United we stand, divided we fall: in-depth proteomic evaluation of the synergistic effect of Mo-CBP3-PepI and Ciprofloxacin against Staphylococcus aureus biofilms.

Staphylococcus aureus forms biofilms, a structure that protects bacterial cells, conferring more resistance to difficult treatment. Synthetic peptides surge as an alternative to overcome the biofilm of multidrug-resistant pathogens. Mo-CBP3-PepI, when combined with Ciprofloxacin, reduced preformed S. aureus biofilm by 50% at low concentrations (0.2 and 6.2 µg. mL-1, respectively). The goal of this study was to evaluate the proteomic profile of biofilms after treatment with the Mo-CBP3-PepI combined with ciprofloxacin. Here, proteomic analysis confirmed with more depth previously described mechanisms and revealed changes in the accumulation of proteins related to DNA and protein metabolism, cell wall biosynthesis, redox metabolism, quorum sensing, and biofilm formation. Some proteins related to DNA and protein metabolism were reduced, while other proteins, like redox system proteins, disappeared in Ciprofloxacin+Mo-CBP3-PepI treatment. Our results indicated a synergistic effect of these two molecules with several mechanisms against S. aureus biofilm and opened new doors for combined treatments with other drugs.

What Do the Transcriptome and Proteome of Menstrual Blood-Derived Mesenchymal Stem Cells Tell Us about Endometriosis?

Given the importance of menstrual blood in the pathogenesis of endometriosis and the multifunctional roles of menstrual mesenchymal stem cells (MenSCs) in regenerative medicine, this issue has gained prominence in the scientific community. Moreover, recent reviews highlight how robust the integrated assessment of omics data are for endometriosis. To our knowledge, no study has applied the multi-omics approaches to endometriosis MenSCs. This is a case-control study at a university-affiliated hospital. MenSCs transcriptome and proteome data were obtained by RNA-seq and UHPLC-MS/MS detection. Among the differentially expressed proteins and genes, we emphasize ATF3, ID1, ID3, FOSB, SNAI1, NR4A1, EGR1, LAMC3, and ZFP36 genes and MT2A, TYMP, COL1A1, COL6A2, and NID2 proteins that were already reported in the endometriosis. Our functional enrichment analysis reveals integrated modulating signaling pathways such as epithelial-mesenchymal transition (↑) and PI3K signaling via AKT to mTORC1 (↓ in proteome), mTORC1 signaling, TGF beta signaling, TNFA signaling via NFkB, IL6 STAT3 signaling, and response to hypoxia via HIF1A targets (↑ in transcriptome). Our findings highlight primary changes in the endometriosis MenSCs, suggesting that the chronic inflammatory endometrial microenvironment can modulate these cells, providing opportunities for endometriosis etiopathogenesis. Moreover, they identify challenges for future research leveraging knowledge for regenerative and precision medicine in endometriosis.

Tryptophan catabolites along the indoleamine 2,3-dioxygenase pathway as a biological link between depression and cancer.

Both depression and cancer are related to a dysregulation of inflammatory and immune pathways. Indeed, depression is associated with increased expression of interferon-γ, interleukin-1ß, and tumor necrosis factor α (TNF-α). In contrast, reductions of the activity of major histocompatibility complex protein molecules - class I and class II and natural killer cells are also observed. Similarly, cancers present elevated levels of TNF-α, reduced major histocompatibility complex class I and II, and natural killer cells. Indoleamine 2,3-dioxygenase (IDO), the rate-limiting enzyme of the tryptophan catabolite (TRYCAT) pathway, is induced by interferon-γ, interleukin-6, TNF-α, and oxidative stress. IDO catabolizes tryptophan, the amino acid precursor of serotonin and melatonin, to the metabolites collectively called TRYCATs. TRYCAT pathway activation is accompanied by downregulation of immune cell proliferation, function, and survival. The increase in IDO activity in tumor microenvironments is related to tumor cell escape from immune surveillance. Despite the evidence of inflammatory mechanisms underlying cancer and depression, it is important to emphasize that both diseases are heterogeneous and, as such, inflammatory mechanisms may not be relevant to all patients. Thus, the purpose of this review is to examine whether detrimental TRYCATs - synthesis of which increases in depression and cancer - are a pathophysiological link between the two diseases, and whether IDO is a potential pharmacological target for the treatment of the comorbid depression and cancer.

Synthetic Peptides Induce Human Colorectal Cancer Cell Death via Proapoptotic Pathways.

Cancer resistance to drugs and chemotherapy is a problem faced by public health systems worldwide. Repositioning antimicrobial peptides could be an efficient strategy to overcome that problem. This study aimed at repurposing antimicrobial peptides PepGAT and PepKAA for cancer treatment. After screening against several cancers, PepGAT and PepKAA presented IC50 values of 125.42 and 40.51 µM at 72 h toward colorectal cancer (CRC) cells. The mechanisms of action revealed that both peptides induced cell cycle arrest in G2/M and drove HCT-116 cells to death by triggering apoptosis. qPCR analysis revealed that peptides modulated gene expression in apoptosis, corroborating the data from caspase 3/7 and flow cytometry experiments. Yet, peptides induced ROS overaccumulation and increased membrane permeabilization, pore formation, and loss of internal content, leading to death. Additionally, peptides were able to inhibit cell invasion. Previous studies from the same group attested to no toxicity to normal human cells. Thus, PepGAT and PepKAA have great potential as anticancer molecules.

A Shortcut from Genome to Drug: The Employment of Bioinformatic Tools to Find New Targets for Gastric Cancer Treatment.

Gastric cancer (GC) is a highly heterogeneous, complex disease and the fifth most common cancer worldwide (about 1 million cases and 784,000 deaths worldwide in 2018). GC has a poor prognosis (the 5-year survival rate is less than 20%), but there is an effort to find genes highly expressed during tumor establishment and use the related proteins as targets to find new anticancer molecules. Data were collected from the Gene Expression Omnibus (GEO) bank to obtain three dataset matrices analyzing gastric tumor tissue versus normal gastric tissue and involving microarray analysis performed using the GPL570 platform and different sources. The data were analyzed using the GEPIA tool for differential expression and KMPlot for survival analysis. For more robustness, GC data from the TCGA database were used to corroborate the analysis of data from GEO. The genes found in in silico analysis in both GEO and TCGA were confirmed in several lines of GC cells by RT-qPCR. The AlphaFold Protein Structure Database was used to find the corresponding proteins. Then, a structure-based virtual screening was performed to find molecules, and docking analysis was performed using the DockThor server. Our in silico and RT-qPCR analysis results confirmed the high expression of the AJUBA, CD80 and NOLC1 genes in GC lines. Thus, the corresponding proteins were used in SBVS analysis. There were three molecules, one molecule for each target, MCULE-2386589557-0-6, MCULE-9178344200-0-1 and MCULE-5881513100-0-29. All molecules had favorable pharmacokinetic, pharmacodynamic and toxicological properties. Molecular docking analysis revealed that the molecules interact with proteins in critical sites for their activity. Using a virtual screening approach, a molecular docking study was performed for proteins encoded by genes that play important roles in cellular functions for carcinogenesis. Combining a systematic collection of public microarray data with a comparative meta-profiling, RT-qPCR, SBVS and molecular docking analysis provided a suitable approach for finding genes involved in GC and working with the corresponding proteins to search for new molecules with anticancer properties.

Modification of resting metabolism, body composition, and muscle strength after resolution of coronavirus disease 2019.

BACKGROUND & AIMS: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection causes changes that can influence human metabolism and modify the distribution of body compartments. We aimed to describe the clinical findings of changes in resting metabolism, muscle strength, and body composition in nonhospitalized patients after being diagnosed with coronavirus disease 2019 (COVID-19). METHODS: Physically active patients were evaluated at a nutrition clinic, and indirect calorimetry (IC) and body composition analysis using portable ultrasound were performed. After a routine appointment, all patients were instructed to inform the staff if they tested positive for SARS-CoV-2 infection. Our sample included individuals diagnosed with COVID-19, confirmed by real-time reverse transcription polymerase chain reaction (RT-PCR), within 7 days of the routine appointment. After an average incubation period of 14-21 days, in which there was no proven transmission of disease by RT-PCR, all of the patients were re-evaluated. RESULTS: A total of 38 volunteers (63.2% female) completed the study and were included in the analysis. The mean age of the participants was 37.3 ± 8.8 years. The comparison between pre- and post-COVID-19 stratified by sex demonstrated significant reduction in the RMR and RMR adjusted for weight (p < 0.0001) for both groups. Regarding body composition, there was a significant increase observed in fat mass in men (p < 0.002) and women (p < 0.01), and a significant reduction observed in fat-free mass (men: p < 0.002; women: p < 0.001) and skeletal muscle mass (men: p = 0.003; women: p < 0.0001). There was a significant difference between the change in the RMR measured by IC (p < 0.0001) and that calculated by the predictive equation of Cunningham (1980) (p < 0.0001), whereas the Harris and Benedict (1918) and Mifflin (1990) equations exhibited no difference. However, the mean difference in RMR between the post- and pre-COVID-19 calculated by the Cunningham equation was -40.4 kcal/day (95% confidence interval [CI]: -56.38 to -24.45), whereas the mean difference measured by IC was -362.3 kcal/day (95% CI: -452.7 to -271.9). CONCLUSION: This study describes the trends in the RMR, and body composition in individuals with COVID-19 who were not hospitalized from the pre-COVID-19 period to the post-COVID-19 period. A significant reduction in resting energy expenditure, and loss of fat-free mass and muscle mass in the post-COVID-19 period were observed in both men and women.

Experimental model of human corpus cavernosum smooth muscle relaxation.

PURPOSE: To describe a technique for en bloc harvesting of the corpus cavernosum, cavernous artery and urethra from transplant organ donors and contraction-relaxation experiments with corpus cavernosum smooth muscle. MATERIALS AND METHODS: The corpus cavernosum was dissected to the point of attachment with the crus penis. A 3 cm segment (corpus cavernosum and urethra) was isolated and placed in ice-cold sterile transportation buffer. Under magnification, the cavernous artery was dissected. Thus, 2 cm fragments of cavernous artery and corpus cavernosum were obtained. Strips measuring 3 x 3 x 8 mm(3) were then mounted vertically in an isolated organ bath device. Contractions were measured isometrically with a Narco-Biosystems force displacement transducer (model F-60, Narco-Biosystems, Houston, TX, USA) and recorded on a 4-channel Narco-Biosystems desk model polygraph. RESULTS: Phenylephrine (1 microM) was used to induce tonic contractions in the corpus cavernosum (3-5 g tension) and cavernous artery (0.5-1 g tension) until reaching a plateau. After precontraction, smooth muscle relaxants were used to produce relaxation-response curves (10(-12) M to 10(-4) M). Sodium nitroprusside was used as a relaxation control. CONCLUSION: The harvesting technique and the smooth muscle contraction-relaxation model described in this study were shown to be useful instruments in the search for new drugs for the treatment of human erectile dysfunction.

Relaxation of rabbit corpus cavernosum smooth muscle and aortic vascular endothelium induced by new nitric oxide donor substances of the nitrosyl-ruthenium complex.

INTRODUCTION: Endothelial dysfunction characterized by endogenous nitric oxide (NO) deficiency made 56% of patients affected with erectile dysfunction decline treatment with PDE-5 inhibitors. New forms of treatment are currently being developed for this group of patients. MATERIALS AND METHODS: The study compared the effect of sodium nitroprusside (SNP) and two substances of the nitrosyl-ruthenium complex, cis-[Ru(bpy)2(SO3)(NO)]PF-6-9 ("FONO1") and trans-[Ru(NH3)4(caffeine)(NO)]C13 ("LLNO1") on relaxation of rabbit corpus cavernosum smooth muscle and aortic vascular endothelium. The samples were immersed in isolated baths and precontracted with 0.1 microM phenylephrine (PE) and the corresponding relaxation concentration/response curves were plotted. In order to investigate the relaxation mechanisms involved, 100 microM ODQ (a soluble guanylate cyclase-specific inhibitor), 3 microM or 10 microM oxyhemoglobin (an extracellular NO scavenger) or 1 mM L-cysteine (a nitrosyl anion-specific scavenger) was added to the samples. RESULTS: All the NO donors tested produced a significant level of relaxation in the vascular endothelium. In corpus cavernosum samples, FONO1 produced no significant effect, but LLNO1 and SNP induced dose-dependent relaxation with comparable potency (pEC50 = 6.14 +/- 0.08 and 6.4 +/- 0.14, respectively) and maximum effect (Emax = 82% vs. 100%, respectively). All NO donors were found to activate soluble guanylate cyclase, since the addition of the corresponding inhibitor (100 microM ODQ) completely neutralized the relaxation effect observed. The addition of oxyhemoglobin reduced the relaxation effect, but did not inhibit it completely. In aortic vascular endothelium 3 microM oxyhemoglobin decreased the relaxation effect by 26% on the average, while 10 microM oxyhemoglobin reduced it by over 52%. The addition of 100 microM L-cysteine produced no significant inhibiting effect. CONCLUSIONS: These results suggest that LLNO1 and FONO1 are potent vasodilators. LLNO1 was shown to induce a significant level of relaxation in rabbit corpus cavernosum. The substances tested were shown to activate soluble guanylate cyclase and release intracellular NO.

Efficacy of the tincture of jalapa in the treatment of functional constipation: a double-blind, randomized, placebo-controlled study.

BACKGROUND: Laxatives are much utilized, but few clinical trials assessed the efficacy of phytotherapics in the functional constipation. AIM: The aim of this study was to evaluate the efficacy of the tincture of jalapa in the treatment of patients with functional constipation. METHODS: Double-blind, randomized, placebo-controlled clinical trial was used in this study. Seventy-six patients were assigned to two treatment groups, jalapa or placebo. The study consisted of three phases: pre-treatment, treatment, and post-treatment, each phase lasting 7 days. The mean frequency of stools, the mean consistency of stools, and the presence of pain and effort to evacuate were assessed. We monitored adverse events before, during, and after the administration of 15 mL of tincture of jalapa or placebo. RESULTS: After treatment, the mean frequency of stools of the jalapa group (0.58 ± 0.25 stools/day; P < 0.0001) was higher than in the placebo group (0.36 ± 0.20 stools/day). In the pre-treatment, stool consistency, according to the Bristol scale, ranged from types 1 to 3 for both groups. The jalapa group showed improved mean consistency of stools (P = 0.0102) after treatment, approximately ranging between types 2 and 4, while the placebo group did not show statistically significant differences (P = 0.1446). The reduction of pain (P = 0.0061) and effort (P = 0.0289) in the jalapa group were statistically significant. Both treatments were well tolerated by the patients. CONCLUSION: The tincture of jalapa was shown to be effective in the acute treatment of functional constipation.

Experimental model of human corpus cavernosum smooth muscle relaxation

PURPOSE: To describe a technique for en bloc harvesting of the corpus cavernosum, cavernous artery and urethra from transplant organ donors and contraction-relaxation experiments with corpus cavernosum smooth muscle. MATERIALS AND METHODS: The corpus cavernosum was dissected to the point of attachment with the crus penis. A 3 cm segment (corpus cavernosum and urethra) was isolated and placed in ice-cold sterile transportation buffer. Under magnification, the cavernous artery was dissected. Thus, 2 cm fragments of cavernous artery and corpus cavernosum were obtained. Strips measuring 3 x 3 x 8 mm3 were then mounted vertically in an isolated organ bath device. Contractions were measured isometrically with a Narco-Biosystems force displacement transducer (model F-60, Narco-Biosystems, Houston, TX, USA) and recorded on a 4-channel Narco-Biosystems desk model polygraph. RESULTS: Phenylephrine (1µM) was used to induce tonic contractions in the corpus cavernosum (3 - 5 g tension) and cavernous artery (0.5 - 1g tension) until reaching a plateau. After precontraction, smooth muscle relaxants were used to produce relaxation-response curves (10-12M to 10-4 M). Sodium nitroprusside was used as a relaxation control. CONCLUSION: The harvesting technique and the smooth muscle contraction-relaxation model described in this study were shown to be useful instruments in the search for new drugs for the treatment of human erectile dysfunction.

Relaxation of rabbit corpus cavernosum smooth muscle and aortic vascular endothelium induced by new nitric oxide donor substances of the nitrosyl-ruthenium complex

INTRODUCTION: Endothelial dysfunction characterized by endogenous nitric oxide (NO) deficiency made 56 percent of patients affected with erectile dysfunction decline treatment with PDE-5 inhibitors. New forms of treatment are currently being developed for this group of patients. MATERIALS AND METHODS: The study compared the effect of sodium nitroprusside (SNP) and two substances of the nitrosyl-ruthenium complex, cis-[Ru(bpy)2(SO3)(NO)]PF-6-9 ("FONO1”) and trans-[Ru(NH3)4(caffeine)(NO)]C13 ("LLNO1”) on relaxation of rabbit corpus cavernosum smooth muscle and aortic vascular endothelium. The samples were immersed in isolated baths and precontracted with 0.1 µM phenylephrine (PE) and the corresponding relaxation concentration/response curves were plotted. In order to investigate the relaxation mechanisms involved, 100 µM ODQ (a soluble guanylate cyclase-specific inhibitor), 3 µM or 10 µM oxyhemoglobin (an extracellular NO scavenger) or 1 mM L-cysteine (a nitrosyl anion-specific scavenger) was added to the samples. RESULTS: All the NO donors tested produced a significant level of relaxation in the vascular endothelium. In corpus cavernosum samples, FONO1 produced no significant effect, but LLNO1 and SNP induced dose-dependent relaxation with comparable potency (pEC50 = 6.14 ± 0.08 and 6.4 ± 0.14, respectively) and maximum effect (Emax = 82 percent vs. 100 percent, respectively). All NO donors were found to activate soluble guanylate cyclase, since the addition of the corresponding inhibitor (100 µM ODQ) completely neutralized the relaxation effect observed. The addition of oxyhemoglobin reduced the relaxation effect, but did not inhibit it completely. In aortic vascular endothelium 3 µM oxyhemoglobin decreased the relaxation effect by 26 percent on the average, while 10 µM oxyhemoglobin reduced it by over 52 percent. The addition of 100 µM L-cysteine produced no significant inhibiting effect. CONCLUSIONS: These results suggest that LLNO1...

Estudo de toxicologia clínica de um fitoterápico a base de associações de plantas, mel e própolis / Clinical toxicology study of a phytomedicine syrup composed of plants, honey and propolis

O estudo objetivou avaliar a segurança clínica do uso de uma formulação fitoterápica composta da associação de Mikania glomerata, Mentha piperita, Eucalyptus globulus e Copaifera multijuga, incorporadas à própolis e mel para tratar doenças respiratórias. O ensaio clínico consistiu de um estudo aberto, com 26 voluntários adultos de ambos os sexos, que receberam quatro doses orais de 15 mL de Saratosse®, durante 28 dias ininterruptos. Os voluntários foram incluídos somente se considerados saudáveis, após exames clínico e complementares que antecederam o estudo. A avaliação laboratorial incluía análises hematológicas, bioquímicas e sorológicas. A mesma avaliação foi repetida após cada semana de tratamento e no pós-estudo, sete dias após a última administração. Os resultados foram submetidos à Análise de Variância (teste de Dunnett), obtendo-se, a menor diferença significante (p < 0,05). O xarope foi bem tolerado. Alguns eventos adversos foram relatados, sendo classificados como possivelmente ou não atribuídos ao fitoterápico. Os índices de hemoglobina, TGO, TGP, creatinina e leucócitos não apresentaram diferenças significativas em relação ao pré-estudo. Todos os parâmetros laboratoriais estiveram dentro das suas respectivas faixas de normalidade. Os exames clínicos, eletrocardiográficos e laboratoriais não evidenciaram sinais de toxicidade nos órgãos e sistemas avaliados.

The aim of this study is to evaluate the safety of the use of a phytomedicine syrup (Saratosse®) composed of several medicinal plants: Mikania glomerata, Mentha piperita, Eucalyptus globulus and Copaifera multijuga, along with honey and propolis to treat respiratory diseases. The clinical trial consisted of an open study with 26 adult volunteers of both sexes, who were given an oral dose of 15 mL of Saratosse® for 28 consecutive days, four times a day. Only volunteers who were found healthy after a clinical and physical examination were included. The laboratory tests included: hematological, biochemical and serological analysis. This evaluation was repeated after each week of treatment and seven days after the last administration. ANOVA analysis (Dunnett's test) showed a significant difference, albeit at low statistical level (p < 0.05). Significant differences to the pre-study were not shown by hemoglobin, SGOT, SGPT, creatinine and leukocytes analysis. Laboratorial tests results were within their maximum and minimum reference values. On the whole the medicine was well tolerated. Some side effects were related, which may or not be attributed to the phytomedicine. Clinical, electrocardiographic and laboratory tests did not show any evidence of toxic signs in the organs and systems studied.

Avaliação da segurança clínica de um fitoterápico contendo Mikania glomerata, Grindelia robusta, Copaifera officinalis, Myroxylon toluifera, Nasturtium officinale, própolis e mel em voluntários saudáveis / Clinical safety evaluation of a phytomedicine containing Mikania glomerata, Grindelia robusta, Copaifera officinalis, Myroxylon toluifera, Nasturtium officinale, honey and propolis in healthy volunteers

O fitoterápico Calmatoss® é um xarope composto por tinturas de guaco (Mikania glomerata), grindélia (Grindelia robusta), copaíba (Copaifera officinalis), bálsamo de Tolú (Myroxylon toluifera), álcoolatura de agrião (Nasturtium officinale), própolis e mel. Sua principal indicação é para o tratamento de afecções do trato respiratório por suas ações antimicrobianas, antitussígenas, expectorantes e broncodilatadora. O objetivo desse estudo foi avaliar a toxicologia clínica dessa preparação em voluntários saudáveis. Para isso realizou-se um ensaio clínico, não controlado com 24 voluntários de ambos os sexos, que receberam ambulatorialmente 15 mL do xarope quatro vezes ao dia, durante 21 dias ininterruptos. Os voluntários foram incluídos no estudo somente quando considerados saudáveis após avaliação clínica, exame físico e exames laboratoriais que antecederam o estudo. O xarope Calmatoss® foi bem tolerado pelos 24 voluntários não apresentando eventos adversos graves. Os exames clínicos, eletrocardiográficos e laboratoriais efetuados antes, durante e após o ensaio não evidenciaram sinais de toxicidade nos diversos órgãos e sistemas avaliados, confirmando a segurança da preparação para utilização em ensaios de eficácia terapêutica.

Calmatoss® is a phytomedicine used in several respiratory tract pathologies treatment composed of seven medicinal plants such as Mikania glomerata, Grindelia robusta, Copaifera officinalis, Myroxylon toluifera, Nasturtium officinale, as well as honey and propolis. The present study investigated the chronic administration of 15 mL Calmatoss® syroup four times a day during 21 days for any toxic effect on healthy volunteers. The clinical trial consisted of an open study with 24 volunteers included in the study only when considered healthy after clinical evaluation, physical examination and laboratory tests, which preceded the study. The laboratory tests included: heamatologic, biochemical and sorologic analysis. This evaluation was repeated after the first, second and third week of treatment and at post-study seven days after the last administration. Calmatoss® was well tolerated by the volunteers. Variations in the laboratory were observed and all of these laboratory changes returned to normal levels during or after the study. In spite of these variations none of the volunteers had their values out of the established normality limits for each parameter. In conclusion, clinical, electrocardiographic and laboratory tests did not show any evidence of toxic signals in the various organs and systems studied.