High Histamine Research Articles

EXTH-65. EXPLORING ANTIHISTAMINES TO TREAT GLIOBLASTOMA

Abstract BACKGROUND Glioblastoma (GBM) is an aggressive and lethal primary malignant brain tumor with a 5-year survival of 6.8%. Interestingly, people with allergies, atopy, or asthma are less likely to develop GBM. A recent study recognized that high histamine receptor 1 (HRH1) expression is inversely related to overall and progression-free survival. METHODS We used a bioinformatics tool, iLINCS, which, after analyzing 99 GBM and 38 healthy samples, identified 36 drugs that can reverse GBM signatures. Based on the concordance score and blood-brain barrier permeability, we selected brompheniramine (Brom), a first-generation HRH1 antagonist, as a potential candidate. HRH1 expression analysis was done on human and mouse GBM cell lines by immunoblotting. The effect of Brom alone or in combination with temozolomide (TMZ) was investigated on proliferation, DNA damage and apoptosis in human (U118 and U251) and three syngeneic (EGFRvIII, p16-/- & GFAP Cre, PTEN+/-, p53R172H+/-, EGFRvIII & GFAP Cre, PTEN-/-, p53R172H-/- & GFAP Cre) cell lines. GBM orthograft mouse models were used to test the efficacy of combination therapy. RESULTS HRH1 is highly expressed in human and syngeneic GBM cells. This expression is further increased in TMZ-resistant cells. Interestingly, the addition of histamine, induced TMZ resistance in GBM cells. TMZ with Brom decreases proliferation in mouse and human GBM cells, and Brom is non-toxic to non-transformed cells. Combination treatment increases DNA damage and decreases the DNA repair mechanism, possibly by downregulating the expression of O6methylguanine-DNA methyltransferase (MGMT). In the GBM orthograft mouse model, the combination therapy reduced tumor volume and significantly increased survival compared to single-agent treatment. CONCLUSIONS Brom is non-toxic; Brom, in combination with TMZ, enhanced the anti-tumor efficacy of TMZ both in in vitro and in vivo GBM models. Further studies in spontaneous mouse models are planned.

The Pathology and Blood Biochemistry of Juvenile Lates calcarifer on Diets Contaminated With Mycotoxins, Histamines and Rancid Fats-A Case Study.

Mycotoxins, originating from contaminated raw materials or suboptimal feed storage, are a growing concern in tropical aquaculture. Common fungi such as Aspergillus spp. and Fusarium spp. produce mycotoxins including aflatoxin, fumonisin, deoxynivalenol and zearalenone. High doses or prolonged exposure (weeks) to low doses of these mycotoxins (< 20 μg/kg) can depress growth, immunity, and cause mortality. This study investigated poor growth and low survival in juvenile Asian seabass, Lates calcarifer fed two different diets (FM40 and ABS3) for 5 weeks. Analysis of these diets revealed high peroxide values, multiple mycotoxins and high histamine levels. Fish fed the FM40 diet, which was contaminated with aflatoxin B1 (13.2 μg/kg), aflatoxin B2 (1.9 μg/kg), deoxynivalenol (29.5 μg/kg), alternariol (2.2 μg/kg), elevated peroxide value (45.91 mEq/kg), and histamine (129.51 mg/kg) developed mild bile duct hyperplasia, depressed total serum proteins (50.40 ± 10.06 g/L), markedly elevated blood potassium (8.2 ± 0.18 mmol/L), and heavy iron deposits in splenic melanomacrophage centres (Perl's stain) indicative of increased haemolysis. The presence of multiple cytotoxic mycotoxins in FM40 diet could explain the increased haemolysis and elevated blood potassium. In contrast, fish fed the ABS3 diet, which had high histamine levels (210.05 mg/kg), exhibited protein-losing nephropathy with multifocal fibrin plugs (Martius scarlet blue stain) indicating acute renal damage, and elevated blood calcium and phosphorus levels. Histamine is metabolised and excreted through the kidneys and known to induce renal arteriolar constriction, disrupt glomerular filtration barrier and increase permeability resulting in protein loss. This study shows that blood biochemistry and histopathology are useful diagnostic tools for assessing the impact of mycotoxins and histamines on fish health.

High histamine levels are associated with acute-on-chronic liver failure and liver-related death in patients with advanced chronic liver disease.

The role of histamine in advanced chronic liver disease (ACLD) is poorly understood. We investigated plasma histamine levels across ACLD stages and their prognostic value. We included patients with evidence of ACLD, defined by portal hypertension (hepatic venous pressure gradient [HVPG] ≥6 mmHg) and/or a liver stiffness measurement by transient elastography ≥10 kPa, who underwent HVPG measurement between 2017 and 2020. Acute-on-chronic liver failure (ACLF) and/or liver-related death were defined as composite endpoint. Of 251 patients, 82.5% had clinically significant portal hypertension (median HVPG: 17 mmHg [interquartile range (IQR) 12-21]) and 135 patients (53.8%) were decompensated at baseline. Median plasma histamine was 8.5 nmol/L (IQR: 6.4-11.5), 37.1% of patients showed elevated values (>9.9 nmol/L). Histamine levels did not differ significantly across Child-Turcotte-Pugh (CTP) stages nor strata of model for end-stage liver disease (MELD) or HVPG. Histamine levels correlated with markers of circulatory dysfunction (i.e. sodium, renin and aldosterone). During a median follow-up of 29.2 months, 68 patients developed ACLF or liver-related death. In univariate as well as in multivariate analysis (adjusting for age, sex, HVPG as well as either MELD, clinical stage, and serum albumin or CTP and serum sodium), elevated histamine levels remained associated with the composite endpoint. CTP-based multivariate model adjusted sub-distribution hazard ratio (asHR): 1.010 (95% CI: 1.004-1.021), p < .001; MELD-based multivariate model asHR: 1.030 (95% CI: 1.017-1.040), p < .001. High levels of histamine were linked to circulatory dysfunction in ACLD patients and independently associated with increased risks of ACLF or liver-related death. Further mechanistic studies on the link between histamine signalling and development of hyperdynamic circulation and ACLF are warranted.

Histochemical and immunohistochemical investigation of the number and localization of mast cells in the feline tongue.

This is the first study to describe the subtypes, number and distribution of mast cells (MC) in cat tongue by histochemical and immunohistochemical methods. Six male adult felines' tongue tissue samples consist of the study's material. Samples were fixed in 10% formaldehyde. MC number and distribution in the feline tongue were assessed using toluidine blue. Also, sections taken from blocks were stained in alcian blue/safranin O (AB/SO) combined dyes to determine the MC subtypes. The Streptavidin biotin complex method using anti-chymase and anti-tryptase primary antibodies was used for immunohistochemistry. Metachromatic MCs were mainly observed in the lamina propria close to the multilayered keratinized stratified squamous epithelium. The high number of MCs in this region may be because the dorsal surface of the tongue plays an essential role in the defence system of tongue tissue and, thus, of the body as a whole. Additionally, the number of MCs stained with AB (+) (1.7 ± 0.08) in the feline tongue was statistically higher than those with SO (+) (0.18 ± 0.02). This might be interpreted as an indication that MC heterogeneity may be due not only to their staining properties but also to their localization. It is also conceivable that the high histamine content may be a factor in this. Tryptase-positive MCs were found in the loose connective tissue around blood vessels, between the glands, as solitary cells, or in groups of several cells. Chymase-positive MCs were observed more individually rather than in groups. Moreover, chymase-positive MCs were detected to be located in the filiform papillae subepithelial and in the blood vessels' immediate vicinity. Animals often lick themselves to clean themselves and promote healing. For this reason, it is very important to protect the tongue, which is in direct contact with the external environment, against foreign agents. Considering both the functional and protective properties of the tongue, we concluded that MCs may play a role in oral cavity immunity and protective effect.

A pilot study on the isolation and identification of histamine-producing bacteria from narrow barred Spanish mackerel and Japanese thread fin bream from Ukkadam fish market, Coimbatore, India

Consumption of marine fishes with high histamine levels due to improper storing leads to cause scombroid or histamine fish poisoning. In spoiled fishes, bacteria begin to break down the histidine into histamine by decarboxylation activity. Previous studies reported that black muscle fishes usually produce large amounts of histamine-producing bacteria (HPB). In this study, two white muscle fishes such as Scomberomorus commerson (Narrow-barred Spanish mackerel) and Nemipterus japonicus (Japanese threadfin bream) were chosen to detect the presence of histamine-producing bacteria (HPB). The fish samples were purchased from Ukkadam fish market in Coimbatore and examined for the occurrence of HPB on Niven agar. The isolated HPB on media were analyzed for identification based on their morphological characteristics, primary test, and biochemical test. Finally, 8 colonies were confirmed as the cause for histamine production of both fishes and among them, 5 belong to the family Enterobacteriaceae and others belong to the family Aeromonadaceae, Morganellaceae, and Enterococcaceae. Our study showed that the histamine-producing bacteria highly reported in dark-muscle fishes were also isolated from the chosen white-muscle fishes. Hence the presence of HPB indicates the improper maintenance of fish in retail fish markets. This considerable incidence of histamine-producing bacteria which on proliferation under suitable conditions may contribute to toxic histamine accumulation in the flesh of fishes.

Molecular Mechanism Biomarkers Predict Diagnosis in Schizophrenia and Schizoaffective Psychosis, with Implications for Treatment.

Diagnostic uncertainty and relapse rates in schizophrenia and schizoaffective disorder are relatively high, indicating the potential involvement of other pathological mechanisms that could serve as diagnostic indicators to be targeted for adjunctive treatment. This study aimed to seek objective evidence of methylenetetrahydrofolate reductase MTHFR C677T genotype-related bio markers in blood and urine. Vitamin and mineral cofactors related to methylation and indolamine-catecholamine metabolism were investigated. Biomarker status for 67 symptomatically well-defined cases and 67 asymptomatic control participants was determined using receiver operating characteristics, Spearman's correlation, and logistic regression. The 5.2%-prevalent MTHFR 677 TT genotype demonstrated a 100% sensitive and specific case-predictive biomarkers of increased riboflavin (vitamin B2) excretion. This was accompanied by low plasma zinc and indicators of a shift from low methylation to high methylation state. The 48.5% prevalent MTHFR 677 CC genotype model demonstrated a low-methylation phenotype with 93% sensitivity and 92% specificity and a negative predictive value of 100%. This model related to lower vitamin cofactors, high histamine, and HPLC urine indicators of lower vitamin B2 and restricted indole-catecholamine metabolism. The 46.3%-prevalent CT genotype achieved high predictive strength for a mixed methylation phenotype. Determination of MTHFR C677T genotype dependent functional biomarker phenotypes can advance diagnostic certainty and inform therapeutic intervention.

Oligomeric Proanthocyanidins Alleviate the Detrimental Effects of Dietary Histamine on Intestinal Health of Juvenile American Eels (Anguilla rostrata)

This study evaluated the oligomeric proanthocyanidins (OPC) on alleviating the detrimental effects in the intestine caused by dietary histamine in juvenile American eels (Anguilla rostrata). A total of 480 fish with a similar body weight of 10.84 ± 0.16 g were randomly divided into four groups, including the control group fed a basal diet, the HIS group fed a diet with a high level of histamine (534 mg/kg), the H + OPC I group fed the high histamine diet with 300 mg/kg OPC, and the H + OPC II group fed the high histamine diet with 600 mg/kg OPC. After the fish were fed the trial diets for 77 days, the intestinal samples were taken, and the related parameters were analyzed. Dietary 300 mg/kg or 600 mg/kg OPC could reverse the decreased activities of lipase, protease, and glutathione peroxidase and the level of total antioxidant capacity; the increased intestinal malondialdehyde and D-lactate acid levels and the activity of diamine oxidase in serum; and the decreased villus height caused by a high level of dietary histamine. There were no significant differences above all the indices between the H + OPC I group and the H + OPC II group. The higher relative abundances of potentially pathogenic bacteria were induced by the high level of dietary histamine. Dietary 300 mg/kg OPC might increase the relative abundance of the potential probiotics and inhibit the colonization of intestinal pathogenic bacteria. The intestinal health status of the H + OPC groups was similar to that of the control group. These results suggested that dietary 300 mg/kg OPC might alleviate the detrimental effects of dietary 534 mg/kg histamine on the intestine health of juvenile American eels by increasing the activity of digestive enzymes, improving the antioxidative potential and barrier function, and beneficially modulating the intestinal microbiota. Dietary 600 mg/kg OPC could not exert further improvement in growth performance and the intestinal health of juvenile American eels.

Ingested histamine and serotonin interact to alter Anopheles stephensi feeding and flight behavior and infection with Plasmodium parasites.

Blood levels of histamine and serotonin (5-HT) are altered in human malaria, and, at these levels, we have shown they have broad, independent effects on Anopheles stephensi following ingestion by this invasive mosquito. Given that histamine and 5-HT are ingested together under natural conditions and that histaminergic and serotonergic signaling are networked in other organisms, we examined effects of combinations of these biogenic amines provisioned to A. stephensi at healthy human levels (high 5-HT, low histamine) or levels associated with severe malaria (low 5-HT, high histamine). Treatments were delivered in water (priming) before feeding A. stephensi on Plasmodium yoelii-infected mice or via artificial blood meal. Relative to effects of histamine and 5-HT alone, effects of biogenic amine combinations were complex. Biogenic amine treatments had the greatest impact on the first oviposition cycle, with high histamine moderating low 5-HT effects in combination. In contrast, clutch sizes were similar across combination and individual treatments. While high histamine alone increased uninfected A. stephensi weekly lifetime blood feeding, neither combination altered this tendency relative to controls. The tendency to re-feed 2weeks after the first blood meal was altered by combination treatments, but this depended on mode of delivery. For blood delivery, malaria-associated treatments yielded higher percentages of fed females relative to healthy-associated treatments, but the converse was true for priming. Female mosquitoes treated with the malaria-associated combination exhibited enhanced flight behavior and object inspection relative to controls and healthy combination treatment. Mosquitoes primed with the malaria-associated combination exhibited higher mean oocysts and sporozoite infection prevalence relative to the healthy combination, with high histamine having a dominant effect on these patterns. Compared with uninfected A. stephensi, the tendency of infected mosquitoes to take a second blood meal revealed an interaction of biogenic amines with infection. We used a mathematical model to project the impacts of different levels of biogenic amines and associated changes on outbreaks in human populations. While not all outbreak parameters were impacted the same, the sum of effects suggests that histamine and 5-HT alter the likelihood of transmission by mosquitoes that feed on hosts with symptomatic malaria versus a healthy host.

FRI-554 - High histamine levels associate with acute-on-chronic liver failure and liver-related death in patients with advanced chronic liver disease

FRI-554 - High histamine levels associate with acute-on-chronic liver failure and liver-related death in patients with advanced chronic liver disease

The induction of histamine synthesis from rat bone marrow derived mast cells with rat stem cell factor

Abstract Mast cells are the main cells to produce histamine regarded as the main factor to induce allergy. To comprehend the allergy by mast cell activation, The mast cells of bone marrow derived mast cells (BMMC) are usually prepared by culturing bone marrow cells including rodent hematopoietic stem cells, in the presence of both IL-3 and stem cell factor (SCF). However, BMMC was reported not to produce the high levels histamine comparable to mature mast cells in host tissue. In this study, rat BMMC (rBMMC) were cultured in 96 well plate to induce diverse conditions in cell numbers per unit culture media, rat IL-3 and rat SCF to find the conditions to synthesize high level histamine. Cells of rat bone marrow cells were cultured in the cell density of 1.1×10 5/ml (low density, lowD) and 5.5 × 10 5/ml (high D), rat IL-3 was added to the levels of 10 ng/ml (low 3) or 50 ng/ml (high 3), Rat SCF was adjusted to the levels of 12.5 (lows), 25 (midS) and 50 (highS) ng/ml. Histamine included in 5×10 4BMMC were extracted by perchloric acid, n-butanol, and heptane. The levels of histamine were measured by ophthalialdehyde solution and read for fluorometric intensity. The levels at culture day 4 (D 4) were in the range of 12~45 ng. The significant elevation was observed at D 11 in the conditions of lowD high3 (153 ng), however, D 8 in highD high3 (253 ng). The average histamine level was 74 ng in the condition of low Dlow 3, but 112 ng in low Dhigh 3 at D 11. That was 106 ng in high Dlow 3 but 183 ng in high Dhigh 3 at culture D 8. These results indicated that rat IL-3 increased the level of histamine synthesis and shorten the histamine production from undifferentiated hematopoietic stem cells. The induction of histamine synthesis from rat bone marrow derived mast cells with rat stem cell factor

Diagnosing gastric ulcers in bottlenose dolphins (Tursiops sp.) using gastroscopy and cytology.

Gastric ulcers have been reported in a range of cetacean species. Bottlenose dolphins (Tursiops spp.), the most common cetacean species held in captivity, are known to experience gastric ulcers in both wild and captive environments. Documented causes of gastric ulceration include bacterial infection by Helicobacter sp., parasitic infections, high dietary histamine and foreign body ingestion. Gastric ulceration without any obvious cause might be related to stress. Currently, the most accurate way to determine the presence of gastric ulcers in captive dolphins is through direct examination of the stomach mucosa using endoscopy (gastroscopy); a procedure that requires substantial animal training and specialised medical equipment. In this study, we investigate whether cytology of the gastric fluid, collected through the less intensive method of intubation, can be used as an alternative to gastroscopy to predict the presence and severity of gastric ulcers in eight captive bottlenose dolphins at uShaka Sea World, South Africa. An ulcer grading scale was developed to quantify the severity of the dolphins' gastric ulcers observed using gastroscopy. Gastric ulcer severity was then compared with the cytological data collected from gastric fluid samples taken during the gastroscopic examinations. The cytological findings were consistent with other studies, but ulcer severity was not found to be linked to the cytological parameters measured. From these results we suggest that routine cytology of the gastric fluid is not a viable alternative to gastroscopy for diagnosing gastric ulcers in bottlenose dolphins.

57 ISCHEMIC HEART SCOMBROID SYNDROME: FURTHER EVIDENCE FROM ALL OVER THE WORLD

Abstract Objective Scombroid syndrome (SS) is a long-standing poisoning illness caused by ingestion of improperly stored fish food (usually tuna fish) contaminated by Gram-negative bacteria. The main pathogenetic feature is toxicity by the high histamine (and/or other toxin) levels within the fish muscle. Scombroid syndrome accounts for approximately 5% of food toxicities at emergency department in the USA. It consists of gastrointestinal symptoms, sickness, at times (head) rash or flushing, peppery taste, hypotension, and other symptoms. Cardiovascular complications are infrequent, but acute coronary syndromes (ACSs) have been described in some patients on admission to hospital. We identified those cases as with ischemic heart scombroid syndrome (IHSS). Methods An accurate literature search for IHSS in case series and single reports, published as by the end of July 2022, was performed. Herein main clinical features are summarized. Results From 1997 to 2022, 18 studies were found describing 25 patients aged 42±15 years, 14 women (56%), likely to have IHSS. Countries of origin are depicted in the Figure nearby. Apart from typical clinical presentation of SS, chest pain, oppression and hypotension were complained by almost all patents. Symptoms and signs of ACS started 10 to 180 minutes from the meal. Four patients (mostly before 2015) required advanced life support or mechanical ventilation. Widespread ST-depression (first in the inferior leads) was the most frequently reported ECG feature (approximately in 90% of cases). Hospitalization was required for unstable patients or those admitted for coronary angiography. However, this latter was performed only in 10 cases (40%), and only in 3 (12%) a culprit lesion was identified and treated. Therapy consisted with antihistamine drugs, fluids and circulatory support, symptomatic drugs, and steroids in some cases. All patients were discharged from hospital with uneventful follow-up. Conclusions To date, 25 published cases of IHSS have been recognized, mostly from Italy, but reports are from all over the world. As in typical SS, myocardial ischemia begins shortly after raw or cocked (tuna) fish ingestion. However, due to unfamiliarity with the syndrome, diagnostic misinterpretation is possible. IHSS can be classified as a transient (usually benign) ACS, caused by coronary vasospasm in response to a nitroxide-mediated histamine-induced tissue hypoperfusion, in a similar way to type-I Kounis syndrome. Based on current knowledge, coronary angiography is mandatory in patients with a history of coronary artery disease and/or allergic diathesis, persistent symptoms or unresponsive to emergency treatment.

Intraoperative chlorhexidine-induced anaphylaxis suggesting an immunoglobulin-E-dependent mechanism indicated by basophil activation tests: two case reports

BackgroundAlthough chlorhexidine allergy has been shown to be mediated by immunoglobulin (Ig) E, few reports investigated the mechanism of chlorhexidine-induced anaphylaxis using basophil activation tests (BATs).Case presentationA 79-year-old man underwent cholecystectomy under general anesthesia. Anaphylaxis was diagnosed based on the clinical symptoms and high serum tryptase and histamine levels. Skin tests showed positive results only for chlorhexidine. Subsequently, BATs demonstrated that the causative agent was likely chlorhexidine. The inhibitory effect of wortmannin, an inhibitor of phosphoinositide 3-kinase, on basophil activation suggested an IgE-dependent mechanism underlying chlorhexidine-induced anaphylaxis.An 89-year-old man underwent inguinal hernioplasty under general anesthesia. Anaphylaxis was diagnosed based on the clinical symptoms and high serum tryptase and histamine levels. Skin tests and BATs with wortmannin were performed, showing similar results to case 1.ConclusionsBATs suggested an IgE-dependent mechanism for chlorhexidine-induced anaphylaxis and might be useful for investigating the mechanisms underlying drug-induced anaphylaxis.

High Dietary Histamine Induces Digestive Tract Oxidative Damage in Juvenile Striped Catfish (Pangasianodon hypophthalmus).

A 56-day feeding trial investigated the effects of dietary histamine on the antioxidant capacity, gastric and intestinal barrier functions, and growth performance of striped catfish (Pangasianodon hypophthalmus). Seven isonitrogenous (34.0% crude protein) and isolipidic (10.5% crude lipid) diets were formulated with supplemental 0, 15, 30, 60, 120, 240, and 480 mg/kg of histamine, named H0, H15, H30, H60, H120, H240, and H480 group, respectively. Results showed that the weight gain rate, specific growth rate, relative intestinal length in the H240 and H480 groups, and the condition factors in the H480 group were significantly lower than those in the H0 group. Intestinal total antioxidant capacity, peroxidase, catalase, superoxide dismutase, glutathione peroxidase, and glutathione reductase activities in the H480 group were significantly lower than those in the H0 group, whereas intestinal malondialdehyde content exhibited the opposite trend. Intestinal complement 3, complement 4, immunoglobulin M, and Recombinant Mucin 2 in the H480 group were significantly lower than those in the H0 group, in contrast to intestinal lipopolysaccharide content. Intestinal IL-10 gene expression in the H480 group was significantly lower than that in the H0 group, whereas the TNF-α, IL-1, IL-6, and IL-8 gene expression exhibited opposite results. Scanning and transmission electron microscopic observation of the gastrointestinal tract revealed severe damage to the gastric mucosa and intestinal epithelium in the H480 group. The abundance of Treponema in the histamine groups was significantly higher than that in the H0 group. These results indicated that high dietary histamine decreases intestinal immunity and antioxidant capacity, inducing digestive tract oxidative damage and ultimately decreasing the growth of striped catfish.

Science, Medicine, and the Anesthesiologist

Science, Medicine, and the Anesthesiologist

The significance of cheese sampling in the determination of histamine concentration: Distribution pattern of histamine in ripened cheeses

Cheeses are becoming a major safety and public health concern: cheeses available in supermarkets occasionally contain high histamine concentrations that can have negative effects on consumer health. In this study, we have attempted to assess the histamine distribution pattern in ripened cheeses, with the purpose of establishing a correct cheese sampling strategy for the quantification of histamine. To this aim, histamine was determined in four distinct areas of twelve long-ripened hard cheeses: the external and internal rind, along with the outer and inner core of the wedge. The concentrations measured were remarkably different: histamine accumulated in the central core, whereas the lowest amount was found in the peripheral rind. To explain this heterogenous distribution, histamine producers were determined in the four areas by identifying the hdc sequences obtained from cheese samples. Non-starter bacteria were identified as main histamine producers; however, these microbiota were homogeneously distributed throughout the wedge. Nevertheless, the analysis of psychochemical properties of the different areas revealed an observable trend: histamine tended to accumulate in the saltier, more humid, and less oxidized areas in a wedge. Overall, this study highlights the significance of a correct sampling strategy when histamine is quantified in cheese.

ANTIBACTERIAL POTENCY OF CELL FREE SUPERNATANT PRODUCED BY LACTIC ACID BACTERIA ISOLATED FROM INDONESIAN FERMENTED FISH PRODUCTS AGAINST HISTAMINE-PRODUCING BACTERIA

The toxicity caused by high histamine content produced by histamine-producing bacteria (HPB) during the fermentation of scombridae fish group based fermented fish products is still a problem that requires a solution. This study aims to explore the potential of secondary metabolites in the form of cell free supernatant (CFS) produced by lactic acid bacteria (LAB) as antibacterial agents against HPB. The LAB was isolated from the fermented fishery products named bekasan, cincalok and fish sauce using MRS-Agar and fermented on MRS-broth for 48 hours. CFS was collected by centrifugation at 15,000 x g for 15 minutes, followed by heating at 100oC for 3 minutes and pH neutralization with 0.1N NaOH. Antimicrobial activity of CFS then tested on HFB namely Morganella morganii TK7, Citrobacter freundii CK1, and Klebsiella sp. CK13.2 (collection of the Laboratory of Fisheries Product Quality and Safety, Department of Fisheries UGM) using the macrodilution method. Isolation of LAB from all three products successfully isolated 34 isolates. The results of the antibacterial activity showed that 4 isolates namely GMCN 1.12, GMBK 2.6, GMBK 2.7, dan GMKI 2.1 were able to inhibit HFB growth more than 70%. The highest activity was shown by GMBK 2.7 which inhibits 98% against Morganella morganii TK7, 99% against Citrobacter freundii CK1, and 84% against Klebsiella sp. CK13.2. The antimicrobial activity was reduced after proteolytic enzymes were added suggesting that the bioactive compound came from peptide-based substances like bacteriocin.

Are there profiles of cheeses with a high GABA and safe histamine content?

Some bacteria decarboxylate amino acids as a means of resisting acid stress. It is a mechanism commonly used by bacteria in fermented dairy products. The decarboxylation of histidine to histamine occurs frequently in some types of cheese, but high concentrations of histamine can be harmful to consumers. Not all amino acid decarboxylation products are, however, undesirable. GABA, the decarboxylation product of glutamate, has positive effects on human health. Although histamine and GABA accumulate in large quantities in many types of cheese, there is little information regarding their relationship, and that which is available is restricted to a few cheese varieties. The present work examines whether cheeses exist with high concentrations of GABA but safe concentrations of histamine. The GABA and histamine contents of 250 cheeses representing 143 European varieties were examined, and the technological, environmental and metabolic (TEM) traits governing their accumulation identified. Two TEM profiles were associated with high GABA contents and safe histamine concentrations. These findings could help the dairy industry select the best technological conditions for making GABA-rich, histamine-safe cheeses. • GABA and histamine were determined in 250 cheeses of 143 different varieties. • GABA (healthy) and histamine (toxic) can accumulate at high concentrations. • A positive correlation exists between GABA and histamine concentration. • Cheeses with high GABA and low histamine contents were identified. • The technological/environmental/metabolic profiles of these cheeses were recorded.

The allergy mediator histamine confers resistance to immunotherapy in cancer patients via activation of the macrophage histamine receptor H1.

Reinvigoration of antitumor immunity remains an unmet challenge. Our retrospective analyses revealed that cancer patients who took antihistamines during immunotherapy treatment had significantly improved survival. We uncovered that histamine and histamine receptor H1 (HRH1) are frequently increased in the tumor microenvironment and induce Tcell dysfunction. Mechanistically, HRH1-activated macrophages polarize toward an M2-like immunosuppressive phenotype with increased expression of the immune checkpoint VISTA, rendering Tcells dysfunctional. HRH1 knockout or antihistamine treatment reverted macrophage immunosuppression, revitalized Tcell cytotoxic function, and restored immunotherapy response. Allergy, via the histamine-HRH1 axis, facilitated tumor growth and induced immunotherapy resistance in mice and humans. Importantly, cancer patients with low plasma histamine levels had a more than tripled objective response rate to anti-PD-1 treatment compared with patients with high plasma histamine. Altogether, pre-existing allergy or high histamine levels in cancer patients can dampen immunotherapy responses and warrant prospectively exploring antihistamines as adjuvant agents for combinatorial immunotherapy.

Flow cytometric analysis of the inhibition of human basophil activation by histamine high dilutions - a replication study

Background: Inhibition of human basophil activation by highly diluted histamine was reported to be a reliable experimental model to examine biological effects of high dilutions. However, independent replications did not always yield concordant results.&#x0D; Aims: We aimed at performing an independent replication of a former study [1] using rigorously controlled experimental conditions to minimise confounding factors.&#x0D; Materials and Methods: In 20 independent experiments, human basophils were treated with highly diluted histamine (15cH, 16cH, corresponding to 10-30-10-32 M) prior to activation by fMLP (formyl-methionyl-leucyl-phenylalanine peptide). Controls were treated with analogously diluted water (15cH, 16cH). The dilutions were prepared freshly for each experiment in deionised water by successive steps of centesimal dilution and agitation (10 s vortex at high speed). Highly diluted samples were blinded and randomised. All samples were set in triplicates. Activated basophils were determined by flow cytometry using anti-CD203c. 20 independent systematic negative control (SNC) experiments were carried out to investigate possible systematic errors. &#x0D; Results: No difference in basophil activation was observed between the highly diluted histamine samples and the highly diluted water controls. There was no evidence for a blood donor specificity of the results. The SNC experiments demonstrated the stability of the test system. Experimental variability within and between experiments was slightly reduced for the highly diluted histamine samples.&#x0D; Discussion: This study was designed as an independent reproduction of a former study [1]. Though we strictly adopted the experimental procedure described in [1], our results do not confirm the large inhibitory effects observed for histamine 15cH and 16cH. This lack of reproducibility might be due to minor differences in the experimental design, such as blinding and randomising of the samples, which we chose to perform in order to reduce the possibility of artifacts but was omitted in the former study.&#x0D; Conclusions: Laboratory independent replication of homeopathic basic research experiments is still a challenge. Assuming that the results formerly obtained with this model were not due to systematic errors, the quest identifying the crucial factors for successful reproducibility is open for future research.&#x0D; Keywords: Human basophils; histamine; high dilutions; flow cytometry&#x0D; Reference:&#x0D; [1] Sainte-Laudy J, Belon P. Improvement of flow cytometric analysis of basophil activation inhibition by high histamine dilutions. A novel basophil specific marker: CD 203c. Homeopathy. 2006;95:3-8.