Histamine Content Research Articles

Analysis of Current Approaches to Monitoring the Content of Depressor Substances

INTRODUCTION. Raw materials of biological origin can contaminate medicines with depressor substances, such as histamine, acetylcholine, bradykinin, serotonin, and prostaglandins. In the Russian Federation, tests for the content of these impurities are conducted in anaesthetised healthy cats in accordance with the General Chapter Test for depressor substances of the State Pharmacopoeia of the Russian Federation. The global trend towards ending animal testing and the search for ways to eliminate animal tests for depressor substances motivate researchers to study international approaches to the determination of histamine-like substances.AIM. This study aimed to analyse Russian and international approaches to the control of biologicals for depressor substances and to consider the applicability of alternative instrument-based physicochemical methods for the determination of histamine-like impurities, with a view to reducing animal testing.DISCUSSION. The rationale for reducing the use of animals as test systems is set forth in Directive 2010/63/EU of the European Parliament and of the Council of 22 September 2010 on the protection of animals used for scientific purposes. In 2023, the European Pharmacopoeia Commission initiated the removal of in vivo tests for histamine-like substances from the European Pharmacopoeia. However, China and India use animal testing as the basic method for assessing the safety of medicines. In the Russian Federation, the control of depressor substances is regulated under General Chapter 1.2.4.0008.18 of the State Pharmacopoeia of the Russian Federation. Since the publication of General Chapter 42-0063-07 Test for histamine in edition XII in 2007, the State Pharmacopoeia of the Russian Federation has included a compendial text providing for a reduction in the number of cats used for testing. At present, Russian scientists are searching for targeted physicochemical and immunochemical methods for the determination of depressor substances. It has been demonstrated that a reasonable approach is to assess the need for testing a particular medicinal product in animals on a case-by-case basis. This approach will help minimise the number of in vivo tests and maintain an adequate quality of medicines at the same time.CONCLUSIONS. Despite the global trend towards ending animal testing, it is still necessary and required to control medicines for the content of depressor substances. Nevertheless, there is no global consensus on the need to use animals for this purpose. The absence of a separate general chapter/monograph, or references to it, will not change the fact that histamine-like substances are controlled worldwide. The decision to exclude in vivo methods from the European Pharmacopoeia necessitates the search for alternative test methods and the implementation of additional manufacturing controls to minimise the contamination risk for finished medicinal products. Studies have shown that high-performance liquid chromatography can determine the content of histamine in a limited amount of a medicinal product.

Efficacy and pharmacodynamics of herbal medicine in patients with metabolic phenotype of osteoarthritis

High prevalence of the metabolic phenotype of osteoarthritis (MPOA) and unsatisfactory methods of its treatment necessitate the development of novel therapeutic approaches for this phenotype of OA. Turmeric preparations have a wide range of biological activity; their use in patients with MPOA can be effective and reduce the drug burden. In the present paper, we review our own published research findings on the effectiveness and pharmacodynamics of turmeric preparations in patients with MPOA. In the two studies, we evaluated the effects of two curcumin-containing parapharmaceuticals: Epigenorm Antivir (EA) and Curcumin. Forty-one women with MFOA were included in both studies. Twenty-three patients received EA at a daily dose of 1000 mg for 12 weeks, and 18 patients received curcumin from Evalar at a dose of 1000 mg for 8 weeks. The endpoints for both studies were VAS pain, dysfunction and other symptoms of OA measured using the KOOS scale. After 12 weeks of EA treatment, there was a 2.5-fold decrease pain levels. The decrease in pain was associated with an improvement in the patient’s daily and sports activities and quality of life. These effect sizes were classified as moderate to large according to Cohen. No adverse events were observed during the period of taking EA. Clinical improvement was associated with a decrease in the content of MS individual components: low-density lipoprotein cholesterol and triglycerides. The treatment caused a decrease in the level of systemic inflammation, as evidenced by a decrease in the concentration of TNFα, histamine, IL-18, C-reactive protein, and neopterin. The concentrations of IL-10 and adiponectin increased after treatment. In another study, treatment with curcumin for 4 weeks had an analgesic effect, improved measures of function and quality of life. Clinical improvement was associated with the reductions in serum levels of a number of proinflammatory cytokines, C-reactive protein, and lipids. Thus, the results of the pilot studies evaluating efficacy, safety, and pharmacodynamics of parapharmaceuticals EA and Curcumin in patients with MFOA indicate pleiotropic effects of the interventions. The findings provide a rationale for conducting larger, controlled, blind, randomized clinical trials.

Pharmacological inhibition of histamine N-methyltransferase extends wakefulness and suppresses cataplexy in a mouse model of narcolepsy.

Histamine, a neurotransmitter, plays a predominant role in maintaining wakefulness. Further, our previous studies showed that histamine N-methyltransferase (HNMT), a histamine-metabolising enzyme, is important for regulating brain histamine concentration. However, the effects of pharmacological HNMT inhibition on mouse behaviour, including the sleep-wake cycle and cataplexy, in a mouse model of narcolepsy have not yet been investigated. In the present study, we investigated the effects of metoprine, an HNMT inhibitor with high blood-brain barrier permeability, in wild-type (WT) and orexin-deficient (OxKO) narcoleptic mice. Metoprine increased brain histamine concentration in a time- and dose-dependent manner without affecting peripheral histamine concentrations. Behavioural tests showed that metoprine increased locomotor activity in both novel and familiar environments, but did not alter anxiety-like behaviour. Sleep analysis showed that metoprine increased wakefulness and decreased non-rapid eye movement (NREM) sleep through the activation of the histamine H1 receptor (H1R) in WT mice. In contrast, the reduction of rapid eye movement (REM) sleep by metoprine occurred independent of H1R. In OxKO mice, metoprine was found to prolong wakefulness and robustly suppress cataplexy. In addition, metoprine has a greater therapeutic effect on cataplexy than pitolisant, which induces histamine release in the brain, and has been approved for patients with narcolepsy. These data demonstrate that HNMT inhibition has a strong effect on wakefulness, demonstrating therapeutic potential against cataplexy in narcolepsy.

Gastrointestinal symptoms in children with mastocytosis

Introduction. Mastocytosis is a heterogeneous group of diseases characterized by the abnormal accumulation of clonal mast cells (MCs) in various tissues and organs, including skin, bone marrow, liver, spleen and lymph nodes. The clinical picture of cutaneous and indolent systemic mastocytosis is formed by a wide range of symptoms associated with activation of mast cells. Single European studies have demonstrated wide variability in the frequency of gastrointestinal symptoms (GI-symptoms)) in children with mastocytosis (from 15 to 50%).Aim. To analyze the frequency of mediator-related GI-symptoms in children with different subtypes and clinical forms of mastocytosis.Materials and methods. A prospective observational study included data from 289 children aged 3 to 17 years who were under observation at the Moscow Scientific and Practical Center of Dermatovenereology and Cosmetology. Symptoms were assessed using the Pediatric grade scale of symptoms of mastocytosis. Clinical manifestations of reactions caused by mast cell degranulation were compared with data from clinical and laboratory studies. The concentrations of tryptase and histamine in the blood serum of patients were determined. The incidence of organomegaly and mesenteric lymphadenitis in children with mediating symptoms and without symptoms was determined using ultrasound.Results. 67 (23.2%) patients had GI symptoms. More than half of the patients (51.6%) indicated abdominal pain, 32.3% reported diarrhea. The severity of pain symptoms correlated with the level of serum tryptase (ρ = 0.564, p < 0.01). The incidence of hepatomegaly and mesadenitis in children with GI- symptoms was 19.4% and 25.4%, respectively. In a comparative analysis in groups of children with and without GI symptoms, the differences in the number of identified cases of organomegaly and mesadenitis were statistically significant (p < 0.001). An increase in histamine levels in the blood was detected in half of the patients with complaints of heartburn and nausea.Conclusion. Our results demonstrated that a study of the level of serum tryptase, ultrasound of the abdominal organs and lymph nodes should be carried out in all children with mediator-related GI-symptoms regardless of the clinical form of mastocytosis.

Ultrasensitive detection of histamine in spoiled meat employing silver nanoparticles decorated Perylene: An experimental-computational conjugation

Meat spoilage has been acquiring increasing attention recently and is directly associated with food safety and human health. Biogenic amines are the spying organic compounds fostered from the microorganism-mediated decarboxylation of amino acids during meat spoilage. Histamine, a biogenic amine acts as a model analyte and is toxic if consumed substantially. It is crucial to monitor histamine levels in meat due to its adverse effects. In this study, a simple and quick fluorescent sensor was fabricated for sensitive and selective detection of histamine. Citrate-capped silver nanoparticles (AgNP) were loaded onto Perylene (PER) to develop a sensing probe that was characterized using UV–visible, FTIR, XRD, and FESEM, and its optical behavior toward histamine was investigated. Moreover, the binding affinity between histamine and PER@AgNP was assessed using a DFT-based computer simulation. Under optimal conditions, the sensor showed linear relationships for histamine concentrations from 25 μM to 3200 μM with LOD 13.52 μM.

Black and white truffles as a source of neuroactive biogenic amines

This study reports the first data about content of serotonin, histamine, and dopamine in black and white truffles fruiting body based on UHPLC-ESI-QqQ-MS/MS. Here we have demonstrated that black (Tuber macrosporum) and white (T. magnatum) truffles are a source of biogenic amines. Also, we demonstrated the absence of correlation between the quality of truffles and the content of biogenic amines.

Chitosan coatings containing thyme essential oil enhance the quality of snakehead (Channa striata) during chilled storage verified by metabolomics approaches.

In the present study, we investigated the preservative effects of chitosan (CS) coatings, with and without thyme essential oil (TEO), combined with vacuum impregnation (VI) on maintaining the quality of snakehead fillets during chilled storage. The results showed that the VI treatment significantly inhibited drip loss, discoloration, microbial proliferation, and the accumulation of biogenic amines (BAs) in the sneakhead fillets. Compared to the control, the fillets treated with VI of 1% (w/w) CS and 1.5% (w/w) TEO (i.e., CSTEO) showed significant reductions in both psychrophiles and mesophiles, with a 2.66 log CFU/g decrease in total viable count (TVC) on day 3 and a 1.89 log CFU/g decline in TVC on day 9, respectively. In addition, the content of histamine and putrescine in the CSTEO groups was maintained at ∼1.14 and 3.23mg/kg during the 12-day chilled storage, respectively. A total of 100 chemical compounds were tentatively identified using untargeted metabolomics approaches. The multivariate analysis further revealed that the combination of VI and CSTEO maintained fish quality mainly through preventing lipid oxidation and protein degradation. Overall, the VI-CSTEO treatment effectively maintained the fish quality during storage at 4°C, with minimum microbial proliferation and accumulation of BAs. PRACTICAL APPLICATION: The preservative effect of chitosan coatings containing thyme essential oil combined with vacuum impregnation on snakehead quality during the 12-day chilled storage was verified, and the underlying mechanisms were deciphered through integrated metabolomics approaches. Our study could provide a promising strategy for the preservation of aquatic products.

Microbiological profile and prevalence of histamine-producing bacteria in fresh sardines stored at different temperatures

Background and Aim: The European pilchard (Sardina pilchardus) is an important fish species for the Moroccan economy in terms of production and export. Biogenic amine histamine is a metabolite produced in the flesh of some fish species after death due to the decarboxylation of free histidine by histaminogenic bacteria. Failure to control the histamine risk in European pilchard may lead to public health and socioeconomic issues. This study aimed to estimate the prevalence of histaminogenic bacteria in association with histamine levels and the growth of microflora in Moroccan sardines (European pilchard). Materials and Methods: We conducted the study by monitoring Moroccan sardines of histamine content and microbiological profile (aerobic plate count [APC], coliforms, and thermo-tolerant coliforms [TTC]) during 6 days of storage at three different temperatures (0°C, 10°C, and ambient temperature [22°C]). The histamine assay was performed using a spectrofluorometric method, and the microbiological identification of histamine-producing bacteria was performed using a combination of biochemical and molecular tests. Results: The histamine content in European pilchard stored at 0°C was negligible. However, high concentrations were observed at 10°C and 22°C. The microbiological profile showed a positive association between microflora counts and histamine content according to storage time. At 0°C, a moderate increase in the APC, a decrease in coliforms, and an absence of TTC were observed. The rapid proliferation of all microflora was observed at 10°C, whereas at 22°C, the proliferation was almost exponential. Bacterial identification revealed the exclusive presence of species belonging to the Enterobacteriaceae family at varying frequencies depending on storage temperature. Morganella morganii and Proteus mirabilis had the highest histamine induction rates in L-histidine-supplemented broth, with 1600 and 255 parts per million (ppm), respectively, after 48-h incubation at 35°C. Klebsiella ozaenae could produce 136 ppm and Serratia plymuthica 115 ppm. Reverse transcription polymerase chain reaction showed positive results for the presence of genes associated with histidine decarboxylase. The hdc genes of M. morganii, P. mirabilis, and K. ozaenae were successfully amplified and exhibited strong similarity with the reference gene of M. morganii. Conclusion: This study describes for the first time the hdc gene in bacteria that form histamine in Moroccan sardines. The results also confirm that respect for the cold chain integrity is a crucial factor in histamine management. This information should help stakeholders in the implementation of sound strategies for managing the hazards associated with seafood and their products. Keywords: Enterobacteriaceae, histamine, Histaminogenic bacteria, polymerase chain reaction, sardine, seafood.

Exposure assessment and semi-quantitative risk analysis of histamine in tuna and tuna-like fish from Indonesia

This study aimed to investigate histamine exposure associated with consumption of fresh tuna and tuna-like species in West Java, and to estimate risk of Scombroid Fish Poisoning (SFP) in Indonesia. A range of species, including tuna (Thunnus spp.), bullet tuna ( Auxis sp.), and skipjack ( Katsuwonus pelamis ) were collected from local markets and fish landing sites. Subsequently, histamine concentrations were determined using NMR analysis and exposure was calculated in mg/day for toddlers, children, and adults. The results showed that skipjack had the highest histamine exposure for all age groups, followed by bullet and regular tuna. The highest EDI for histamine was from skipjack consumption, accounting for 38.67; 37.77 and 20.74 percentage of exposure for toddlers, children and adults, respectively. These values are below the defined thresholds levels (ARfD), indicating no potential risk of acute health effect. Cooked bullet tuna and skipjack were estimated to cause similar illnesses, accounting for 6–7 cases per 100,000 individuals, which was higher than cooked tuna at 1–2 cases per 100,000 individuals. Considering the preparation of raw tuna in restaurants following Good Hygienic Practices (GHP), the predicted annual cases decreased significantly to 4–5 cases per million individuals. This risk estimation only considered histamine levels in fresh fish, without including data from fish preparation. Therefore, further studies were recommended to estimate the risk level in raw/fresh tuna and similar species before consumption.

Highly sensitive biosensors for real-time monitoring of histamine at acupoint PC6 in rats based on graphene-modified acupuncture needles.

Acupoints are the local initial response sites of acupuncture therapeutic effects. As a biomarker, histamine is released into the acupoint region and plays its role concurrently as acupuncture needles are inserted into acupoints. Hence, real-time monitoring of histamine at acupoints is important to elucidate the effectiveness of the acupoint-activation process in acupuncture. Therefore, we developed highly sensitive acupuncture/Au particles/graphene biosensors by electrodeposition, brushing, and annealing methods based on bare acupuncture needles. We achieved a histamine detection limit of approximately 4.352 (±3.419) × 10-12 mol L-1 and good sensitivity of approximately 6.296 (±3.873) μA μM-1, with satisfactory specificity, repeatability, and stability in vitro, rendering them more competitive and suitable for real-time monitoring in vivo without causing additional damage. Subsequently, we conducted real-time histamine monitoring at non-acupoint and acupoint PC6 in rats, respectively. Our results showed minimal changes at the non-acupoint, whereas a trend of initial increase followed by a decrease was observed at acupoint PC6. The change in histamine concentration at acupoint PC6 reflected its involvement in the acupoint-activation procedure. Moreover, its peak position at ∼18 min could provide guidance for optimizing needle retaining time for maximum therapeutic effect. This work presents the first real-time in vivo monitoring of histamine at acupoints with high sensitivity and underscores the specificity of histamine release between non-acupoint and acupoint PC6, demonstrating great potential for elucidating the acupoint-activation mechanisms in acupuncture. Additionally, this work expands the application of nanomaterials in the integration of medicine and engineering, which is an important aspect of the future development of materials science.

Classification of Histamine Content in Fish Using Near-Infrared Spectroscopy and Machine Learning Techniques

Near-infrared (NIR) spectroscopy has emerged as a popular technique for assessing food quality due to its advantages over complex chemical analysis methods. However, the application of NIR spectroscopy for evaluating fish quality based on histamine content has not been extensively explored. This study investigates the use of NIR spectroscopy in combination with machine learning (ML) techniques to classify fish samples into two safety classes, Safe and Unsafe, based on their histamine content. A comprehensive NIR dataset comprising 11,360 spectra collected at eight distinct positions within the fish body was obtained from 284 fish samples of mackerel, tuna, and pompano species. ML experiments were conducted to classify fish samples based on whether their histamine content exceeded the permissible limit of 100 ppm. To address class imbalance and optimize ML models, various data pre-processing and feature extraction techniques as well as ML algorithms were explored. The results demonstrated that utilizing NIR data specifically obtained from the tail’s flesh, a specific location within the fish, yielded superior models for fish safety classification. A feature extraction method employing pre-processed NIR spectra and their second derivatives, combined with an optimized convolutional neural network architecture, outperformed traditional ML classifiers with an accuracy of approximately 93%.

Gelatin-kappa-carrageenan-based edible film incorporated with ethanol extracts from Premna microphylla Turcz leaves for preservation of sailfish fillets

This study proposes to develop edible films (EF) by modification of gelatin/kappa (κ)-carrageenan (KCAR) as a base formulation while incorporating the Premna microphylla Turcz leaf (PMTL) extracts. PMTLs were extracted via distilled water or 95% ethanol (1:20 ratio, w/v) to investigate the content of bioactive substances and evaluate their antioxidant and antimicrobial capacities. The PMTL ethanol extract (PEE) showed satisfactory antimicrobial and antioxidant performance compared to the water extract (PWE). Therefore, different ratios of PEE (0, 5, 10, and 15%) were incorporated into the basic formulation of EF supplemented with the plasticizing agent glycerol (0.5 and 1%) to prolong the shelf-life of sailfish fillets (SF). Incorporating PEE into EF formulations may change the color from transparent to yellow-green, which improves EF's moisture content, solubility, swelling degree, and hydrophilicity. PEE's total phenolic content (TPC) and total flavonoid content (TFC) provided EF with antioxidant capacity, causing EF with predictable mitigation of sharp variation in pH, total plate count, thiobarbituric acid reactive substances (TBARS) value, and histamine content during the storage period of SF (12 days at 4 °C). Altogether, this study indicated that EFs manufactured by PEE would be potential candidates for application in food preservation to improve food safety.

Microstereolithography-fabricated microneedles for fluid sampling of histamine-contaminated tuna

A custom-designed microneedle sampling system was prepared using dynamic mask microstereolithography; this sampling system was used for determination of histamine content in fresh, histamine-spiked, and spoiled tuna flesh. Lateral flow (test strip) assays were successfully utilized in the microneedle sampling system to assess histamine content. Good agreement was noted between data obtained from the microneedle sampling system and a commercially available histamine detection kit. A discrepancy was noted in the results from the microneedle sampling system and the commercially available histamine detection kit at low (negative) levels of histamine. There was an improvement in the agreement between the microneedle sampling system and the commercially available histamine detection kit at higher histamine levels. The results, which showed an improvement in the test duration and the amount of reagent needed for histamine detection, indicate the promise of printed microneedle sampling systems for histamine detection in seafood samples and other types of food testing.

Application of near-infrared spectroscopy as at line method for the evaluation of histamine in tuna fish (Thunnus albacares)

The assessment of histamine levels in fishery products has emerged as a paramount issue in the context of global food safety, given its profound implications for human health and the consequential impact on food quality and trade. Histamine intoxication, stemming from the ingestion of foods containing heightened histamine levels, results from the bacterial decarboxylation of histidine under conditions of improper handling, processing, or storage. This study endeavors to provide a thorough examination of histamine contamination in frozen-thawed tuna (Thunnus albacares) samples, employing an integrated approach that combines near-infrared spectroscopy (NIRS) with advanced machine learning techniques. One hundred and one samples were considered, and a systematic fortification process was applied to obtain samples with 4 histamine concentrations (0; 50; 150; 250 mg/kg); the fortification levels were confirmed by the LC-MS/MS analysis. Subsequently, NIRS spectra were collected and chemometric analyses, including modified partial least squares regression (MPLS) and support vector machine (SVM), were employed for quantitative and qualitative evaluation, respectively. Histamine quantification through MPLS utilizing the full spectrum exhibited good predictive performance in cross-validation and in hold-out validation (R2CV = 0.88; R2P = 0.74, respectively), confirming the potential of NIRS for estimating histamine levels in tuna. SVM classification models, both binary (presence/absence) and multiclass (four levels), demonstrated high accuracy (100% and 93%, respectively). The study highlights the effectiveness of NIRS combined with machine learning for rapid and accurate histamine detection in frozen-thawed tuna, offering a non-destructive, environmentally friendly alternative to traditional methods. This approach holds significant promise for food business operators and regulatory authorities, enhancing product safety, quality control, and decision-making processes related to histamine contamination in the seafood industry.

Valorization of refined sardine oil in canned Sardina pilchardus

This study aimed to use refined sardine oil as a covering juice in canned Sardina pilchardus. The oil’s fatty acid profile, acidity, peroxide value, and p-anisidine value were analyzed. The biochemical composition, histamine content, and bacteriological stability of the canned sardines were determined. The results showed that canned sardines contained 51.40 g of moisture, 27.87 g of fat, 17.91 g of protein, and 1.03 g of salt per 100 g net weight. The fatty acid composition included 10.35% polyunsaturated fatty acids, 8.86% saturated fatty acids, and 6.27% monounsaturated fatty acids, with the most abundant fatty acids being eicosapentaenoic acid (EPA), palmitic acid, oleic acid, myristic acid, and docosahexaenoic acid (DHA). This recipe provided 8.88 g/100 g (net weight) of EPA and DHA. These findings support the interest in using refined fish oil to enrich foods with essential marine fatty acids, highly recommended for their health benefits and leading to encouraging prospects for sardine canneries to develop new and nutritious value products.

Valorization of refined sardine oil in canned Sardina pilchardus

This study aimed to use refined sardine oil as a covering juice in canned Sardina pilchardus. The oil’s fatty acid profile, acidity, peroxide value, and p-anisidine value were analyzed. The biochemical composition, histamine content, and bacteriological stability of the canned sardines were determined. The results showed that canned sardines contained 51.40 g of moisture, 27.87 g of fat, 17.91 g of protein, and 1.03 g of salt per 100 g net weight. The fatty acid composition included 10.35% polyunsaturated fatty acids, 8.86% saturated fatty acids, and 6.27% monounsaturated fatty acids, with the most abundant fatty acids being eicosapentaenoic acid (EPA), palmitic acid, oleic acid, myristic acid, and docosahexaenoic acid (DHA). This recipe provided 8.88 g/100 g (net weight) of EPA and DHA. These findings support the interest in using refined fish oil to enrich foods with essential marine fatty acids, highly recommended for their health benefits and leading to encouraging prospects for sardine canneries to develop new and nutritious value products.

DETECTION OF THE VITAL ORIGIN OF THE STRANGULATION MARK IN MECHANICAL ASPHYXIA AS AN INDICATOR OF THE DEVELOPMENT OF FORENSIC MEDICINE

Death by mechanical asphyxia occupies one of the leading positions in the structure of violent deaths. In the structure of mechanical asphyxia, one of the most common is mechanical strangulation, which includes hanging, ligature strangulation and manual strangulation. Solving the problem of the ante-mortem nature of mechanical asphyxia remains relevant in the practical work of forensic experts. In the case of mechanical asphyxia by strangulation, the most common and effective methods are the study of changes in the area of local injuries on the neck, such as the ligature mark during hanging. Aim of the study. To shed light on the process of developing methods for determining the ante-mortem nature of mechanical strangulation asphyxia by studying changes in the area of the ligature mark and to determine effective ways of solving this problem at the current stage of development of forensic medicine. Methods. Analysis and synthesis of information from literary sources available in online databases. Results. The use of macroscopic and histological methods to study changes in the area of the ligature mark may not be sufficient to resolve the issue of ante-mortem formation of injuries in the neck area during hanging, due to the extremely short period of time that elapses between the onset of the asphyxia process and the onset of death. The time interval between the onset of asphyxia and death is often insufficient for the development of a specific reactive cellular and even vascular response. Immunohistochemical examination, aimed at identifying initial reactive signs in the area of injury, is more effective in establishing ante-mortem status during extremely short time intervals between the cause of injury and the onset of death. In the case of mechanical asphyxia by strangulation, significant results have been obtained in the study of ligature marks by immunohistochemical markers such as tryptase, IL-15, CD-15, MHC-II, CD1a, AQP1, AQP3, fibronectin D, P-selectin. In addition, biochemical methods for the detection of physiologically active substances, which allow the detection of early signs of reactive changes in the skin due to injury, such as the study of the concentration of histamine, serotonin, heparin, prostaglandins F and E, as well as the ratio of certain ions, can be highly effective in diagnosing the ante-mortem nature of the ligature mark. Conclusion. The use of immunohistochemical research methods to confirm the ante-mortem formation of a ligature mark is based on the detection of markers of the inflammatory reaction, which are released before the appearance of any vascular and cellular reactions, allowing the detection of the first signs of the inflammatory reaction even within a very short period of time between the cause of injury and the onset of death. Thus, the use of immunohistochemical methods can be effective in confirming the ante-mortem nature of mechanical asphyxia, especially in cases where the use of standard histological staining methods has been ineffective. The detection of certain biochemical markers in the skin from the area of the ligature mark may be considered a promising, rapid and effective method to confirm the ante-mortem nature of strangulation asphyxia. This direction requires further research to improve its effectiveness and can be used in forensic practice to confirm the ante-mortem formation of the ligature mark.

Butyrate Increases Heparin Synthesis and Storage in Human Mast Cells.

Sulphated glycosaminoglycans (GAGs) such as heparin are a major component of mast cell granules and form the matrix within which biogenic mediators are stored. Since GAGs released from mast cells also play an important role in helminth expulsion, understanding GAG storage can offer new insights into mast cell function. Sodium butyrate (NaBu), a short-chain fatty acid, causes ultrastructural changes within the granules of human mast cells (HMC-1) and increases their histamine content. Therefore, we hypothesized that NaBu treatment would also modify the storage of polysaccharides such as GAGs. NaBu (1 mM) significantly increased GAG content and granularity in a time- and concentration-dependent manner without affecting cell viability and metabolic activity. NaBu increased the expression of enzymes associated with heparin biosynthesis (GLCE, NDST1, NDST2, HS6ST1, and GALT1) in a time-dependent manner. A cholesteryl butyrate emulsion (CholButE) increased heparin content after 24 and 48 h and modestly altered the expression of genes involved in heparin biosynthesis. Similar to NaBu, CholButE reduced cell proliferation without significantly altering viability or metabolic activity. These data show that butyrate increases the synthesis and storage of heparin in human mast cells, perhaps by altering their metabolic pathways.

Influence of the Gold Nanoparticle Size on the Colorimetric Detection of Histamine.

Histamine is a well-known biogenic amine (BA) that is often associated with allergic reactions and is a significant cause of foodborne illnesses resulting from the consumption of spoiled food. Detecting histamine is essential for maintaining food safety standards and preserving the quality. In this work, we developed a simple, low-cost, and rapid colorimetric method for detecting histamine. Gold nanoparticles (AuNPs) of different sizes (16, 25, and 40 nm) were synthesized by using the citrate reduction method. The particle size was controlled by adjusting the precursor molar ratio (MR), with smaller ratios leading to larger particles and a red-shift in the surface plasmon resonance (SPR) peak (520, 524, and 528 nm). The nanoparticles were allowed to interact with increasing concentrations of histamine (ranging from 1 to 100 ppm), and the changes in the absorbance spectra and color of the solution were monitored. AuNP aggregation was induced by interaction with histamine through amino and imidazole groups that will coordinate with the AuNP's surface via electrostatic and hydrogen-bonding interactions, causing the solution to turn blue from red. The size variations of AuNPs significantly affected the colorimetric response to histamine. Among the varied sizes, 25 nm AuNPs exhibited the lowest detection limit of 0.72 μM and a linear detection range of 1-10 ppm. Notably, this sensor offered rapid detection (under 1 min) and a remarkable selectivity toward histamine analyte, highlighting its potential for practical applications.

Inhibitory potential of natural antimicrobial compounds against histamine-forming lactic acid bacteria

This work evaluated the potential of natural antimicrobial compounds to prevent histamine formation by inhibiting the microbial growth of Lentilactobacillus parabuchneri and Limosilactobacillus reuteri, bacteria selected for their ability to produce histamine in cheese. Of the evaluated compounds, eugenol, carvacrol, thymol and cinnamaldehyde were the most effective in inhibiting histamine-forming lactic acid bacteria and, therefore, in reducing histamine generation, even at concentrations below the minimum bactericidal concentration (MBC). Carvacrol significantly impacted both microbial viability and the metabolism of bacteria with the inhibition of decarboxylase activity. In the case of L. parabuchneri, the use of 0.7 mg/mL carvacrol (MBC/2) resulted in an 88% reduction in histamine content, and for L. reuteri a 96% reduction was observed by using 0.65 mg/(MBC/2). Therefore, using essential oil components during food processing and preservation would be a good strategy to prevent histamine accumulating in cheese and to, consequently, mitigate its adverse effects.