RESUMO
Snakebite accidents, neglected tropical diseases per the WHO, pose a significant public health threat due to their severity and frequency. Envenomation by Bothrops genus snakes leads to severe manifestations due to proteolytic enzymes. While the antibothropic serum produced by the Butantan Institute saves lives, its efficacy is limited as it fails to neutralize certain serine proteases. Hence, developing new-generation antivenoms, like monoclonal antibodies, is crucial. This study aimed to explore the inhibitory potential of synthetic peptides homologous to the CDR3 regions of a monoclonal antibody targeting a snake venom thrombin-like enzyme (SVTLE) from B. atrox venom. Five synthetic peptides were studied, all stable against hydrolysis by venoms and serine proteases. Impressively, four peptides demonstrated uncompetitive SVTLE inhibition, with Ki values ranging from 10-6 to 10-7 M. These findings underscore the potential of short peptides homologous to CDR3 regions in blocking snake venom toxins, suggesting their promise as the basis for new-generation antivenoms. Thus, this study offers potential advancements in combatting snakebites, addressing a critical public health challenge in tropical and subtropical regions.
Assuntos
Anticorpos Monoclonais , Bothrops , Peptídeos , Serina Proteases , Animais , Anticorpos Monoclonais/química , Anticorpos Monoclonais/farmacologia , Peptídeos/química , Peptídeos/farmacologia , Serina Proteases/química , Serina Proteases/metabolismo , Antivenenos/química , Antivenenos/imunologia , Antivenenos/farmacologia , Regiões Determinantes de Complementaridade/química , Venenos de Crotalídeos/antagonistas & inibidores , Venenos de Crotalídeos/imunologia , Venenos de Crotalídeos/enzimologia , Venenos de Crotalídeos/química , Sequência de Aminoácidos , Inibidores de Serina Proteinase/química , Inibidores de Serina Proteinase/farmacologiaRESUMO
Horses are seasonal polyoestrous animals, and the photoperiod is the main factor modulating their reproductive activity. There is no consensus on the andrological and biochemical factors that influence breeding seasonality. To assess the involvement of climate in reproduction, Mangalarga Marchador stallions were monitored over 1 year regarding semen quality and seminal plasma proteome. Here, we show that kallikrein (KLKs) proteoforms in seminal plasma are involved in climate conditioning of reproduction. During the breeding season, greater abundance and different types of KLKs occurred simultaneously to lower sperm motility, greater semen volumes and higher concentrations of glucose and cholesterol. Considering that vasodilation due to activation of the kallikrein-kinin system and the consequent inhibition of the renin-angiotensin system may be associated with lower sperm motility, unravelling the involvement of KLK proteoforms in reproductive seasonality is a priority in horse breeding.
Assuntos
Análise do Sêmen , Motilidade dos Espermatozoides , Cavalos , Masculino , Animais , Motilidade dos Espermatozoides/fisiologia , Calicreínas , Sêmen/fisiologia , Reprodução/fisiologia , Espermatozoides/fisiologiaRESUMO
Canastra artisanal Minas cheese samples were collected in Minas Gerais - Brazil. The samples were evaluated in order to observe the presence of antimicrobial peptides during 30â¯days of ripening. Soluble peptides extracted from the cheeses were fractionated by reverse phase liquid chromatography and their fractions evaluated for inhibitory action of E. coli. Fractions containing antimicrobial activity were analyzed by MALDI-TOF/TOF and then peptides were sequenced and identified using MASCOT Daemon coupled with UniProt database. The identified peptides were then validated by SCAFFOLD application. The peptides present in fractions with antimicrobial activity were RPKHPIKHQ, RPKHPIKHQG, RPKHPIKHQGLPQ and RPKHPIKHQGLPQE, HQPHQPLPPT and MHQPHQPLPPT. Peptide sequences PKHPIKHQ, RPKHPIKHQG, RPKHPIKHQGLPQ and RPKHPIKHQGLPQE were originated from αs1-casein and are their fragments belonging to Isracidine, which in turn is a well known antimicrobial peptide. The HQPHQPLPPT and MHQPHQPLPPT peptides were related to ß-casein and were isolated in other studies, but their biological activities are still unknown.
Assuntos
Antibacterianos/análise , Caseínas/análise , Queijo/análise , Queijo/microbiologia , Escherichia coli/metabolismo , Peptídeos/análise , Peptídeos/metabolismoRESUMO
In this work we analyzed protein expression in the Aedes aegypti midgut during the larval (fourth instar, L4), pupal, and adult stages [including newly emerged (NE), sugar-fed (SF) and blood-fed (BF) females]. Two-dimensional electrophoresis showed 13 spots in the midgut of larvae, 95 in the midgut of pupae, 90 in the midgut of NE, and 76 in the midgut of SF or BF females. In the larval midguts, high serpin expression was noted, while in the pupae, protein abundance was lower than in the NE, SF, and BF females. The spots related to proteins linked to energy production, protein metabolism, signaling, and transport were highly expressed in the NE stage, while spots related proteins involved in translation were abundant in SF and BF females. The differential abundance of proteins in the midgut of A. aegypti at different developmental stages supports the necessity for midgut development during immature stage followed by the necessity of proteins related to digestion in adults.
Assuntos
Aedes/embriologia , Trato Gastrointestinal/embriologia , Trato Gastrointestinal/metabolismo , Mucosa Intestinal/metabolismo , Larva/crescimento & desenvolvimento , Proteômica , Aedes/crescimento & desenvolvimento , Animais , Metabolismo Energético/fisiologia , Feminino , Larva/metabolismo , Transporte Proteico/fisiologia , Proteínas/genética , Proteínas/metabolismo , Pupa/metabolismo , Serpinas/biossíntese , Transdução de SinaisRESUMO
Bumblebees are widely distributed across the world and have great economic and ecological importance as pollinators in the forest as well as in agriculture. The insect midgut consists of three cell types, which play various important roles in digestion, absorption, and hormone production. The present study characterized the anterior and posterior midgut regions of the bumblebee, Bombus morio. The digestive, regenerative and endocrine cells in the midgut showed regional differences in their number, nuclear size, as well as the size of the striated border. Ultrastructurally, the digestive cells contained many mitochondria and long microvilli; however, in the anterior midgut region, these cells showed dilated basal labyrinths with a few openings for the hemocoel, whereas the labyrinths of the basal posterior region remained inverse characteristics. Thus, the characterization of the midgut of B. morio supported an ecto-endoperitrophic circulation, contributing to a better understanding of the digestive process in this bee.