A Closer Look at Histamine in Drosophila.

The present work intends to provide a closer look at histamine in Drosophila. This choice is motivated firstly because Drosophila has proven over the years to be a very simple, but powerful, model organism abundantly assisting scientists in explaining not only normal functions, but also derangements that occur in higher organisms, not excluding humans. Secondly, because histamine has been demonstrated to be a pleiotropic master molecule in pharmacology and immunology, with increasingly recognized roles also in the nervous system. Indeed, it interacts with various neurotransmitters and controls functions such as learning, memory, circadian rhythm, satiety, energy balance, nociception, and motor circuits, not excluding several pathological conditions. In view of this, our review is focused on the knowledge that the use of Drosophila has added to the already vast histaminergic field. In particular, we have described histamine's actions on photoreceptors sustaining the visual system and synchronizing circadian rhythms, but also on temperature preference, courtship behavior, and mechanosensory transmission. In addition, we have highlighted the pathophysiological consequences of mutations on genes involved in histamine metabolism and signaling. By promoting critical discussion and further research, our aim is to emphasize and renew the importance of histaminergic research in biomedicine through the exploitation of Drosophila, hopefully extending the scientific debate to the academic, industry, and general public audiences.

The role of Carcinine treatment on glico-lipidic imbalance of patients with altered blood glucose pattern.

Objective: Several studies support the active role of Carcinine, an L-carnosine metabolite, in insulin resistance and dyslipidaemia, to modulate the insulinemic/glycaemic profile and fat metabolism. Materials and methods: 100 (50 women and 50 men) volunteers, aged between 40 and 85 years with a body mass index (BMI) between 25 and 34,9 kg/m2, spontaneously addressed to our "Second Opinion Medical Consulting Network" (Modena, Italy) between 2019 and 2020, were included in this anecdotal, observational, retrospective trial. The aim of the study was to find an adequate possibly natural treatment for unbalanced insulin resistance pattern notwithstanding ongoing administration of statins, and hypoglycaemic chemical agents in healthy overweight/obese subjected. All the patients were divided in two groups: 1) the first group included 50 patients that were admi-nistered with a specific galenic nutraceutical product containing 20 mg of carcinine, and 2) the second group included 50 patients, which were administered with lithothamnion calcareum alga (190 mg) and three-time day for two months. The waist circumference, glycaemia, homeostasis model asses-sment (HOMA-IR), glycated haemoglobin, total cholesterol values were detected at time 0, and time 1 (after treatment). At the same time, the pre versus post treatment, Advanced Glycation End products (AGEs), that play an important role in the development of diabetic va-scular complications, were instrumentally measured at time 1 and 2. Results: After 60 mg/day of Carcinina treatment, glycaemia (p=0,001), glycated haemoglobin (p<0,001), total cholesterol (p<0,003), serum insulin (p<0.05) were significantly reduced, respect to placebo period. Abdominal circumference (p<0.2), HOMA index (p<0.03) progressively were reduced as well. No cardiovascular risk and untoward effects were observed at the prescribed dosages. The AGE reader test showed a statistically meaningful reduced risk due to reduced amount. Conclusions: Carcinine, at the daily dose of 60 mg/day, was able to modify, safely, the AGEs that induced cardiovascular risk, the waist circumference, and some glycolipid-metabolic parameters in over-weight/obese patients with altered blood glucose pattern, improving significantly the impending metabolic syndrome.

Whole-cell biotransformation for large scale production of carcinine in Escherichia coli.

Carcinine is a natural imidazole-containing peptide derivative. It is widely used in the cosmetics industry as anti-aging supplement with antioxidant, anti-glycation and glycation reversal functions, and it also has a notable pharmacological effect as anti-tumor drug and in protection against retinopathy. However, a technological method for synthesis and production of carcinine has not been established. In this study, a whole-cell transformation system converting ß-alanine and histamine to carcinine by the enzymes Ebony and phosphopantetheine transferase (Sfp) has been developed. The results revealed that the catalytic efficiency of the strain containing the fusion protein of Ebony and Sfp (Sfp-glycine-serine-glycine-Ebony, SGE) in Escherichia coli W3110 (WSGE strain) is significantly higher (7.45 mM) than the combinatorial strain of pET28a-ebony and pACYCDuet-sfp in E. coli BL21(DE3) (BSE strain) (2.17 mM). Under the optimal reaction conditions (25 â„ƒ, pH 7.0, 12.5 g/L wet cells, 20 mM ß-alanine and 40 mM histamine), the carcinine can be quickly synthesized within 24 h up to a concentration of 22.63 mM. To achieve a continuous and efficient conversion of the precursors, a batch-feeding catalysis was designed. With this system, ß-alanine (40 mM) and histamine (40 mM) could be completely transformed to carcinine (40.34 mM) in 36 h with a productivity of 0.204 g/L h reaching a titer of 7.34 g/L. Hence, the batch-feeding whole-cell biocatalysis is a promising technology for the high yield production of carcinine which can promote the industrial production of carcinine.

Location and functions of Inebriated in the Drosophila eye.

Histamine (HA) is a neurotransmitter in arthropod photoreceptors. It is recycled via conjugation to ß-alanine to form ß-alanylhistamine (carcinine). Conjugation occurs in epithelial glia that surround photoreceptor terminals in the first optic neuropil, and carcinine (CA) is then transported back to photoreceptors and cleaved to liberate HA and ß-alanine. The gene Inebriated (Ine) encodes an Na+/Cl--dependent SLC6 family transporter translated as two protein isoforms, long (P1) and short (P2). Photoreceptors specifically express Ine-P2 whereas Ine-P1 is expressed in non-neuronal cells. Both ine1 and ine3 have significantly reduced head HA contents compared with wild type, and a smaller increase in head HA after drinking 1% CA. Similarly, uptake of 0.1% CA was reduced in ine1 and ine3 mutant synaptosomes, but increased by 90% and 84% respectively for fractions incubated in 0.05% ß-Ala, compared with wild type. Screening potential substrates in Ine expressing Xenopus oocytes revealed very little response to carcinine and ß-Ala but increased conductance with glycine. Both ine1 and ine3 mutant responses in light-dark phototaxis did not differ from wild-type. Collectively our results suggest that Inebriated functions in an adjunct role as a transporter to the previously reported carcinine transporter CarT.

Quantification of Histamine and Carcinine in Drosophila melanogaster Tissues.

Histamine is a neurotransmitter crucial to the visual processing of Drosophila melanogaster. It is inactivated by metabolism to carcinine, a ß-alanyl derivative, and the same enzyme that controls that process also converts dopamine to N-ß-alanyl-dopamine. Direct detection of histamine and carcinine has not been reported in single Drosophila brains. Here, we quantify histamine, carcinine, dopamine, and N-ß-alanyl-dopamine in Drosophila tissues by capillary electrophoresis coupled to fast-scan cyclic voltammetry (CE-FSCV). Limits of detection were low, 4 ± 1 pg for histamine, 10 ± 4 pg for carcinine, 2.8 ± 0.3 pg for dopamine, and 9 ± 3 pg for N-ß-alanyl-dopamine. Tissue content was compared in the brain, eyes, and cuticle from wild-type (Canton S) and mutant (tan(3) and ebony(1)) strains. In tan(3) mutants, the enzyme that produces histamine from carcinine is nonfunctional, whereas in ebony(1) mutants, the enzyme that produces carcinine from histamine is nonfunctional. In all fly strains, the neurotransmitter content was highest in the eyes and there were no strain differences for tissue content in the cuticle. The main finding was that carcinine levels changed significantly in the mutant flies, whereas histamine levels did not. In particular, tan(3) flies had significantly higher carcinine levels in the eyes and brain than Canton S or ebony(1) flies. N-ß-Alanyl-dopamine was detected in tan(3) mutants but not in other strains. These results show the utility of CE-FSCV for sensitive detection of histamine and carcinine, which allows a better understanding of their content and metabolism in different types of tissues to be obtained.

Patenting strategies, the authentication US fiscal methodology, discovery and development of imidazole-containing peptide compounds with free-radical scavenging and transglycating properties acting as targeted drug regulators and homeostatic agents with diverse therapeutic activities for pharmacy of diabetes and metabolic diseases.

INTRODUCTION: Diabetes mellitus is the seventh-leading cause of death in the US and diabetic complications are interlaced with specific diverse microvascular and macrovascular pathologies resulting from hyperglycemia. The society should expand prowess and patenting of biotechnology to cure disease and complications. AREAS COVERED: The work summarizes biological activities of patented carnosine mimetics resistant in formulations to enzymatic hydrolysis with human carnosinases that are acting as a universal form of antioxidant, deglycating and transglycating agents that inhibit sugar-mediated protein crosslinking, chelate or inactivate a number of transition metal ions (including ferrous and copper ions), possess lipid peroxidase type of activity and protection of antioxidant enzymes from inactivation. L-Carnosine released systemically from N-acetylcarnosine lubricant eye drops or from skeletal muscle during exercise is transported into hypothalamic tuberomammillary nucleus-histamine neurons and hydrolyzed. The resulting L-histidine is subsequently converted into histamine acting as metabolic fuel feeding for the hypothalamic histaminergic system. This mechanism is responsible for the effects of L-carnosine on autonomic neurotransmission and physiological function of pancreas, stimulating in vivo regeneration of insulin-producing ß cells. EXPERT OPINION: Therapeutic benefits for imidazole-containing antioxidants (nutraceutical non-hydrolyzed carnosine, carcinine, D-carnosine, ophthalmic prodrug N-acetylcarnosine, leucyl-histidylhydrazide and patented formulations thereof) are an essential part of diabetes treatment.

The detox strategy in smoking comprising nutraceutical formulas of non-hydrolyzed carnosine or carcinine used to protect human health.

The increased oxidative stress in patients with smoking-associated disease, such as chronic obstructive pulmonary disease, is the result of an increased burden of inhaled oxidants as well as increased amounts of reactive oxygen species generated by various inflammatory, immune and epithelial cells of the airways. Nicotine sustains tobacco addiction, a major cause of disability and premature death. In addition to the neurochemical effects of nicotine, behavioural factors also affect the severity of nicotine withdrawal symptoms. For some people, the feel, smell and sight of a cigarette and the ritual of obtaining, handling, lighting and smoking a cigarette are all associated with the pleasurable effects of smoking. For individuals who are motivated to quit smoking, a combination of pharmacotherapy and behavioural therapy has been shown to be most effective in controlling the symptoms of nicotine withdrawal. In the previous studies, we proposed the viability and versatility of the imidazole-containing dipeptide-based compounds in the nutritional compositions as the telomere protection targeted therapeutic system for smokers in combination with in vitro cellular culture techniques being an investigative tool to study telomere attrition in cells induced by cigarette smoke (CS) and smoke constituents. Our working therapeutic concept is that imidazole-containing dipeptide-based compounds (non-hydrolyzed carnosine and carcinine) can modulate the telomerase activity in the normal cells and can provide the redox regulation of the cellular function under the terms of environmental and oxidative stress and in this way protect the length and the structure of telomeres from attrition. The detoxifying system of non-hydrolyzed carnosine or carcinine can be applied in the therapeutic nutrition formulations or installed in the cigarette filter. Patented specific oral formulations of non-hydrolyzed carnosine and carcinine provide a powerful manipulation tool for targeted therapeutic inhibition of cumulative oxidative stress and inflammation and protection from telomere attrition associated with smoking. It is demonstrated in this work that both non-hydrolyzed carnosine and carcinine are characterized by greater bioavailability than pure l-carnosine subjected to enzymatic hydrolysis with carnosinase, and perform the detoxification of the α,ß-unsaturated carbonyl compounds present in tobacco smoke. We argue that while an array of factors has shaped the history of the 'safer' cigarette, it is the current understanding of the industry's past deceptions and continuing avoidance of the moral implications of the sale of products that cause the enormous suffering and death of millions that makes reconsideration of 'safer' cigarettes challenging. In contrast to this, the data presented in the article show that recommended oral forms of non-hydrolyzed carnosine and carcinine protect against CS-induced disease and inflammation, and synergistic agents with the actions of imidazole-containing dipeptide compounds in developed formulations may have therapeutic utility in inflammatory lung diseases where CS plays a role.