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Testing for inactivation of pertussis toxin and reversion to toxicity in aP vaccines has historically relied on the murine histamine sensitization test, that lacks mechanistic understanding, suffers from standardization problems and is associated with severe animal suffering. Though the regulatory requirements for in vivo testing of acellular pertussis (aP) vaccine products have been waived in Europe, it is still common practice globally. Easy and quantitative in vitro methods are therefore urgently needed. One of the alternatives under development is our reporter cell line - CHO-CRE cells - that carries a cAMP-reporter construct. After exposure to pertussis toxin, cells are stimulated with a low concentration of forskolin to allow detection of pertussis toxin dependent changes in intracellular cAMP levels. Here, the results of two prevalidation studies with purified pertussis toxin and pertussis toxin spiked aP vaccines are described that were performed according to the principles of the ICH Q2(R1) guidelines for a content assay. We confirmed the assay's specificity, accuracy, precision, linearity and range. The cAMP-PTx reporter assay allows for objective, reliable and quantitative assessment of pertussis toxin levels in aP vaccines and can thereby boost broad and global replacement of the histamine sensitization test. [ABSTRACT FROM AUTHOR]

Introduction: Endometrioid endometrial cancer ranks among the most prevalent malignant tumors affecting women in developed nations, with its incidence steadily rising. Over the past decade, its frequency currently reaching 79 cases per 100,000 women in Europe. Histamine, a biogenic amine, exhibits immunomodulatory properties and serves as the primary mediator of severe and acute inflammatory reactions, as well as immediate hypersensitivity reactions. Histamine operates via four distinct receptor subtypes: HRH1, HRH2, HRH3, and HRH4, with its expression documented across various cancer types. Aim: This study aimed to assess alterations in the expression pattern of genes associated with the histaminergic system in boThendometrial tissue samples and whole blood derived from women diagnosed wiThendometrioid endometrial cancer. Material and methods: The study cohort comprised 30 women diagnosed wiThendometrioid endometrial cancer who underwent hysterectomy. Within this group, 15 cases were classified as G1 (well-differentiated), 8 cases as G2 (moderately differentiated), and 7 cases as G3 (poorly differentiated). The control cohort comprised 30 women with no neoplastic changes during routine gynecological examinations. Molecular analysis encompassed microarray analysis of mRnAs associated with the histaminergic system, reverse-transcription quantitative polymerase chain reaction (RT-qPCR), and enzymelinked immunosorbent assay (ELISA). Results: Among 65 mRnAs associated with the histaminergic system, 10 exhibited differentiations between tissue and blood samples obtained from endometrioid endometrial cancer patients compared to the control group (p<0.05). mRnA histamine receptor 1,3 (HRH1, HRH3), and solute carrier family 22 member 3 (SLC23A2) showed differentiation in endometrioid endometrial cancer samples irrespective of their comparison to the control or G group. Conclusions: The findings reveal a complex architecture of the histaminergic system. The identified mRnA targets hold promise for molecularly targeted therapies in the context of endometrioid endometrial cancer. Furthermore, these results may offer insights into assessing the severity of endometrioid endometrial cancer. [ABSTRACT FROM AUTHOR]

Abstract: Seventy-eight salted fish products originating from Turkey and European countries were analyzed for their content of biogenic amines (BA), water activity (aw), salt and moisture, and their microbiological counts. About 10% of the samples exceeded regulated histamine levels established by the FDA and/or EU authorities. The highest histamine value was found for brined anchovy as 422 ppm. Among the other BAs, the highest level was obtained for tyramine as 524 ppm. The low aw (<0.9) values were obtained with samples containing water phase salt (%WPS) content over 20%. Brined and dry salted products had higher %WPS than lakerda, which is a Turkish and Greek salted fish product. Despite of salt used as preservative agent in fish processing, it is found that salting alone is not enough to guarantee low levels of histamine and other BAs. This study indicated that commercial salted fish products, particularly lakerda, may still present health risk for BAs depending on processing technique, handling and storage conditions of raw material, and during processing. Therefore, processors and regulatory bodies are advised to re-evaluate their productions for safe handling measures. [Copyright &y& Elsevier]

Abstract: The histamine sensitisation test (HIST) for pertussis toxin is currently an official batch release test for acellular pertussis containing combination vaccines in Europe and North America. However, HIST, being a lethal endpoint assay, often leads to repeated tests due to large variations in test performance. Although a more precise HIST test based on measurement of temperature reduction after the histamine challenge is used in Asian countries, this test still uses animals. An in vitro test system based on a combination of enzyme coupled-HPLC and carbohydrate-binding assays with results analysed by a mathematical formula showed a good agreement with the in vivo HIST results based on measurement of temperature reduction after histamine challenge. The new in vitro test system was shown to be a potential alternative to the current in vivo HIST. [Copyright &y& Elsevier]

Tumour necrosis factor (TNF)-based isolated limb perfusion (ILP) is an approved and registered treatment for sarcomas confined to the limbs in Europe since 1998, with limb salvage indexes of 76%. TNF improves drug distribution in solid tumours and secondarily destroys the tumour-associated vasculature (TAV). Here we explore the synergistic antitumour effect of another vasoactive agent, histamine (Hi), in doxorubicin (DXR)-based ILP and evaluate its antivascular effects on TAV. We used our well-established rat ILP model for in vivo studies looking at tumour response, drug distribution and effects on tumour vessels. In vitro studies explored drug interactions at cellular level on tumour cells (BN-175) and Human umbilical vein endothelial cells (HUVEC). There was a 17% partial response and a 50% arrest in tumour growth when Hi was combined to DXR, without important side effects, against 100% progressive disease with DXR alone and 29% arrest in tumour growth for Hi alone. Histology documented an increased DXR leakage in tumour tissue combined to a destruction of the TAV, when Hi was added to the ILP. In vitro no synergy between the drugs was observed. In conclusion, Hi is a vasoactive drug, targeting primarily the TAV and synergises with different chemotherapeutic agents. [ABSTRACT FROM AUTHOR]

The aim of the research was the assessment of 8 biogenic amines occurrence (BA) in ciders. Forty samples with ethanol content ≤4.5% v/v (LC ; low alcohol content) and 34 samples with ethanol content > 4.5% v/v (HC ; high alcohol content) manufactured in the Central Europe region were tested. The cider samples were immediately analyzed after purchase and at the end of the best before date. The most abundant BA across all ciders was tyramine, followed by putrescine and cadaverine (p < 0.05). Samples reported low levels of tryptamine, spermidine, spermine, histamine and phenylethylamine. No cider at the end of the best before date had a sum of BA below <5 mg l−1. Moreover, 67% of the samples at the beginning of the storage period and 47% of the ciders after the best before date presented a total BA content in the range of 5–20 mg l−1. A total number of 14% of the samples immediately after purchase and 31% of samples at the end of the best before date showed a BA concentration in the range of 20–50 mg l−1. The BA content of 16 samples was >50 mg l−1 at the end of the best before date. However, one LC cider and two HC products displayed a BA sum of nearly 120 mg l−1. In general, higher concentrations of tyramine, cadaverine and putrescine were detected in LC samples. All in all, with the prolonging of storage the BA concentration increased. • Eight biogenic amines were identified in 74 cider samples by HPLC. • Ciders with low alcohol content presented higher level of biogenic amines. • The most frequently detected biogenic amine was tyramine, putrescine and cadaverine. • Sixteen ciders presented biogenic amine content higher than 50 mg l−1. • For all ciders higher values of biogenic amines were detected at the end of the best before date. [ABSTRACT FROM AUTHOR]

The objective of the current study was the evaluation of 8 biogenic amines (BA) occurrence in beer samples (115 samples in total) manufactured in microbreweries of the Central Europe region in relation to the progress of the storage period (at 20 ± 2 °C). The examined beer samples were divided into 3 main groups according to their extract of original wort value (EOW): (i) Light craft beer (EOW ≤ 10; 12 samples in total), (ii) Lager craft beer (11 ≤ EOW ≤ 12; 65 samples in total), (iii) Special craft beer (EOW ≤ 13; 38 samples in total). The tested craft beer samples were analyzed immediately after purchase and at the end of the best before date. Furthermore, the most frequently detected BA was tyramine. In addition, other abundant monitored BA were putrescine and cadaverine. Moreover, concentrations of histamine above 20 mg/l were detected in lager craft beer and special craft beer samples (at the end of the best before date). On the whole, with the progress of the storage time the BA concentration increased. Thereafter, more than 30% of the tested samples presented total BA content in the range of 50–100 mg/l. However, 18% of the examined craft beer samples had a total amount of BA higher than 100 mg/l. • Biogenic amine occurrence in Czech craft beers was monitored. • With the progress of the storage the biogenic amine content increased. • The most frequently detected biogenic amine was tyramine. • 18% of the beers presented biogenic amine values higher than 100 mg/l. [ABSTRACT FROM AUTHOR]

The aim of the study is to model concentrations of selected biogenic amines in various fish species (Atlantic salmon, Atlantic cod, striped catfish) bought in retail stores in Central Europe. Since the data contains non-detectable values, statistical methods for left-censored values from the exponential and Weibull distributions are applied and used to evaluate and compare the amount of biogenic amines in fish samples. Moreover, a risk of exceeding certain limits of biogenic amine concentrations to protect human health is determined. There are relatively high concentrations of putrescine, cadaverine and histamine in almost all fish species. Moreover, there was a significant difference in mean concentrations (distributions of concentrations, respectively) of histamine, tyramine and spermidine among the species. • Use of censored distributions for modelling data with non-detectable values. • High concentrations of putrescine, cadaverine and histamine in almost all species. • Significant difference in concentrations of histamine, tyramine and spermidine. • High probabilities of exceeding specified limit values of putrescine concentration. [ABSTRACT FROM AUTHOR]

Accurate and precise methods of skin testing are essential for "in vivo" standardization of extracts from the same and different sources, for diagnostic efficacy as well as an important prognostic factor for evaluating the risk and benefit of immunotherapy. Skin test methods such as parallel line bioassay and ID50EAL which are based on the intradermal erythema response and yield the most sensitive, accurate, and precise bioassays as well as requiring the fewest injections and replicates should be preferentially selected for clinical use. Differentiating characteristics of parallel line bioassay and the ID50EAL method are presented. Proficiency test methods for assessing puncture and intradermal skin test methodologies should be made part of the clinicians' training and be incorporated into clinical practice in conjunction with standardized allergenic extracts. Assignment of Allergy Units to reference extracts based on accurate and precise "in vivo" methods such as the ID50EAL and to test extracts based on "in vitro" assays predictive of their "in vivo" relative potency as determined by parallel-line bioassay should facilitate the safe and effective clinical use of allergenic extracts.