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At present, there is a lack of clinical evidence about the impact and long-term durability of the immune response induced by the third dose of mRNA vaccines. In this study, we followed up the B cell compartment behavior in a cohort of immunocompetent individuals three and six months after the third dose of vaccine. During this period, some subjects contracted the virus. In uninfected vaccinated subjects, we did not report any changes in serum spike-specific IgG levels, with a significant reduction in IgA. Instead, subjects recovered from natural infection showed a significant increase in both specific IgG and IgA. Moreover, we showed a time-related decrease in IgG neutralizing potential to all SARS-CoV-2 variants of concern (VOC) in uninfected compared to recovered subjects, who displayed an increased neutralizing ability, particularly against the omicron variant. Finally, we underlined the presence of a pool of SARS-CoV-2-specific B cells in both groups that are prone to respond to restimulation, as demonstrated by their ability to differentiate into plasma cells and to produce anti-SARS-CoV-2-specific immunoglobulins. These data lead us to assert the long-term effectiveness of the BNT162b2 vaccine in contrasting the severe form of the pathology and prevent COVID-19-associated hospitalization.

Solid organ transplant recipients (SOTRs) show higher rates of COVID-19 breakthrough infection than the general population, and nowadays, vaccination is the key preventative strategy. Nonetheless, SOTRs show lower vaccine efficacy for the prevention of severe COVID-19. Moreover, the emergence of new SARS-CoV-2 variants of concern has highlighted the need to improve vaccine-induced immune responses by the administration of repeated booster doses. In this study, we analyzed the humoral and cellular responses in a cohort of 25 SOTRs, including 15 never-infected SOTRs who received the fourth dose of the mRNA vaccine and 10 SOTRs who contracted SARS-CoV-2 infection after the third dose. We analyzed the serum IgG and IgA levels through CLIA or ELISA, respectively, and the Spike-specific T cells by ELISpot assay. We report a significant increase in anti-Spike IgG and no differences in IgA secretion in both groups of patients before and after the booster dose or the natural infection. Still, we show higher IgA levels in recovered SOTRs compared to the fourth dose recipients. Conversely, we show the maintenance of a positive Spike-specific T-cell response in SOTRs who received the fourth dose, which, instead, was significantly increased in SOTRs who contracted the infection. Our results suggest that the booster, either through the fourth dose or natural infection, in vulnerable poor responder SOTRs, improves both humoral and cellular-specific immune responses against SARS-CoV-2.

To prevent PGI (Protected Geographical Indication) cherry tomato of Pachino (Sicily, Italy) from frauds, an alternative method, which includes chemometric treatments, was proposed. The content of 32 inorganic elements (macro-micronutrients and lanthanides) present in 16 PGI and 24 not PGI cherry tomato samples cv Naomy, and in 16 PGI and 8 not PGI soil samples, was determined by Inductively Coupled Plasma - Mass Spectrometer (ICP-MS). To identify the elements able to differentiate PGI and not PGI cherry tomato samples, Principal Components Analysis (PCA) and Canonical discriminant analysis (CDA) were performed. The first two principal components (PC1-PC2) explain a total variance of 71,41% between PGI and not PGI group, whereas CDA showed Zn, Cd, Mn and Ca as inorganic markers able to correctly classify the 100% of samples. Furthermore, with a translocation factor (K), evaluated in soil/plant chain, the comparison of absorption trends for PGI and not PGI samples was realized., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Background: Several studies have indicated that anti-SARS-CoV-2 mRNA vaccinations are less effective in inducing robust immune responses among solid organ transplant recipients (SOTRs) compared with the immunocompetent. The third dose of vaccine in SOTRs showed promising results of immunogenicity, even though clinical studies have suggested that immunocompromised subjects are less likely to build a protective immune response against SARS-CoV-2 resulting in lower vaccine efficacy for the prevention of severe COVID-19. Methods: Serological IgG and IgA were analyzed through CLIA or ELISA, respectively, while Spike-specific T cells were detected by ELISpot assay after the second and third dose of vaccine in 43 SOTRs. Results: The third dose induced an improvement in antibody response against SARS-CoV-2. We also reported a strong correlation between specific humoral and cellular responses after the third dose, even though we did not see significant changes in the magnitude of the SARS-CoV-2-specific T cell response. SOTRs who contracted the SARS-CoV-2 infection after the third dose, despite eliciting a positive IgG response, failed to mount an anti-Spike-S1 IgA response, both after the third dose and after SARS-CoV-2 infection. Conclusions: We can conclude that serum IgA detection can be helpful, along with IgG detection, for the evaluation of vaccine efficacy, principally in fragile subjects at high risk of infection.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which causes coronavirus disease 2019 (COVID-19), is modifying human activity all over the world with significant health and economic burden. The advent of the SARS-CoV-2 pandemic prompted the scientific community to learn the virus dynamics concerning transmissibility, epidemiology, and usefulness of vaccines in fighting emerging health hazards. Pieces of evidence suggest that the first and second doses of mRNA vaccines induce a significant antibody response in vaccinated subjects or patients who recovered from SARS-CoV-2 infection, demonstrating the importance of the previously formed memory. The aim of this work has been to investigate the effects of BNT162b2 Pfizer-BioNTech mRNA-based vaccine booster dose in a cohort of 11 uninfected immunocompetent (ICs), evaluating the humoral and cellular responses, with more carefulness on memory B and T cells. Our findings underscore the potential benefit of the third dose of mRNA vaccine on the lifespan of memory B and T cells, suggesting that booster doses could increase protection against SARS-CoV-2 infection., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Busà, Sorrentino, Russelli, Amico, Miceli, Miele, Di Bella, Timoneri, Gallo, Zito, Di Carlo, Conaldi and Bulati.)

Endotoxin content is a critical factor that affects the safety of biological pharmaceutical products. International pharmacopoeias describe several reference methods to determine endotoxin levels in advanced therapy medicinal product (ATMP) preparations. Administration of ATMPs must be done as rapidly as possible to ensure complete viability and potency of the cellular product. To evaluate the endotoxin content in the shortest time possible, we chose to validate an alternative method based on the use of the Charles River Portable Testing System (PTS) and FDA-approved cartridges, compliant with the requirements of the European Pharmacopoeia and providing results in <20 min. Here, we describe a unique and complete validation approach for instrument, personnel, and analytical method for assessment of endotoxins in ATMP matrices. The PTS system provides high sensitivity and fast quantitative results and uses less raw material and accessories compared with compendial methods. It is also less time consuming and less prone to operator variability. Our validation approach is suitable for a validated laboratory with trained personnel capable of conducting the ATMP release tests, and with very low intra-laboratory variability, and meets the criteria required for an alternative approach to endotoxin detection for in-process and product-release testing of ATMPs., Competing Interests: The authors declare no competing interests., (© 2021 The Author(s).)

We constrained the origin and genetic environment of modern iron ooids (sand-sized grains with a core and external cortex of concentric laminae) providing new tools for the interpretation of their fossil counterparts as well as the analogous particles discovered on Mars. Here, we report an exceptional, unique finding of a still active deposit of submillimetric iron ooids, under formation at the seabed at a depth of 80 m over an area characterized by intense hydrothermal activity off Panarea, a volcanic island north of Sicily (Italy). An integrated analysis, carried out by X-ray Powder Diffraction, Environmental Scanning Electron Microscopy, X-ray Fluorescence and Raman spectroscopy reveals that Panarea ooids are deposited at the seafloor as concentric laminae of primary goethite around existing nuclei. The process is rapid, and driven by hydrothermal fluids as iron source. A sub-spherical, laminated structure resulted from constant agitation and by degassing of CO 2 -dominated fluids through seafloor sediments. Our investigations point the hydrothermal processes as responsible for the generation of the Panarea ooids, which are neither diagenetic nor reworked. The presence of ooids at the seawater-sediments interface, in fact, highlights how their development and growth is still ongoing. The proposed results show a new process responsible for ooids formation and gain a new insight into the genesis of iron ooids deposits that are distributed at global scale in both modern and past sediments.

The subject of this present work is a group of nine historical pictures shot in Palermo by the Sicilian photographer E. Interguglielmi in 1912. They are nine matte-collodion prints mounted on the original cardboard supports and all of them show foxing stains affecting the paper surface. In order to characterise the chemical composition of the supports and investigate foxing spots, non-destructive and micro-destructive analysis were carried out. X-rays fluorescence (XRF) analysis was used to characterise the elemental composition of all the mounting boards, allowing a comparison between the foxing spots and non-affected areas. Laser-Induced Breakdown Spectroscopy was used to investigate the presence of lower atomic number elements, not detectable by XRF, while SEM imaging allowed the investigation of surface appearance and nature of original paper samples from the cardboards.

This case series evaluated the healing of deep intrabony defects treated with a combination of enamel matrix derivative and autologous particulate bone harvested from the buccal and lingual/palatal cortical plate with a Piezosurgery device. A total of 15 defects with a predominantly one- or two-wall component were consecutively treated in 15 patients with advanced chronic periodontitis. In all selected sites, the three-wall component was ≤ 25% of the total defect depth. Clinical and radiographic parameters were recorded at baseline and 12 and 24 months postoperatively. All defects showed favorable clinical and radiographic outcomes at the 24-month follow-up. The probing depth reduction was 4.4 ± 1.6 mm, and more than 50% of the defects presented clinical attachment level gain of at least 5 mm. The bone fill was 3.1 ± 1.6 mm.

The geological, biological and geochemical features of a particular field of hydrothermal vents, discovered in the Panarea Volcanic Complex during a research survey carried out in 2015, are described for the first time. The site, located at 70-80 m depth off the South-western coast of the islet of Basiluzzo, was named Smoking Land for the presence of a large number of wide and high active chimneys and was characterized in terms of dissolved benthic fluxes, associated macrofauna and megafauna communities and preliminary mineralogy and geochemistry of chimney structures. On the whole field, a total of 39 chimneys, different in size and shape, were closely observed and described; 14 of them showed emission of low temperature hydrothermal fluids of marine origin characterized by acidified chemical conditions. The CTD and benthic chamber measurements highlighted that the Smoking Land is able to form a sea water bottom layer characterized by variable acidity and high DIC and trace elements concentrations; these characteristics weaken moving away from the chimney mouths. The SEM-EDS analysis of the collected solid samples revealed a chimney structure principally composed by amorphous and low crystalline Fe-oxyhydroxides of hydrothermal origins. The ROV explorations revealed a wide coverage of red algae (Peyssonnelia spp.) colonized by the green algae Flabiella petiolata and by suspension feeders, mainly sponges, but also bryozoans, and tubicolous polychaetes. Although novent-exclusive species were identified, the benthic communities found in association to the chimneys included more taxa than those observed in the surrounding no-vent rocky areas. These first findings evidence a submarine dynamic habitat where geological, chemical and biological processes are intimately connected, making the Smoking Land an important site in terms of marine heritage that should be safeguarded and protected.

The human EBV-transformed lymphoblastoid cell line (LCL), obtained by infecting peripheral blood monocular cells with Epstein-Barr Virus, has been extensively used for human genetic, pharmacogenomic, and immunologic studies. Recently, the role of exosomes has also been indicated as crucial in the crosstalk between EBV and the host microenvironment. Because the role that the LCL and LCL exosomal cargo might play in maintaining persistent infection, and since little is known regarding the non-coding RNAs of LCL, the aim of our work was the comprehensive characterization of this class of RNA, cellular and viral miRNAs, and cellular lncRNAs, in LCL compared with PBMC derived from the same donors. In this study, we have demonstrated, for the first time, that all the viral miRNAs expressed by LCL are also packaged in the exosomes, and we found that two miRNAs, ebv-miR-BART3 and ebv-miR-BHRF1-1, are more abundant in the exosomes, suggesting a microvescicular viral microRNA transfer. In addition, lncRNA profiling revealed that LCLs were enriched in lncRNA H19 and H19 antisense, and released these through exosomes, suggesting a leading role in the regulation of the tumor microenvironment., (Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.)

Myelin basic protein (MBP) is one of the main protein components of central nervous system that as shown in our work, under appropriate condition, forms spherulites. The structural and morphological features of these elements and the ability of trehalose to decrease or completely avoid their formation have been clarified with different but complementary techniques. The FTIR spectra provided compelling evidence for the presence of a small amount of elements organized in secondary structures such as helices and sheets. Polarized optical microscope observations show the presence of spherulites with an average size ranged from 41 to 61 μm, characterized by non-birifrangent core. The fluorescence data supply useful informations that are consistent with the presence of a molecular exclusion effect, following interaction of the disaccharide with protein that involves the environment around the single tryptophan residue. Moreover, study of protein conformational states by SDS-PAGE, let us to state that trehalose completely avoid autocatalytic cleavage properties of MBP up to 4 days of incubation at 37 °C and pH 7.4., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Tunicates comprising a wide variety of different species synthesize antimicrobial peptides as important effector molecules of the innate immune system. Recently, two putative gene families coding for antimicrobial peptides were identified in the expressed sequence tag database of the tunicate Ciona intestinalis. Two synthetic peptides representing the cationic core region of one member of each of the families displayed potent antibacterial and antifungal activities. Moreover, the natural peptides were demonstrated to be synthesized and stored in distinct hemocyte types. Here, we investigated the presence of these natural peptides, namely Ci-MAM-A and Ci-PAP-A, in the tunic of C. intestinalis considering that the ascidian tunic is a body surface barrier exposed to constant microbial assault. Furthermore, as the tunic may represent a major route of entry for pathogen invasion after its damage we monitored the location of these peptides upon a local inflammatory-like reaction induced by injection of foreign cells. Using immunocytochemistry and electron microscopy both peptides were localized to the tunic and were massively present in granulocytes of inflamed tissue. Conclusively, antimicrobial peptides may constitute a chemical barrier within the tunic of urochordates.

Since glucocorticoids have a role in maintaining the homeostatic status in fish, in the present paper mRNA expression (in situ hybridization) and tissue immunohistochemical localization of a glucocorticoid receptor (DlGR1) in several Dicentrarchus labrax organs are reported. Riboprobe and specific antibodies were prepared by using the DlGR1 that has been previously cloned and sequenced from peritoneal cavity leukocytes. Both mRNA and receptor were identified in head kidney, spleen, gills, intestine, heart and liver tissues. The functional roles of DlGR1 localization are discussed.

Flavonoids are natural substances with a lot of biological activities, including the antioxidant one. Their use in pharmaceutical field is, however, limited by their aqueous insolubility. As the formation of the inclusion complexes can improve their solubility in water, the flavonoids hesperetin, hesperidin, naringenin and naringin have been complexed with beta-cyclodextrin (beta-CD) by the coprecipitation method and studied in solution and in solid state by NMR, FT-IR, differential scanning calorimetry and X-ray techniques. The effects of complexation on the chemical shifts of the internal and external protons of beta-CD in the presence of each flavonoid were observed.

The extravasation of metastatic cells is regulated by molecular events involving the initial adhesion of tumor cells to the endothelium and subsequently the migration of cells in the host connective tissue. E-selectin on endothelial cells and sialyl Lewis X carbohydrate component on tumor cells are mainly involved in the adhesion of colon carcinoma cells to the endothelium of target organ. Interaction of T84 colon cancer cells to purified E-selectin in vitro caused an increase in the tyrosine phosphorylation of a number of proteins as well as the modulation of cellular properties correlated to the metastatic phenotype. Specifically, E-selectin-stimulated actin reorganization, increased collagenase secretion, and induced cell migration. Treatment of T84 cells with herbimycin A inhibited cell adhesion as well as selectin-induced increase of cell migration, and cytoskeleton assembly. Our data demonstrate that binding of cancer cells to E-selectin starts signal transduction pathways which may affect the tumor metastatic abilities., ((c) 2002 Elsevier Science (USA).)

Background: Serum IgD and IgE levels were measured in children with atopic asthma and in control Group in order to determine their relationship with clinical status., Methods: Samples of venous blood (of 5 cc) were drawn from 25 asthmatic children (Group A) and 25 healthy children (Group B) at the moment of first diagnosis (T0), after 6 months (T180) and after 18 months (T540). To measure IgD, an ELISA assay based on the sandwich principle was used., Results: At T0, IgD were significantly higher in Group A (182.7 5+/-88.18 IU/ml) in comparison with Group B (69.58+/-4.93 IU/ml, p<0.0001); IgD levels decreased in Group A at T540., Conclusions: In conclusion, a significant increase of IgD levels observed in children at first signs of asthma and the following normalization of these same levels after 18 months, may represent a non specific response or an attempt of the organism to block asthma, favouring therefore immunologic tolerance.

The formation of inclusion complexes between beta-cyclodextrin with the two beta-blockers, atenolol and celiprolol, have been studied in the aqueous environment and in the solid state by nuclear magnetic resonance (NMR) spectroscopy, X-ray, differential scanning calorimetry (DSC) and scanning electron microscopy (SEM) techniques. The magnitude of the chemical shifts of the interior and exterior beta-cyclodextrin protons in the presence of each beta-blocker indicated that these are included within the beta-cyclodextrin cavity. In aqueous solution they form 1:1 complexes. In the solid state the formation of the beta-cyclodextrin/atenolol (celiprolol) complexes is confirmed by X-ray, DSC and SEM, also employed to characterize pure substances and their physical mixtures.

1. Patch-clamp technique was used in primary cultures of cerebellar granule neurons to study the modulation of the cyclothiazide analogue (IDRA21) and of the diazoxide derivative (IDRA 5) on KA-evoked currents. 2. The dose-response of kainic acid (KA) reveals an EC50=90 microM and an Hill coefficient of 1.3. IDRA 21 and cyclothiazide potentiate KA-evoked current in a dose dependent way, being cyclothiazide more potent but less efficacious than IDRA 21. Conversely IDRA 5 acts as a negative modulator of KA evoked -current. 3. Application of IDRA 21 and cyclothiazide results in a current potentiation of 125+/-18% and 80+/-12% respectively, while IDRA 5 decreases KA-current (-21+/-5%). Coapplication of cyclothiazide and IDRA 21 produces a potentiation of 110+/-17%, suggesting a competition of the two drugs for the same site. 4. In the same experimental model we studied the ability of IDRA compounds of promoting toxicity through AMPA-receptor activation. Under basal conditions AMPA treatment (50 microM for 1 hour) results in a negligible excitotoxicity. 5. In contrast similar treatment with AMPA + IDRA 21 (1 mM) or + IDRA 5 (1 mM) or + cyclothiazide (100 microM) induces citotoxicity. The neurotoxic damage induced by IDRA 21 and cyclothiazide is blocked by GYKI 53655 (50 microM) and by NBQX (10 microM). Interestingly GYKI and NBQX are ineffective in reducing IDRA 5 toxicity.

Following an injuring factor such as the injection of erythrocytes into the tunic of Ciona intestinalis, an inflammatory-like response occurs and blood cells are massively involved in these mechanisms. Electron microscope observations illustrate the infiltration and migration of blood cells throughout the mantle epithelium into the tunic and show several prominent morphological changes., (Copyright 2000 Academic Press.)

The pseudo-first order rate constant for the hydrolysis of cinnoxicam as a function of temperature was obtained by variable-temperature kinetic experiments. The method used is on a generalization of non-isothermal analysis, and takes advantage of the capabilities of modern data collection and processing systems. A spectrophotometric method under non isothermal conditions was carried out. The results obtained are identical to those obtained under the same conditions by using traditional constant-temperature kinetic runs. This provides the possibility of reducing the amount of time spent and chemicals usually used in collecting kinetic data in mechanistic studies in solution by an order of magnitude.

A combination of thin-layer chromatography, high-performance liquid chromatography, UV spectroscopy and NMR spectrometry techniques for analysis of flavonoid content of Guiera senegalensis J.F.Gmel. (Combretaceae) leaves is described. Four components, catechin, myricitrin, rutin and quercetin were evidenced and determined by HPLC.

Electron microscopic studies on the encapsulation induced by erythrocyte injection into the tunic of the ascidian Ciona intestinalis were carried out. The observations reported in the present paper complete the description previously given of capsule architecture and contribute to the characterization of the cells involved in the inflammatory reaction. The inflamed area is surrounded by an ample and peculiar "three-layered coat" respectively composed of flattened and packed extratunical hemocytes, the monolayered epithelium, and a layer of intratunical electron-dense particles. The latter are also clustered, variously arranged, and distributed in the tunic ground substance. The epithelial cells appear to be undergoing an active secretory phase; in several regions they also show discontinuities and organule and surface changes. The infiltrating hemocytes, mainly globular and unilocular granulocytes, appear frequently to be degranulating and in relation with electron-dense particles, net-like fibrous materials, and fine membranes. The involvement of morula-shaped granulocytes is discussed, as well as possible relationships with the melanization process, and finally analogies in the structural organization with the inflammatory reactions induced in other invertebrates. A schematic drawing, based on all available observations of the capsule architecture, is presented in order to reconstruct the entire inflamed area and illustrate the relative fine features.

A cyclodextrin-modified micellar capillary electrophoretic method was developed for the chiral separation of 2-methyltaurine. The racemic mixture was separated after derivatization with dansyl chloride. The enantiomeric separation of the dansyl derivative was done using electrolyte solutions containing beta-cyclodextrin (beta-CD) or different ratios of beta- and gamma-CD. The mixture of beta- and gamma-CD optimally resolved the compound studied.

Previous studies on the ascidian Ciona intestinalis have shown that an encapsulation response is experimentally induced by inserting vertebrate erythrocytes into the tunic, which initiates a massive inflammatory cell infiltration to isolate the injured area. Several hemocytes contribute to capsule formation, destruction of the foreign cells, tunic regeneration, and wound healing. The fine features of some inflammatory cell types are described although the complete capsular structure is not yet reported. Accordingly, the present investigation further examines various aspects of this cellular reaction against erythrocytes and, for the first time, presents the involvement of extratunical circulating hemocytes and mantle epithelium in capsule formation.

Studies aided by transmission electron microscopy were made in order to evaluate the occurrence and efficiency of the human follicular fluid in the activation of the processes leading to the acrosome reaction; quantitative, qualitative and morphological data and relative evaluations are here presented. The effects of the human follicular fluid were compared with those determined by other types of treatment, semen untreated, Pellet-Swim-up, and Centrifugation on Discontinuous Percoll Gradient. The transmission electron microscope observations permitted to evaluate the percentage of sperm with an activation of the acrosome reaction in the different groups. The analysis of the data showed a statistically relevant difference (P < 0.001) among the first group (Control group) and the SU group, the MP group and the hFF group, regarding the sperm with AR activation. The sperm treatment with 50% diluted hFF represents an important option to the preparation protocols of the semen samples useful for ART.

The influence of 11 newly synthesized 2,6-bis(1,1-dimethylethyl)phenol derivatives substituted in the 4 position as measured on the carrageenan paw edema assay in Sprague-Dawley rats, was studied using indomethacin as a reference drug. Furthermore we studied the possible interference of few of these compounds on the calcium binding sites by using the specific ligand [3H]-PN 200-110 in "in vitro" experiments. As far as regard the antiinflammatory activity only the compounds 2b, 2j and 2k, dosed at 20 mg/Kg/os, exerted an inhibitory effect on paw edema which was practically equal, after 6 h, to that of indomethacin (approximately 30%) dosed at 2.5 mg/Kg. The compound 2k, however, showed, in comparison with indomethacin and the other new tested compounds, a longer lasting effect, reaching, after 8 h, a 56.7% inhibition of the edema. Finally the above mentioned compounds, when tested alone or in combination with nitrendipine, did not exert any displacing activity on [3H]-PN 200-110 binding to synaptosomal membranes. It is noteworthy however that compound 2e, which incidentally was inactive as antiinflammatory agent, showed a negative allosteric modulatory activity on the ability of nitrendipine to displace [3H]-PN 200-110 binding.

In bovine retinal pigment epithelium membranes we have found three hydrolases which were active against trans-retinyl palmitate. This was possible by assaying different subcellular fractions as a function of pH in the range 3-9. Detection of these activities has been favored by the use in the enzyme assay of Triton X-100, which has an activating effect up to a concentration of 0.03% at a detergent-protein ratio of about 1.5-3.0. Apparent kinetic parameters for the retinyl ester hydrolases have been determined after a study of the optimization of assay conditions. Vmax values for hydrolases acting at pH 4.5, 6.0, and 7.0 were, respectively, 156, 55, and 70 nmol/h/mg. To identify the subcellular site for these hydrolytic activities, assays of marker enzymes from various organelles in each subcellular preparation were carried out, demonstrating the lysosomal origin of the pH 4.5 retinyl ester hydrolase and the microsomal origin of the pH 6.0 retinyl ester hydrolase and suggesting that the pH 7.0 retinyl ester hydrolase originates from the Golgi complex.

The scavenging activity of some isosteric antiinflammatory heteoarylalkanoic acids with respect to oxygen-free radicals has been studied. All the compounds were found to inhibit the depolymerization of hyaluronic acid due to enzymatically or chemically generated hydroxyl radicals. The reduced consumption of oxygen by xanthine oxidase indicates a cotemporaneous phenomenon of enzymatic inhibition.

Erythrocytes injected into the tunic of Ciona intestinalis induced an inflammatory-like reaction which can cause lysis of the neighboring tissue. Electron microscopy observations of the granulocytes of the injured tunic show lysosomic figures and degranulations features suggesting granulocyte involvement in the destructive process.

The antitumor activity of the methyl-6-chloro-4H-1,2,4-benzothiadiazin-3-yl-S-carbamothioate+ ++-S,S,-dioxide (1) and the methyl-4H-1,2,4-benzothiadiazin-3-yl-carbamodithioate-S,S-di oxide (2), has been evaluated. Compound 2 passed activity criteria for leukemia P388 and was also moderately active against transplanted mouse tumor L1210 leukemia and M5076 sarcoma.

The concentration and the distribution of mercury in six different brain areas of developing rats (21 and 60 days of age) exposed to methylmercury (MMC) during prenatal life were determined by using pressure decomposition of the tissues and cold-vapor atomic absorption spectrometry. The distribution of mercury in brain samples showed that the metal distributes to all brain areas, but with different levels. The amount of mercury in the brain areas was about 10-100 times higher, depending on the tested area, in MMC exposed rats than in control, at day 21 of age, while it was practically equal to controls at 60 days of age. These data seem to be of importance in order to correlate the presence of mercury in the brain and its distribution to brain areas after a single exposure to MMC with transient or permanent changes in specific neurotransmitter system and altered behaviors.

The rates of reaction toward cysteine of a series of carbamimidothioic acid phenylmethylesters halides were investigated in phosphate buffer pH 7.4 at 37 degrees C under pseudo-first-order conditions. The reaction involves nucleophilic attack of cysteine on the C-S bond of isothiouronium mojety and corresponding benzenemethanethiols were detected as product of the reaction. Apparent first-order rate constants were determined.

In this study we tested the potential taurine-antagonistic properties of three rigid analogs of taurine, 3-amino- (1), 3-hydrazino- (2) and 3-aminomethyl-1,2,4-benzothiadiazine-1,1-dioxide (3), which were prepared in our laboratory, using TAG (6-aminomethyl-3-methyl-1,2,4-benzothiadiazine-1,1- dioxide) (4), the only antagonist of taurine so far available, as reference compound in "in vivo" and "in vitro" experiments. Some physicochemical properties of (1), (2) and (3) were studied and the synthesis of TAG (4) was improved with a new preparative method. A dose-effect study performed by injecting intracerebroventricularly (1), (2) and (3) showed that these compounds have none of exciting effects exerted by the high doses of TAG (4). (1) and (3) as well as TAG (4), were found to antagonize the controlateral turning induced by the intracerebro injection of taurine and to potentiate the sedative effect of diazepam. We failed to find specific binding for taurine in different brain synaptic membrane preparations using 3H-taurine as radioligand and taurine, (1) and (3) as binding displacer. (1), (3) and TAG (4) however were found to antagonize the inhibitory effect of taurine on 3H-diazepam binding. These results seem to indicative that at least (1) and (3), which were more extensively studied than (2) because of their better solubility, are taurine antagonists with an apparent better selectivity than TAG (4).

A series of carbamimidothioic acid phenylmethyl ester salts and their N,N'-tetramethyl derivatives were synthesized, many of which exert an interesting inhibiting action on Gram-positive and Gram-negative bacteria and also on fungi. This activity is noteworthy in view of the large number of strains antagonized; the (3,4,5-trichlorophenyl) methyl ester chloride of carbamimidothioic acid appeared to be the best term of the series. The possible influence on this activity of decomposition kinetics to the corresponding mercapto-derivatives was investigated and compared with that of a series of N,N'-tetramethyl derivatives of previously studied carbamimidothioic acid phenyl ester salts.

A series of N-arylglycine derivatives and their thioisosters (1-10) was synthesized and evaluated as in vitro aldose reductase inhibitors. Among these, (6-chloro-1,2,4-benzothiadiazine-1,1-dioxide-3-yl)-2-propanoic acid (5) shows an inhibitory activity (IC50:5.1 microM) comparable to Alrestatin.

A series of (1,2,4-benzothiadiazin-1,1-dioxide-3-yl)acetic acids was prepared so that antiinflammatory activity could be tested. Some of the compounds synthetized showed marked activity against plantar edema induced by carrageenin and/or dextran. It was found that in this series antiinflammatory activity is related to the nature of the substituents on the benzene ring as well as their number, dihalogen substitution being the most favourable modification and dichlorosubstitution in particular giving the compounds most active against both types of edema.

A series of 3,4-dihydro derivatives of 6-chloro-, 7-chloro-, 5,7-dichloro-, 6,7-dichloro-, 5,7-dibromo-, 5-nitro-7-chloro-, 7-nitro-, 7-amino-1,2,4-benzothiadiazine-1,1-dioxide, various substituted on the heterocyclic carbon, was prepared and tested for cardiovascular activity. It was found that in this series of compounds cardiac activity predominates and is exclusively of the depressant type. Bradycardial activity seems to be affected both by the nature of the substituent on the heterocyclic carbon atom and by the substituents on the benzene ring. An alkyl chain of three carbon atoms, normal or alpha-ethylsubstituted, on C3 and the presence of a chlorine atom in positions 6 or 7 seem to be the most significant structural features for this activity. Some of the substances tested showed a pressor effect (hypotension and/or increase in differential pressure).

A series of quaternary ammonium salts of 3-alkylaminoderivatives of 1,2,4-benzothiadiazine-1,1-dioxide were prepared and tested for cardiovascular activity. Almost all the substances tested proved to have some measure of depressant activity on arterial pressure and the most interesting compounds in this respect were (XII), (XIII), (XV) and (XIX) for which hypotensive values between 60 and 70% with respect to the base level were recorded. Compound (XV) also caused increase in the differential pressure and decrease of cardiac frequency. The later however was increased by some other compounds [(XII), (XVIII)]. The structure-activity relationships are discussed taking into account also the activity of the parent tertiary amines.

A series of 3-(4H-1,2,4-benzothiadiazine-1,1-dioxide-3-yl-thio)-propanoic acids was synthesized in order to study their possible antiinflammatory in comparison with the corresponding lower homologues and isosters previously studied. Most of the compounds proved effective against carrageenan-induced plantar oedema. The possible influence of some chemico-physical parameters on the occurrence of this biological activity was investigated.

A series of S-Aryl(tetramethyl)isothiouronium salts were prepared and evaluated in vitro for antimicrobial activity. Some compounds revealed interesting inhibiting action on Gram-positive bacteria which is noteworthy in view of the large number of strains antagonized and of the low MIC values. The possible influence of decomposition kinetics to the corresponding mercaptoderivatives was intestigated and compared with those of salts previously studied.

A series of 2-nitro and 2-aminobenzenesulfon-N-acylamides variously substituted on the benzene ring were prepared and tested for anticonvulsant activity (electro shock, pentamethylenetetrazole). Some of the compounds tested gave some measure of protection against convulsions induced by electric shock. The importance of the trifluoromethyl substituent in this effect was demonstrated.

A series of 3-(4H-1,2,4-benzothiadiazine-1,1-dioxide-3-ylamino)-propanoic acids was synthesized in order to study their possible antiinflammatory activity and to compare it with the corresponding homologous or isoster series previously studied. Of particular interest is parent compound 1, which is unsubstituted in the benzenoid moiety and exerts a very marked inhibitory effect on carrageenan-induced plantar oedema greater than that observed for any of the terms of the other series hitherto studied. The possible influence of some chemico-physical parameters on the occurrence of this antiinflammatory activity is investigated.

A series of carboxy- and carbomethoxyalkyl derivatives of 3-amino-1,2,4-benzothiaidazin-1,1-dioxide either substituted or unsubstituted in the benzene ring [compounds (I leads to XVIII)] was prepared and tested for cardiovascular activity. It was found that the introduction of a carboxy-or carbalkoxy-group in the omega position of 3-alkylamino derivatives of 1,2,4-benzothiadiazine-1,1-dioxide usually causes marked decrease or abolition of the cardiovascular activity shown by the parent compounds. The negative effect on the pressor trace (hypotension and increase in differential pressure) is more frequent and significant than that on bradycardial activity.