21.6% oligoarticular, 31.9% RF-negative polyarthritis, 34.6% other categories) and 100 healthy children, were enrolled in six centres. The JAMAR components discriminated well healthy subjects from JIA patients except for the health-related quality of life psychosocial health subscales. All JAMAR components revealed good psychometric performances. In conclusion, the Romanian version of the JAMAR is a valid tool for the assessment of children with JIA and is suitable for use both in routine clinical practice and clinical research.

Starch from the Sago (Metroxylon sagu) Palm Tree-Properties, Prospects, and Challenges as a New Industrial Source for Food and Other Uses.

The common industrial starches are typically derived from cereals (corn, wheat, rice, sorghum), tubers (potato, sweet potato), roots (cassava), and legumes (mung bean, green pea). Sago (Metroxylon sagu Rottb.) starch is perhaps the only example of commercial starch derived from another source, the stem of palm (sago palm). Sago palm has the ability to thrive in the harsh swampy peat environment of certain areas. It is estimated that there are about 2 million ha of natural sago palm forests and about 0.14 million ha of planted sago palm at present, out of a total swamp area of about 20 million ha in Asia and the Pacific Region, most of which are under- or nonutilized. Growing in a suitable environment with organized farming practices, sago palm could have a yield potential of up to 25 tons of starch per hectare per year. Sago starch yield per unit area could be about 3 to 4 times higher than that of rice, corn, or wheat, and about 17 times higher than that of cassava. Compared to the common industrial starches, however, sago starch has been somewhat neglected and relatively less attention has been devoted to the sago palm and its starch. Nevertheless, a number of studies have been published covering various aspects of sago starch such as molecular structure, physicochemical and functional properties, chemical/physical modifications, and quality issues. This article is intended to piece together the accumulated knowledge and highlight some pertinent information related to sago palm and sago starch studies.

Modelling the genetics and demography of step cline formation: gastropod populations preyed on by experimentally introduced crabs.

Whether a prey population goes extinct or adapts in response to an invading predator may depend on the number of contiguous populations that experience increased predation. We created invaded snail populations by building shelters for predatory shore crabs on a rocky intertidal bench. The crabs preyed selectively on thin-shelled snails tethered next to the shelters but did not prey on those more than 2 m away. This caused strong directional selection for increased shell thickness in populations close to the shelters but did not change selection in those farther away. The field experiment was used to parameterize a new individual-based quantitative genetic model that included demography. In the model a detectable step cline in shell thickness evolved rapidly even though the region of increased predation was shorter than Slatkin's characteristic length. The cline's step size in the model was similar to that measured in the field 10 years after the experiment began.

Rapid elimination of Helicobacter pylori and reduction of histocompatibility leucocyte antigen-DR expression 12 h after a single dose of omeprazole, amoxycillin and metronidazole triple therapy.

BACKGROUND: Studies of eradication of Helicobacter pylori and subsequent resolution of H. pylori-related gastritis, have focused mainly on medium and long-term change following eradication therapies. Results from those studies have shown that both acute and chronic inflammatory changes found in gastric mucosa eventually return to normal. However, the early events in the stomach, particularly the effects on bacterial density and acute inflammatory markers of anti-H. pylori treatment, are largely unknown. The objective of this study was therefore to examine changes in the number of H. pylori, and the severity of gastric mucosal inflammation in the gastric biopsy specimens of patients before (0 h group, n = 14) and 12 h (12 h group, n = 14) after initiating anti-H. pylori treatment. METHODS: Biopsies were assessed, either quantitatively or semi-quantitatively, for the presence of H. pylori, neutrophils, mast cells, intraepithelial lymphocytes and the expression of histocompatibility leucocyte antigen (HLA)-DR by gastric epithelium and the results were compared between groups. RESULTS: Median H. pylori scores were 5 (range 2-5) and 0 (range 0-2) in biopsies from untreated and 12 h post-treatment groups, respectively (P < 0.001). In most 12 h post-treatment biopsies, H. pylori organisms could not be identified. There was a significant reduction in HLA-DR expression by gastric epithelium (median 3.5 with range 2-4 at 0 h group vs median 2 with range 0-4, P < 0.05), but there was no significant difference in the number of intraepithelial lymphocytes, CD3+ cells, mast cells or the distribution and density of neutrophils (all P > 0.05). Furthermore, the severity of gastritis as scored with the Sydney system was similar in both untreated and treated groups. CONCLUSIONS: The results of this study indicate that elimination of H. pylori organisms and resolution of some inflammatory markers occurs as early as 12 h following a single dose of omeprazole 40 mg, amoxycillin 1.0 g and metronidazole 400 mg, which suggests that rational therapeutic strategies with shorter duration using the currently available drugs may be possible.

Extreme hypoventilation reduces ventilator-induced lung injury during ventilation with low positive end-expiratory pressure in saline-lavaged rabbits.

OBJECTIVE: To compare the degrees of ventilator-induced lung injury caused by two ventilation protocols. DESIGN: Randomized trial. SETTING: University animal laboratory. SUBJECTS: Sixteen New Zealand white rabbits. INTERVENTIONS: After five sequential saline lung lavages, eight pairs of anesthetized rabbits were allocated randomly to receive either of two ventilation protocols for 4 hrs during neuromuscular blockade. Both groups received 3 cm H2O of positive end-expiratory pressure and 100% oxygen. Control group animals received an estimated tidal volume of 12 mL/kg, an inspiratory time of 0.7 sec, and a ventilatory rate adjusted for a PaCO2 of 35 to 45 torr (4.7 to 6 kPa). Study group animals were ventilated through an intratracheal catheter, with inspiratory time of 1.5 secs, ventilatory rate of 20 breaths/min, and peak inspiratory pressure of 4 to 8 cm H2O, adjusted to maintain PaCO2 at 150 to 250 torr (20 to 33 kPa). MEASUREMENTS AND MAIN RESULTS: Arterial blood gases were measured every 30 mins. After 4 hrs, a final lung lavage was performed. Physiologic parameters, cell counts and protein concentration in the final lavage, and lung histology were compared between groups. The alveolar-arterial oxygen tension gradient was higher in the study group over the first 1.5 hrs, but the time profile showed significantly (p = .001) greater improvement in the study group. After 4 hrs, the mean alveolar-arterial oxygen tension gradient was lower in the study group (94 torr [12.5 kPa] vs. 201 torr [26.8 kPa]). The increase in neutrophil count from the initial to the final lung lavage was lower in the study group (0.27 x 10(7) cells/L vs. 2.01 x 10(7) cells/L, p = .037), as was the absolute value of the neutrophil count in the final lavage (1.33 x 10(7) cells/L vs. 3.02 x 10(7) cells/L, p = .04). The median hyaline membrane score was lower in the study group (0.5 vs. 3.0) but the difference was not statistically significant. CONCLUSION: These findings suggest that a very low tidal volume reduces ventilator-induced lung injury in saline-lavaged rabbits during ventilation at low lung volume.

37 kBq 14C-urea breath test and gastric biopsy analyses of H. pylori infection.

BACKGROUND: The treatment of H. pylori-associated gastroduodenal disease is increasingly aimed at bacterial eradication which requires follow-up assessment of therapeutic effectiveness and re-infection. A simplified 37 kBq 14C-urea breath test for H. pylori infection has been developed. METHODS: The 37 kBq 14C-urea breath test was compared with biopsy urease (CLO) and histological analyses of gastric-biopsies obtained from 63 patients undergoing endoscopy. RESULTS: The 30-min breath test correlated closely with biopsy findings, had a sensitivity of 100%, a specificity of 95% and a positive predictive value of 92%. CONCLUSIONS: The simplified, low-dose, 14C-urea breath test is a convenient, low-cost, transportable means of facilitating the management of H. pylori-associated diseases.

Pressure-limited ventilation with permissive hypercapnia and minimum PEEP in saline-lavaged rabbits allows progressive improvement in oxygenation, but does not avoid ventilator-induced lung injury.

OBJECTIVE: To determine whether pressure-limited intermittent mandatory ventilation with permissive hypercapnia and positive end-expiratory pressure (PEEP) titrated to arterial oxygen tension (PaO2) prevents or reduces acute lung injury, compared to conventional ventilation, in saline-lavaged rabbits. DESIGN: Prospective randomised trial. SETTING: University animal laboratory. SUBJECTS: 18 New Zealand White rabbits. INTERVENTIONS: Following five sequential saline lung lavages, anaesthetised rabbits were randomly allocated in pairs to receive either of two ventilation protocols using intermittent mandatory ventilation. The study group had peak inspiratory pressure limited to 15 cm H2O and arterial partial pressure of carbon dioxide (PaCO2) was allowed to rise. The control group received 12 ml/kg tidal volume with rate adjusted for normocarbia. PEEP and fractional inspired oxygen (FIO2) were adjusted to maintain, PaO2 between 8 and 13.3 kPa (60 and 100 mm Hg) using a predetermined protocol. At 10 h or following death, lung lavage was repeated and lung histology evaluated. MEASUREMENTS AND MAIN RESULTS: The mean increase in lavage cell counts and protein concentration and hyaline membrane scores were not significantly different between the groups. Oxygenation progressively improved more in the study group (p = 0.01 vs control for PaO2/FIO2 ratio and alveolar-arterial oxygen tension gradient (AaDO2)). PEEP was similar and the mean airway pressure higher in the control group, suggesting that this probably resulted from less ventilator-induced injury in the study group. Four deaths occurred in the control group (three due to pneumothorax and one to hypoxaemia) and none in the study group (p = 0.08). CONCLUSIONS: This ventilatory protocol may have failed to prevent lung overdistension or it may have provided insufficient PEEP to prevent injury in this model; PEEP greater than the lower inflection point of the pressure-volume curve has been shown to prevent injury almost entirely.

Small bowel ischaemia in Fabry's disease.

A patient with previously diagnosed Fabry's disease and a long history of post-prandial abdominal pain died following small bowel infarction. Post-mortem demonstrated Fabry's type deposits in the small vessels and nerves supplying the bowel but in addition, a localized atheromatous stenosis of the superior mesenteric artery. In retrospect, his terminal illness and possibly his chronic symptoms were related to the latter finding. Angioplasty to the superior mesenteric artery may have been of benefit. Mesenteric angiography should be considered in patients with chronic post-prandial pain because large vessel disease may coexist with other a priori pathologies and is imminently treatable.

Deaths from electricity.

This paper reviews the circumstances of 95 fatalities from electrical injuries. Eighty-nine were accidental, four were suicides and two occurred during autoerotic electrical stimulation. Forty-nine of the accidental fatalities occurred at work, Twenty-eight in the home and twelve in the course of outside recreational activities. In many accidents the circumstances were distressingly similar and included: (1) Contact with overhead distribution lines by a length of conductor such as a yacht mast or crane. (2) Faulty wiring or electrical repairs performed by unqualified people. (3) Badly deteriorated cords, plugs and occasionally appliances. (4) Failure to use isolating transformers when indicated. Deaths involving children are a particular cause of concern. Nine fatalites involved children under the age of five years who contacted inadequately protected wires.