Environmental Homeopathy: Homeopathic Potencies Regulate the Toxicity and Growth of Raphidiopsis raciborskii (cyanobacteria) and can be Tracked Physico-Chemically. Part 1: Biological Results.

INTRODUCTION: Cyanobacteria are microorganisms found in many parts of the world and several genera, such as Raphidiopsis raciborskii, are producers of cyanotoxins. Homeopathic potencies have been found to modulate toxicity in different biological models, and the present study endeavors to discover whether this might also be the case with cyanobacteria. OBJECTIVES: Our objective was to investigate the possible effects of homeopathic potencies on the resilience of Artemia franciscana (brine shrimp) embryos to saxitoxin (STX; cyanotoxin) and on controlling the growth of R. raciborskii in vitro. METHOD: A. franciscana cysts were cultivated in seawater in 96-well plates to evaluate the hatching rate and vitality, plus the gene expression of heat shock proteins (HSPs), after being challenged with R. raciborskii extract containing 2.5 µg/L of STX and treated with different homeopathic potencies. Untreated wells were used as controls ("base-line"). Potencies were chosen from a screening process based on seven selected homeopathic preparations according to the similitude of STX symptoms (Sulphur, Zincum metallicum, Nitric acidum, Plumbum metallicum, Mercurius solubilis, Phosphoric acidum, Isotherapic from R. raciborskii extract; all at 6cH, 30cH and 200cH). Cultures of R. raciborskii maintained in an artificial seawater medium were equally treated with screened homeopathic potencies selected from the same list but specifically for their growth control as a function of time. RESULTS: A 15% lower rate of hatching of A. franciscana cysts was observed after treatment with Nitric acidum 6cH in comparison with baseline (p = 0.05). A complete toxicity reversal was seen after treatment with Isotherapic 200cH, with a 23-fold increase of Hsp 26 gene expression (p = 0.023) and a 24-fold increase of p26 gene expression (p ≤ 0.001) in relation to baseline. Nitric acidum 200cH and Mercurius solubilis 30cH limited the exponential growth of cyanobacteria up to 95% and 85% respectively (p ≤ 0.003) in relation to baseline. Succussed water presented only a transitory 50% inhibition effect. CONCLUSION: Isotherapic 200cH improved A. franciscana bioresilience to STX; Nitric acidum 200cH and Mercurius solubilis 30cH showed the optimal performance on limiting R. raciborskii growth. The results point to the potential of homeopathic potencies to mitigate environmental problems related to water quality.

Environmental Homeopathy: Homeopathic Potencies Regulate the Growth and Toxicity of Raphidiopsis raciborskii (cyanobacteria) and Can be Tracked Physico-Chemically. Part 2: Physico-chemical Results.

INTRODUCTION: The control of cyanobacterial toxicity and growth by homeopathic potencies was described in Part 1 of this two-part report. Here, a parallel approach characterized the physico-chemical features of the potencies used and the liquid media treated with them, correlating these results with their respective biological effects. OBJECTIVES: Our objective was to establish if physico-chemical parameters can track homeopathic potencies in seawater or artificial seawater medium (ASM)-1 and to discover whether these parameters correlate with previously described biological effects. METHOD: Artemia franciscana (brine shrimp) cysts were cultivated in seawater challenged with Raphidiopsis raciborskii extract and treated with different homeopathic potencies chosen from a screening process. Cultures of R. raciborskii maintained in ASM-1 were also treated with previously screened homeopathic potencies, and their growth was monitored as a function of time. The physico-chemical properties of the treated media (seawater or ASM-1) were evaluated by their interaction with solvatochromic dyes and changes in pH, conductivity and temperature. RESULTS: Coumarin 7 was found to be a marker for Nitric acidum 6cH and Isotherapic (R. raciborskii extract) 200cH in seawater (analysis of variance [ANOVA], p = 0.0015). Nile red was found to be a marker for Nitric acidum 200cH and Mercurius solubilis 30cH in ASM-1 (ANOVA, p ≤ 0.001). An increase in pH of ASM-1 and endothermic effects were observed after these treatments (two-way ANOVA, p = 0.0001). Seawater and ASM-1 to which potencies had been added were also subjected to a constant unidirectional 2,400 Gauss static magnetic field and found to have enhanced effects on the solvatochromic dyes tested. CONCLUSION: Homeopathic potencies were specifically traceable in aqueous media using solvatochromic dyes, especially when the samples were subjected to a magnetic field. Results from monitoring other physical parameters, such as pH and temperature, were less specific in relation to potency tracking. However, potency-induced endothermic effects might provide valuable thermodynamic data relating to the nature of potencies.

Ultraviolet C as a method of disinfecting medical silicone used in facial prostheses: An in vitro study - Part 2.

STATEMENT OF PROBLEM: Disinfection is an important factor in preserving facial prostheses and maintaining tissue health. However, whether disinfection with ultraviolet C is an effective disinfection method is unclear. PURPOSE: The purpose of this in vitro study was to evaluate the effectiveness of irradiation with different exposure durations of an ultraviolet-C light-emitting diode in the disinfection of the silicone (A-588-1; Factor II) used in facial prostheses. MATERIAL AND METHODS: A total of 216 specimens were prepared, contaminated by multispecies biofilm, and divided into 9 groups (n=24) for different treatments: chlorhexidine 0.12% (G CHG), ultraviolet-C light-emitting diode for 5 minutes (G UVC5), ultraviolet-C light-emitting diode for 10 minutes (G UVC10), ultraviolet-C light-emitting diode for 20 minutes (G UVC20), their respective untreated controls (Gcontrol CHG, Gcontrol UVC5, Gcontrol UVC10, Gcontrol UVC20), and dimethyl sulfoxide (G DMSO) as the negative control. Cell viability was measured by using the methyl tetrazolium salt (MTT) method. Two statistical analyses were performed. First, a 2×3 ANOVA was carried out to compare the control groups (Gcontrol UVC5, Gcontrol UVC10, and Gcontrol UVC20) and the experimental groups of UV-C LED light with different exposure durations (G UVC5, G UVC10, and G UVC20). The second analysis was performed using generalized linear models to compare the optical density of the groups (G UVC5, G UVC10, G UVC20, G CHG, and G DMSO). RESULTS: Cell viability results demonstrated a microbial reduction after exposure to the ultraviolet-C light-emitting diode for 20 minutes (G UVC20) compared with untreated controls (P<.05). The 5- and 10-minute exposures were statistically similar to their respective control groups (P>.05). The 20 minutes exposure had the lowest average optical density value, being statistically different from the 5-minute exposure (P<.05). A 20-minute exposure to the ultraviolet-C light-emitting diode (G UVC20) was similarly effective when compared with the standard disinfection treatment (G CHG) and dimethyl sulfoxide (G DMSO) (P>.05). CONCLUSIONS: Irradiation with an ultraviolet-C light-emitting diode for 20 minutes decreased the in vitro microbial cell viability on the medical silicone used in facial prostheses.

Highly Diluted Glyphosate Mitigates Its Effects on Artemia salina: Physicochemical Implications.

Glyphosate is an herbicide widely used in agriculture but can present chronic toxicity in low concentrations. Artemia salina is a common bio-indicator of ecotoxicity; it was used herein as a model to evaluate the effect of highly diluted-succussed glyphosate (potentized glyphosate) in glyphosate-based herbicide (GBH) exposed living systems. Artemia salina cysts were kept in artificial seawater with 0.02% glyphosate (corresponding to 10% lethal concentration or LC10) under constant oxygenation, luminosity, and controlled temperature, to promote hatching in 48 h. Cysts were treated with 1% (v/v) potentized glyphosate in different dilution levels (Gly 6 cH, 30 cH, 200 cH) prepared the day before according to homeopathic techniques, using GBH from the same batch. Controls were unchallenged cysts, and cysts treated with succussed water or potentized vehicle. After 48 h, the number of born nauplii per 100 µL, nauplii vitality, and morphology were evaluated. The remaining seawater was used for physicochemical analyses using solvatochromic dyes. In a second set of experiments, Gly 6 cH treated cysts were observed under different degrees of salinity (50 to 100% seawater) and GBH concentrations (zero to LC 50); hatching and nauplii activity were recorded and analyzed using the ImageJ 1.52, plug-in Trackmate. The treatments were performed blind, and the codes were revealed after statistical analysis. Gly 6 cH increased nauplii vitality (p = 0.01) and improved the healthy/defective nauplii ratio (p = 0.005) but delayed hatching (p = 0.02). Overall, these results suggest Gly 6cH treatment promotes the emergence of the more GBH-resistant phenotype in the nauplii population. Also, Gly 6cH delays hatching, another useful survival mechanism in the presence of stress. Hatching arrest was most marked in 80% seawater when exposed to glyphosate at LC10. Water samples treated with Gly 6 cH showed specific interactions with solvatochromic dyes, mainly Coumarin 7, such that it appears to be a potential physicochemical marker for Gly 6 cH. In short, Gly 6 cH treatment appears to protect the Artemia salina population exposed to GBH at low concentrations.

Prenatal restraint stress downregulates the hypothalamic kisspeptidergic system transcripts genes, reduces the estrogen plasma levels, delayed the onset of puberty, and reduced the sexual behavior intensity in female rats.

AIMS: This study investigated the possible relationships between the expression of the Kiss1 and Gpr54 gene expressions and the pituitary-gonadal hormones with the female onset of puberty and sexual behavior. The Kiss1 and Gpr54 gene expressions were examined because they are critical to controlling the hypothalamic activation of GnRH neurons and, in turn, the pituitary-gonadal hormones related to the early onset of puberty and sexual behavior. Further, it was evaluated that the pituitary and gonadal hormones involved in the vaginal opening and the expression of sexual behavior. METHODS: Pregnant rats exposed to PRS from gestation days 17 to 20 were evaluated for maternal and open-field behaviors. The maternal behavior was analyzed because it may alter brain sexual organization affecting the pups development. It was observed in female pups the physical and development and, in adult age, the open-field behavior, the anxiety-like behavior, the estrous cycle, the sexual behavior, the serum FSH, LH, estrogen, progesterone, and testosterone levels, and the gene expression of kisspeptin protein (Kiss1) and Gpr54 in the hypothalamus. RESULTS: the maternal and open-field behaviors were unaffected. In the F1 generation, PRS reduced weight at weaning, delayed the day of the vaginal opening and reduced the intensity of lordosis, the estrogen levels, and the Kiss1 and Gpr54 gene expression. These effects were attributed to hypothalamic kisspeptidergic system downregulation of transcripts genes and the reduced estrogen levels affected by the PRS.

Bacterial infiltration and detorque at the implant abutment morse taper interface after masticatory simulation.

This study evaluated the bacterial infiltration and the detorque of indexed and non-indexed abutments of Morse taper implants (MTI) after mechanical cycling (MC). 40 MTI were distributed into four groups: IIA (indexed implant abutments); NIIA (non-indexed implant abutments); IIAMC (indexed implant abutments submitted to MC); NIIAMC (non-indexed implant abutments submitted to MC), which were carried out under one million 5 Hz frequency and 3 Bar pressure. After mechanical cycling, all groups were immersed in a bacterial solution in Brain Heart Infusion Agar. After detorque, the bacteria infiltration was evaluated by counting the colony-forming units. For the bacterial infiltration, analysis was applied to the Kruskal-Wallis test (p = 0.0176) followed by Dunn's test. For the detorque analysis, the two-way repeated-measures ANOVA was applied, followed by the Tukey's test (p < 0.0001). Bacteria infiltration was highly observed in NIIA (p = 0.0027) and were absent in IIAMC and NIIAMC. The detorque values for IIA (19.96Ncm ± 0.19Ncm), NIIA (19.90Ncm ± 0.83Ncm), and NIIAMC (19.51Ncm ± 0,69Ncm) were similar and remained close to the initial value, while IIAMC (55.2Ncm ± 2.36Ncm) showed an extremely significant torque value increase (p < 0.0001). The mechanical cycling resulted in mechanical sealing of the implant-abutment interface, preventing bacterial infiltration in the indexed and non-indexed specimens, and increasing the detorque strength in the group of indexed abutments.

Pilot project. Resveratrol intake by physical active and sedentary older adult women and blood pressure.

OBJECTIVE: Although the use of resveratrol is widespread, there is a lack of studies concerning its use in older adults who practice regular physical activity. The present study aimed to evaluate how resveratrol influenced anthropometric parameters, heart measures, blood analytes in women participants, aged 60 to 80 years old, distributed in four groups consisting of those who regularly exercise or not, and those who were under resveratrol treatment or not. The older adult women exercised in a Community Center. METHODS: A prospective, comparative clinical pilot study was carried out based on convenience non-probability sampling. Resveratrol 300 mg/daily/60 days was orally-administered to 43 participants that were regularly accompanied and checked for anthropometrical parameters, heart measures, blood analytes, leukogram, and plaquetogram indices. RESULTS: The effect of resveratrol and exercise was not significant in most of the parameters evaluated in the study. Nonetheless, both systolic (p = 0.0120) and diastolic (p = 0.0241) blood pressures (BP) were elevated in sedentary older adult women in comparison with older women that regularly exercised. However, they remained within the normal BP range considered for the age. Mean corpuscular volume (MCH; p = 0.0198) and red blood cell distribution width (RDW; p < 0.0001) were improved in the groups receiving resveratrol and exercises compared to the sedentary group receiving resveratrol. The sedentary group receiving resveratrol showed diminished segmented cells (p = 0.0459), basophils (p < 0.0001), and lymphocytes (p < 0.0001). CONCLUSIONS: Present findings showed that resveratrol consumption for sedentary older women could lead to dysregulated blood pressure. Although resveratrol consumption is indicated to treat inflammation, its use must be discussed with a health professional and not be inadvertently self-administered due to significant alterations in blood pressure in older adult women, primarily if not related to exercise practice.

Environmental parameters modulate lead chloride toxicity on Artemia salina

Different environmental conditions can influence the effects of toxic agents on living beings. Recently, a series of experiments performed in Artemia salina submitted to different kinds of intoxication have shown that both, isotherapic and succussed watercan change Artemia salina ́s bio resilience at different levels. Moreover, it seems to vary according to the circalunar cycle. Objective:To verify if circalunar phases and water agitation can modify the toxicity of lead chloride on Artemia salina in vitro. Methodology:Artemia salina cysts were exposed to seawater containing 0.04% of lead chloride (equal to EC10 or 10% effective concentration, previously determined in a pilot study) in 96-well culture plates. Thirty-six experimental repetitions were performed in four series to observe the possible effects of adding stirred water, the so-called succussed water, and the moon phases. The hatched cysts were recorded after 48 hours using a digital microscope (1000x magnification) to identify the hatching percentage and the viability and mobility of the born nauplii. Results:The exposition of cysts to PbCl2 led to an increase in the hatching rate, and it was more evident during the full moon (p = 0.00014) The addition of succussed water into the seawater medium reduced this effect to the baseline levels. An increase in mobility was seen in nauplii born from exposed cysts during the full moon (p = 0.00077), but this effect was not affected by the treatment with succussed water. Discussion:Although the effects of lead chloride EC10 on the increase of nauplii hatching were expected, two environmental variables changed the sensitivity of cysts to this harmful stimulus. The circalunar cycle varied the hatching rate according to the moon phase, even in laboratory conditions, and the addition of succussed water into the medium reduced the hatching rate, even with different intensities according to the moon phase. The organization of nano and microbubbles generated after the succussion of water could be related to this protective effect and can explain, at least partially the effects of high diluted preparations on this biological context. Conclusion:Environmental factors, such as the circalunar cycle and products of water agitation, can modulate the adaptative control of hatching in Artemia salina exposed to lead chloride at EC10.

Characterization of physicochemical markers for homeopathic medicines and biological supernatant samples

Solvatochromic dyes are probes to detect variations on the dipole moment of solvents after the insertion of homeopathic potencies. Recent studies have shown they can be useful tools in laboratory and field studies to detect the activity of homeopathic remedies.Objective: Determine whether solvatochromic dyes can be a diagnostic tool for cells infected by different agents and/or markers to identify the activity of homeopathic medicines. Methods: Ethilicum1cH, Siliceaterra6, 30, 200cH; Zincummetallicum6, 30, 200cH and Phosphorus6, 30 and 200cH were analyzed by pouring the samples (in a 1:60 rate) into a series of seven dyes (rhodamine, ET 33, ET 30, coumarin 7, NN DMIA, Nile red, methylene violet) diluted in absolute ethanol using pre-established working concentrations. Oscillations of dye absorbance were observed at visible light spectrophotometry according to the remedy and potency. Water and succussed water were used as controls. In a second moment, the absorbance profile of the remedies will be compared with those of biological samples (supernatants) and checked with the biological effect previously obtained from each treatment.Supernatants of RAW 264.7 macrophages stimulated by Calmette-Guérin bacilli (BCG) or infected with Encephalitozoon cuniculiwill be analyzed. Results: Preliminary results have shown that Siliceaterra6cH, Phosphorus30 and 200cH and Zincummetallicum6, 30 and 200cH reduced the absorbance of methylene violet (p=0.01). Repetitions and analysis of supernatants are expected to be performed in the next steps of the study. Future perspectives: Establish a pattern of reactivity of the studied medicines with different dyes and the putative relation with the corresponding supernatants, as an attempt to obtain a "physicochemical signature" for each kind of infection and/or treatment.

Mercury chloride toxicity attenuation of the Brine Shrimp Artemia Salinaafter treatment with Mercurius corrosivusas isotherapic

Isotherapics preparedfromtoxic substances have been described as attenuation factors for heavy metal intoxicationin aquatic animals. Herein, Artemia salinaand mercury chloride were usedas a model to identify treatment-related bioresilience. The aim was to describe the effects of Mercurius corrosivus(MC) in different potencies on Artemia salinacyst hatching and on mercury bioavailability. Artemia salinacysts were exposed to 5.0 µg/mL of mercury chloride during the hatching phase. MC6cH, 30cH, and 200cHwere prepared and poured into artificial seawater. Different controls were used (nonchallenged cysts and challenged cysts treated with water, succussed water, and Ethilicum 1cH). Four series of nine experiments were performed for4 weeks to evaluate the percentage of cyst hatchingconsidering all moon phases. Soluble total mercury (THg) levels and precipitated mercury content were also evaluated. Solvatochromic dyes were used to check for eventual physicochemical markers of MCbiological activity. Two-way analysis of variance (ANOVA) with mixed modelswas used for evaluating the effect of different treatments andthe simultaneous influence of the moon phases on the cystshatching rate, at both observation times (24 and 48 hours).When necessary, outliers were removed, using the Tukeycriterion.Thelevel of significance αwas set at 5%. Significant delay (p<0.0001) in cyst hatching was observed after treatment with MC30cH, compared with the controls. An increase inTHg concentration in seawater (p<0.0018) and of chlorine/oxygen ratio (p<0.0001) in suspended micro-aggregateswas also seen, with possiblerelation with mercury bioavailability. Specific interaction of MC30cH with the solvatochromic dye ET33 (p<0.0017) was found. The other observed potencies of Mercurius corrosivus6 and 200 cH were not significant in relation to the observed groups.The results werepostulated as being protective effects of MC30cH on Artemia salina, by improving its bioresilience.

Plant extract incorporated into glass ionomer cement as a photosensitizing agent for antimicrobial photodynamic therapy on Streptococcus mutans.

BACKGROUND: A plant extract (EB) incorporated into glass ionomer cement (GIG) could be a potential photosensitizer for Antimicrobial PDT (aPDT) against caries-microorganisms, replacing methylene blue (MB), due to the presence of chlorophyll. GIC + EB + aPDT could be an therapeutic alternative to dentin decontamination and sealing, allowing reduction of operative time. OBJECTIVE: Evaluate Dioscorea altissima (EB) incorporated into GIC as a photosensitizer for aPDT against Streptococcus mutans. METHODS: Groups (n = 24; ntotal = 192): G1-GIC; G2-GIC + LASER; G3-GIC/EB; G4-GIC/EB + LASER; G5-GIC+MB; G6-GIC + aPDT; G7-GIC/EB + MB; and G8 - GIC/EB+aPDT. In aPDT groups, MB was the photosensitizer. In LASER groups, MB was not used. The irradiation protocol was 660 nm/100 mW/5 J/150 J/cm²/50 s, with a 5-min pre-irradiation time for the MB groups. Antibacterial assays were carried out in 24-well microplates. The wells were completed with one milliliter of a S. mutans in BHI at 1.3 × 108 CFU/mL suspension. After incubation, PDT or laser was performed. After MTT bacteria viability test, the data were submitted to the Kolgomorov-Smirnoff normality test, followed by one-way ANOVA and Tukey's posterior test, α < 0.05. RESULTS: Group G6 showed significant inhibition (p < 0.001), followed by groups G4, G5, G7, and G8, which did not show significant differences among them (p > 0.05). Groups G2 and G3 also showed similar results (p > 0.05) and were the least active compared to the others. CONCLUSIONS: EB potentiated the antimicrobial action of GIC against S. mutans and laser irradiation over GIC/EB presented better antimicrobials results. The results indicate that EB could be a potential photosensitizer for aPDT.

Bioresilience to Mercury Chloride of the Brine Shrimp Artemia Salina after Treatment with Homeopathic Mercurius Corrosivus.

INTRODUCTION: Finding solutions to mitigate the impact of pollution on living systems is a matter of great interest. Homeopathic preparations of toxic substances have been described in the literature as attenuation factors for intoxication. Herein, an experimental study using Artemia salina and mercury chloride was developed as a model to identify aspects related to bioresilience. AIMS: The aim of the study was to describe the effects of homeopathic Mercurius corrosivus (MC) on Artemia salina cysts hatching and on mercury bioavailability. METHODS: Artemia salina cysts were exposed to 5.0 µg/mL of mercury chloride during the hatching phase. MC potencies (6cH, 30cH, and 200cH) were prepared in sterile purified water and poured into artificial sea water. Different controls were used (non-challenged cysts and challenged cysts treated with water, succussed water, and Ethilicum 1cH). Four series of nine experiments were performed to evaluate the percentage of cyst hatching. Soluble total mercury (THg) levels and precipitated mercury content were also evaluated. Solvatochromic dyes were used to check for eventual physicochemical markers of MC biological activity. RESULTS: Significant delay (p < 0.0001) in cyst hatching was observed only after treatment with MC 30cH, compared with controls. This result was associated with an increase of THg concentration in water (p = 0.0018) and of chlorine/oxygen ratio (p < 0.0001) in suspended micraggregates, suggesting changes in mercury bioavailability. A specific interaction of MC 30cH with the solvatochromic dye ET33 (p = 0.0017) was found. CONCLUSION: Changes in hatching rate and possible changes in Hg bioavailability are postulated as protective effects of MC 30cH on Artemia salina, by improving its natural bioresilience processes.

Ultraviolet C as a method of disinfecting medical silicone used in facial prostheses: An in vitro study.

STATEMENT OF PROBLEM: Hygiene and disinfection are important factors for preserving facial prostheses and supporting tissue health. However, a method that does not accelerate degradation or color change is necessary. PURPOSE: The purpose of this in vitro study was to evaluate the effectiveness of irradiation with ultraviolet C light-emitting diode (UV-C LED) light in the disinfection and initial color stability of the silicone (A-588-1; Factor II) used in facial prostheses. MATERIAL AND METHODS: One hundred and twenty specimens were made, contaminated by multispecies biofilm, and divided into 5 groups (n=24) with different treatments: control, distilled water, 0.12% chlorhexidine, UV-C LED light, and dimethyl sulfoxide (DMSO) as the negative control. Cell viability was measured by the methyl tetrazolium salt method. Statistical analysis was performed by generalized linear models. Additional descriptive analysis was performed for color analysis by using 16 silicone specimens made with light and dark intrinsic coloring in 4 groups (controls and treatments n=4) submitted to UV-C LED light. The ΔE of the specimens was obtained by CIEDE200. RESULTS: The results of cell viability demonstrated a statistically significant difference among the groups (P<.001), with a microbial reduction after UVC-LED exposure compared with the control group. Regarding the color, the groups presented an average ΔE (light 0.205 and dark 0.308) compatible with visually imperceptible changes (light <0.7 and dark <1.2). CONCLUSIONS: Irradiation with UV-C LED light decreased the in vitro microbial cell viability of the medical silicone used in facial prostheses, demonstrating initial color stability.

Luffa operculata at a late period of gestation dysregulates melatonin and cytokines interfering with weight of dams and their male offspring.

ETHNOPHARMACOLOGICAL RELEVANCE: The tea made with the fruits of Luffa operculata (L.) Cogn. (Cucurbitaceae; EBN) is popularly used as abortive. AIM OF THE STUDY: The present work aimed at accessing how the exposition of female Wistar rats to 1.0 mg/kg of EBN (experimental group, EG), or distilled water (control group, CG), by gavage, at gestational days (GD) 17-21 interfered with the reproductive performance, and with dams' behavior after weaning. MATERIALS AND METHODS: At post-natal day 2 (PND2), the number of male and female pups was evaluated, as well as their weight. After weaning (PND21), dams were euthanized, and their liver and kidneys were removed for histological and biochemical analyses, while the blood was used in the evaluation of cytokines IL-1α, IL-1ß, IL-6 and TNF-α, corticosterone, adrenocorticotrophic hormone, melatonin, AST, ALT and creatinine levels. RESULTS AND DISCUSSION: Dams that were treated with EBN showed an anxiety-like behavior, weight loss at the end of gestation and weight gain at weaning, accompanied with a significant decrease in pro-inflammatory cytokines and in the melatonin level. No significant histological or biochemical alterations have occurred in the liver or kidneys. The number of female pups was significantly higher in the EG. The male pups showed weight gain at PND60. CONCLUSION: The presence of cucurbitacins is probably involved in the dysregulations that were found, due to their polycyclic steroid triterpene structure.

Vertical exposition to Luffa operculata extract deregulates behavior and hypothalamus neurotransmitters in juvenile rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Luffa operculata (L.) Cogn (Cucurbitaceae) is a traditional plant popularly used in the abortion induction, against sinusitis and is toxic. AIM OF THE STUDY: To verify the influence of the aqueous extract obtained from the dry fruit of L. operculata (BNE) on the male rats vertically exposed to a subabortive dose of BNE, by evaluating alterations in behavior and neurochemical features in hypothalamus, striatum and frontal cortex, at a juvenile age, after receiving a stress challenge given by the use of the "New York subway stress" technique (NYS). MATERIALS AND METHODS: Pregnant female rats (F0 generation) received 1.0 mg/kg BNE, or distilled water (100 mL/kg), by gavage, between gestation days GD17 and GD21. The pups were weaned at PND21 and were kept up to PND60 (juvenile age) in controlled environmental conditions. Four groups were obtained: control (CG), experimental (EG), stress control (SCG) and stress experimental (SEG) After being stressed, the animals were behavioral screened for in the open field (OF) and in light-dark box (LDB) apparatuses. They were euthanized, and the liver, kidneys and brain were removed for both macroscopic and microscopic analyses, and for quantification of vanillylmandelic acid (VMA), norepinephrine (NE), dopamine (DA) and its metabolite, 3,4-dihydroxyphenylacetic acid (DOPAC) and the serotonin (5-HT) and its metabolite 5-hydroxyindolylacetic acid (5-HIAA) were accessed in the hypothalamus, frontal cortex and striatum. RESULTS AND DISCUSSION: although most of the behavior changes were due to the stress challenge, the rats spent more time in the dark side of the LDB and were less likely to explore the light side, indicating that the treatment with BNE induced to fear. Interferences of BNE over behavior were due to impairment of VMA, NE, 5-HT and DA and increasing of DOPAC in the hypothalamus, and an increase of 5-HIAA in the frontal cortex, indicating alterations in the hypothalamic-hypophysis-adrenal axis (HHAA). No macroscopic or histopathological changes were observed in the liver, kidneys, or brain, although GFAP was diminished in the SCG, as expected for stressed rats. CONCLUSION: the vertical exposition of juvenile rats to BNE led to the manifestation of fear and to a down regulation of the hypothalamic-hypophysis-adrenal axis.

Susceptibility of virulent and resistant Escherichia coli strains to non-polar and polar compounds identified in Microplumeria anomala.

BACKGROUND AND AIM: Escherichia coli is one of the main pathogens responsible for veterinary and human infections, and it is associated with significant economic losses in the livestock, as it causes severe diseases to humans, particularly in children. For that reason, there is a need for introducing new drugs to treat E. coli diseases. The Brazilian species richness is a source of potential new antibacterial natural products. The study aimed at the biological and chemical investigation of the organic extract obtained from the stem of Microplumeria anomala (Apocynaceae), EB127, as it was identified as a potential source of new antibacterial compounds to be used in Veterinary. MATERIALS AND METHODS: The antibacterial activity was evaluated by disk diffusion and microdilution assays; chromatography, nuclear magnetic resonance spectrometry, and mass spectrometry were used in the isolation and identification of compounds. RESULTS: EB127 showed activity against E. coli ATCC25922, and against three E. coli strains that were isolated from frigarte's cloaca, named 31/1A, 35A, and 51A. Lupeol, 3-acetyl-11-oxo-ß-amyrin, 3-acetyl-11-oxo-α-amyrin, sitosterol, stigmasterol, 3ß,7α-dihydroxy-cholest-5-ene, 3ß-hydroxy-cholest-5-en-7-one, and 3ß-hydroxy-cholest-5,22-dien-7-one were identified in fraction Hex/CHCl3, while loganin, loganic acid, methylanomaline, and anomaline were all identified in EB127 and protocatechuic acid hexoside, ferulic acid, secoxyloganin, feruloylquinic acid, vanillic acid hexoside, protocatechuic acid-4-O-ß-hexoside, and rosmarinic acid were tentatively identified in fraction 10%ACN/H2O. E. coli 51A (virulent/non-resistant) showed sensitivity to the antibacterial action of fraction Hex/CHCl3 which contains alkaloids, triterpenes, and steroids, while E. coli 35A (resistant/non-virulent) were more susceptible to 10%ACN/H2O, which contains iridoids as loganin and loganic acid, and glycosylated and non-glycosylated caffeic acids. CONCLUSION: Fraction 10%ACN/H2O is of interest in pursuing new drugs to treat resistant E. coli, in veterinary. All compounds were isolated from the plant for the first time and have shown potential as new antibacterial natural products from Amazon plants to be used in veterinary and human diseases.

Interaction between Solvatochromic Dyes and Water Sampled from a Natural Source Treated with High Dilutions of Phosphorus.

BACKGROUND: Highly diluted and succussed solutions interact with solvatochromic dyes, indicating that changes in solvent and solute polarity could be related to their mechanism of action. It is not known, however, how the activity associated with succussed high dilutions is transferred to untreated water and what the limits of this process are. AIMS: The aims of the present study were to ascertain whether a succussed high dilution of phosphorus (1.5 × 1-59 M; Phos 30cH) seeded into a natural water source that fed a fjord and two connected lakes could propagate itself through the lake system (total volume 2200 m3) and, moreover, whether the process could be tracked using solvatochromic dyes. METHODS: Samples of water were collected before and after seeding, at different times and places throughout the lake system. Controls comprised water taken from an untreated and adjacent, but independent, lake (1385 m3). RESULTS: Water samples taken up to 72 hours after the source treatment produced significant increases (p ≤ 0.03) in the absorbance of the solvatochromic dye methylene violet (MV), while samples from the control lake produced no changes. CONCLUSIONS: The study indicates that activity associated with Phos 30c can propagate itself through large volumes of water, causing changes throughout a whole connected lake system, and that these changes can be tracked using the solvatochromic dye MV. This in turn means the use of homeopathic medicines in large volumes of drinking water, in farming and ecological contexts, now has the potential to be assessed with physico-chemical monitoring.

In Vitro Analysis of the Implant-Abutment Interface Connection and Bacterial Infiltration in Two Extraoral Implant Models.

PURPOSE: To compare the connection microgaps and the bacterial infiltration of implant-abutment interfaces of two extraoral implant models. MATERIALS AND METHODS: Two implant models were used: the inner connection and the flush connection types. The implant-abutment microgaps of five sets of each extraoral implant were evaluated with scanning electron microscopy. Eleven additional sets of each model design were immersed in Staphylococcus aureus cultures for 24 hours, and samples were obtained from the external surface and from the implant's internal chamber to quantify the colony-forming units. RESULTS: Scanning electron microscopy analysis showed that microgaps of the flush connection were smaller compared with the inner connection (P < .0001), and that bacterial counts were higher at the inner connection compared with the flush connection (P < .0001). CONCLUSION: Within the limitations of this experimental study, it can be concluded that the flush connection model presented a smaller microgap and fewer bacterial colonies compared with the inner connection model.

Characterization of Antimonium crudum Activity Using Solvatochromic Dyes.

BACKGROUND: The mechanism by which highly diluted and agitated solutions have their effect is still unknown, but the development in recent years of new methods identifying changes in water and solute dipole moments is providing insights into potential modes of action. OBJECTIVE: The objective of the current study was to compare the biological effects of Antimonium crudum (AC) previously obtained by our group and already described in the literature with now measurable physico-chemical effects on solvatochromic dyes. METHODS: Different dilutions of AC and succussed water have been characterized with respect to their effect on the visible spectra of the solvatochromic dyes methylene violet (MV), a pyridinium phenolate (ET33), and a dimethylamino naphthalenone (BDN) compared with in-vitro action against Leishmania amazonensis-infected macrophages. RESULTS: Dye responses varied according to the dye used and the level of AC dilution and results were found to corroborate previously published in-vivo and in-vitro effects of AC. In addition, a very significant enhancement in the absorbance increase of MV was seen using the supernatant from AC 200cH-treated cells (15%; p < 0.0001) over that seen with AC 200cH itself (4%; p = 0.034), suggesting the amplification of ultra-high dilution effects by biological systems. Furthermore, supernatants from AC-treated cells increased the range of dilutions of AC that were capable of producing effects on the spectra of MV. The effect of AC dilutions on dye ET33 was eliminated by a weak electric current passed through potency solutions. CONCLUSION: The data confirm a correspondence between the biological effects of dilutions of AC in-vitro and physico-chemical effects on solvatochromic dyes as measured by changes in their visible spectra. Results also indicate high dilutions of AC are sensitive to exposure to electric currents and biological systems.

Amazon climatic factors driving terpene composition of Iryanthera polyneura Ducke in terra-firme forest: A statistical approach.

Iryanthera polyneura (Myristicaceae) is popularly known as cumala-colorada, and can be found in the Amazon rain forest. The present study aimed the evaluation of the chemical composition of the essential oils (EOs) obtained from the leaves of I. polyneura throughout a two-year period in order to correlate chemical findings with seasonality and climatic variation. Leaves from I. polyneura were collected 15 times from three different individuals, identified as 22EO, 80EO and 530EO, between the years of 2009 and 2011. The EOs were obtained and tested by gas-chromatography-mass spectrometry techniques. ANOVA and multivariate analyses were used to assess the relationship between the percentage of terpenes and seasonal/climatic variations. Fifty-nine compounds were detected in the EOs, of which 44 were identified (74.5%). The main components of the EOs were spathulenol (6.42 ± 1.02%), α-cadinol (5.82 ± 0.40%) and τ-muurolol (5.24 ± 0.03%). Higher levels of spathulenol were observed in 22EO during rainy season, while τ-muurolol occurred in high amounts during the dry season, as did α-cadinol in 22EO and 80EO. Correspondence analysis revealed a distinction between two groups of EOs based on seasonality, whereas a canonic correspondence analysis and ANOVA determined how the major compounds were related to both seasonality and climatic factors. Non-metric multidimensional scaling in association with an analysis of similarities showed that the abundance and composition of terpenes in the samples varied moderately among the three Iryanthera individual plants. Present findings have shown that there is variation in the occurrence of the major compounds spathulenol, τ-muurolol and α-cadinol produced by I. polyneura and that the pattern of variation is related to season and climatic changes.