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3-bromine carbazole (3-BCZ) represents a group of emerging aromatic disinfection byproducts (DBP) detected in drinking water; however, limited information is available regarding its potential cardiotoxicity. To assess its impacts, zebrafish embryos were exposed to 0, 0.06, 0.14, 0.29, 0.58, 1.44 or 2.88 mg/L of 3-BCZ for 120 h post fertilization (hpf). Our results revealed that ≥1.44 mg/L 3-BCZ exposure induced a higher incidence of heart malformation and an elevated pericardial area in zebrafish larvae; it also decreased the number of cardiac muscle cells and thins the walls of the ventricle and atrium while increasing cardiac output and impeding cardiac looping. Furthermore, 3-BCZ exposure also exhibited significant effects on the transcriptional levels of genes related to both cardiac development (nkx2.5, vmhc, gata4, tbx5, tbx2b, bmp4, bmp10, and bmp2b) and cardiac function (cacna1ab, cacna1da, atp2a1l, atp1b2b, atp1a3b, and tnnc1a). Notably, N-acetyl-L-cysteine, a reactive oxygen species scavenger, may alleviate the failure of cardiac looping induced by 3-BCZ but not the associated cardiac dysfunction or malformation; conversely, the aryl hydrocarbon receptor agonist CH131229 can completely eliminate the cardiotoxicity caused by 3-BCZ. This study provides new evidence for potential risks associated with ingesting 3-BCZ as well as revealing underlying mechanisms responsible for its cardiotoxic effects on zebrafish embryos., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Conventional water treatment practices utilizing chemical disinfection, especially chlorination, are considered generally effective in producing microbiologically safe drinking water. However, protozoan pathogens such as oocysts of Cryptosporidium parvum are very resistant to chlorine, which has led to consideration of alternative disinfectants for their control. Free bromine, HOBr, has not been evaluated extensively as an alternative halogen disinfectant for inactivation of Cryptosporidium parvum in drinking water or reclaimed water for non-potable uses. Bromine is a versatile disinfectant consisting of different chemical forms with persistent microbicidal efficacy under varied water quality conditions and is effective against a range of waterborne microbes of health concern. The objectives of this study are to (1) compare the efficacy of free bromine to free chlorine at similar concentrations (as milligrams per liter) for disinfection of Cryptosporidium parvum oocysts, Bacillus atrophaeus spores, and MS2 coliphage in a model buffered water and (2) evaluate the kinetics of inactivation of these microorganisms using appropriate disinfection models. Overall, at a target concentration of ∼5 mg/L, bromine averaged 0.6 log (73.8%) reductions of C. parvum oocyst infectivity after 300 min (CT: 1166 min·mg/L) and produced up to a 0.8 log reduction disinfectant activity. An ∼5.0 mg/L chlorine dose increased oocyst infectivity by only 0.4 log (64%) after 300 min (CT: 895 min·mg/L). Bacillus atrophaeus spores and MS2 coliphage treated with bromine and chlorine were reduced by 4 log 10 (99.99%) for both disinfectants over the duration of the experiments.

We present the synthesis and characterization of organic Salphen compounds containing bromine substituents at the para/ortho-para positions, in their symmetric and non-symmetric versions, and describe the X-ray structure and full characterization for the new unsymmetrical varieties. We report for the first time antiproliferative activity in metal-free brominated Salphen compounds, by evaluations in four human cancer cell lines, cervix (HeLa), prostate (PC-3), lung (A549) and colon (LS 180) and one non-cancerous counterpart (ARPE-19). We assessed in vitro cell viability against controls using the MTT assay ((3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide)) and determined the concentration required for 50 % growth inhibition (IC 50 ), together with their selectivity vs. non-cancerous cells. We found promising results against prostate (9.6 μM) and colon (13.5 μM) adenocarcinoma cells. We also found a tradeoff between selectivity (up to 3-fold vs. ARPE-19) and inhibition, depending upon the symmetry and bromine-substitution of the molecules, showing up to 20-fold higher selectivity vs. doxorubicin controls., (© 2023 The Authors. Chemistry & Biodiversity published by Wiley-VHCA AG, Zurich, Switzerland.)

2,2-bis(6-bromo-1H-indol-3-yl) ethanamine, a marine bisindole alkaloid, showed anticancer property in several tumor cell lines thanks to the presence of a 3,3'-diindolylmethane scaffold. Here, the modifications in its chemical structure into alkaloid-like derivatives, have been evaluated, to investigate changes in its biological activities. Three derivatives have been considered and their potential apoptotic action has been evaluated through morpho-functional analyses in a human cancer cell line. Apoptosis appears strongly decreased in the derivatives without the bromine atoms (1) and in those where the bromine atoms have been substituted with fluorine atoms (2). On the contrary, the methylation of indole NH (3) does not alter the alkaloid apoptotic activity that occurs through mitochondria involvement supported by cardiolipin peroxidation and dysfunctional mitochondria presence. This manuscript highlights the alkaloid derivative cytotoxic effect, which is strictly correlated to the presence of N-methylated bisindole alkaloid and bromine atoms, conditions which assure to maintain the pro-apoptotic activity. Since molecular therapies, by targeting mitochondria pathways, have shown positive outcomes against several cancer cells, the alkaloid with bisindole methylated scaffold and the two bromine atoms can be considered a promising candidate to develop new derivatives with strong anticancer property. RESEARCH HIGHLIGHTS: 2,2-bis(6-bromo-1H-indol-3-yl) ethanamine is an alkaloid known for its anticancer properties. Morpho-functional analyses evaluated cytotoxicity of its synthetic derivatives in tumor cells. Anticancer properties depend on the presence of bisindole scaffold and the two bromine units., (© 2022 Wiley Periodicals LLC.)

Inflammation is the most common cause of most acute and chronic debilitating diseases. Towards unveiling novel therapeutic options for patients with such complications, N‑bromotaurine (TauNHBr) has emerged as a potential anti‑inflammatory agent; however, its therapeutic efficacy is hindered due to its relatively poor stability. To address this challenge, the present study focused on examining the effects of a stable active bromine compound, named bromamine T (BAT). The present study examined the protective properties of BAT against lipopolysaccharide (LPS)‑mediated inflammation in vitro , by using LPS‑stimulated murine J774.A1 macrophages (Mφs), as well as in vivo , by using a murine LPS‑mediated air‑pouch model. Additionally, its efficacy was compared with that of taurine, a known potent anti‑inflammatory molecule. In LPS‑stimulated J774A.1 Mφs, BAT and taurine were very effective in reducing the secretion of pro‑inflammatory mediators. The in vitro experiments indicated that LPS‑mediated inflammation was attenuated due to the protective properties of BAT and of taurine, probably through the inhibition of phosphorylated p65 NF‑κB subunit (Ser 536) nuclear translocation. The in vivo experiments also revealed that BAT and taurine inhibited LPS‑mediated inflammation by reducing total cell/polymorphonuclear cell (PMN) infiltration in the air‑pouch and by decreasing pouch wall thickness. The analysis of exudates obtained from pouches highlighted that the inhibitory effects of BAT and taurine on the secretion of pro‑inflammatory cytokines were similar to those observed in vitro . Notably, the effect of BAT at the highest concentration tested was superior to that of taurine at the highest concentration. Taken together, the findings of the present study indicate that BAT prevents the LPS‑induced inflammatory response both in vitro and in vivo .

Exposure to excessive or uncontrolled stress is a major factor associated with various diseases including posttraumatic stress disorder (PTSD). The consequences of exposure to trauma are affected not only by aspects of the event itself, but also by the frequency and severity of trauma reminders. It was suggested that in PTSD, hippocampal-dependent memory is compromised while amygdala-dependent memory is strengthened. Several lines of evidence support the role of the endocannabinoid (eCB) system as a modulator of the stress response. In this study we aimed to examine cannabinoids modulation of the long-term effects (i.e., 1 month) of exposure to a traumatic event on memory and plasticity in the hippocampus and amygdala. Following exposure to the shock and reminders model of PTSD in an inhibitory avoidance light-dark apparatus rats demonstrated: (i) enhanced fear retrieval and impaired inhibitory extinction (Ext), (ii) no long-term potentiation (LTP) in the CA1, (iii) impaired hippocampal-dependent short-term memory in the object location task, (iv) enhanced LTP in the amygdala, and (v) enhanced amygdala-dependent conditioned taste aversion memory. The cannabinoid CB1/2 receptor agonist WIN55-212,2 (0.5mg/kg, i.p.) and the fatty acid amide hydrolase (FAAH) inhibitor URB597 (0.3mg/kg, i.p.), administered 2 hr after shock exposure prevented these opposing effects on hippocampal- and amygdala-dependent processes. Moreover, the effects of WIN55-212,2 and URB597 on Ext and acoustic startle were prevented by co-administration of a low dose of the CB1 receptor antagonist AM251 (0.5mg/kg, i.p.), suggesting that the preventing effects of both drugs are mediated by CB1 receptors. Exposure to shock and reminders increased CB1 receptor levels in the CA1 and basolateral amygdala 1 month after shock exposure and this increase was also prevented by administering WIN55-212,2 or URB597. Taken together, these findings suggest the involvement of the eCB system, and specifically CB1 receptors, in the opposite effects of severe stress on memory and plasticity in the hippocampus and amygdala., (© 2017 Wiley Periodicals, Inc.)

Antidepressant activity of N-methyl-D-aspartate (NMDA) receptor antagonists and negative allosteric modulators (NAMs) has led to increased investigation of their behavioral pharmacology. NMDA antagonists, such as ketamine, impair cognition in multiple species and in multiple cognitive domains. However, studies with NR2B subtype-selective NAMs have reported mixed results in rodents including increased impulsivity, no effect on cognition, impairment or even improvement of some cognitive tasks. To date, the effects of NR2B-selective NAMs on cognitive tests have not been reported in nonhuman primates. The current study evaluated two selective NR2B NAMs, CP101,606 and BMT-108908, along with the nonselective NMDA antagonists, ketamine and AZD6765, in the nonhuman primate Cambridge Neuropsychological Test Automated Battery (CANTAB) list-based delayed match to sample (list-DMS) task. Ketamine and the two NMDA NR2B NAMs produced selective impairments in memory in the list-DMS task. AZD6765 impaired performance in a non-specific manner. In a separate cohort, CP101,606 impaired performance of the nonhuman primate CANTAB visuo-spatial Paired Associates Learning (vsPAL) task with a selective impairment at more difficult conditions. The results of these studies clearly show that systemic administration of a selective NR2B NAM can cause transient cognitive impairment in multiple cognitive domains.

Point-of-use (POU) gravity-driven household water purifiers have been proven to be a simple, low-cost and effective intervention for reducing the impact of waterborne diseases in developing countries. The goal of this study was to compare commonly used water disinfectants for their feasibility of adoption in low-cost POU water purifiers. The potency of each candidate disinfectant was evaluated by conducting a batch disinfection study for estimating the concentration of disinfectant needed to inactivate a given concentration of the bacterial strain Escherichia coli ATCC 11229. Based on the concentration of disinfectant required, the size, weight and cost of a model purifier employing that disinfectant were estimated. Model purifiers based on different disinfectants were compared and disinfectants which resulted in the most safe, compact and inexpensive purifiers were identified. Purifiers based on bromine, tincture iodine, calcium hypochlorite and sodium dichloroisocyanurate were found to be most efficient, cost effective and compact with replacement parts costing US$3.60-6.00 for every 3,000 L of water purified and are thus expected to present the most attractive value proposition to end users.

Objectives: The activity of oxidants, such as halogens and active halogen compounds, decreases generally in the presence of proteinaceous material. A quantification of consumption effects was performed to judge the suitability of different representatives as antiseptics and their compatibility with pharmaceutical additives., Methods: An iodometric approach served to assess the temporal loss of oxidation capacity in the presence of peptone and fetal calf serum (FCS). The tested agents comprised active halogen compounds, well-known and in particular novel chloramine-based agents indicated for the topical treatment of infections., Key Findings: The decrease in oxidation capacity was higher in the presence of FCS than of peptone and correlated with the reactivity of the oxidants in both cases. The highest consumption rates were for active bromine compounds followed by hypochlorous acid and heterocyclic chlorimides, such as dichloro-isocyanuric acid, while N-chlorotaurine and related amine-based analogues were least consumed. The pH dependence was only remarkable for chloramine T., Conclusions: The observed consumption effects are the result of the differing oxidizing (chlorinating) potencies. Since consumption and irritation are founded on the very same reactions, representatives with low reactivity (N-chloro amino acids) are regarded as more tolerable and retain more oxidative capacity, which provides a more sustained antimicrobial activity., (© 2012 The Authors. JPP © 2012. Royal Pharmaceutical Society.)

Molecular charge-transfer complexes of the tetramethylethylenediamine (TMEDA) with picric acid (Pi-OH), benzene-1,4-diol (QL), tin(IV) tetrachloride (SnCl(4)), iodine, bromine, and zinc chloride (ZnCl(2)) have been synthesized and investigated by elemental and thermal analysis, electronic, infrared, Raman and proton-NMR, energy-dispersive X-ray spectroscopy, X-ray powder diffraction and positron annihilation lifetime spectroscopy, and scanning electron microscopy. In this work, three types of acceptors π-acceptors (Pi-OH and QL), σ-acceptors (iodine and bromine), and vacant orbital acceptors (SnCl(4) and ZnCl(2)) were covered. The results of elemental analysis indicated that the CT complexes were formed with ratios 1:1 and 1:2 for QL, SnCl(4), and ZnCl(2) acceptors and iodine, Pi-OH, and Br(2) acceptors, respectively. The type of chelating between the TMEDA donor and the mentioned acceptors depends upon the behavior of both items. The positron annihilation lifetime parameters were found to be dependent on the structure, electronic configuration, and the power of acceptors. The correlation between these parameters and the molecular weight and biological activities of studied complexes was also observed. Regarding the electrical properties, the AC conductivity and the dielectric coefficients were measured as a function of frequency at room temperature. The TMEDA charge-transfer complexes were screened against antibacterial (Escherichia coli, Staphylococcus aureus, Bacillus subtilis, and Pseudomonas aeruginosa) and antifungal (Aspergillus flavus and Candida albicans) activities., (Copyright © 2012 Elsevier B.V. All rights reserved.)

The Naturally occurring novel cyclohexanonyl bromophenol 2(R)-2-(2,3,6-tribromo-4,5-dihydroxybenzyl)cyclohexanone (4) was synthesized as a racemic compound. Cyclohexylphenyl methane derivatives (10-17) with Br, OMe, CO, and OH were also obtained. Inhibition of four human carbonic anhydrase (hCA, EC 4.2.1.1) isozymes I, II, IV, and VI, with compounds 2-4, 8, and 10-26 was investigated. These compounds were found to be promising carbonic anhydrase inhibitors and some of them showed interesting inhibitory activity. Some of the compounds investigated here showed effective hCA inhibitory activity, and might be used as leads for generating novel carbonic anhydrase inhibitors which are valuable drug candidates for the treatment of glaucoma, epilepsy, gastric and duodenal ulcers, neurological disorders, and osteoporosis., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

A series of bromo-retrochalcones was designed, synthesized and evaluated as PTP1B inhibitors based on licochalcone A and E. Compounds 6, 12, 13, 14, 25, 36, 37, 39, and 41 showed potent inhibitory effects against PTP1B, and compound 37, the most potent among the series, had an IC(50) value of 1.9 μM, about two-fold better than that of the positive control, ursolic acid., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Contaminated drinking water is responsible for causing diarrheal diseases that kill millions of people a year. Additionally, toxin-producing blue-green algae associated with diarrhea and neurologic effects continues to be an issue for many drinking water supplies. Disinfection has been used to reduce these risks. A novel gravity-fed household drinking water system with canisters containing N-halamine bromine or chlorine media was challenged with MS2 bacteriophage and microcystin. Chlorine and bromine systems were effective against this virus, with an mean +/- SE reduction of 2.98 +/- 0.26 log(10) and 5.02 +/- 0.19 log(10), respectively. Microcystin toxin was reduced by 27.5% and 88.5% to overall mean +/- SE concentrations of 1,600 +/- 98 ng/L and 259 +/- 50 ng/L for the chlorine and bromine canisters, respectively. Only the bromine units consistently produced microcystin effluent < 1,000 ng/L (the World Health Organization recommended level) when challenged with 2,500 ng/L and consistently surpassed the U.S. Environmental Protection Agency virus reduction goal of 99.99%.

In this study, polyacrylamide gel (PAG) dosimetry is used to quantitatively assess the efficiency of radiation sensitizers. The local dose enhancement caused by the K-edge absorption of certain atoms such as bromine and iodine can be employed to increase the damage to neighboring molecules and cells. Clonogenic assays can assess the radiation survival of cells to evaluate the efficiency of radiation sensitizers, but this technique requires reliable cell growth in culture media and is time consuming. Our purpose is to use PAG dosimetry to investigate the sensitizing potential of radiation sensitizers such as iodinated compounds. Incorporation of iodinated radiation sensitizers such as NaI and an iodinated contrast agent leads to a quantifiable dose enhancement ratio. When irradiated at low energy (approximately 40 keV), the dose enhancement ratio of the iodinated contrast agent at concentrations of 0.01 (3.5 mg ml(-1)), 0.05 (6 mg ml(-1)) and 0.1 (12 mg ml(-1)) M are 1.16 +/- 0.02, 1.39 +/- 0.03 and 1.82 +/- 0.04, respectively. No dose enhancement was observed when the samples were irradiated with 1.25 MeV gamma photons.

The aim of this study was to determine the biological activity of 4 steroidal derivatives (9a, 9b and 10a, 10b) prepared from the commercially available 17alpha acetoxyprogesterone, where 9a, 9b, have the Delta(4)-3-oxo structure and 10a and 10b an epoxy group at C-4 and C-5. These steroids were tested as inhibitors of 5alpha-reductase enzyme, which is present in androgen-dependent tissues and converts testosterone to its more active reduced metabolite dihydrotestosterone. The pharmacological effect of these steroids was demonstrated by the significant decrease of the weight of the prostate gland of gonadectomized hamsters treated with testosterone plus finasteride or with steroids 10a and 10b. For the studies in vitro the IC(50) values were determined by measuring the steroid concentration that inhibits 50% of the activity of-5alpha-reductase. In this study we also determined the capacity of these steroids to bind to the androgen receptor present in the rat prostate cytosol. The results from this work indicated that compounds 9a, 9b, 10a, and 10b inhibited the 5alpha reductase activity with IC(50) values of 360, 370, 13 and 4.9 nM respectively. However these steroids did not bind to the androgen receptors since none competed with labeled mibolerone. Steroid 10b, an epoxy steroidal derivative containing bromine atom in the ester moiety, was the most active inhibitor of 5alpha-reductase enzyme, present in human prostate homogenates with an IC(50) value of 4.9 nM and also showed in vivo pharmacological activity since it decreased the weight of the prostate from hamsters treated with testosterone in a similar way as finasteride.

The importance of DNA supercoiling in transcriptional regulation has been known for many years, and more recently, transcription itself has been shown to be a source of this superhelicity. To mimic the effect of transcriptionally induced negative superhelicity, the G-quadruplex/i-motif-forming region in the c-Myc promoter was incorporated into a supercoiled plasmid. We show, using enzymatic and chemical footprinting, that negative superhelicity facilitates the formation of secondary DNA structures under physiological conditions. Significantly, these structures are not the same as those formed in single-stranded DNA templates. Together with the recently demonstrated role of transcriptionally induced superhelicity in maintaining a mechanosensor mechanism for controlling the firing rate of the c-Myc promoter, we provide a more complete picture of how c-Myc transcription is likely controlled. Last, these physiologically relevant G-quadruplex and i-motif structures, along with the mechanosensor mechanism for control of gene expression, are proposed as novel mechanisms for small molecule targeting of transcriptional control of c-Myc.

Aims: In vitro experiments were undertaken to evaluate biocide formulations commonly used in cooling water systems against protozoa previously isolated from cooling towers. The investigations evaluated the efficacy of these formulations against amoebic cysts and trophozoites., Methods and Results: Laboratory challenges against protozoa isolated from cooling towers using chlorine, bromine and isothiazolinone biocides showed that all were effective after 4 h. The presence of molybdate and organic phosphates resulted in longer kill times for bromine and isothiazolinones. All treatments resulted in no detectable viable protozoa after 4 h of exposure., Conclusions: The chemical disinfection of planktonic protozoa in cooling water systems is strongly influenced by the residence time of the formulation and less so by its active constituent. Bromine and isothiazolinone formulations may require higher dosage of concentrations than currently practiced if used in conjunction with molybdate- and phosphate-based scale/corrosion inhibitors., Significance and Impact of the Study: Cooling water systems are complex microbial ecosystems in which predator-prey relationships play a key role in the dissemination of Legionella. This study demonstrated that at recommended dosing concentrations, biocides had species-specific effects on environmental isolates of amoebae that may act as reservoirs for Legionella multiplication in cooling water systems.

Magnaporthe grisea is a fungal pathogen of rice that forms appressoria that penetrate the outer cuticle of the rice plant. Data from recent studies indicate that M. grisea isocitrate lyase (ICL), a key enzyme in the glyoxylate cycle, is highly expressed during appressorium-mediated plant infection. Bromophenols isolated from the red alga Odonthalia corymbifera exhibited potent ICL inhibitory activity and blocked appressoria formation by M. grisea in a concentration-dependent manner. In addition, these compounds protected the rice plants from infection by M. grisea. Rice plants infected with wild-type M. grisea Guy 11 exhibited significantly lower disease severity with bromophenol treatment than without, and the treatment effect was comparable to the behavior of the Deltaicl knockout mutant I-10. The protective effect of bromophenols and their strong inhibition of appressorium formation on rice plants suggest that ICL inhibitors may be promising candidates for crop protection, particularly to protect rice plants against M. grisea.

Schizosaccharomyces pombe Rng2 is an IQGAP protein that is essential for the assembly of an actomyosin ring during cytokinesis. Rng2 contains an amino-terminal calponin-homology (CH) domain, 11 IQ repeats and a RasGAP-homology domain. CH domains are known mainly for their ability to bind F-actin, although they have other ligands in vivo and there are only few examples of actin-binding single CH domains. The structures of several CH domains have already been reported, but this is only the third report of an actin-binding protein that contains a single CH domain (the structures of calponin and EB1 have been reported previously). The 2.21 A resolution crystal structure of the amino-terminal 190 residues of Rng2 from Br- and Hg-derivatives includes 40 residues (150-190) carboxyl-terminal to the CH domain that resemble neither the extended conformation seen in utrophin, nor the compact conformation seen in fimbrin, although residues 154-160 form an unstructured coil which adopts a substructure similar to dystrophin residues 240-246 in the carboxyl-terminal portion of the CH2 domain. This region wraps around the stretch of residues that would be equivalent to the proposed actin-binding site ABS1 and ABS2 from dystrophin. This distinctive feature is absent from previously published CH-domain structures. Another feature revealed by comparing the two derivatives is the presence of two loop conformations between Tyr92 and Arg99.

The influx of chlorides into Trichoderma viride vegetative submerged mycelium was measured by means of the radionuclide (36)Cl(-). It was found that the (36)Cl(-) influx was time-dependent (the steady-state was established with t(1/2 )= 25 min at 25 degrees C), pH-dependent (with pH optimum between 4-5.5), temperature-dependent (at about 15 degrees C), and concentration-dependent (K(M)(Cl(-))) = 47.6 +/- 4.2 micromol x l(-1); J(max) = 11.5 +/- 0.7 pmol(Cl(-)) x min(-1). mg(dry mass) (-1)). The (36)Cl(-) influx was inhibited by Br(-) but not F(-), I(-), SO(4)(2-), HPO(3)(2-) and HCO(3)(-). The presence of vanadate (P-type ATPase inhibitor) moderately stimulated the (36)Cl(-) influx but the presence valinomycin (electrogenic K(+) ionophore), salicylate (known to release Ca(2+) from Trichoderma viride internal stores) were without effect on the (36)Cl(-) influx. The results suggest that the (36)Cl(-) influx is mediated by a carrier and that the transport is electroneutral, probably Cl(-)/OH(-) antiport., (Copyright 2004 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)

Objective: Previous studies have reported the link between residual microbial contamination of toothbrushes and periodontal diseases. The goal of this pilot study was to evaluate the effects of an antimicrobial additive (Microban) to toothbrushes on residual retention of periodontal pathogens., Methodology: Twenty patients had one side of their mouths brushed with a toothbrush containing the antimicrobial agent (experimental side), and the other side with a toothbrush containing no agent (control). Toothbrushes were air-dried (25 degrees C) for four or 24 hours. Toothbrush heads were vortexed and cultured for Prevotella species (Ps), Porphyromonas gingivalis (Pg), Actinobacillus actinomycetemcomitans (Aa), and non-specific colony-forming units (NS). The plates were incubated and counted. Means and standard deviations were calculated, and data were analyzed using a series of t-tests (paired and unpaired) and Wilcoxon matched-pairs signed-rank test., Results: No significant inter- or intra-group differences in mean counts were found; however, when four-hour and 24-hour data for Aa, Pg, or NS were combined, experimental counts were lower than controls in 39/50 (78%) of the matched pairs (Wilcoxon signed-rank test p = 0.01)., Conclusion: Toothbrushes containing the antimicrobial additive showed lower microbial counts than those without, but between-group means were not statistically significant.

Statement of Problem: Contamination of removable prostheses with microorganisms, particularly Candida albicans, is a common clinical problem. Microban, a broad-spectrum antimicrobial containing triclosan, recently has been proposed to inhibit microbial growth., Purpose: This study aimed to determine whether the addition of Microban to PermaSoft denture liner prevents the growth of C albicans and affects the cytotoxicity of the PermaSoft material., Material and Methods: Experimental specimen disks (5 x 1 mm each) with and without incorporated Microban were fabricated aseptically (n = 6) against polyester film to produce a smooth surface. To assess the cytotoxic effect of Microban, the MTT assay was used. To determine the effect of Microban on the growth of C albicans, disks were placed in Transwell dishes, covered with Sabouraud's broth containing an ATCC strain of C albicans, and incubated at 37 degrees C for 24 hours. Wells containing fluorocarbon resin disks or broth alone served as controls. The disks were rinsed to remove unattached C albicans and then sonicated in sterile water to remove surface organisms. Serial dilutions of the water extracts were plated on Sabouraud's agar and returned to the incubator for 24 hours. Colonies were counted with a Brunswick Colony Counter. Growth of C albicans in the internal aspects of the specimens was determined in a manner as previously described, with the exception that the specimens were sonicated to remove surface organisms, minced, and sonicated once more before making serial dilutions. The results were compared with ANOVA and Tukey intervals (alpha=.05)., Results: The number of colonies formed ranged from 17 to 31 x 10(5) (mean = 23 +/- 4 x 10(5)) and 14 to 69 x 10(5) (mean = 32 +/- 20 x 10(5)) for the PermaSoft with and without Microban groups, respectively. There was no statistically significant difference between PermaSoft with and without Microban., Conclusion: The addition of Microban did not significantly alter the cytotoxicity of the PermaSoft denture lining material or reduce the adherence of viable C albicans to the surface of PermaSoft material after 24 hours.

The ability of bromine and rat liver microsomes (RLM) to convert organophosphorus (OP) protoxicants to esterase inhibitors was determined by measuring acetylcholinesterase (AChE) and neuropathy target esterase (NTE) inhibition. Species specific differences in susceptibility to esterase inhibition were determined by comparing the extent of esterase inhibition observed in human neuroblastoma cells and hen, bovine, and rodent brain homogenates. OP protoxicants examined included tri-o-tolyl phosphate (TOTP), O-ethyl O-p-nitrophenyl phenylphosphonothioate (EPN), leptophos, fenitrothion, fenthion, and malathion. Bromine activation resulted in greater AChE inhibition than that produced by RLM activation for equivalent concentrations of fenitrothion, malathion, and EPN. For EPN and leptophos, bromine activation resulted in greater inhibition of NTE than RLM. Only preincubation with RLM activated TOTP; resultant inhibition of AChE was less in hen brain (13 +/- 3%) than in neuroblastoma cells (73 +/- 1%) at 10(-6) M. In contrast, 10(-6) M RLM-activated TOTP produced more inhibition of hen brain NTE (89 +/- 6%) than NTE of human neuroblastoma cells (72 +/- 7%). Human neuroblastoma cells and brain homogenates from hens, the accepted animal model for study of OP-induced neurotoxicity, were relatively similar in sensitivity to esterase inhibition. Homogenates from hens were more sensitive to NTE inhibition induced by phenyl saligenin phosphate (PSP), an active congener of TOTP, than were homogenates from less susceptible species (mouse, rat, bovine). AChE of hen brain homogenates was also more sensitive than homogenates from other species to malaoxon, the active form of malathion.

The paper presents and discusses a novel on-line real-time non-destructive continuous-flow system for biocide testing on industrial biofilms. This laboratory system is capable of monitoring changes in growth, accumulation and respiratory activity of biofilms in response to biocidal treatment. The system incorporates a fouling monitor for continuous measuring of the rate of biofilm accumulation (heat transfer resistance), a sensor for monitoring of microbial activity (oxygen meter for monitoring the rate of biofilm respiratory activity), and subsystems necessary for microbial life support and control of operation parameters. Examples of system operation and testing of oxidizing and non-oxidizing biocides are presented.

Stimulus-induced changes in free cytosolic Ca2+ in different types of plant cells have been monitored with the aid of fluorescent calcium indicator dyes. However, there is no simple and convenient method for introducing these dyes into the plant cell cytoplasm. This paper reports tests of different procedures for loading either free fluorescent dyes or their acetoxymethyl esters (Fluo-3 and Fluo-3/AM, respectively) into Sinapis alba root tissue. Loading of Fluo-3 was pH and temperature dependent. Moreover, in the presence of beta-escin (saponin) in the loading medium very high fluorescent signals in root tissues were observed. The highest signals were recorded when tissue was loaded in a medium containing 0.1% beta-escin, at pH 5.0 and 30 degrees C. Only very weak fluorescence signals were found in mustard roots loaded with Fluo-3/AM. Acidity and temperature of the medium had no significant effect on the process. However, addition of eserine, a cholinesterase inhibitor led to a dramatic increase in fluorescence in the root cells. On the basis of these observations rapid and efficient methods of loading both Fluo-3 and Fluo-3/AM into mustard root tissues are proposed.

1. Chloride currents through the membrane of rat psoas muscle fibre segments were investigated with a double Vaseline gap under conditions minimizing the currents of other ion species. 2. In Cl(-)-free solutions a time- and voltage-independent conductance of 1.1 +/- 0.4 microS was observed. 3. As with intact fibres, the steady-state Cl- conductance was 2.5 +/- 0.9 mS cm-2; the halide selectivity was Cl- > Br- > I-, and Cl- currents were completely blocked by 0.1 mM 9-anthracene carboxylic acid (9-AC). 4. Voltage steps from -85 mV to between -125 and +55 mV elicited currents with deactivation upon hyperpolarization and activation upon depolarization. Activation was fitted with two exponentials, the smaller time constant increasing from 37.5 ms at +55 mV to 67.0 ms at -5 mV, the larger time constant (450 ms) being independent of potentials more positive than -5 mV. The two deactivation time constants ranged between 30.6 (-105 mV) and 99.3 ms (-35 mV), and 139.4 (-105 mV) and 738.5 ms (-35 mV). 5. The activation curve was fitted with a Boltzmann distribution (half-maximum, -39 mV; slope at inflexion point, 1/17.2 mV). Deactivation was incomplete. At very negative potentials about one-quarter of the maximum number of channels were open. 6. When tested with 5 and 61 mM intracellular Cl- concentration ([Cl-]i) the kinetic parameters were not different. 7. During depolarizations lasting > 5 s, activation was followed by a decline. With progressively longer prepulses going positive to the reversal potential and test pulses going negative, the responses to test and prepulses decreased with similar time constants, suggesting a real inactivation process.

Characteristics of the ion transport of the ascending thin limb of Henle's loop (ATL) were reviewed with special reference to the regulatory mechanisms of the putative Cl- channel. The ATL is highly permeable to NaCl, moderately permeable to urea, and virtually impermeable to water. The Cl- permeability is extremely high, having saturation kinetics and being inhibited by halogens. The Cl- conductances exist in series in both apical and basolateral membranes. They are inhibited by anion transport inhibitors including DIDS, phloretin, and NPPB. The Cl- conductance is pH sensitive and activated by Ca2+. NEM exhibits dual action on this conductance: irreversible inhibition and reversible stimulation. This highly effective Cl- transport system is favorable for the operation of the countercurrent multiplication system in the renal medulla.

T1 phages with and without 5-bromo-2'-deoxyuridine (BrdU) labelling were irradiated in solids with monochromatic X-rays at 12.40 and 13.51 keV, below and just above the K-absorption edge of bromine (13.47 keV) in vacuum and wet states. Irradiated phages were assayed on uracil-DNA glycosylase (Udg) deficient (ung-1) and sufficient (ung+) host strains of Escherichia coli, in order to investigate the nature of the lethal damage induced by Auger cascade following X-ray absorption at the K-shell of bromine as a key atom. The results were: (1) An Auger-specific enhancement (1.15) was observed only when BrdU-labelled phages were irradiated in the wet state and assayed on ung+ host cells. (2) A Udg-specific enhancement was observed only for BrdU-labelled phages, not for unlabelled phages. (3) The sensitivities of BrdU-unlabelled phages were almost the same, despite the irradiation states and strains of the host cell, indicating that this sensitivity was a common fraction of the sensitivity under all conditions. (4) The lethal damage for the T1 phage was categorized into four fractions according to the sensitivities under different conditions: the general fraction was defined as being the sensitivity of unlabelled phages (G-fraction); BrdU-specific, but unrecognizable by Udg (B-fraction); Udg specific, but not Auger-specific (U-fraction); and Auger-specific (A-fraction). (5) Although the so-called indirect action of water radicals increased only the G-fraction by about three-fold, the B- and U-fractions were not affected by any change in the irradiation states, thus indicating that these two fractions were caused by the so-called direct action.

The effect of adding Br or Zn supplementation to the dialysate on the concentrations of Br and Zn in the blood of hemodialysed patients, is investigated. Patients with end-stage renal failure on hemodialysis show an abnormal trace element pattern. Our patients showed lowered serum Br and Zn concentrations. Four patients were subjected to dialysates with varying Br content. The impact on their serum and packed cells concentrations was evaluated. The supplementation resulted in an increase in the concentrations in serum and in packed cells. The Br concentration in serum and packed cells closely followed the dialysate content. In order to restore the Zn concentration to the normal level a ZnCl2 solution was added to the dialysate of 4 other patients. Zn accumulated in the patient as a consequence of its diffusion against the concentration gradient.

Six duocarmycins have been discovered during our search for new antitumor antibiotics and they showed extremely potent cytotoxic activity with IC50 values of 10(-12) M-10(-9) M on HeLa S3 cell. Three different producing strains isolated from soils were taxonomically assigned as Streptomyces. Duocarmycin A was unstable in culture broth, so improved culture conditions were designed to produce a high titer of duocarmycins B1, B2, C1 and C2 which are halogenated seco-compounds of duocarmycin A. Duocarmycin SA, one of the most potent cytotoxic agents yet discovered, was shown to be more stable in culture media than duocarmycin A, despite the structural similarity on their spirocyclopropylhexadienone moiety. In contrast to the duocarmycin A fermentation, no halogenated seco-compounds of duocarmycin SA were detected in culture broth supplemented with Br- or Cl-. All duocarmycins could be produced using one producing strain with improved media and culture conditions.

Despite many studies suggesting the role of enolpyruvate as a bound intermediate in the pyruvate kinase reaction, direct evidence for it has been lacking. By use of a combination of chemical trapping and isolation of a derivative, significant amounts of enzyme-bound enolpyruvate have now been demonstrated. The method distinguishes enolpyruvate. It is based on reaction of bromine with enolpyruvate in acid, derivatization of formed bromopyruvate with thionitrobenzoate, and resolution by reversed-phase HPLC of the thioether derivative. As little as 10 pmol of the thioether derivative could be quantitated reliably. With this method, the internal equilibria, including the E.ATP.enolpyruvate intermediate, have been determined. Enzyme-enolpyruvate concentration was shown to be pH-dependent. Phosphoenolpyruvate also reacts with bromine to form bromopyruvate. To quantitate enolpyruvate specifically in a background of phosphoenolpyruvate, advantage was taken of phosphoenolpyruvate's much greater stability in acid. When bromide/was added 10 min after the acid quench, ketonization of enolpyruvate was complete, and only phosphoenolpyruvate was measured. Enolpyruvate is thus determined by difference between the bromopyruvate measured with and without delayed bromine addition.

1. Myeloperoxidase catalysed the H2O2-supported oxidation of aminopyrine in the presence of Cl-, generating the aminopyrine cation radical (AP+.). The rate of AP+. formation was determined by monitoring the absorbance at 565 nm. The pH optimum of the reaction was around 5.0. The rate of AP+. formation increased with increasing concentration of aminopyrine. Inhibition by excess H2O2 was seen at pH 4.5-5.5. 2. When Cl- was replaced by Br-, the rate of AP+. formation increased significantly, and inhibition by H2O2 became less evident and was observed only at pH 5.5. The rate of chemical oxidation of aminopyrine by HOCl was much slower than that by Br2. 3. Compared with the chlorination of monochlorodimedone (MCD), the reaction between aminopyrine and HOCl, produced by the enzymic peroxidation of Cl-, is rate-limiting in the myeloperoxidase-catalysed oxidation of aminopyrine. The differences in kinetic behaviour between the myeloperoxidase-catalysed chlorination of MCD and oxidation of aminopyrine are explained by the low reactivity of HOCl towards aminopyrine.

Eosinophils (EOs) participate in a variety of inflammatory states characterized by endothelial cell damage, such as vasculitis, pneumonitis, and endocarditis. We find that 100 U/ml TNF-alpha/cachectin (TNF), a concentration attainable in the blood of humans with parasitic infestations, stimulates highly purified populations of EOs to damage human umbilical vein endothelial cells (HUVEC), a model of human endothelium. This TNF-dependent EO cytotoxicity is strongly inhibited by heparin and methyprednisolone but unaffected by the platelet-activating factor antagonist BN52012 or scavengers of superoxide anion and H2O2, superoxide dismutase and catalase. However, addition of a physiologically relevant concentration of Br- (100 microM) enhances EO/TNF damage to HUVEC, implicating the possible participation of EO peroxidase (EPO) in the killing mechanism. EOs adherent to FCS-coated plastic wells more than double their production of superoxide anion and the cytotoxic EPO-derived oxidant HOBr when exposed to TNF, showing that TNF activates the respiratory burst of EOs attached to a "physiologic" surface. Unlike PMNs, EOs were not irreversibly activated to kill unopsonized endothelium by previous exposure to TNF, and did not degranulate or upregulate CR3 expression as detected by Mo1 in the presence of 100 U/ml TNF. HUVEC exposed 18 h to TNF were considerably more susceptible to lysis by PMA-activated EOs and reagent H2O2, demonstrating a direct effect of TNF upon endothelium, perhaps through inhibition of antioxidant defenses. These findings suggest that abnormally elevated serum levels of TNF may provoke EOs to damage endothelial cells and thereby play a role in the pathogenesis of tissue damage in hypereosinophilic states.

The acute cytogenetic effects of potassium bromate (KBrO3) on rat bone marrow cells in vivo were studied. The incidence of chromosome aberrations in bone marrow cells increased rapidly, reaching a maximum level 12 h after intraperitoneal injection and decreased within 24 h. Dose-response relationships were obtained for both intraperitoneal and oral administrations.

A continuous flow type apparatus is used for investigation of the chlorine and bromide activity in vitro on the cysts of two free-living amoeba strains : Naegleria gruberi and Acanthamoeba polyphaga. The Naegleria cysts are sensitive more to chlorine than to bromine ; those of Acanthamoeba are perfectly resistant to these disinfectants in the limit of the available concentrations in swimming-pools. The sensibility to these disinfectants stays the same at 25 degrees C and 35 degrees C.

Human kidney cells in culture and cells of mouse sarcoma-180 were allowed to incorporate bromine into their DNA. Cultured cells with and without incorporated BUdR were irradiated with electromagnetic radiations ranging in energy from 12 keV X-rays to 60Co gamma-rays to find out whether or not there exists any energy dependence of the bromine dose enhancement ratio BER. Such a dependence should show in the immediate neighbourhood of the K-absorption edge of bromine (13.5 keV). Any influence of the Auger effect triggered in bromine by external irradiation should show by a significant increase of the BER for energies rising from slightly below to slightly above the bromine K-edge. Values of D37, D0 and the extrapolation numbers of the cell survival curves served as biological endpoints. Measured values of BER ranged from 1.12-2.00 without any significant dependence energy. A weakly pronounced peak was found for 50 kV X-rays of 26 keV mean energy. Sarcoma-180 were irradiated with 14 kEV X-rays and 60C gamma-rays. BUdR was administered i.v., i.p. and directly into the tumours in quantities of up to 1 ml of a 10(-3) M solution. Tumour regression caused by the irradiations served as a measure of the radiation effects with and without incorporated BUdR. No significant dose enhancement effect of bromine was observed in this case. Within error limits, the combination of BUdR with 14 keV X-rays proved more effective than BUdR combined with 60Co gamma-rays.

Organ cultures of the guinea pig hypothalamo-neurohypophysial complex synthesize the octapeptide hormone, vasopressin, a specific product of the neurosecretory cells of the supraoptic nucleus. Inhibitors of both protein and RNA synthesis (cycloheximide and bromotubercidin respectively) were found to block vasopressin biosynthesis. In the presence of bromotubercidin, the apparent half-time of decline in the rate of hormone biosynthesis was about 28 h. Colchicine inhibited the distal transport of vasopressin into the posterior pituitary. Ultrastructural studies on colchicine-treated cultures indicated the neuronal stalks were intact and that neurotubules were still present. The narcotic drug, levorphanol at 10-7 M and 10-9 M was found to inhibit RNA synthesis by 20 percent. At these concentrations it had no demonstrable effect on vasopressin synthesis. Cultures established from animals that had been rendered tolerant to narcotics also had no observable alterations in vasopressin biosynthesis, although the initial pituitary vasopressin content of these cultures was reduced by about 35 percent. Various pharmacologic and biologic compounds were tested for their effects on vasopressin biosynthesis in organ cultures. Dibutyryl cyclic AMP, estradiol-17beta, nicotine, nerve growth factor (NGF), and pineal extract all had no effects under the present experimental regimen. Medium conditioned by the presence of fetal hypothalami of 40-55 days gestation produced a 2-4 fold increase in vasopressin biosynthesis in cultures established from adult animals. Medium conditioned by fetal cerebral cortex, liver, or hypothalamic tissue from fetuses of less than 33 days gestation did not have this stimulatory effect.

The principle of Le Chatelier was used in demonstrating that 3-chloro-4,4-dimethyl-2-oxazolidinone (compound 1) itself kills Staphylococcus aureus rather than the very small amount of free chlorine in hydrolysis equilibrium with compound 1. On the other hand, when the N-bromo analog of compound 1 (compound 1B) was used as the disinfectant, the mixture of combined compound 1B and free bromine formed in the hydrolysis equilibrium provided disinfection. When the hydrolysis equilibrium for 1B was suppressed to the level at which a negligible amount of free bromine remained in solution, combined compound 1B was much more efficacious than combined compound 1 at killing S. aureus.