Guamara and Cocuixtle: Source of Proteases for the Transformation of Shrimp By-Products into Hydrolysates with Potential Application.

Since the fruits of Bromelia pinguin and Bromelia karatas are rich in proteases, the aim of this research was to optimize the hydrolysis process of cooked white shrimp by-products due to the effect of these proteases. A robust Taguchi L16' design was used to optimize the hydrolysis process. Similarly, the amino acid profile by GC-MS and antioxidant capacity (ABTS and FRAP) were determined. The optimal conditions for hydrolysis of cooked shrimp by-products were pH 8.0, 30 °C, 0.5 h, 1 g of substrate and 100 µg/mL of B. karatas, pH 7.5, 40 °C, 0.5 h, 0.5 g substrate and 100 µg/mL enzyme extract from B. pinguin and pH 7.0, 37 °C, 1 h, 1.5 g substrate and 100 µg/mL enzyme bromelain. The optimized hydrolyzates of B. karatas B. pinguin and bromelain had 8 essential amino acids in their composition. The evaluation of the antioxidant capacity of the hydrolyzates under optimal conditions showed more than 80% inhibition of in ABTS radical, B. karatas hydrolyzates had better higher ferric ion reduction capacity with 10.09 ± 0.02 mM TE/mL. Finally, the use of proteolytic extracts from B. pinguin and B. karatas to optimize hydrolysis process allowed obtaining hydrolyzates of cooked shrimp by-products with potential antioxidant capacity.

Edible pectin film added with peptides from jackfruit leaves obtained by high-hydrostatic pressure and pepsin hydrolysis.

Jackfruit (Artocarpus heterophyllus Lam.) is an evergreen tree that produces a high waste of leaves. This study evaluated the obtention of peptides from jackfruit leaves using pancreatin and pepsin, their antifungal activity, and their effect on pectin films. The protein content was 7.64 ± 0.12 g/100 g of jackfruit fresh leaves. Pancreatin produced a higher yield than pepsin in the obtention of peptides (p ≤ 0.05). However, peptides obtained after 2 h by pepsin hydrolysis (Pep-P) had six essential amino acids and inhibited > 99% of mycelial growth and spore germination of Colletotrichum gloeosporioides. Pectin films with Pep-P showed a slight brown color, lower thickness, water vapor permeability, and moisture content, as well as higher thermal stability and better inhibition properties against C. gloeosporioides than pectin films without Pep-P (p ≤ 0.05). Pectin films added with Pep-P from jackfruit leaf could be a green alternative to anthracnose control in tropical fruits.

Prevalence of antibiotic resistance and virulent factors in nosocomial clinical isolates of Pseudomonas aeruginosa from Panamá

ABSTRACT Background: Pseudomonas aeruginosa is an important causative agent of nosocomial infections. As pathogen, P. aeruginosa is of increasing clinical importance due to its ability to develop high-level multidrug resistance (MDR). Methods: The aim of the present study was to better understand the intrinsic virulence of circulating strains of Pseudomonas aeruginosa, by surveying and characterizing the antibiotic resistance profiles and prevalence of virulence factors in 51 clinical isolates of P. aeruginosa obtained from children admitted to Hospital del Niño-Panamá during the period of October 2016 until March 2017. Antimicrobial susceptibilities were assessed by determining the minimum inhibitory concentration for 12 antibiotics against P. aeruginosa clinical isolates using the VITEK system (https://www.biomerieux.com). Additionally, all isolates were examined by Polymerase Chain Reaction (PCR) for the presence of components of the MexAB-OprM efflux pump system (mexABR) and pyoverdine receptor genes and betalactamases resistance genes (ESBL) using gene-specific primers. Results: A total of 51 pyoverdine producing clinical isolates were analyzed, all of which expressed resistance genes such as genes of the MexAB-OprM efflux pump system (mexABR) and pyoverdine receptor genes (fpvA). Out of 51 MDR isolates, 22 were ESBL producers. The most common ESBL gene was blaTEM expressed by 43% of the isolates. The isolates tested in this study showed increased resistance to antibiotics in the following categories: (i) penicillins (ampicillin (69%), piperacillin (22%); (ii) pyrimethamines (trimethoprim, 65%); (iii) nitrofurans (nitrofurantoin, 63%), and (iv) third-generation cephalosporin cefotaxime (53%). These results underscore a high prevalence of MDR amongst clinical isolates from Panama. Conclusions: The present study indicates that prevalence of BlaTEM-carrying strains is increasing with subsequent multidrug resistance in Panamá and as well reported worldwide. The virulent factors identified in this study provide valuable information regarding the prevalence of resistance genes and their potential impact on treatments that exploit the unique physiology of the pathogen. To prevent further spread of MDR, the proportions of resistant strains of Pseudomonas aeruginosa should be constantly evaluated on healthcare institutions of Panamá. More importantly, this information can be used to better understand the evolution and dissemination of strains hoping to prevent the development of resistance in Pseudomonas aeruginosa. Future studies quantifying the expression of these virulent genes will emphasize on the acquisition of multidrug resistance.

Prevalence of antibiotic resistance and virulent factors in nosocomial clinical isolates of Pseudomonas aeruginosa from Panamá.

BACKGROUND: Pseudomonas aeruginosa is an important causative agent of nosocomial infections. As pathogen, P. aeruginosa is of increasing clinical importance due to its ability to develop high-level multidrug resistance (MDR). METHODS: The aim of the present study was to better understand the intrinsic virulence of circulating strains of Pseudomonas aeruginosa, by surveying and characterizing the antibiotic resistance profiles and prevalence of virulence factors in 51 clinical isolates of P. aeruginosa obtained from children admitted to Hospital del Niño-Panamá during the period of October 2016 until March 2017. Antimicrobial susceptibilities were assessed by determining the minimum inhibitory concentration for 12 antibiotics against P. aeruginosa clinical isolates using the VITEK system (https://www.biomerieux.com). Additionally, all isolates were examined by Polymerase Chain Reaction (PCR) for the presence of components of the MexAB-OprM efflux pump system (mexABR) and pyoverdine receptor genes and betalactamases resistance genes (ESBL) using gene-specific primers. RESULTS: A total of 51 pyoverdine producing clinical isolates were analyzed, all of which expressed resistance genes such as genes of the MexAB-OprM efflux pump system (mexABR) and pyoverdine receptor genes (fpvA). Out of 51 MDR isolates, 22 were ESBL producers. The most common ESBL gene was blaTEM expressed by 43% of the isolates. The isolates tested in this study showed increased resistance to antibiotics in the following categories: (i) penicillins (ampicillin (69%), piperacillin (22%); (ii) pyrimethamines (trimethoprim, 65%); (iii) nitrofurans (nitrofurantoin, 63%), and (iv) third-generation cephalosporin cefotaxime (53%). These results underscore a high prevalence of MDR amongst clinical isolates from Panama. CONCLUSIONS: The present study indicates that prevalence of BlaTEM-carrying strains is increasing with subsequent multidrug resistance in Panamá and as well reported worldwide. The virulent factors identified in this study provide valuable information regarding the prevalence of resistance genes and their potential impact on treatments that exploit the unique physiology of the pathogen. To prevent further spread of MDR, the proportions of resistant strains of Pseudomonas aeruginosa should be constantly evaluated on healthcare institutions of Panamá. More importantly, this information can be used to better understand the evolution and dissemination of strains hoping to prevent the development of resistance in Pseudomonas aeruginosa. Future studies quantifying the expression of these virulent genes will emphasize on the acquisition of multidrug resistance.

Characterization of submicron emulsion processed by ultrasound homogenization to protect a bioactive extract from sea grape (Coccoloba uvifera L.).

In this study, the stability of a submicron emulsion to protect an extract obtained from sea grape fruit (Coccoloba uvifera L.) was evaluated. Extract characterization by MS-HPLC revealed the presence of 3 anthocyanins (cyanidin 3-glucoside, malvidin 3-glucoside, and delphinidin 3-glucoside), the content of total phenols was 263.86 ± 1.86 mg gallic acid equivalent/100 g, with an antioxidant capacity determined by ABTS and DPPH of 128.95 ± 1.00 and 26.18 ± 0.60 µg Trolox equivalents/mL, respectively. A submicron emulsion (0.424 µm) by Ultrasound with monomodal distribution, stable over time and low viscosity (1.94 mPa s) classified as a shear-thinning fluid was obtained. The thermogravimetric analysis (TGA) demonstrated the stability of the C. uvifera extract in the emulsion, which is thermostable (212 °C). These emulsions can be added into a beverage as a nutraceutical, dried for later use as pills or incorporated in foods.

A mosquito juvenile hormone binding protein (mJHBP) regulates the activation of innate immune defenses and hemocyte development.

Insects rely on the innate immune system for defense against pathogens, some aspects of which are under hormonal control. Here we provide direct experimental evidence showing that the juvenile hormone-binding protein (mJHBP) of Aedes aegypti is required for the regulation of innate immune responses and the development of mosquito blood cells (hemocytes). Using an mJHBP-deficient mosquito line generated by means of CRISPR-Cas9 gene editing technology we uncovered a mutant phenotype characterized by immunosuppression at the humoral and cellular levels, which profoundly affected susceptibility to bacterial infection. Bacteria-challenged mosquitoes exhibited significantly higher levels of septicemia and mortality relative to the wild type (WT) strain, delayed expression of antimicrobial peptides (AMPs), severe developmental dysregulation of embryonic and larval hemocytes (reduction in the total number of hemocytes) and increased differentiation of the granulocyte lineage. Interestingly, injection of recombinant wild type mJHBP protein into adult females three-days before infection was sufficient to restore normal immune function. Similarly, injection of mJHBP into fourth-instar larvae fully restored normal larval/pupal hemocyte populations in emerging adults. More importantly, the recovery of normal immuno-activation and hemocyte development requires the capability of mJHBP to bind JH III. These results strongly suggest that JH III functions in mosquito immunity and hemocyte development in a manner that is perhaps independent of canonical JH signaling, given the lack of developmental and reproductive abnormalities. Because of the prominent role of hemocytes as regulators of mosquito immunity, this novel discovery may have broader implications for the understanding of vector endocrinology, hemocyte development, vector competence and disease transmission.

Pre-exposure to non-pathogenic bacteria does not protect Drosophila against the entomopathogenic bacterium Photorhabdus.

Immune priming in insects involves an initial challenge with a non-pathogenic microbe or exposure to a low dose of pathogenic microorganisms, which provides a certain degree of protection against a subsequent pathogenic infection. The protective effect of insect immune priming has been linked to the activation of humoral or cellular features of the innate immune response during the preliminary challenge, and these effects might last long enough to promote the survival of the infected animal. The fruit fly Drosophila melanogaster is a superb model to dissect immune priming processes in insects due to the availability of molecular and genetic tools, and the comprehensive understanding of the innate immune response in this organism. Previous investigations have indicated that the D. melanogaster immune system can be primed efficiently. Here we have extended these studies by examining the result of immune priming against two potent entomopathogenic bacteria, Photorhabdus luminescens and P. asymbiotica. We have found that rearing D. melanogaster on diet containing a non-pathogenic strain of Escherichia coli alone or in combination with Micrococcus luteus upregulates the antibacterial peptide immune response in young adult flies, but it does not prolong fly life span. Also, subsequent intrathoracic injection with P. luminescens or P. asymbiotica triggers the Immune deficiency and Toll signaling pathways in flies previously exposed to a live or heat-killed mix of the non-pathogenic bacteria, but the immune activation fails to promote fly survival against the pathogens. These findings suggest that immune priming in D. melanogaster, and probably in other insects, is determined by the type of microbes involved as well as the mode of microbial exposure, and possibly requires a comprehensive and precise alteration of immune signaling and function to provide efficient protection against pathogenic infection.

Activation of mosquito complement antiplasmodial response requires cellular immunity.

The mosquito complement-like system is a major defense mechanism that limits Plasmodium infection. Ookinete midgut invasion results in irreversible damage to invaded cells and triggers epithelial nitration and complement activation. Several lines of evidence suggest that hemocytes participate in early antiplasmodial responses that target ookinetes, but their role remains unclear. The fate of hemocytes in response to Plasmodium infection was investigated by labeling this cell population in vivo. We found that midgut nitration triggers the local release of hemocyte-derived microvesicles (HdMv) into the basal labyrinth of the midgut. Several different strategies, such as gene silencing, immune priming, or systemic injection of polystyrene beads, were used to either enhance or reduce HdMv release. We provide direct experimental evidence that contact of hemocytes with the nitrated midgut basal surface triggers HdMv release and that this response is necessary for effective activation of mosquito complement. Our studies suggest that hemocyte-derived microvesicles may deliver some critical factor(s) that promote activation of thioester-containing protein 1, a key effector of the mosquito antiplasmodial immunity.

TGF-ß signaling regulates resistance to parasitic nematode infection in Drosophila melanogaster.

Over the past decade important advances have been made in the field of innate immunity; however, our appreciation of the signaling pathways and molecules that participate in host immune responses to parasitic nematode infections lags behind that of responses to microbial challenges. Here we have examined the regulation and immune activity of Transforming Growth Factor-beta (TGF-ß) signaling in the model host Drosophila melanogaster upon infection with the nematode parasites Heterorhabditis gerrardi and H. bacteriophora containing their mutualistic bacteria Photorhabdus. We have found that the genes encoding the Activin and Bone Morphogenic Protein (BMP) ligands Dawdle (Daw) and Decapentaplegic (Dpp) are transcriptionally induced in flies responding to infection with the nematode parasites, containing or lacking their associated bacteria. We also show that deficient Daw or Dpp regulates the survival of D. melanogaster adults to the pathogens, whereas inactivation of Daw reduces the persistence of the nematodes in the mutant flies. These findings demonstrate a novel role for the TGF-ß signaling pathways in the host anti-nematode immune response. Understanding the molecular mechanisms of host anti-nematode processes will potentially lead to the development of novel means for the efficient control of parasitic nematodes.

[Laparoscopic cholecystectomy in pregnant patient: experience in hospital General de Mexico in a period of 5 years (2008-2012)]. / Colecistectomía laparoscópica en pacientes embarazadas: experiencia del Hospital General de México 2008-2012.

BACKGROUND: Cholecystitis is the second cause of abdominal pain during pregnancy. 1-8 of 10,000 requiring surgery, being performed in the first and second quarter laparoscopically. 100% of patients with cholecystitis, about 12% are associated with pancreatitis with high rates of maternal and fetal morbidity and mortality. OBJECTIVE: To evaluate advantages--disadvantages of maternal-fetal pregnancy laparoscopic cholecystectomy and its preventive character avoiding cases of pancreatitis. We analyze the results obtained in Perinatology Service in General Hospital of Mexico (2008 to 2012) comparing them with the current literature. MATERIAL AND METHODS: A retrospective, cross sectional, descriptive. Analyzing the following variables: maternal age, gestational age, number of gestations, surgical technique, and postoperative complications trans, maternal and perinatal morbidity, gallbladder colic episodes prior, liver ultrasound report--bile ducts, tocolytic management. RESULTS: 20 laparoscopic cholecystectomies were performed in pregnant patients. Maternal age 21-38 years, mostly multigesta. 5 patients was performed at weeks 9, 14, 20 and 25 between the SDG and 1 at 27.5 SDG.Vesicular colicky eight previous USG mostly with gallstones.Two cases of mild acute pancreatitis satisfactorily resolved. No trans or postoperative complications. Open technique for performing pneumoperitoneum (Hasson). Tocolytic management indomethacin in 100% of cases. CONCLUSIONS: The results obtained are consistent with the current literature, confirming that laparoscopic cholecystectomy is the best treatment option with minimal fetal maternal morbidity, reducing the incidence of pancreatitis and maternal- fetal consequences.

Síndrome de Evans Fisher asociado con esclerodermia / Fisher Evans syndrome associated with scleroderma

El síndrome de Evans Fisher, descrito por primera vez en 1951, es un desorden autoinmune caracterizado por la presencia simultánea o secuencial de anemia hemolítica, trombocitopenia inmune y, en ocasiones, neutropenia inmune; con una prueba de antiglobulina directa positiva. Puede ser de causa primaria o secundaria a otras condiciones, como el lupus eritematoso sistémico, los síndromes linfoproliferativos o inmunodeficiencias primarias. Es muy rara su asociación con la esclerodermia. Con el término esclerodermia, que en sentido literal significa piel dura, se designa un grupo de enfermedades y síndromes que tienen como característica común la induración y el engrosamiento cutáneos. Se caracteriza por la presencia de un depósito excesivo de los componentes del tejido conjuntivo, expresado en forma de fibrosis hística, y por alteraciones estructurales del lecho vascular. Con un cuadro clínico muy amplio, afecta fundamentalmente la piel y ciertos órganos internos, como tubo digestivo, pulmón, corazón y riñón. Se presenta una paciente femenina de 75 años de edad, piel negra, con antecedentes de hipertensión arterial, diabetes mellitus tipo 2, cardiopatía isquémica y esclerodermia, esta última diagnosticada seis meses antes de su ingreso. Acudió por decaimiento marcado, palidez cutáneo-mucosa intensa y petequias generalizadas. En los estudios realizados se detectó anemia y trombocitopenia severas, reticulocitosis, prueba de antiglobulina directa positiva e hipercelularidad medular con hiperplasia severa de los sistemas megacariopoyético y eritropoyético. Se diagnosticó un síndrome de Evans Fisher y se trató con esteroides e inmunomoduladores; se logró la mejoría clínica y la remisión hematológica(AU)

The Evans syndrome, first described in 1951, is an autoimmune disorder characterized by the simultaneous or sequential development of hemolytic anemia and immune thrombocytopenia or immune neutropenia. It may be of primary origin or secondary to other diseases or conditions such as systemic lupus erythematosus, lymphoproliferative disorders or primary immunodeficiencies. Its association with scleroderma is considered very rare. The word scleroderma, which literally means hard skin, designates a group of diseases and syndromes of common feature in induration and thickening the skin. It is characterized by the presence of excessive deposition of connective tissue components, expressed as histic fibrosis, and structural alterations of the vascular bed. With a broad clinical view, it primarily affects the skin and certain internal organs such as gastrointestinal tract, lung, heart and kidney. We present a 75 year-old female, black skin, with a history of hypertension, type 2 diabetes, ischemic heart disease and scleroderma, the latter diagnosed six months before admission. The patient referred marked weakness, pale skin and generalized petechiae. The complete blood count detected severe anemia, thrombocytopenia and reticulocytosis. Other studies showed positive direct Coombs test and severe hypercellularity. Evans Fisher syndrome was diagnosed and treated with steroids and immunomodulators; clinical improvement and hematologic remission was achieved(AU)

Síndrome de Evans Fisher asociado con esclerodermia / Fisher Evans syndrome associated with scleroderma

El síndrome de Evans Fisher, descrito por primera vez en 1951, es un desorden autoinmune caracterizado por la presencia simultánea o secuencial de anemia hemolítica, trombocitopenia inmune y, en ocasiones, neutropenia inmune; con una prueba de antiglobulina directa positiva. Puede ser de causa primaria o secundaria a otras condiciones, como el lupus eritematoso sistémico, los síndromes linfoproliferativos o inmunodeficiencias primarias. Es muy rara su asociación con la esclerodermia. Con el término esclerodermia, que en sentido literal significa piel dura, se designa un grupo de enfermedades y síndromes que tienen como característica común la induración y el engrosamiento cutáneos. Se caracteriza por la presencia de un depósito excesivo de los componentes del tejido conjuntivo, expresado en forma de fibrosis hística, y por alteraciones estructurales del lecho vascular. Con un cuadro clínico muy amplio, afecta fundamentalmente la piel y ciertos órganos internos, como tubo digestivo, pulmón, corazón y riñón. Se presenta una paciente femenina de 75 años de edad, piel negra, con antecedentes de hipertensión arterial, diabetes mellitus tipo 2, cardiopatía isquémica y esclerodermia, esta última diagnosticada seis meses antes de su ingreso. Acudió por decaimiento marcado, palidez cutáneo-mucosa intensa y petequias generalizadas. En los estudios realizados se detectó anemia y trombocitopenia severas, reticulocitosis, prueba de antiglobulina directa positiva e hipercelularidad medular con hiperplasia severa de los sistemas megacariopoyético y eritropoyético. Se diagnosticó un síndrome de Evans Fisher y se trató con esteroides e inmunomoduladores; se logró la mejoría clínica y la remisión hematológica

The Evans syndrome, first described in 1951, is an autoimmune disorder characterized by the simultaneous or sequential development of hemolytic anemia and immune thrombocytopenia or immune neutropenia. It may be of primary origin or secondary to other diseases or conditions such as systemic lupus erythematosus, lymphoproliferative disorders or primary immunodeficiencies. Its association with scleroderma is considered very rare. The word scleroderma, which literally means hard skin, designates a group of diseases and syndromes of common feature in induration and thickening the skin. It is characterized by the presence of excessive deposition of connective tissue components, expressed as histic fibrosis, and structural alterations of the vascular bed. With a broad clinical view, it primarily affects the skin and certain internal organs such as gastrointestinal tract, lung, heart and kidney. We present a 75 year-old female, black skin, with a history of hypertension, type 2 diabetes, ischemic heart disease and scleroderma, the latter diagnosed six months before admission. The patient referred marked weakness, pale skin and generalized petechiae. The complete blood count detected severe anemia, thrombocytopenia and reticulocytosis. Other studies showed positive direct Coombs test and severe hypercellularity. Evans Fisher syndrome was diagnosed and treated with steroids and immunomodulators; clinical improvement and hematologic remission was achieved

Molecular mechanisms of aging and immune system regulation in Drosophila.

Aging is a complex process that involves the accumulation of deleterious changes resulting in overall decline in several vital functions, leading to the progressive deterioration in physiological condition of the organism and eventually causing disease and death. The immune system is the most important host-defense mechanism in humans and is also highly conserved in insects. Extensive research in vertebrates has concluded that aging of the immune function results in increased susceptibility to infectious disease and chronic inflammation. Over the years, interest has grown in studying the molecular interaction between aging and the immune response to pathogenic infections. The fruit fly Drosophila melanogaster is an excellent model system for dissecting the genetic and genomic basis of important biological processes, such as aging and the innate immune system, and deciphering parallel mechanisms in vertebrate animals. Here, we review the recent advances in the identification of key players modulating the relationship between molecular aging networks and immune signal transduction pathways in the fly. Understanding the details of the molecular events involved in aging and immune system regulation will potentially lead to the development of strategies for decreasing the impact of age-related diseases, thus improving human health and life span.

A novel method for infecting Drosophila adult flies with insect pathogenic nematodes.

Drosophila has been established as an excellent genetic and genomic model to investigate host-pathogen interactions and innate immune defense mechanisms. To date, most information on the Drosophila immune response derives from studies that involve bacterial, fungal or viral pathogens. However, immune reactions to insect parasitic nematodes are still not well characterized. The nematodes Heterorhabditis bacteriophora live in symbiosis with the entomopathogenic bacteria Photorhabdus luminescens, and they are able to invade and kill insects. Interestingly, Heterorhabditis nematodes are viable in the absence of Photorhabdus. Techniques for infecting Drosophila larvae with these nematodes have been previously reported. Here, we have developed a method for infecting Drosophila adult flies with Heterorhabditis nematodes carrying (symbiotic worms) or lacking (axenic worms) their associated bacteria. The protocol we present can be readily adapted for studying parasitic strategies of other insect nematodes using Drosophila as the host infection model.

Insect immune responses to nematode parasites.

Host innate immunity plays a central role in detecting and eliminating microbial pathogenic infections in both vertebrate and invertebrate animals. Entomopathogenic or insect pathogenic nematodes are of particular importance for the control of insect pests and vectors of pathogens, while insect-borne nematodes cause serious diseases in humans. Recent work has begun to use the power of insect models to investigate host-nematode interactions and uncover host antiparasitic immune reactions. This review describes recent findings on innate immune evasion strategies of parasitic nematodes and host cellular and humoral responses to the infection. Such information can be used to model diseases caused by human parasitic nematodes and provide clues indicating directions for research into the interplay between vector insects and their invading tropical parasites.