Alzheimer’s disease (AD) is a neurodegenerative disorder characterized by extracellular accumulation of amyloid-β (Aβ) peptides in the brain. This study demonstrates the direct association of the disposable pipette extraction (DPX) with tandem mass spectrometry (MS/MS) for the analysis of Aβ peptides in cerebrospinal fluid (CSF) samples. Different parameters were optimized in order to improve detectability in the MS/MS including mobile phase percentage of ammonium hydroxide, mobile phase flow rate and acquisition mode. Also, this method used an electrospray ionization (ESI) low-flow probe and direct infusion of an organic solution in the MS/MS. The DPX-MS/MS method showed adequate linearity for determining Aβ peptides in CSF-linearity ranged from 0.1 to 1.5 ng mL 1. The coefficients of determination were higher than 0.99; the precision coefficient of variation (CV) ranged from 0.3 to 12.7%; and the accuracy relative standard deviation (RSD) ranged from -13.6 to 13.2%. Alzheimers Alzheimer s AD (AD amyloidβ amyloid β (Aβ brain DPX (DPX MSMS MS (MS/MS CSF (CSF samples hydroxide mode Also ESI (ESI lowflow low DPXMS/MS DPXMSMS CSFlinearity 01 0 1 0. 15 5 0.99 099 99 CV (CV 03 3 12.7% 127 12 7 RSD (RSD 13.6 136 13 6 -13. 132 2 13.2% DPXMS 0.9 09 9 12.7 13. -13 13.2 12. -1 -

Resumen Introducción. En becerras prepúberes, la Kisspeptina-10 ha estimulado la secreción de las hormonas luteinizante (LH) y folículo estimulante (FSH), y en vacas posparto amamantando ha estimulado la secreción de LH sin evidencia de inducir la ovulación. Objetivo. Inducir la ovulación con Kisspeptina-10 u hormona liberadora de gonadotropinas (GnRH), en vacas productoras de carne en condiciones subtropicales de México. Materiales y métodos. El estudio se realizó en la región nororiente del estado de Puebla durante los años 2020 al 2021, con 252 vacas cruzadas Bos taurus x Bos indicus. El protocolo de sincronización estral consistió en la aplicación de un dispositivo intravaginal impregnado con progesterona (1,9 g) y la inyección intramuscular de 2 mg de benzoato de estradiol (día 0). El día siete se removió el dispositivo intravaginal y de inmediato las vacas fueron inyectadas vía intramuscular con 0,150 mg de D-cloprostenol. Al día siguiente (día ocho), las vacas fueron inyectadas vía intramuscular con 1 mg de benzoato de estradiol. Entre 52 y 56 horas posteriores al retiro del dispositivo intravaginal (día nueve), las vacas se inyectaron vía intramuscular con GnRH o Kisspeptina-10. Las vacas se asignaron a cuatro tratamientos: T1, 100 µg de GnRH (control positivo), T2, T3 y T4 500 µg, 1000 µg y 1500 µg de Kisspeptina-10, respectivamente. La ovulación se detectó mediante ultrasonografía a los cuatro días post-tratamiento. La variable de respuesta evaluada fue la tasa de ovulación. Los datos se analizaron mediante modelos lineales generalizados y se aplicó una función liga logit. Resultados. Las tasas de ovulación obtenidas con T1 y T4 fueron iguales estadísticamente (99,9 %; p>0,05) y mayores (p<0,05) que las logradas con T2 (98,5 %) y T3 (88,4 %), estas dos últimas fueron iguales estadísticamente (p>0,05). Conclusión. La Kisspeptina-10 1500 µg fue tan efectiva como la GnRH para inducir la ovulación.

Abstract Introduction. In prepubertal heifers, Kisspeptin-10 has stimulated the secretion of luteinizing hormone (LH) and follicle-stimulating hormone (FSH), and in postpartum lactating cows, it has stimulated LH secretion without evidence of inducing ovulation. Objective. To induce ovulation using Kisspeptin-10 or gonadotropin-releasing hormone (GnRH) in beef-producing cows under subtropical conditions in Mexico. Materials and methods. The study was conducted in the northeastern region of the state of Puebla uring the years 2020 to 2021, with 252 Bos taurus x Bos indicus cows. The estrous synchronization protocol involved the insertion of an intravaginal device impregnated with progesterone (1.9 g) and intramuscular injection of 2 mg of estradiol benzoate (day 0). On day seven, the intravaginal device was removed, and cows were immediately intramuscularly injected with 0.150 mg of D-cloprostenol. On the following day (day eight), Kisspeptin-10 was as effective as GnRH in inducing ovulation. 56 hours after the removal of the intravaginal device (day nine), cows were intramuscularly injected with GnRH or Kisspeptin-10. Cows were assigned to four treatments: T1, 100 µg of GnRH (positive control), T2, T3, and T4 with 500 µg, 1000 µg, and 1500 µg of Kisspeptin-10, respectively. Ovulation was detected via ultrasonography four days post-treatment. The response variable evaluated was the ovulation rate. Data were analyzed using generalized linear models with a logit link function. Results. Ovulation rates obtained with T1 and T4 were statistically equal (99.9%; p>0.05) and higher (p<0.05) than those achieved with T2 (98.5%) and T3 (88.4%), with the latter two being statistically equal (p>0.05). Conclusion. Kisspeptin-10 1500 µg was as effective as GnRH in inducing ovulation.

Resumen: Introducción : La artritis reumatoide (AR) es una enfermedad inflamatoria crónica, autoinmune, caracterizada por poliartritis crónica, aditiva, simétrica, que puede cursar con manifestaciones extraarticulares (MExA) asociadas a una mayor morbimortalidad. Objetivo: de describir las MExA más frecuentes en AR. Métodos: se realizó un estudio descriptivo, transversal y retrospectivo de revisión de historias clínicas de pacientes diagnosticados con AR, durante el periodo diciembre 2011-enero 2022. Resultados: Participaron 150 pacientes con AR, con una edad promedio de 53,7±12,5 años, el sexo predominante fue el femenino con 84,6%, el tiempo de evolución de la AR fue de 7,2±8,9 años; en cuanto a las características serológicas, 91,1% tenía Factor reumatoideo positivo y 76,9% tenía anticuerpos contra péptidos cíclicos citrulinados positivo. Tanto al ingreso, como en la última consulta los pacientes presentaron alguna manifestación extraarticular (MExA), 61,3% y 70%, respectivamente, siendo las más frecuentes la anemia de los trastornos crónicos (al ingreso 44,6% y en última consulta 50,6%), seguida de nódulos reumatoideos (al ingreso con 8% y en la última consulta 9,3%). Conclusiones: Las MExA se presentaron en 70% de los pacientes, siendo las más frecuentes la anemia y los nódulos reumatoideos. Estos datos muestras los cambios que han tenido la frecuencia de estas MExA en esta enfermedad a través del tiempo.

Abstract: Introduction: Rheumatoid arthritis (RA) is a chronic inflammatory, autoimmune disease, characterized by chronic, additive, symmetrical polyarthritis, which can present with extra-articular manifestations (ExAM), associated with greater morbidity and mortality. Objective: to describe the most frequent ExAM in RA. Methods: a descriptive, cross-sectional and retrospective study was carried out to review the medical records of patients diagnosed with RA, during the period December 2011-January 2022. Results: 150 patients with RA participated, with an average age of 53.7±12.5 years, the predominant sex was female with 84.6%, the evolution time of RA was 7.2±8.9. years; Regarding serological characteristics, 91.1% had positive Rheumatoid Factor and 76.9% had positive antibodies against cyclic citrullinated peptides. Both upon admission and at the last consultation, patients presented some extra-articular manifestation (ExAM), 61.3% and 70%, respectively, with the most frequent being chronic anemia (at admission 44.6% and at last consultation 50.6%), followed by rheumatoid nodules (at admission with 8% and at the last consultation 9.3%). Conclusions: ExAM occurred in 70% of patients, the most frequent being anemia and rheumatoid nodules. These data show the changes in the frequency of ExAM in the disease over time.

Abstract Background: Heart failure (HF) is a condition associated with high morbidity and mortality. Its early diagnosis in primary health care (PHC) represents a substantial challenge, considering its non-specific clinical manifestations and the limitations on timely access to diagnostic techniques. Objective: To evaluate the prevalence of HF, characterize subtypes and determine the prognosis of patients consulting in PHC for dyspnea Edema of the lower extremities. Methods: Prospective study in 340 patients who consulted in PHC, without previous diagnosis of HF. Clinical evaluation, electrocardiogram, NT-proBNP point-ofcare and echocardiography with telematic interpretation by cardiologists were performed. Using the HFA-PEFF and H2FPEF algorithms patients were classified as: 1) HF with reduced ejection fraction (HFREF); 2) HF with preserved ejection fraction (HFPEF) and 3) No HF. Actuarial survival analyses were performed. Results: We observed a prevalence of HFREF of 8%, high probability of HFPEF by HFA-PEFF in 42% and by H2FPEF in 8%. Intermediate probability of HFPEF, requiring complementary study, was observed in 47% of patients with HFA-PEFF and 76% of patients with H2FPEF (p<0.05). Overall survival at 36 months was 90±2% and cardiovascular survival at 36 months was 95±1%. Using HFA-PEFF, patients with HF presented lower overall survival compared to patients with no HF (HR 2.3, 95%CI 1.1-4.9; p=0.029). We did not observe mortality differences with H2FPEF. Conclusions: In patients consulting for dyspnea and/or lower extremity edema at PHC and undergoing evaluation with NT-proBNP and echocardiography, we observed a HF prevalence of 50%. HF classification through HFA-PEFF was associated with lower survival rates.

Resumen Introducción: La insuficiencia cardíaca (IC) tiene alta morbilidad y mortalidad. Su diagnóstico temprano en atención primaria de salud (APS) es un reto dada la baja especificidad de sus criterios clínicos y las limitaciones en acceso a técnicas diagnósticas. Objetivo: Analizar la prevalencia de IC, subtipos y pronóstico de pacientes con disnea y/o edema de extremidades inferiores que consultan en APS. Metodología: Se trata de un estudio prospectivo de 340 pacientes en APS, sin diagnóstico previo de IC. Se realizó una evaluación clínica, electrocardiograma, NT-proBNP “point-of-care”, ecocardiografía con interpretación telemática por cardiólogos. Utilizando los algoritmos HFA-PEFF y H2FPEF se clasificaron los pacientes como :1) IC con fracción de eyección (FE) reducida (ICFER); 2) IC con FE preservada (ICFEP) y 3) pacientes sin diagnóstico de IC. Se efectuó un análisis de sobrevida de los diferentes grupos. Resultados: La prevalencia de ICFER fue 8%, ICFEP por HFA-PEFF 42% y por H2FPEF 8%. Los algoritmos sugieren efectuar un estudio complementario en el 47% con HFA-PEFF y 76% con H2FPEF (p<0.05). La sobrevida global a 36 meses fue 90±2% y cardiovascular 95±1%. Usando HFA-PEFF, los pacientes con IC tuvieron menor sobrevida que aquellos sin IC (HR 2.3, IC95% 1.14.9; p=0.029). No hubo diferencias de mortalidad con H2FPEF. Conclusiones: En pacientes de APS que consultan por disnea y/o edema de extremidades inferiores sometidos a evaluación con NT-proBNP y ecocardiografía, se observó una prevalencia de IC de hasta 50%, 8% de ICFER y 42% de ICFEP. La caracterización de IC utilizando HFA-PEFF está asociada al pronóstico vital.

ABSTRACT Antimicrobial peptides are small molecules that exhibit strong antagonistic activity against pathogenic microorganisms and are synthesized by a wide variety or organisms, including probiotic bacteria. The aim of the study was to identify peptides with antimicrobial activity in the Enterococcus faecium IP5-2a strain previously isolated from the intestinal tract of Nile tilapia (Oreochromis niloticus). Cellular and extracelullar proteins extraction was performed, followed by 1D SDS-PAGE gel electrophoresis and MALDI-TOF/TOF mass spectrometry. The results revealed peptides with antimicrobial activity, including P54 (SagA) protein, a bacteriocin-like protein and a lantibiotic. These proteins play an important role in the antagonistic capacity of the E. faecium IP52a strain, allowing it to be considered as a potential probiotic in the aquaculture industry.

RESUMEN Los péptidos antimicrobianos son moléculas pequeñas que poseen una fuerte actividad antagónica frente a microorganismo patógenos y son sintetizados por una amplia variedad de organismos, entre ellos, bacterias probióticas. El objetivo del estudio fue identificar péptidos con actividad antimicrobiana en la cepa Enterococcus faecium IP52a previamente aislada del tracto intestinal de tilapia del Nilo. Se realizó extracción de proteínas celulares y extracelulares, electroforesis en gel SDS-PAGE 1D y espectrometría de masas MALDI-TOF/TOF. Los resultados mostraron péptidos con actividad antimicrobiana, entre ellos la proteína P54 (SagA), una proteína similar a bacteriocina y un lantibiótico. Estas proteínas juegan un rol importante en la capacidad antagónica de la cepa E. faecium IP52a, permitiendo considerarla como un potencial probiótico en la industria acuícola.

Abstract Antimicrobial peptides (AMPs) are small groups of proteins obtained from plants and animals. AMPs participate in the immune response, as they provide a quick line of defense against infections, while others may be related to the plant's defense against certain pests and pathogens. The objective of the present study was to evaluate an inhibitory activity of fractions obtained from immature fruits of Capsicum chinense Jacq. (accession UENF 1755) on trypsin, chymotrypsin and α-amylase families and on yeast growth. The peptides were obtained from the immature fruits using saline extraction. The extract was semi-purified by DEAE-Sepharose chromatography into two fractions: D1 (non-retained fraction) and D2 (retained fraction), and analyzed using SDS-tricine-gel electrophoresis. The antifungal activity of these fractions was tested on Candida albicans, Candida buinensis, Candida parapsilosis and Candida tropicalis. To elucidate the antimicrobial mechanism of these fractions, membrane permeabilization and endogenous reactive oxygen species (ROS) induction assays were performed. The fractions were also tested for inhibition of trypsin, chymotrypsin and α-amylase enzymes. The two fractions, D1 and D2, inhibited yeast growth at a concentration of 100 μg.mL-1, promoted membrane permeabilization and caused an increase in the induction of endogenous ROS in C. albicans and C. tropicalis. Both these fractions were able to inhibit trypsin and α-amylase enzyme, while only D1 inhibited chymotrypsin activity. Thus, D1 was found to possess a greater antifungal and enzymatic inhibition activity on trypsin, chymotrypsin and α-amylase. (AMPs animals response infections plant s pathogens Jacq accession 1755 αamylase α amylase extraction semipurified semi purified DEAESepharose DEAE Sepharose D nonretained non retained fraction fraction, , SDStricinegel SDS tricine gel electrophoresis buinensis tropicalis (ROS performed enzymes 10 μg.mL1, μgmL1 μgmL μg.mL 1, μg mL 1 μg.mL-1 C enzyme Thus αamylase. amylase. 175 mL1 μg.mL1 μg.mL- 17

Abstract Natural products have been studied as potential anthelmintic drugs. This study aimed to evaluate the effects of the polypeptides/proteins exuded from Mimosa caesalpiniifolia mature seeds on the model nematode Caenorhabditis elegans. After seed immersion for 24 h at 25 ºC in sodium phosphate buffer, pH 7.0, containing 0.15 M NaCl and 1.0 M MgSO4, the constituent polypeptides/proteins were fractionated using ammonium sulfate precipitation followed by ultrafiltration and their effects on C. elegans motility were assessed. Moreover, we evaluated by Atomic Force Microscopy (AFM) whether exposition of the nematode to the polypeptide fraction McSEX10-2 could cause any alteration on the animal’s external surface (cuticle). The whole seed exudates (SEX) from the M. caesalpiniifolia mature seeds (named McSEX) and its polypeptide/protein derived fractions, termed McSEX90%/80°C, McSEX30-10, and McSEX10-2, inhibited the C. elegans motility with an IC50 of 15.18, 12.59, 9.74 and 1.34 μg mL-1.The McSEX10-2 fraction, which encompassed the polypeptides with the molecular mass varying from 10 kDa to 2 kDa, was the most effective. In addition, McSEX10-2 altered the ultrastructure and the mechanical properties of the C. elegans cuticle. McSEX10-2, in particular, can be used as a model to bioinspire structure-based design of new peptides that could control, alone or in combination with other drugs, economically important parasitic nematodes. drugs polypeptidesproteins proteins buffer 70 7 0 7.0 015 15 0.1 1 1. MgSO4 MgSO C assessed Moreover AFM (AFM McSEX102 McSEX McSEX10 McSEX10- animals animal s cuticle . (cuticle) SEX (SEX named polypeptideprotein protein fractions McSEX9080C McSEXC McSEX90 80 McSEX90%/80°C McSEX3010, McSEX3010 McSEX30 10, McSEX30-10 McSEX102, 2, IC IC5 1518 18 15.18 1259 12 59 12.59 974 9 74 9.7 134 34 1.3 mL1.The mL1The mLThe mL 1.The effective addition particular structurebased structure based control nematodes 7. 01 0. McSEX1 (cuticle McSEX9 8 McSEX301 McSEX3 McSEX30-1 151 15.1 125 5 12.5 97 9. 13 3 mL1 1The McSEX30- 15. 12.

Three-dimensional structures of proteins are intimately linked to their functions, therefore understanding their conformation in solution is essential. While nuclear magnetic resonance spectroscopy and X-ray crystallography are widely employed for protein structural determination, their limitations make the process challenging and expensive. Theoretical calculations of chemical shifts present a potential complement to experimental techniques, facilitating the study of protein structures. This investigation aims to assess the applicability of chemical shift calculations in analyzing three-dimensional structures of peptides, focusing on the tryptophan zipper 1 peptide as a model. Furthermore, a mutated variant of this peptide was proposed to evaluate the stability of its structural elements under sequence modifcations. Through calculations, a potential structural alteration in the β-turn region of the mutant peptide compared to tryptophan zipper 1 was identifed. This research demonstrates the potential of using computational approaches to complement experimental methods in studying protein structures and their functional implications. Threedimensional Three dimensional functions essential Xray X ray determination expensive techniques threedimensional three peptides model Furthermore modifcations βturn β turn identifed implications

Abstract Glioblastoma multiforme is a tumor of the central nervous system. Focal Adhesion Kinase (FAK) and αB-crystalline are two proteins involved in glioblastoma development. In this study, we investigated whether the FAK/αB-crystalline interaction is important for glioblastoma cells, we aimed to investigate the interaction of these two proteins in the glioblastoma multiforme cell line U87-MG. Two peptides named FP01 peptide (derived from αB-crystalline) and FP02 peptide (derived from FAK) were synthesized for this study. Treatment of U87-MG with the peptides FP01 and FP02 in the concentration at 50 µM reduced the viability cellular to around 41% and 51%, respectively. Morphological alterations in the cells treated with the peptides when compared to the control were observed. This study suggests that the interaction between FAK and αB-crystalline is important for the viability of glioblastoma cells. system (FAK αBcrystalline αB crystalline development FAK/αBcrystalline FAKαBcrystalline FAK/αB U87MG. U87MG UMG U87 MG. MG U FP FP0 derived 5 41 51 51% respectively observed FAKαB U8 4

Abstract Background: The bioactive peptides derived from snake venoms of the Viperidae family species have been promising as therapeutic candidates for neuroprotection due to their ability to prevent neuronal cell loss, injury, and death. Therefore, this study aimed to evaluate the cytoprotective effects of a synthetic proline-rich oligopeptide 7a (PRO-7a; <EDGPIPP) from Bothrops jararaca snake, on oxidative stress-induced toxicity in neuronal PC12 cells and astrocyte-like C6 cells. Methods: Both cells were pre-treated for four hours with different concentrations of PRO-7a, submitted to H2O2-induced damage for 20 h, and then the oxidative stress markers were analyzed. Also, two independent neuroprotective mechanisms were investigated: a) L-arginine metabolite generation via argininosuccinate synthetase (AsS) activity regulation to produce agmatine or polyamines with neuroprotective properties; b) M1 mAChR receptor subtype activation pathway to reduce oxidative stress and neuron injury. Results: PRO-7a was not cytoprotective in C6 cells, but potentiated the H2O2-induced damage to cell integrity at a concentration lower than 0.38 μM. However, PRO-7a at 1.56 µM, on the other hand, modified H2O2-induced toxicity in PC12 cells by restoring cell integrity, mitochondrial metabolism, ROS generation, and arginase indirect activity. The α-Methyl-DL-aspartic acid (MDLA) and L-NΩ-Nitroarginine methyl ester (L-Name), specific inhibitors of AsS and nitric oxide synthase (NOS), which catalyzes the synthesis of polyamines and NO from L-arginine, did not suppress PRO-7a-mediated cytoprotection against oxidative stress. It suggested that its mechanism is independent of the production of L-arginine metabolites with neuroprotective properties by increased AsS activity. On the other hand, the neuroprotective effect of PRO-7a was blocked in the presence of dicyclomine hydrochloride (DCH), an M1 mAChR antagonist. Conclusions: For the first time, this work provides evidence that PRO-7a-induced neuroprotection seems to be mediated through M1 mAChR activation in PC12 cells, which reduces oxidative stress independently of AsS activity and L-arginine bioavailability. Background loss injury death Therefore prolinerich proline rich PRO7a PROa PRO (PRO-7a <EDGPIPP EDGPIPP stressinduced induced PC PC1 astrocytelike astrocyte like C Methods pretreated pre treated PRO7a, 7a, H2O2induced HOinduced H2O2 H O 2 h analyzed Also investigated Larginine L arginine (AsS b M Results 038 0 38 0.3 μM However 156 1 56 1.5 µM hand metabolism αMethylDLaspartic α Methyl DL aspartic MDLA (MDLA LNΩNitroarginine NΩ Nitroarginine LName, LName Name , (L-Name) NOS, NOS (NOS) Larginine, arginine, PRO7amediated PROamediated DCH, DCH (DCH) antagonist Conclusions time PRO7ainduced PROainduced bioavailability HO H2O 03 3 0. 15 5 1. (L-Name (NOS amediated (DCH ainduced

ABSTRACT Lectins are proteins that recognize and bind to carbohydrates in a reversible and specific manner. In this work, a lectin from Crotalaria incana L. seeds was purified by Sephadex G-50 affinity chromatography. The purified lectin was named CiL and presented affinity towards D-mannose, D-glucose, D-galactose, α-methyl-D-mannoside and derivatives. CiL was stable over a wide range of temperatures and pH values, and it was divalent cation-dependent. SDS-PAGE analysis indicated that CiL is composed of two subunits with apparent masses of 29 and 30 kDa. The amino acid sequence of five tryptic peptides was obtained through mass spectrometry. Partial primary structure data indicated the similarity between CiL and lectins from Phaseolus vulgaris, Cladrastis kentukea, Lens culinaris, Pisum sativum, Crotalaria pallida and C. juncea. CiL showed no toxicity to Artemia salina nauplii at the concentration of 2 mg/mL, thus reinforcing the potential of this protein for further studies in other biological models and elucidation of possible effects. manner work L G50 G 50 G-5 chromatography Dmannose, Dmannose D mannose, mannose D-mannose Dglucose, Dglucose glucose, glucose D-glucose Dgalactose, Dgalactose galactose, galactose D-galactose αmethylDmannoside α methyl mannoside derivatives values cationdependent. cationdependent cation dependent. dependent cation-dependent SDSPAGE SDS PAGE 3 kDa spectrometry vulgaris kentukea culinaris sativum C juncea mgmL mg mL mg/mL effects G5 5 G-

This study evaluated the impact of in silico simulated gastrointestinal digestion (GID) of four quinoa globulins on the potential to release ACE and DPP-IV inhibitor peptides (antihypertensive and antidiabetic properties, respectively), as well as performed a molecular docking study to evaluate the interactions produced in the peptide-enzyme complexes. In silico GID performed on quinoa globulins resulted in the formation of amino acids as well as peptides with two to five residues. The peptides PSF, IPG, CSG, SPR, CSPG and PPN stood out for their high bioactivity scores (> 0.6), for not showing toxicity, as well as presenting potential inhibitory properties to both ACE and DPP-IV enzymes evaluated by ToxinPred, PeptideRanker and BioPep tools, respectively. The molecular docking analysis allowed highlighting that all peptides interacted with the enzymes, finding favorable binding energy values, different number and type of interactions, either at the level of the enzyme active sites or not, characteristics that together would define the potential of the established interaction of the complexes formed. The results, at the level of a first screening, support that GID of quinoa globulins can give rise to peptides with both antihypertensive and antidiabetic properties, requiring further in vitro and in vivo studies. (GID DPPIV DPP IV respectively, respectively , respectively) peptideenzyme peptide residues PSF IPG CSG SPR > ( 0.6, 06 0.6 0 6 0.6) toxicity ToxinPred tools values formed results screening studies 0.

ABSTRACT Objective: To assess the efficacy of bioactive glass, self-assembling peptide, and ozone-remineralizing agents on the artificial carious lesion. Material and Methods: On the extracted 60 premolar teeth, an artificial carious lesion/demineralization was created. Later, the remineralization of demineralized teeth was done with respective remineralizing agents (Group A: Calcium sodium phosphosilicate (bioactive glass), Group B: Self-assembling peptide, Group C: Ozone remineralizing agents and Group D (Control): De ionized water. The degree of demineralization and remineralization were evaluated using the Vickers Hardness Number. Results: There was a decrease in microhardness from baseline to demineralization in all the groups, and this reduction was found to be statistically considerable. After the remineralization of demineralized samples with respective remineralizing agents, there was an increase in microhardness of 312.38, 276.67, and 254.42 in groups A, B, and C, respectively. In contrast, in Group D, there were no changes. Conclusion: Bioactive glass and self-assembling peptides had higher remineralizing capacities, which can be used to treat early carious lesions. Objective selfassembling self assembling peptide ozoneremineralizing ozone lesion Methods 6 lesiondemineralization created Later A , glass) B Selfassembling Self C Control (Control) water Number Results considerable 31238 312 38 312.38 27667 276 67 276.67 25442 254 42 254.4 respectively contrast changes Conclusion capacities lesions (Control 3123 31 3 312.3 2766 27 276.6 2544 25 4 254. 312. 2 276.

Abstract The skin of anuran species is a protective barrier against predators and pathogens, showing also chemical defense by substances that represent a potential source for bioactive substances. This review describes the current chemical and biological knowledge from the skin secretions of Leptodactylidae species, one of the most diverse neotropical frog families. These skin secretions reveal a variety of substances such as amines (12), neuropeptides (16), and antimicrobial peptides (72). The amines include histamine and its methylated derivatives, tryptamine derivatives and quaternary amines. The peptides of Leptodactylidae species show molecular weight up to 3364 Da and ocellatins are the most reported. The peptides exhibit commonly glycine (G) or glycine-valine (GV) as C-terminal amino acids, and the most common N-terminal amino acids are glutamic acid (E), lysine (K), and valine (V). The substances from Leptodactylidae species have been evaluated against pathogenic microorganisms, particularly Escherichia coli and Staphylococcus aureus, and the most active peptides showed MIC of 1-15 µM. Furthermore, some compounds showed also pharmacological properties such as immunomodulation, treatment of degenerative diseases, anticancer, and antioxidant. Currently, only 9% of the species in this family have been properly studied, highlighting a large number of unstudied species such as an entire subfamily (Paratelmatobiinae). The ecological context, functions, and evolution of peptides and amines in this family are poorly understood and represent a large field for further exploration. pathogens families 12, 12 , (12) 16, 16 (16) 72. 72 . (72) 336 reported G (G glycinevaline GV (GV Cterminal C terminal Nterminal N E, E (E) K, K (K) V. V (V) microorganisms aureus 115 1 15 1-1 µM Furthermore immunomodulation diseases anticancer antioxidant Currently 9 studied Paratelmatobiinae. Paratelmatobiinae (Paratelmatobiinae) context functions exploration (12 (16 7 (72 33 (E (K (V 11 1- (Paratelmatobiinae (1 (7 3 (