In vitro bactericidal activity of a carbohydrate polymer with zinc oxide for the treatment of chronic wounds.

Introduction. Biological adhesives and effective topical therapeutic agents that improve wound healing are urgently required for the treatment of chronic ulcers. A biodegradable adhesive based on a carbohydrate polymer with zinc oxide (CPZO) was shown to possess anti-inflammatory activity and enhance wound healing, but its bactericidal activity was unknown.Aim. To investigate the bactericidal activity of CPZO against bacteria commonly present as infectious agents in chronic wounds.Methodology. We examined the bactericidal activity of CPZO against three biofilm-producing bacteria (Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa) through three strategies: bacterial suspension, biofilm disruption and in vitro wound biofilm model.Results. In suspension cultures, CPZO had direct, potent bactericidal action against S. aureus within 24 h, whereas E. coli took 7 days to be eliminated. By contrast, P. aeruginosa survived up to 14 days with CPZO. CPZO had biofilm disruption activity against clinical isolates of S. aureus in the anti-biofilm test. Finally, in the in vitro wound biofilm model, CPZO dramatically reduced the bacterial viability of S. aureus and P. aeruginosa.Conclusions. Together with its previously shown anti-inflammatory properties, the bactericidal activity of CPZO gives it the potential to be a first-line therapeutic option for chronic various ulcers and, possibly, other chronic ulcers, preventing or controlling microbial infections, and leading to the healing of such complicated chronic ulcers.

Prevalence of inhibitors and clinical characteristics in patients with haemophilia in a middle-income Latin American country.

INTRODUCTION: Development of inhibitors is the most serious complication in patients with haemophilia (PWH). The prevalence of inhibitors in patients with severe haemophilia A (HA) is approximately 25%-30%. Inhibitor prevalence differs among populations. Some studies report a prevalence of almost twice in Hispanic as compared to Caucasian patients. Most data available, on the prevalence of inhibitors and their predisposing factors, originate from centres in developed countries. AIM: Establish the prevalence of inhibitors of FVIII and FIX in Mexico. METHODS: This was an observational, cross-sectional and descriptive study. The records of all patients diagnosed with haemophilia A (HA) or B (HB), with and without inhibitors, were included. Clinical and demographical characteristics of patients with inhibitors were assessed. Statistical analysis was performed using IBM SPSS version 22. The Ethics Committees of the various participating institutions approved this study. RESULTS: A total of 1455 patients from the 20 participating centres were recruited, from which 1208 (83.02%) had HA and 247 (16.97%) were diagnosed with HB. The presence of inhibitors in severe HA was reported in 93/777(11.96%), and 10/162 (6.17%) in severe HB. Of them, 91.7% exhibited high titres in HA and 100% in HB. CONCLUSION: In Mexico, the general prevalence of inhibitors varies considerably among centres. This study established a basis of comparison for future development and advances in the treatment and follow-up of patients. These findings also augment our understanding of risk factors related to inhibitor development.

Elucidation of the Amygdalin Pathway Reveals the Metabolic Basis of Bitter and Sweet Almonds (Prunus dulcis).

Almond (Prunus dulcis) is the principal Prunus species in which the consumed and thus commercially important part of the fruit is the kernel. As a result of continued selection, the vast majority of almonds have a nonbitter kernel. However, in the field, there are trees carrying bitter kernels, which are toxic to humans and, consequently, need to be removed. The toxicity of bitter almonds is caused by the accumulation of the cyanogenic diglucoside amygdalin, which releases toxic hydrogen cyanide upon hydrolysis. In this study, we identified and characterized the enzymes involved in the amygdalin biosynthetic pathway: PdCYP79D16 and PdCYP71AN24 as the cytochrome P450 (CYP) enzymes catalyzing phenylalanine-to-mandelonitrile conversion, PdUGT94AF3 as an additional monoglucosyl transferase (UGT) catalyzing prunasin formation, and PdUGT94AF1 and PdUGT94AF2 as the two enzymes catalyzing amygdalin formation from prunasin. This was accomplished by constructing a sequence database containing UGTs known, or predicted, to catalyze a ß(1→6)-O-glycosylation reaction and a Basic Local Alignment Search Tool search of the draft version of the almond genome versus these sequences. Functional characterization of candidate genes was achieved by transient expression in Nicotiana benthamiana Reverse transcription quantitative polymerase chain reaction demonstrated that the expression of PdCYP79D16 and PdCYP71AN24 was not detectable or only reached minute levels in the sweet almond genotype during fruit development, while it was high and consistent in the bitter genotype. Therefore, the basis for the sweet kernel phenotype is a lack of expression of the genes encoding the two CYPs catalyzing the first steps in amygdalin biosynthesis.

Synteny-Based Development of CAPS Markers Linked to the Sweet kernel LOCUS, Controlling Amygdalin Accumulation in Almond (Prunus dulcis (Mill.) D.A.Webb).

The bitterness and toxicity of wild-type seeds of Prunoideae is due to the cyanogenic glucoside amygdalin. In cultivated almond (Prunus dulcis (Mill.) D.A. Webb), a dominant mutation at the Sk locus prevents amygdalin accumulation and thus results in edible sweet kernels. Here, we exploited sequence similarity and synteny between the genomes of almond and peach (Prunus persica (L.) Batsch) to identify cleaved amplified polymorphic sequence (CAPS) molecular markers linked to the Sk locus. A segregant F1 population was used to map these markers on the Sk genomic region, together with Sk-linked simple sequence repeat (SSR) markers previously described. Molecular fingerprinting of a cultivar collection indicated the possibility to use CAPS polymorphisms identified in this study in breeding programs arising from different parental combinations. Overall, we highlight a set of codominant markers useful for early selection of sweet kernel genotypes, an aspect of primary importance in almond breeding. In addition, by showing collinearity between the physical map of peach and the genetic map of almond with respect to the Sk genomic region, we provide valuable information for further marker development and Sk positional cloning.

ß-Glucosidase activity in almond seeds.

Almond bitterness is the most important trait for breeding programs since bitter-kernelled seedlings are usually discarded. Amygdalin and its precursor prunasin are hydrolyzed by specific enzymes called ß-glucosidases. In order to better understand the genetic control of almond bitterness, some studies have shown differences in the location of prunasin hydrolases (PH, the ß-glucosidase that degrades prunasin) in sweet and bitter genotypes. The aim of this work was to isolate and characterize different PHs in sweet- and bitter-kernelled almonds to determine whether differences in their genomic or protein sequences are responsible for the sweet or bitter taste of their seeds. RNA was extracted from the tegument, nucellus and cotyledon of one sweet (Lauranne) and two bitter (D05-187 and S3067) almond genotypes throughout fruit ripening. Sequences of nine positive Phs were then obtained from all of the genotypes by RT-PCR and cloning. These clones, from mid ripening stage, were expressed in a heterologous system in tobacco plants by agroinfiltration. The PH activity was detected using the Feigl-Anger method and quantifying the hydrogen cyanide released with prunasin as substrate. Furthermore, ß-glucosidase activity was detected by Fast Blue BB salt and Umbelliferyl method. Differences at the sequence level (SNPs) and in the activity assays were detected, although no correlation with bitterness was found.

Cyanogenic Glucosides and Derivatives in Almond and Sweet Cherry Flower Buds from Dormancy to Flowering.

Almond and sweet cherry are two economically important species of the Prunus genus. They both produce the cyanogenic glucosides prunasin and amygdalin. As part of a two-component defense system, prunasin and amygdalin release toxic hydrogen cyanide upon cell disruption. In this study, we investigated the potential role within prunasin and amygdalin and some of its derivatives in endodormancy release of these two Prunus species. The content of prunasin and of endogenous prunasin turnover products in the course of flower development was examined in five almond cultivars - differing from very early to extra-late in flowering time - and in one sweet early cherry cultivar. In all cultivars, prunasin began to accumulate in the flower buds shortly after dormancy release and the levels dropped again just before flowering time. In almond and sweet cherry, the turnover of prunasin coincided with increased levels of prunasin amide whereas prunasin anitrile pentoside and ß-D-glucose-1-benzoate were abundant in almond and cherry flower buds at certain developmental stages. These findings indicate a role for the turnover of cyanogenic glucosides in controlling flower development in Prunus species.

Recent advancements to study flowering time in almond and other Prunus species.

Flowering time is an important agronomic trait in almond since it is decisive to avoid the late frosts that affect production in early flowering cultivars. Evaluation of this complex trait is a long process because of the prolonged juvenile period of trees and the influence of environmental conditions affecting gene expression year by year. Consequently, flowering time has to be studied for several years to have statistical significant results. This trait is the result of the interaction between chilling and heat requirements. Flowering time is a polygenic trait with high heritability, although a major gene Late blooming (Lb) was described in "Tardy Nonpareil." Molecular studies at DNA level confirmed this polygenic nature identifying several genome regions (Quantitative Trait Loci, QTL) involved. Studies about regulation of gene expression are scarcer although several transcription factors have been described as responsible for flowering time. From the metabolomic point of view, the integrated analysis of the mechanisms of accumulation of cyanogenic glucosides and flowering regulation through transcription factors open new possibilities in the analysis of this complex trait in almond and in other Prunus species (apricot, cherry, peach, plum). New opportunities are arising from the integration of recent advancements including phenotypic, genetic, genomic, transcriptomic, and metabolomics studies from the beginning of dormancy until flowering.

Further experimental studies on a biodegradable adhesive for protection of colorectal anastomosis.

BACKGROUND AND AIMS: Anastomotic leaks (AL) continue to be a devastating complication after colorectal surgery. The purpose of this experimental study was to confirm if Pebisut® applied to intestinal suture lines increases tensile strength in the critical days of healing and to evaluate its anti-inflammatory properties. METHODS: Bursting pressures and histological evaluation of suture lines in dogs were performed, comparing the burst strength with collagen or fatty tissue patches with/without Pebisut® (patent granted in the USPTO 8,252.333, 26.01.2006, in the European Union 2,062,602, 01.12.2010, in Canada 2,661,686, 21.08.2007 and in Mexico P.C.T./MX/a/2009/001737). RESULTS: Pebisut® significantly increases burst strength in suture lines in long-term procedures with both collagen and fat pad patches. The adhesive penetrates rapidly into the suture lines, sealing them and progressing towards the intestinal lumen, disappearing in 14-20 days. It was well tolerated without any evidence of "foreign body" reaction. CONCLUSIONS: Application of the biodegradable adhesive Pebisut® is easy, well tolerated by mammalian tissues and consistently increases the burst strength of suture lines. Therefore, it may provide more tensile strength in anastomosis and help protect AL, one of the most serious complications in gastrointestinal surgery. If this experimental finding could be translated to clinical surgery, the protection of colorectal anastomosis could be beneficial to patients. Additionally, this could also have a positive impact on the economic expenditures of healthcare systems and patients.

Potent anti-inflammatory activity of carbohydrate polymer with oxide of zinc.

Pebisut is a biological adhesive composed of naturally occurring carbohydrates combined with zinc oxide (ZnO) initially used as a coadjutant for healing of anastomoses. Likewise some works demonstrated that carbohydrate complexes exerts anti-inflammatory activity and it is widely known that ZnO modulate inflammation. However, the direct effects of Pebisut on isolated cells and acute inflammatory responses remained to be investigated. The present study evaluated anti-inflammatory effect of Pebisut using lipopolysaccharide (LPS) stimulated human mononuclear cells, chemotaxis, and cell infiltration in vivo in a murine model of peritonitis. Our data show that human cells treated with different dilutions of Pebisut release less IL-6, IL-1 ß , and IL-8 after LPS stimuli compared with the control treated cells. In addition, Pebisut lacked chemotactic activity in human mononuclear cells but was able to reduce chemotaxis towards CCL2, CCL5, and CXCL12 that are representative mononuclear cells chemoattractants. Finally, in a murine model of peritonitis, we found less number of macrophages (F4/80(+)) and T lymphocytes (CD3(+)) in peritoneal lavages from animals treated with Pebisut. Our results suggest that Pebisut has anti-inflammatory activity, which might have a beneficial effect during anastomoses healing or wounds associated with excessive inflammation.

A new biodegradable adhesive for protection of intestinal anastomoses. Preliminary communication.

BACKGROUND AND AIMS: Anastomotic leaks continue to be a devastating complication for patients and surgeons worldwide. The few surgical adhesives available to date have not achieved the desired clinical results. The purpose of this experimental study was to determine if Pebisut® applied to intestinal suture lines provides increased resistance and protection during the critical days of healing. METHODS: Intestinal lesions were caused in rats and dogs and a new biodegradable adhesive (Pebisut®) (patent granted in the European Union 07808494.4-1219, 01.12.2010, in Mexico P.C.T./MX/a/2009/001737, 16.02.2009, pending in the U.S.P.T.O. 60/762,136, 26.01.2006) was applied to compare the resistance of suture lines using bursting pressures and histologically. RESULTS: Under acute and chronic conditions, Pebisut® strengthened and made the suture lines more resistant, while histologically penetrating and sealing them. The adhesive disappears within 2-3 weeks and is well tolerated by the intestinal tissues. CONCLUSIONS: This biodegradable adhesive provides greater resistance, temporarily protects suture lines and may prevent anastomotic leaks.

[Occurrence of Amrineus cocofolius Flechtmann (Prostigmata: Eriophyidae) in coconut fruits (Cocos nucifera L.) in Cuba]. / Ocorrência de Amrineus cocofolius Flechtmann (Prostigmata: Eriophyidae) em frutos de coqueiro (Cocos nucifera L.) em Cuba.

The presence of the eriophyid mite, Amrineus cocofolius Flechtmann, was confirmed in association with equatorial necrotic bands on the coconut fruit epidermis, in different growth areas in the Provinces of La Habana, Granma y Guantánamo, Cuba, from February 2003 to March 2004.

Ocorrência de Amrineus cocofolius Flechtmann (Prostigmata: Eriophyidae) em frutos de coqueiro (Cocos nucifera L.) em Cuba / Occurrence of Amrineus cocofolius Flechtmann (Prostigmata: Eriophyidae) in coconut fruits (Cocos nucifera L.) in Cuba

A presença do ácaro eriofiídeo Amrineus cocofolius Flechtmann foi constatada em associação com bandas necróticas equatoriais na epiderme dos frutos de coqueiros, em diferentes plantações nas províncias de La Habana, Granma e Guantánamo, Cuba, de fevereiro de 2003 a março de 2004.

The presence of the eriophyid mite, Amrineus cocofolius Flechtmann, was confirmed in association with equatorial necrotic bands on the coconut fruit epidermis, in different growth areas in the Provinces of La Habana, Granma y Guantánamo, Cuba, from February 2003 to March 2004.

Large duodenal villous adenoma requiring head of the pancreas and pylorus-preserving total duodenectomy.

Villous adenomas of the duodenum (VAD) are infrequent lesions of the gastrointestinal tract but have a high risk of recurrence and malignancy. For these reasons and its specific topographic location, the surgical treatment of VAD is still controversial. Herein we present a case of large VAD located in the second duodenal portion that was successfully treated with a head of the pancreas, pylorus-preserving total duodenectomy (PPTD). PPTD should be an excellent option in patients with large adenomas because it allows preservation of the pancreas, gastrointestinal function is maintained, the possibility of a recurrence and of an invasive carcinoma of the ampulla is eliminated, and finally because it permits an adequate endoscopic follow-up. PPTD should not be used in the presence of malignancy and/or high-grade dysplasia.

Corrección del reflujo gastroesofágico en obesidad mórbida, con banda gástrica ajustable sueca / Correction of gastroesophageal reflux in morbid obesity, with swedish gastric band

La obesidad mórbida es una afección clínica frecuente, la cual representa un factor de riesgo para varias entidades clínicas que incrementan la morbilidad y mortalidad de la misma. En los últimos años el tratamiento quirúrgico ha sido una excelente alternativa, gracias a los resultados obtenidos por cirugía laparoscópica y entre los diferentes procedimientos existentes, la banda gástrica ajustable ha jugado un importante papel. La obesidad mórbida está asociada con varios desordenes clínicos, entre estos el reflujo gastroesofágico en el cual el tratamiento médico por si solo no es efectivo para su corrección definitiva, por lo que se ha recurrido a la cirugía. Objetivos: Comparar el índice de reflujo gastroesofágico, previo a la colocación de la banda gástrica ajustable y posterior a su colocación. Materiales y métdos: Se realiza estudio prospectivo, longitudinal y analítico en el cual se estudiaron 10 pacientes con obesidad mórbida asociados a reflujo gastroesofágico severo a quienes se les colocó banda gástrica ajustable. Resultados: Se obtuvieron adecuados resultados en la obesidad mórbida, y completa ausencia de los síntomas de reflujo gastroesofágico, demostrando por clínica, endoscopía, manometría y pH metría de 24 horas pos operatoria la corrección del reflujo gastroesofágico patológico, al igual que la normalización de la presión del esfínter esofágico inferior. Con lo cual confirmamos la hipótesis, de que la banda gástrica ajustable, no solo disminuye la obesidad mórbida, sino corrige el reflujo gastroesofágico

Cirugia laparoscopica para reflujo gastroesofágico / Laparoscopic surgery for gastroesophageal reflux

La funduplicación de Nissen es una de las operaciones para reflujo gastroesofágico de mayor aceptación; sin embargo, es una operación con importante morbilidad para una patología benigna. Esta operación, se ha venido haciendo en los últimos años mediante cirugía laparoscópica con menores problemas que la cirugía clásica abierta. Presentamos una experiencia preliminar de 44 casos resueltos con este abordaje en el que tenemos un 94.8 por ciento de pacientes con alivio de los síntomas y mínimas complicaciones. Solo 2 pacientes requirieron una segunda cirugía. La presión final del esfinter fue de 25 mmH y la longitud de la funduplicatura fue de 3.06 cms

Reparación laparoscópica de los defectos herniarios inguino-crurales: propuesta de clasificación (informe preliminar) / Laparoscopic hernia repair

De febrero de 1992 a diciembre de 1994 se intervinieron 138 pacientes con un total de 179 hernias. Se efectuaron diferentes tipos de reparación dependiendo del defecto a tratar; a) ligadura del defecto en el cuello del saco invertido; b) disección peritoneal y sutura del defecto; c) disección peritoneal, sutura del defecto y colocación de malla y d) disección peritoneal y colocación de polipropileno para cubrir la región. Ochenta y tres fueron hombres y 55 mujeres, con edad mínima de 8 años y máxima de 91, promedio 39 años, sin selección de casos. Los tipos de reparación fueron dependiendo de su apariencia endoscópica. Las indirectas con orificio interno menor a un centímetro de diámetro(23) fueron tratadas con cierre del orificio del saco herniario invertido, (19) indirectas con orificio entre 1 y 2 centímetros de diámetro con disección del peritoneo y aproximación con sutura del tracto ileopúbico al arco del transverso, y las indirectas de mas de 2 cm. de diámetro del orificio interno (89), así como las directas de cualquier tamaño (17), hernias de pantalón (15), femorales (2), las que tenían deslizamiento de vísceras (6) y las recidivantes (8) con disección del peritoneo y aproximación con sutura en los casos que era posible hacerlo sin tensión y aplicación de malla. La reintegración normal a las actividades fueantes de una semana en el 90/100 de los casos. La recidiva fue de 1.1/100. Se presentaron 2 casos de meralgia parestésica, tres de hematoma resuelto espontáneamente, y una necrosis de grasa sin recidiva de la hernia, que se reexploró. No se registró mortalidad. De acuerdo a los anteriores resultados clasificamos las hernias con base en su apariencia endoscópica para individualizar el tratamiento y evaluar los resultados a largo plazo en: Tipo Ia, indirectas con orificio interno de menos de 1 cm de diámetro. Ib indirectas con orificio interno entre 1 y 2 cm. de diámetro y Ic indirectas con orificio interno de más de 2 cm. de diámetro. Tipo IIa las directas de cualquier tamaño. IIb las hernias en pantalón, IIc hernias femorales. Tipo III hernias con deslizamiento. Tipo IV recidivantes

Cateterización percutánea y corrección de una fístula duodenal lateral / Percutaneous catheterization and repair of a lateral duodenal fistula

Se informa de un caso clínico que sufrió como complicación de cirugía bilio-pancreática, una fístula duodenal lateral. El manejo conservador de nutrición parenteral, reemplazo hidroelectrolítico adecuado, etc. no produjo cese del drenaje de dicha fístula apareciendo entonces una complicación infecciosa de la alimentación parenteral que por otra parte tuvo que ser suspendida al no conterse con otras venas adecuadas para su utilización. Antes de proceder con la reintervención quirúrgica y la colocación de yeyunostomías distales, se solicitó al Departamento de Radiología la cateterización distal, percutánea, transduodenal, utilizando el trayecto fistuloso con una sonda que nos permitiera administrar la alimentación enteral sin residuo. Esto fue logrado pero además, sorpresivamente, la instalación de la sonda permitió un taponamiento adecuado de dicho orificio fistuloso lo cual llevó al tratamiento conservador a un resultado exitoso