Antifungal and Antiparasitic Activities of Metallocene-Containing Fluconazole Derivatives.

The search for new anti-infectives based on metal complexes is gaining momentum. Among the different options taken by researchers, the one involving the use of organometallic complexes is probably the most successful one with a compound, namely, ferroquine, already in clinical trials against malaria. In this study, we describe the preparation and in-depth characterization of 10 new (organometallic) derivatives of the approved antifungal drug fluconazole. Our rationale is that the sterol 14α-demethylase is an enzyme part of the ergosterol biosynthesis route in Trypanosoma and is similar to the one in pathogenic fungi. To demonstrate our postulate, docking experiments to assess the binding of our compounds with the enzyme were also performed. Our compounds were then tested on a range of fungal strains and parasitic organisms, including the protozoan parasite Trypanosoma cruzi (T. cruzi) responsible for Chagas disease, an endemic disease in Latin America that ranks among some of the most prevalent parasitic diseases worldwide. Of high interest, the two most potent compounds of the study on T. cruzi that contain a ferrocene or cobaltocenium were found to be harmless for an invertebrate animal model, namely, Caenorhabditis elegans (C. elegans), without affecting motility, viability, or development.

Human Embryonic Stem Cell-Derived Immature Midbrain Dopaminergic Neurons Transplanted in Parkinsonian Monkeys.

Human embryonic stem cells (hESCs) differentiate into specialized cells, including midbrain dopaminergic neurons (DANs), and Non-human primates (NHPs) injected with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine develop some alterations observed in Parkinson's disease (PD) patients. Here, we obtained well-characterized DANs from hESCs and transplanted them into two parkinsonian monkeys to assess their behavioral and imaging changes. DANs from hESCs expressed dopaminergic markers, generated action potentials, and released dopamine (DA) in vitro. These neurons were transplanted bilaterally into the putamen of parkinsonian NHPs, and using magnetic resonance imaging techniques, we calculated the fractional anisotropy (FA) and mean diffusivity (MD), both employed for the first time for these purposes, to detect in vivo axonal and cellular density changes in the brain. Likewise, positron-emission tomography scans were performed to evaluate grafted DANs. Histological analyses identified grafted DANs, which were quantified stereologically. After grafting, animals showed signs of partially improved motor behavior in some of the HALLWAY motor tasks. Improvement in motor evaluations was inversely correlated with increases in bilateral FA. MD did not correlate with behavior but presented a negative correlation with FA. We also found higher 11C-DTBZ binding in positron-emission tomography scans associated with grafts. Higher DA levels measured by microdialysis after stimulation with a high-potassium solution or amphetamine were present in grafted animals after ten months, which has not been previously reported. Postmortem analysis of NHP brains showed that transplanted DANs survived in the putamen long-term, without developing tumors, in immunosuppressed animals. Although these results need to be confirmed with larger groups of NHPs, our molecular, behavioral, biochemical, and imaging findings support the integration and survival of human DANs in this pre-clinical PD model.

Terapia de extinción y estimulación magnética transcraneal para el tratamiento de trastornos de ansiedad / Extinction therapy and transcranial magnetic stimulation for the treatment of anxiety disorders

Resumen Objetivo: Describir los factores que pueden determinar la reducción de los síntomas en el trastorno de ansiedad generalizada y trastorno por estrés postraumático, mediante estimulación magnética transcraneal en combinación con terapia de extinción. Material y Métodos: Se realizó una búsqueda en bases de datos (Cochrane, EBSCO, Pubmed, Sciencedirect y Wiley), con las palabras clave "transcranial magnetic stimulation", "human", "fear extinction". Los criterios de selección incluyen estudios en humanos, tratamientos con terapia de extinción y EMT, en donde se registre la conductancia de la piel como variable de respuesta. Resultados: Existe poca investigación que cumpla con los criterios de la presente revisión bibliográfica. Se obtuvieron 5 artículos enfocados en el tratamiento de síntomas como el miedo y la recurrencia de recuerdos traumáticos. Los protocolos de estimulación son heterogéneos, la frecuencia de estimulación va de 1 Hz a 30 Hz. La estimulación de alta frecuencia fue la más utilizada. La duración máxima de los efectos reportados fue de 1 mes. Conclusiones: La EMT junto con la terapia de extinción como tratamiento para TEPT y TAG es un campo de estudio que requiere de más investigación. Los resultados sobre su eficacia no son concluyentes, el tamaño de muestra es pequeño y es necesario identificar qué protocolos son eficaces a largo plazo. Los estudios clínicos con pacientes que presenten estos trastornos son relevantes para conocer los efectos de aquellos protocolos que han sido exitosos en pacientes sanos (condicionados al miedo).

Abstract Objective: To describe the factors that can determine the reduction of symptoms in generalized anxiety and posttraumatic stress disorders by transcranial magnetic stimulation in combination with extinction therapy. Material and methods: A bibliographic review was conducted in databases (Cochrane, EBSCO, PubMed, ScienceDirect y Wiley), using the keywords: "transcranial magnetic stimulation", "human" and "fear extinction". A selection of clinical trials that used extinction therapy plus TMS and the skin conductance as variable quantified was made. Results: Five articles focused on the treatment of symptoms, like fear and recurrence of traumatic memories were obtained. There is little research on the topic. Stimulation protocols are heterogeneous between studies (stimulation frequency ranges from 1 to 30 Hz). Most of the studies reviewed reported the use of high-frequency stimulation. The maximum duration of therapeutic effects reported was one month. Conclusions: TMS and extinction therapy as a treatment for PTSD and GAD has a growing research field. Effectiveness results are not conclusive, sample sizes are small, and studies do not focus on which protocols are effective in the long-term. New studies that include patients with diagnosed PTSD and GAD are relevant to assess the protocols that have already been successful in healthy patients (fear-conditioned).

Platinum and Palladium Organometallic Compounds: Disrupting the Ergosterol Pathway in Trypanosoma cruzi.

Current treatment for Chagas' disease is based on two drugs, Nifurtimox and Benznidazol, which have limitations that reduce the effectiveness and continuity of treatment. Thus, there is an urgent need to develop new, safe and effective drugs. In previous work, two new metal-based compounds with trypanocidal activity, Pd-dppf-mpo and Pt-dppf-mpo, were fully characterized. To unravel the mechanism of action of these two analogous metal-based drugs, high-throughput omics studies were performed. A multimodal mechanism of action was postulated with several candidates as molecular targets. In this work, we validated the ergosterol biosynthesis pathway as a target for these compounds through the determination of sterol levels by HPLC in treated parasites. To understand the molecular level at which these compounds participate, two enzymes that met eligibility criteria at different levels were selected for further studies: phosphomevalonate kinase (PMK) and lanosterol 14-α demethylase (CYP51). Molecular docking processes were carried out to search for potential sites of interaction for both enzymes. To validate these candidates, a gain-of-function strategy was used through the generation of overexpressing PMK and CYP51 parasites. Results here presented confirm that the mechanism of action of Pd-dppf-mpo and Pt-dppf-mpo compounds involves the inhibition of both enzymes.

Transcriptomic analysis of the adaptation to prolonged starvation of the insect-dwelling Trypanosoma cruzi epimastigotes.

Trypanosoma cruzi is a digenetic unicellular parasite that alternates between a blood-sucking insect and a mammalian, host causing Chagas disease or American trypanosomiasis. In the insect gut, the parasite differentiates from the non-replicative trypomastigote forms that arrive upon blood ingestion to the non-infective replicative epimastigote forms. Epimastigotes develop into infective non-replicative metacyclic trypomastigotes in the rectum and are delivered via the feces. In addition to these parasite stages, transitional forms have been reported. The insect-feeding behavior, characterized by few meals of large blood amounts followed by long periods of starvation, impacts the parasite population density and differentiation, increasing the transitional forms while diminishing both epimastigotes and metacyclic trypomastigotes. To understand the molecular changes caused by nutritional restrictions in the insect host, mid-exponentially growing axenic epimastigotes were cultured for more than 30 days without nutrient supplementation (prolonged starvation). We found that the parasite population in the stationary phase maintains a long period characterized by a total RNA content three times smaller than that of exponentially growing epimastigotes and a distinctive transcriptomic profile. Among the transcriptomic changes induced by nutrient restriction, we found differentially expressed genes related to managing protein quality or content, the reported switch from glucose to amino acid consumption, redox challenge, and surface proteins. The contractile vacuole and reservosomes appeared as cellular components enriched when ontology term overrepresentation analysis was carried out, highlighting the roles of these organelles in starving conditions possibly related to their functions in regulating cell volume and osmoregulation as well as metabolic homeostasis. Consistent with the quiescent status derived from nutrient restriction, genes related to DNA metabolism are regulated during the stationary phase. In addition, we observed differentially expressed genes related to the unique parasite mitochondria. Finally, our study identifies gene expression changes that characterize transitional parasite forms enriched by nutrient restriction. The analysis of the here-disclosed regulated genes and metabolic pathways aims to contribute to the understanding of the molecular changes that this unicellular parasite undergoes in the insect vector.

Copper(I)-Thiosemicarbazone Complexes with Dual Anticancer and Antiparasitic Activity.

Four new Cu(I) complexes of the general formula [Cu(PP)(LL)][BF4 ], in which PP is a phosphane ligand (triphenylphosphane or 1,2-bis(diphenylphosphano)ethane (dppe)) and LL is a bioactive thiosemicarbazone ligand (4-(methyl)-1-(5-nitrofurfurylidene)thiosemicarbazone) or 4-(ethyl)-1-(5-nitrofurfurylidene)thiosemicarbazone) were synthesized and fully characterized by classical analytical and spectroscopic methods. The anti-trypanosome and anticancer activities were investigated in vitro on Trypanosoma cruzi and in two human cancer cell lines (ovarian OVCAR3 and prostate PC3). To test the selectivity toward parasites and cancer cells, the cytotoxicity on normal monkey kidney VERO and human dermal fibroblasts HDF cells was also evaluated. The new heteroleptic complexes were more cytotoxic on T. cruzi and chemoresistant prostate PC3 cells than the benchmark drugs nifurtimox and cisplatin. The compounds also showed a high level of cellular internalization by the OVCAR3 cells and, in particular, those containing the dppe phosphane showed activation of the cell death mechanism via apoptosis. On the other hand, the production of reactive oxygen species induced by these complexes was not evident.

Multifunctional organometallic compounds for the treatment of Chagas disease: Re(I) tricarbonyl compounds with two different bioactive ligands.

Chagas' disease (American Trypanosomiasis) is an ancient and endemic illness in Latin America caused by the protozoan parasite Trypanosoma cruzi. Although there is an urgent need for more efficient and less toxic chemotherapeutics, no new drugs to treat this disease have entered the clinic in the last decades. Searching for metal-based prospective antichagasic drugs, in this work, multifunctional Re(I) tricarbonyl compounds bearing two different bioactive ligands were designed: a polypyridyl NN derivative of 1,10-phenanthroline and a monodentate azole (Clotrimazole CTZ or Ketoconazol KTZ). Five fac-[Re(CO)3(NN)(CTZ)](PF6) compounds and a fac-[Re(CO)3(NN)(KTZ)](PF6) were synthesized and fully characterized. They showed activity against epimastigotes (IC50 3.48-9.42 µM) and trypomastigotes of T. cruzi (IC50 0.61-2.79 µM) and moderate to good selectivity towards the parasite compared to the VERO mammalian cell model. In order to unravel the mechanism of action of our compounds, two potential targets were experimentally and theoretically studied, namely DNA and one of the enzymes involved in the parasite ergosterol biosynthetic pathway, CYP51 (lanosterol 14-α-demethylase). As hypothesized, the multifunctional compounds shared in vitro a similar mode of action as that disclosed for the single bioactive moieties included in the new chemical entities. Additionally, two relevant physicochemical properties of biological interest in prospective drug development, namely lipophilicity and stability in solution in different media, were determined. The whole set of results demonstrates the potentiality of these Re(I) tricarbonyls as promising candidates for further antitrypanosomal drug development.

New multifunctional Ru(II) organometallic compounds show activity against Trypanosoma brucei and Leishmania infantum.

Human African trypanosomiasis (sleeping sickness) and leishmaniasis are prevalent zoonotic diseases caused by genomically related trypanosomatid protozoan parasites (Trypanosoma brucei and Leishmania spp). Additionally, both are co-endemic in certain regions of the world. Only a small number of old drugs exist for their treatment, with most of them sharing poor safety, efficacy, and pharmacokinetic profiles. In this work, new multifunctional Ru(II) ferrocenyl compounds were rationally designed as potential agents against these trypanosomatid parasites by including in a single molecule 1,1'-bis(diphenylphosphino)ferrocene (dppf) and two bioactive bidentate ligands: 8-hydroxyquinoline derivatives (8HQs) and polypyridyl ligands (NN). Three [Ru(8HQs)(dppf)(NN)](PF6) compounds were synthesized and fully characterized. They showed in vitro activity on bloodstream Trypanosoma brucei (IC50 140-310 nM) and on Leishmania infantum promastigotes (IC50 3.0-4.8 µM). The compounds showed good selectivity towards T. brucei in respect to J774 murine macrophages as mammalian cell model (SI 15-38). Changing hexafluorophosphate counterion by chloride led to a three-fold increase in activity on both parasites and to a two to three-fold increase in selectivity towards the pathogens. The compounds affect in vitro at least the targets of the individual bioactive moieties included in the new chemical entities: DNA and generation of ROS. The compounds are stable in solution and are more lipophilic than the free bioactive ligands. No clear correlation between lipophilicity, interaction with DNA or generation of ROS and activity was detected, which agrees with their overall similar anti-trypanosoma potency and selectivity. These compounds are promising candidates for further drug development.

Do bioactive 8-hydroxyquinolines oxidovanadium(IV) and (V) complexes inhibit the growth of M. smegmatis?

The antiproliferative effects of four series of VIVO- and VVO-based compounds containing 8-hydroxyquinoline ligands on the bacterium Mycolicibacterium smegmatis (M. smeg) were investigated. The effects on M. smeg were compared to the antiproliferative effects on the protozoan parasite Trypanosoma cruzi (T. cruzi), the causative agent for Chagas disease. In this study, we investigate the speciation of these compounds under physiological conditions as well as the antiproliferative effects on the bacterium M. smeg. We find that the complexes are more stable the less H2O is present, and that the stability increases in lipid-like environments. Only one heteroleptic complex and two homoleptic complexes were found to show similar antiproliferative effects on M. smeg as reported for T. cruzi so the responses generally observed by M.smeg. is less than observed by the pathogen. In summary, we find that M. smeg is more sensitive to the detailed structure of the V-complex but overall these complexes are less effective against M. smeg compared to T. cruzi.

Comparative analysis of striatal [18 F]FDOPA uptake in a partial lesion model of Parkinson's disease in rats: Ratio method versus graphical model.

Animal models of Parkinson's disease are useful to evaluate new treatments and to elucidate the etiology of the disease. Hence, it is necessary to have methods that allow quantification of their effectiveness. [18 F]FDOPA-PET (FDOPA-PET) imaging is outstanding for this purpose because of its capacity to measure changes in the dopaminergic pathway noninvasively and in vivo. Nevertheless, PET acquisition and quantification is time-consuming making it necessary to find faster ways to quantify FDOPA-PET data. This study evaluated Male Wistar rats by FDOPA, before and after being partially injured with 6-OHDA unilaterally. MicroPET scans with a duration of 120 min were acquired and Patlak reference plots were created to estimate the influx constant Kc in the striatum using the full dynamic scan data. Additionally, simple striatal-to-cerebral ratios (SCR) of short static acquisitions were computed and compared with the Kc values. Good correlation (r > 0.70) was obtained between Kc and SCR, acquired between 80-120 min after FDOPA administration with frames of 10 or 20 min and both methods were able to separate the FDOPA-uptake of healthy controls from that of the PD model (SCR -28%, Kc -71%). The present study concludes that Kc and SCR can be trustfully used to discriminate partially lesioned rats from healthy controls.

Application of microwave plasma atomic emission spectrometry in bioanalytical chemistry of bioactive rhenium compounds.

Five newly synthetized fac-Re(I) tricarbonyl compounds were explored as prospective antitrypanosomal agents. The biological activity of the whole series was evaluated preliminarily against the epimastigote form of Trypanosoma cruzi. All compounds showed activity against epimastigotes with IC50 values in the low micromolar range. The most active compound [fac-Re(I)(CO)3(tmp)(CTZ)](PF6), with CTZ = clotrimazole and tmp = 3,4,7,8-tetramethyl-1,10-phenantroline, showed good selectivity towards the parasites and thus was selected to carry out further metallomic studies. For this task, a newly bioanalytical method based on microwave plasma atomic emission spectrometry (MP-AES) was developed and validated. The accuracy of the method was ensured by testing a certified reference material. Results of rhenium elemental analysis by MP-AES agreed with the proposed formula of the studied compounds, contributing to the overall validation of the method, which was then applied to evaluate the percentage of rhenium uptaken by the parasites and the association of the compounds with parasite biomacromolecules. Metallomics results showed low total rhenium percentage uptaken by parasites (∼1.2%) and preferential accumulation in the soluble proteins fraction (∼82.8%). Thus, the method based on MP-AES turned out to be an economical and green alternative for metallomics studies involving potential rhenium metallodrugs. Moreover, a comparison against rhenium determination by electrothermal atomic absorption spectrometry (ET-AAS) was included.

To Resurface or Not to Resurface the Patella in Total Knee Arthroplasty, That Is the Question: A Meta-Analysis of Randomized Controlled Trials.

Background and Objetives: Currently, total knee arthroplasty is one of the most common surgeries, increasing with the increase in life expectancy. Whether or not to replace the patella has been a subject of debate over the years, remaining in controversy and without reaching a consensus. Over the years, different meta-analyses have been carried out in order to provide evidence on the subject, although, in recent times, there have not been many new studies in this regard. Therefore, it is considered necessary that the latest works form part of a new meta-analysis. Materials and Method: We searched the literature using PUBMED, SCOPUS, the Cochrane database and VHL from 2010 to 2020. The search terms used were "patellar" AND "resurfacing" OR "Replacement" and "no resurfacing" OR "no replacement". A meta-analysis was performed with Stata software (Stata version 15.1). Forest plots were generated to illustrate the overall effect of knee arthroplasty interventions. Results: As a result, it was obtained that there is a significantly higher risk of suffering AKP in the non-resurfacing group, in addition to a significant increase in the risk of undergoing a reoperation in the non-resurfacing group. On the other hand, significant differences were obtained in favor of the resurfacing group in both the clinical and Feller KSS, with functional KSS being inconclusive. After analyzing different variables throughout the literature, it does seem clear that the non-resurfacing group may present a higher risk of reoperation than the resurfacing group. Conclusions: For all these reasons, we think that, although it does seem that not replacing the patella can precipitate a reoperation, it is not clear whether this reoperation is a direct consequence of not having replaced the patella. Therefore, in our opinion, the treatment must be individualized for each patient.

Preclinical Studies and Drug Combination of Low-Cost Molecules for Chagas Disease.

Chagas disease is caused by the protozoan Trypanosoma cruzi (T. cruzi). It remains the major parasitic disease in Latin America and is spreading worldwide, affecting over 10 million people. Hundreds of new compounds with trypanosomicidal action have been identified from different sources such as synthetic or natural molecules, but they have been deficient in several stages of drug development (toxicology, scaling-up, and pharmacokinetics). Previously, we described a series of compounds with simple structures, low cost, and environmentally friendly production with potent trypanosomicidal activity in vitro and in vivo. These molecules are from three different families: thiazolidenehydrazines, diarylideneketones, and steroids. From this collection, we explored their capacity to inhibit the triosephosphate isomerase and cruzipain of T. cruzi. Then, the mechanism of action was explored using NMR metabolomics and computational molecular dynamics. Moreover, the mechanism of death was studied by flow cytometry. Consequently, five compounds, 314, 793, 1018, 1019, and 1260, were pre-clinically studied and their pharmacologic profiles indicated low unspecific toxicity. Interestingly, synergetic effects of diarylideneketones 793 plus 1018 and 793 plus 1019 were evidenced in vitro and in vivo. In vivo, the combination of compounds 793 plus 1018 induced a reduction of more than 90% of the peak of parasitemia in the acute murine model of Chagas disease.

Heteroleptic Oxidovanadium(V) Complexes with Activity against Infective and Non-Infective Stages of Trypanosoma cruzi.

Five heteroleptic compounds, [VVO(IN-2H)(L-H)], where L are 8-hydroxyquinoline derivatives and IN is a Schiff base ligand, were synthesized and characterized in both the solid and solution state. The compounds were evaluated on epimastigotes and trypomastigotes of Trypanosoma cruzi as well as on VERO cells, as a mammalian cell model. Compounds showed activity against trypomastigotes with IC50 values of 0.29-3.02 µM. IN ligand and the new [VVO2(IN-H)] complex showed negligible activity. The most active compound [VVO(IN-2H)(L2-H)], with L2 = 5-chloro-7-iodo-8-hydroxyquinoline, showed good selectivity towards the parasite and was selected to carry out further biological studies. Stability studies suggested a partial decomposition in solution. [VVO(IN-2H)(L2-H)] affects the infection potential of cell-derived trypomastigotes. Low total vanadium uptake by parasites and preferential accumulation in the soluble proteins fraction were determined. A trypanocide effect was observed when incubating epimastigotes with 10 × IC50 values of [VVO(IN-2H)(L2-H)] and the generation of ROS after treatments was suggested. Fluorescence competition measurements with DNA:ethidium bromide adduct showed a moderate DNA interaction of the complexes. In vivo toxicity study on C. elegans model showed no toxicity up to a 100 µM concentration of [VVO(IN-2H)(L2-H)]. This compound could be considered a prospective anti-T. cruzi agent that deserves further research.

Effect of extremely low frequency magnetic fields on oxidative balance in rat brains subjected to an experimental model of chronic unpredictable mild stress.

BACKGROUND: There has been an increasing interest in researching on the effects of extremely low-frequency magnetic fields on living systems. The mechanism of action of extremely low-frequency magnetic fields on organisms has not been established. One of the hypotheses is related to induce changes in oxidative balance. In this study, we measured the effects of chronic unpredictable mild stress induced-oxidative balance of rat's brain exposed to extremely low-frequency magnetic fields. METHODS: A first experiment was conducted to find out if 14 days of chronic unpredictable mild stress caused oxidative unbalance in male Wistar rat's brain. Catalase activity, reduced glutathione concentration, and lipoperoxidation were measured in cerebrum and cerebellum. In the second experiment, we investigate the effects of 7 days extremely low-frequency magnetic fields exposure on animals stressed and unstressed. RESULTS: The main results obtained were a significant increase in the catalase activity and reduced glutathione concentration on the cerebrum of animals where the chronic unpredictable mild stress were suspended at day 14 and then exposed 7 days to extremely low-frequency magnetic fields. Interestingly, the same treatment decreases the lipoperoxidation in the cerebrum. The stressed animals that received concomitant extremely low frequency magnetic fields exposure showed an oxidative status like stressed animals by 21 days. Thus, no changes were observed on the chronic unpredictable mild stress induced-oxidative damage in the rat's cerebrum by the extremely low-frequency magnetic field exposure together with chronic unpredictable mild stress. CONCLUSIONS: The extremely low-frequency electromagnetic field exposure can partially restore the cerebrum antioxidant system of previously stressed animals.

Antarctic Rahnella inusitata: A Producer of Cold-Stable ß-Galactosidase Enzymes.

There has been a recent increase in the exploration of cold-active ß-galactosidases, as it offers new alternatives for the dairy industry, mainly in response to the current needs of lactose-intolerant consumers. Since extremophilic microbial compounds might have unique physical and chemical properties, this research aimed to study the capacity of Antarctic bacterial strains to produce cold-active ß-galactosidases. A screening revealed 81 out of 304 strains with ß-galactosidase activity. The strain Se8.10.12 showed the highest enzymatic activity. Morphological, biochemical, and molecular characterization based on whole-genome sequencing confirmed it as the first Rahnella inusitata isolate from the Antarctic, which retained 41-62% of its ß-galactosidase activity in the cold (4 °C-15 °C). Three ß-galactosidases genes were found in the R. inusitata genome, which belong to the glycoside hydrolase families GH2 (LacZ and EbgA) and GH42 (BglY). Based on molecular docking, some of these enzymes exhibited higher lactose predicted affinity than the commercial control enzyme from Aspergillus oryzae. Hence, this work reports a new Rahnella inusitata strain from the Antarctic continent as a prominent cold-active ß-galactosidase producer.

Natural Pigments of Bacterial Origin and Their Possible Biomedical Applications.

Microorganisms are considered one of the most promising niches for prospecting, production, and application of bioactive compounds of biotechnological interest. Among them, bacteria offer certain distinctive advantages due to their short life cycle, their low sensitivity to seasonal and climatic changes, their easy scaling as well as their ability to produce pigments of various colors and shades. Natural pigments have attracted the attention of industry due to an increasing interest in the generation of new products harmless to humans and nature. This is because pigments of artificial origin used in industry can have various deleterious effects. On this basis, bacterial pigments promise to be an attractive niche of new biotechnological applications, from functional food production to the generation of new drugs and biomedical therapies. This review endeavors to establish the beneficial properties of several relevant pigments of bacterial origin and their relation to applications in the biomedical area.

Can changes in implant macrogeometry accelerate the osseointegration process?: An in vivo experimental biomechanical and histological evaluations.

OBJECTIVES: The propose was to compare this new implant macrogeometry with a control implant with a conventional macrogeometry. MATERIALS AND METHODS: Eighty-six conical implants were divided in two groups (n = 43 per group): group control (group CON) that were used conical implants with a conventional macrogeometry and, group test (group TEST) that were used implants with the new macrogeometry. The new implant macrogeometry show several circular healing cambers between the threads, distributed in the implant body. Three implants of each group were used to scanning electronic microscopy (SEM) analysis and, other eighty samples (n = 40 per group) were inserted the tibia of ten rabbit (n = 2 per tibia), determined by randomization. The animals were sacrificed (n = 5 per time) at 3-weeks (Time 1) and at 4-weeks after the implantations (Time 2). The biomechanical evaluation proposed was the measurement of the implant stability quotient (ISQ) and the removal torque values (RTv). The microscopical analysis was a histomorphometric measurement of the bone to implant contact (%BIC) and the SEM evaluation of the bone adhered on the removed implants. RESULTS: The results showed that the implants of the group TEST produced a significant enhancement in the osseointegration in comparison with the group CON. The ISQ and RTv tests showed superior values for the group TEST in the both measured times (3- and 4-weeks), with significant differences (p < 0.05). More residual bone in quantity and quality was observed in the samples of the group TEST on the surface of the removed implants. Moreover, the %BIC demonstrated an important increasing for the group TEST in both times, with statistical differences (in Time 1 p = 0.0103 and in Time 2 p < 0.0003). CONCLUSIONS: Then, we can conclude that the alterations in the implant macrogeometry promote several benefits on the osseointegration process.

High Throughput Approaches to Unravel the Mechanism of Action of a New Vanadium-Based Compound against Trypanosoma cruzi.

Treatment for Chagas disease, a parasitosis caused by Trypanosoma cruzi, has always been based on two drugs, nifurtimox and benznidazole, despite the toxic side effects described after prolonged prescription. In this work, we study a new prospective antitrypanosomal drug based on vanadium, here named VIVO(5Brsal)(aminophen). We found a good IC50 value, (3.76 ± 0.08) µM, on CL Brener epimastigotes. The analysis of cell death mechanism allowed us to rule out the implication of a mechanism based on early apoptosis or necrosis. Recovery assays revealed a trypanostatic effect, accompanied by cell shape and motility alterations. An uptake mostly associated with the insoluble fraction of the parasites was deduced through vanadium determinations. Concordantly, no drastic changes of the parasite transcriptome were detected after 6 h of treatment. Instead, proteomic analysis uncovered the modulation of proteins involved in different processes such as energy and redox metabolism, transport systems, detoxifying pathways, ribosomal protein synthesis, and proteasome protein degradation. Overall, the results here presented lead us to propose that VIVO(5Brsal)(aminophen) exerts a trypanostatic effect on T. cruzi affecting parasite insoluble proteins.

Comparative high-throughput analysis of the Trypanosoma cruzi response to organometallic compounds.

There is an urgent need to develop new drugs against Chagas' disease. In addition, the mechanisms of action of existing drugs have not been completely worked out at the molecular level. High throughput approaches have been demonstrated to be powerful tools not only for understanding the basic biology of Trypanosoma cruzi, but also for the identification of drug targets such as proteins or pathways that are essential for parasite infection and survival within the mammalian host. Here, we have applied these tools towards the discovery of the effects of two organometallic compounds with trypanocidal activity, Pd-dppf-mpo and Pt-dppf-mpo, on the transcriptome and proteome of T. cruzi epimastigotes. These approaches have not yet been reported for any other prospective metal-based anti T. cruzi drug. We found differentially expressed transcripts and proteins in treated parasites. Pd-dppf-mpo treatment resulted in more modulated transcripts (2327 of 10 785 identified transcripts) than Pt-dppf-mpo treatment (201 of 10 773 identified transcripts) suggesting a mechanism of action for Pd-dppf-mpo at the transcriptome level. Similar numbers of differentially expressed proteins (342 and 411 for Pd-dppf-mpo and Pt-dppf-mpo respectively) were also observed. We further functionally categorized differentially expressed transcripts and identified cellular processes and pathways significantly impacted by treatment with the compounds. Transcripts involved in DNA binding, protein metabolism, transmembrane transport, oxidative defense, and the ergosterol pathways were found to be modulated by the presence of the compounds. Our transcriptomic dataset also contained previously validated essential genes. These data allowed us to hypothesize a multimodal mechanism of action for the trypanocidal activity of Pd-dppf-mpo and Pt-dppf-mpo, and a differential contribution of the metal moiety of each compound.