Insights into the hydrolytic activity of Asclepias fruticosa L. protease.

OBJECTIVE: To determine the enzymatic properties of asclepain f, a plant cysteine protease isolated and purified from the latex of Asclepias fruticosa, and to investigate its potential application to hydrolyze soybean proteins. RESULTS: Kinetic parameters were determined by hydrolysis of p-Glu-Phe-Leu-p-nitroanilide (PFLNA). The Km value for asclepain f was 6 to 8 times higher than those achieved for papain, bromelain and ficin, the main plant cysteine proteases. Asclepain f showed 12 cut-off points toward the oxidized B chain insulin, revealing that the enzyme possesses broad substrate specificity. The cut specificity was governed by the presence of hydrophobic residues (F, L, V) in the P2 position. Asclepain f was able to selectively hydrolyze soybean proteins at pH 10, employing an enzyme/substrate ratio of 0.2% (w/w). The enzymatic hydrolysis allowed a strong increase in the solubility, water and oil holding capacity. CONCLUSIONS: Asclepain f was revealed as a successful enzyme for biocatalysis of protein hydrolysis processes at alkaline pH. This new plant protease has a broad substrate specificity and is capable of selectively degrading the fractions of soy proteins and improving its functional properties.

A Bowman-Birk protease inhibitor purified, cloned, sequenced and characterized from the seeds of Maclura pomifera (Raf.) Schneid.

MAIN CONCLUSION: A new BBI-type protease inhibitor with remarkable structural characteristics was purified, cloned, and sequenced from seeds of Maclura pomifera , a dicotyledonous plant belonging to the Moraceae family. In this work, we report a Bowman-Birk inhibitor (BBI) isolated, purified, cloned, and characterized from Maclura pomifera seeds (MpBBI), the first of this type from a species belonging to Moraceae family. MpBBI was purified to homogeneity by RP-HPLC, total RNA was extracted from seeds of M. pomifera, and the 3'RACE-PCR method was applied to obtain the cDNA, which was cloned and sequenced. Peptide mass fingerprinting (PMF) analysis showed correspondence between the in silico-translated protein and MpBBI, confirming that it corresponds to a new plant protease inhibitor. The obtained cDNA encoded a polypeptide of 65 residues and possesses 10 cysteine residues, with molecular mass of 7379.27, pI 6.10, and extinction molar coefficient of 9105 M-1 cm-1. MpBBI inhibits strongly trypsin with K i in the 10-10 M range and was stable in a wide array of pH and extreme temperatures. MpBBI comparative modeling was applied to gain insight into its 3D structure and highlighted some distinguishing features: (1) two non-identical loops, (2) loop 1 (CEEESRC) is completely different from any known BBI, and (3) the amount of disulphide bonds is also different from any reported BBI from dicot plants.

Latex peptidases of Calotropis procera for dehairing of leather as an alternative to environmentally toxic sodium sulfide treatment.

Dehairing of crude leather is a critical stage performed at the beginning of its processing to obtain industrially useful pieces. Tanneries traditionally apply a chemical process based on sodium sulfide. Since this chemical reactive is environmentally toxic and inefficiently recycled, innovative protocols for reducing or eliminating its use in leather depilation are welcomed. Therefore, latex peptidases from Calotropis procera (CpLP) and Cryptostegia grandiflora (CgLP) were assayed for this purpose. Enzyme activity on substrates representative of skin such as hide powder azure (UHPA), elastin (UE), azocollagen (UAZOCOL), keratin (UK), and epidermis (UEP) was determined, while depilation activity was assayed on cow hide. Only CpLP was active against keratin (13.4 UK) and only CgLP was active against elastin (0.12 UE). CpLP (93.0 UHPA, 403.6 UAZOCOL, 36.3 UEP) showed higher activity against the other substrates than CgLP (47.6 UHPA, 261.5 UAZOCOL, 8.5 UEP). In pilot assays, CpLP (0.05% w/v with sodium sulfite 0.6% w/v as activator) released hairs from cow hide pieces. Macroscopic and microscopic analyses of the hide revealed that the dehairing process was complete and the leather structure was preserved. The proteolytic system of C. procera is a suitable bioresources to be exploited by tanneries.

Surgical Techniques for Displaced Radial Neck Fractures: Predictive Factors of Functional Results.

BACKGROUND: Fractures of the radial neck represent about 1% of all childhood fractures and 5% to 10% of childhood traumatic lesions involving the elbow. Management of these fractures in children is still controversial. Intramedullary percutaneous nail reduction (Métaizeau technique) is considered the most effective surgical technique because of its excellent results and easy learning curve. Complications may arise, however, especially in Böhler technique, in which a percutaneous pin is placed over the radial head. When this technique does not provide correct reduction, open reduction must be performed. Because open reduction is traditionally associated with a high risk of complications, however, its use is restricted to severely displaced fractures and only when the percutaneous techniques have failed or their application contraindicated because of associated injuries to the distal radius. METHODS: In this retrospective study, we evaluated 51 children between the ages of 6 and 15 years who presented to our institution from 1996 to 2012 with Métaizeau-modified Judet grades 3, 4a, and 4b radial head fractures. The surgical techniques used were closed reduction and casting under general anesthesia (n=7), closed reduction and intramedullary nailing using Métaizeau technique (n=27), and Métaizeau technique and open reduction with intramedullary nailing (n=17). Functional results of the 3 surgical techniques were evaluated using the Mayo Elbow Performance Score (MEPS) and compared by modified Judet classification using χ analysis. RESULTS: No statistic significant association was found between type of surgery and final MEPS was observed (P=0.110). However, a significant association was found between initial modified Judet grade and final MEPS. CONCLUSIONS: In the present study, final functional outcome seems to be not affected by open reduction but was significantly associated with initial modified Judet grade. LEVEL OF EVIDENCE: Level III-retrospective study comparing closed and open reduction techniques, performed at the same institution.

Production, characterization, and identification using proteomic tools of a polygalacturonase from Fusarium graminearum.

Since enzymatic degradation is a mechanism or component of the aggressiveness of a pathogen, enzymatic activities from a Fusarium graminearum isolate obtained from infected wheat spikes of Argentina Pampa region were studied in order to understand the disease progression, tending to help disease control. In particular, the significance of the study of polygalacturonase activity is based on that such activity is produced in the early stages of infection on the host, suggesting a crucial role in the establishment of disease. In this sense, polygalacturonase activity produced by this microorganism has been purified 375 times from 2-day-old culture filtrates by gel filtration and ion-exchange chromatography successively. The purified sample showed two protein bands in sodium dodecyl sulfate-polyacrylamide gels, with a molecular mass of 40 and 55 kDa. The protein bands were identified as an endopolygalacturonase and as a serine carboxypeptidase of F. graminearum, respectively, by peptide mass fingerprinting (matrix-assisted laser desorption/ionization time-of-flight (MALDI TOF/TOF) fragment ion analysis). The pattern of substrate degradation analyzed by thin layer chromatography confirmed the mode of action of the enzyme as an endopolygalacturonase. High activity of the polygalacturonase against polygalacturonic acid was observed between 4 and 6 of pH, and between 30 and 50 °C, being 5 and 50 °C the optimum pH and temperature, respectively. The enzyme was fully stable at pH 5 for 120 min and 30 °C and sensible to the presence of some metal ions. This information would contribute to understand the most favorable environmental conditions for establishment of the disease.

Trigger thumb treatment approach: Results of a survey of EPOS members.

Purpose: The aim of this study is to analyze the clinical reality of pediatric trigger thumb among members of the European Paediatric Orthopaedic Society. Methods: A 35-question survey was sent to all European Paediatric Orthopaedic Society members, focusing on surgeon's profile and experience, trigger thumb diagnostic and trigger thumb treatment approach. Descriptive statistics were performed. Results: A total of 99 responses were analyzed. Most important factor considered in the therapeutic approach was the presence of a locked interphalangeal joint (52%). Regarding treatment, 41.4% opt for conservative approach as the first line of treatment, while 30.3% consider surgery and 28.3% just observe. Nevertheless, 76% mention to treat conservatively their patients and 99% to consider surgery at some stage. Regarding surgical technique, 96% do it open and 56% consider 2 years as the minimum age for procedure. Most surgeons perform this procedure in out-patient care (87%), don't administer prophylactic antibiotherapy (80%), use a tourniquet (75%), don't use any postoperative immobilization (64%), and report complications related to surgery (64%), mainly recurrence/incomplete division (59%) and superficial wound infection (30%). Conclusion: Our study shows a significant variability in the initial management of pediatric trigger thumb with a predominance of conservative management, followed by surgery and observation without treatment. The disparity in treatment options and timing, or waiting times before moving on to different therapeutic options, shows us that this is a controversial issue and that the development of prospective randomized controlled studies is needed to analyze the different treatment methods and determine which ones give the best outcomes.

Trapeziometacarpal Dislocations in Pediatric Age, Is There a Better Treatment? Series of Cases and a Systematic Review.

(1) Background: Dislocations of the trapeziometacarpal joint (TMC) are uncommon in children and adolescents. Only a few isolated cases are reported in the literature. Therapeutic guidance is minimal and inconclusive. (2) Methods: The authors present four patients treated for this unusual lesion. We evaluated the evolution according to treatment, age, patient activity, and quickDASH. Despite the clear limitation of the small number of patients, it is relevant to try to better understand this lesion and its evolution. A systematic review of the literature was also conducted. (3) Results: This is the largest published series of TMC dislocations in children and adolescents. Patients included a 12-year-old girl treated conservatively with a poor quickDASH; a 9-year-old girl treated surgically with the Eaton-Littler technique for a new dislocation with a partially modified quickDASH; a 13-year-old boy with two necessary closed reductions for a new dislocation and a very good final quickDASH; and a 12-year-old boy treated with closed reduction and percutaneous fixation with excellent final results with quickDASH. (4) Conclusions: In the absence of scientific evidence, conservative treatment and ligament reconstruction did not provide good functionality. In contrast, closed reduction with percutaneous fixation provided excellent results. Therefore, the authors would recommend closed reduction and percutaneous needle fixation as an elective method to treat TMC dislocations in pediatric and adolescent patients.

Reverse Ishiguro Extension Block Technique as an Alternative for Irreducible Osseous Mallet Finger.

Subacute or late-presenting unstable osseous mallet finger might be hard to reduce and, therefore, remain subluxed when using the traditional Ishiguro technique. In such cases, we suggest it is best to prioritize correction of joint subluxation over step-by-step adherence to the traditional Ishiguro technique. Specifically, we contend that carrying out the procedure in reverse order typically results in an easier and more stable reduction of both joint and fracture.

Lateralized overgrowth as a guiding sign of abdominal neoplasms for pediatric orthopedic surgeons.

OBJECTIVES: This study aims to increase the awareness of the association between lateralized overgrowth (LO) and abdominal tumor among the pediatric orthopedic community and to evaluate its incidence in our center. PATIENTS AND METHODS: Between January 1997 and December 2021, a total of 166 patients with Wilms tumors and hepatoblastomas were retrospectively analyzed. Data including age, sex, initial clinical signs (hematuria, abdominal mass with or without general discomfort), type of asymmetric regional body overgrowth (isolated or in relation with any syndrome), and tumor stage at diagnosis were recorded. In addition, age at which asymmetric regional body overgrowth was described and age at the time of tumor diagnosis were noted. RESULTS: Of a total of 166 patients, 133 were diagnosed with Wilms tumors (nephroblastomas) and 33 were diagnosed with hepatoblastomas. In 94% of the cases, the initial clinical signs were an abdominal mass and/or hematuria. Overall, five (3%) patients presented with LO. Four patients with Wilms tumor presented it at the initial clinical examinations. In three of these cases (2.3%), we found it isolated and, in the remaining patient (0.75%), it was associated with Beckwith-Wiedemann spectrum. Only one patient affected from hepatoblastoma (3%) presented with an isolated LO at the time of tumor diagnosis. CONCLUSION: Our study results show an incidence of LO in relation to intra-abdominal tumors of 3%. The latest updates recommend genetic testing to identify subgroups with a higher risk for tumor development that are more likely to benefit from tumor protocol surveillance.

Glioblastoma Cells Counteract PARP Inhibition through Pro-Survival Induction of Lipid Droplets Synthesis and Utilization.

Glioblastoma multiforme (GBM) is the most common and aggressive primary brain tumor in adults. Poly (ADP-ribose) polymerase inhibitors (PARPi) represent a new class of anti-neoplastic drugs. In the current study, we have characterized the mechanism by which glioblastoma cells evade the effect of PARPi as anti-tumor agents. We have found that suppression of PARP activity exerts an anti-stemness effect and has a dual impact on autophagy, inducing its activation in the first 24 h (together with down-regulation of the pro-survival mTOR pathway) and preventing autophagosomes fusion to lysosomes at later time-points, in primary glioma cells. In parallel, PARPi triggered the synthesis of lipid droplets (LDs) through ACC-dependent activation of de novo fatty acids (FA) synthesis. Notably, inhibiting ß-oxidation and blocking FA utilization, increased PARPi-induced glioma cell death while treatment with oleic acid (OA) prevented the anti-glioma effect of PARPi. Moreover, LDs fuel glioma cells by inducing pro-survival lipid consumption as confirmed by quantitation of oxygen consumption rates using Seahorse respirometry in presence or absence of OA. In summary, we uncover a novel mechanism by which glioblastoma escapes to anti-tumor agents through metabolic reprogramming, inducing the synthesis and utilization of LDs as a pro-survival strategy in response to PARP inhibition.

Poland syndrome: a new classification system based on a retrospective analysis of 74 cases.

LEVEL OF EVIDENCE: III.

[Dairy matrix: nutritional and sanitary benefits of the interrelation between its nutrients]. / Matriz láctea: beneficios nutricionales y sanitarios de la interrelación entre sus nutrientes.

INTRODUCTION: The effect of nutrients on health has been studied traditionally. From this point of view, milk and dairy products contribute to maintaining and improving nutritional status. Due to its high content of some nutrients such as saturated fat, among others, dairy consumption has been linked to the appearance of several diseases such as obesity or cardiovascular disease. However, food cannot be classified as good or bad based on its nutrient content. On the contrary, those interactions between nutrients and other bioactive compounds that occur in the food matrix must also be taken into account, as they can lead to different results than those expected derived from their consumption. In the case of dairy products, the scientific evidence indicates that dairy matrix could play an important role in the prevention of several non-communicable diseases.

INTRODUCCIÓN: Tradicionalmente se ha estudiado el efecto de los nutrientes en la salud. Desde este punto de vista, la leche y los derivados lácteos son alimentos que contribuyen a mantener y mejorar el estado nutricional. Por su elevado contenido en algunos nutrientes como la grasa saturada, entre otros, el consumo de lácteos se ha relacionado con la aparición de diversas enfermedades como la obesidad o la enfermedad cardiovascular. Sin embargo, los alimentos no se pueden clasificar como buenos o malos por su contenido en nutrientes, sino que también hay que tener en cuenta que en la matriz alimentaria se producen interacciones entre los nutrientes y otros compuestos bioactivos que pueden hacer que el consumo de dichos alimentos tenga resultados distintos a los esperados. En el caso de los lácteos, la evidencia científica señala que la matriz láctea podría jugar un papel importante frente a la prevención de diversas enfermedades crónicas no transmisibles.

[Importance of nutrition during pregnancy. Impact on the composition of breast milk]. / Importancia de la nutrición durante el embarazo. Impacto en la composición de la leche materna.

INTRODUCTION: The consumption of a healthy diet is essential during the preconception period, pregnancy and lactation to guarantee maternal-fetal and newborn health. It is important to take into account when configuring the maternal diet that inadequate intakes of vitamins and minerals, as well as a high consumption of foods or beverages with refined sugar increase the incidence of low birth weight infants, while the adequate consumption of micronutrients and carbohydrates (mainly integral) can decrease it. Prenatal consumption of fish is also associated with less intrauterine growth retardation. Maternal nutritional deficits that lead to intrauterine growth retardation can alter the expression of some genes, causing abnormal programming in organ and tissue development. In response, the fetus adapts to this situation of scarcity and may have difficulties adapting to an abundant consumption of food after birth, increasing its propensity to suffer from cardiovascular and metabolic diseases in adult life. After delivery, the nutrition of the infant must be guaranteed through breast milk. There is a relationship between the nutritional status of the mother and the composition of the mother's milk and, therefore, in the supply of nutrients to the infant, which may condition their health. Despite the great importance of an adequate maternal nutritional state, serum deficiencies in vitamins A, E, C, B2, B1, calcium and zinc are observed in pregnant mothers and breast milk, being important to identify and prevent these imbalances before conception and during pregnancy and lactation.

INTRODUCCIÓN: El consumo de una dieta saludable es esencial durante el periodo preconcepcional, embarazo y lactancia para garantizar la salud maternofetal y del neonato. Es importante tener en cuenta al configurar la dieta materna que ingestas inadecuadas de vitaminas y minerales, así como un elevado consumo de alimentos o bebidas con azúcar refinada, aumentan la incidencia de recién nacidos con bajo peso, mientras que el consumo adecuado de micronutrientes y de hidratos de carbono (principalmente integrales) pueden disminuirla. También el consumo prenatal de pescado se asocia con menor retraso del crecimiento intrauterino. Los déficits nutricionales maternos que conducen a un retraso del crecimiento intrauterino pueden alterar la expresión de algunos genes ocasionando una programación anormal en el desarrollo de órganos y tejidos. Como respuesta, el feto se adapta a esta situación de escasez y puede tener dificultad de adaptación ante un consumo abundante de alimentos después del nacimiento, lo cual aumenta su propensión al padecimiento de enfermedades cardiovasculares y metabólicas en la vida adulta. Después del parto, la nutrición del lactante debe estar garantizada mediante la leche materna. Existe una relación entre el estado nutritivo de la madre y la composición de la leche materna y, por lo tanto, el aporte de nutrientes al lactante, lo cual puede condicionar su salud. Pese a la gran importancia que tiene un adecuado estado nutritivo materno, se observan niveles séricos deficitarios en vitaminas A, E, C, B2, B1, calcio y zinc en madres gestantes y en leche materna, por lo que es importante identificar y prevenir estos desequilibrios antes de la concepción y durante el embarazo y la lactancia.

Seymour fracture: Better do not underestimate it.

OBJECTIVES: This study aims to analyze the functional results, management, and complications of acute Seymour fracture treatment and to generalize the understanding of Seymour fractures, as well as awareness about its controversial treatment and critical sequelae. PATIENTS AND METHODS: Between January 1994 and December 2019, a total of 29 patients (20 males, 9 females; mean age: 7.9±3.9 years; range, 1 to 15 years) who presented within the first 24 h of injury and were diagnosed with Seymour fractures and treated in the emergency setting were retrospectively analyzed. Clinical and radiological data were collected from medical records at the time of diagnosis and during follow-up, within a week after the treatment and in the visits required until fracture healing, and no sequelae were observed. In the event of complications, a minimum of one year of follow-up was carried out. Radiographs were taken of the anteroposterior and lateral views during each visit. RESULTS: The mean follow-up was 10.8±8.6 (range, 2 to 36) months. Surgical treatment in the operating room was performed in 24 (82.7%) patients using a single longitudinal Kirschner wire (K-wire) fixation through the distal phalanx and the distal interphalangeal joint in 21 patients. Non-operative treatment based on closed reduction and splinting was performed in five (17.3%) patients. There was no statistically significant difference in the final passive range of motion and physeal growth arrest in relation to the use or non-use of K-wires. The use of antibiotics in any of the three possible administrations (intravenous antibiotic regimen, intravenous and later oral antibiotic at-home or oral antibiotics), in relation to the non-use of antibiotics seemed to be a protective factor against infections (odds ratio=0.04; 95% confidence interval: 0.006-0.2; p=0.001). CONCLUSION: The identification of Seymour fractures is crucial for applying the correct treatment and reducing the risk of complications, such as osteomyelitis and physeal alterations. Based on our study results, we can suggest that the use of an antibiotic regimen causes a lower risk of infections in acute Seymour fractures. The prompt identification of these fractures with a standardized protocol covering irrigation, debridement, reduction, fixation, and prophylactic antibiotics is needed to avoid complications.

Sequencing and characterization of asclepain f: the first cysteine peptidase cDNA cloned and expressed from Asclepias fruticosa latex.

Asclepain f is a papain-like protease previously isolated and characterized from latex of Asclepias fruticosa. This enzyme is a member of the C1 family of cysteine proteases that are synthesized as preproenzymes. The enzyme belongs to the alpha + beta class of proteins, with two disulfide bridges (Cys22-Cys63 and Cys56-Cys95) in the alpha domain, and another one (Cys150-Cys201) in the beta domain, as was determined by molecular modeling. A full-length 1,152 bp cDNA was cloned by RT-RACE-PCR from latex mRNA. The sequence was predicted as an open reading frame of 340 amino acid residues, of which 16 residues belong to the signal peptide, 113 to the propeptide and 211 to the mature enzyme. The full-length cDNA was ligated to pPICZalpha vector and expressed in Pichia pastoris. Recombinant asclepain f showed endopeptidase activity on pGlu-Phe-Leu-p-nitroanilide and was identified by PMF-MALDI-TOF MS. Asclepain f is the first peptidase cloned and expressed from mRNA isolated from plant latex, confirming the presence of the preprocysteine peptidase in the latex.

[Nutritional problematic related to frailty and sarcopenia in older]. / Problemática nutricional relacionada con la fragilidad y la sarcopenia en personas de edad avanzada.

INTRODUCTION: Introduction: frailty and sarcopenia are prevalent conditions in older adults which diagnosis and treatment often overlap. Objective: to analyze the nutritional problematic related to frailty and sarcopenia in older adults. Methods: a literature review, about particularities of frailty and sarcopenia, diagnosis and nutritional treatment, has been carried out. Results: frailty is a geriatric syndrome that can be identified by at least three of the following criteria: weight loss, exhaustion, reduced physical activity, increased walk time, low grip strength. Sarcopenia is a muscular disease characterized by low muscle mass and reduced muscle strength as the main determinant. Both conditions can benefit from adequate protein intake (1-1.5 g/kg/day) and regular physical activity. Conclusion: identification, prevention, monitoring and nutritional treatment of frailty and sarcopenia in older adults is important because of its high prevalence. The main goal is to maintain the muscular integrity and minimize muscle loss in older adults. A healthy aging includes regular physical activity and the fulfillment of the protein requirements of this population.

INTRODUCCIÓN: Introducción: la fragilidad y la sarcopenia son dos condiciones altamente prevalentes en el adulto mayor que se solapan en diferentes vertientes del diagnóstico y del tratamiento. Objetivos: analizar la problemática nutricional relacionada con la fragilidad y la sarcopenia en personas de edad avanzada. Métodos: se ha realizado una revisión bibliográfica de las particularidades de la fragilidad y de la sarcopenia, así como del diagnóstico y del tratamiento nutricional. Resultados: la fragilidad es un síndrome geriátrico que puede identificarse si se tienen al menos tres de los siguientes criterios: pérdida de peso, fuerza de presión de la mano disminuida, velocidad de la marcha lenta, actividad física reducida y agotamiento. La sarcopenia es una enfermedad muscular que se caracteriza por una masa muscular baja y una fuerza muscular reducida como determinantes principales. Ambas condiciones pueden verse beneficiadas por un consumo adecuado de proteínas (1-1,5 g/kg/día) y una actividad física de resistencia regular. Conclusión: la identificación, prevención, seguimiento y tratamiento nutricional de la fragilidad y la sarcopenia en adultos mayores es de vital importancia dada su elevada prevalencia. El objetivo es mantener la integridad y minimizar la pérdida muscular en los adultos mayores. Un envejecimiento saludable incluye la realización de actividad física regular y el cumplimiento de los requerimientos proteicos de esta población.

[Nutritional intervention in the control of gallstones and renal lithiasis]. / Intervención nutricional en el control de la colelitiasis y la litiasis renal.

INTRODUCTION: Cholelithiasis and kidney stones are frequent pathologies in developed countries. Gallstones can be pigmentary, cholesterol (75%) or mixed. Age, female sex, obesity, rapid weight loss, consumption of refined sugars, saturated fat, iron deficiency, vitamin D and low intake of fiber and vitamin C are factors associated with an increased risk of cholelithiasis. On the other hand, the intake of ω-3 fatty acids, oleic acid, calcium, magnesium, fiber, fruits and vegetables, dairy products, nuts, coffee, moderate consumption of alcohol, vitamin C supplements, physical exercise and a regular diet have a protective paper. The most frequent kidney stones are calcium oxalate followed by mixed ones (oxalate and calcium phosphate), struvite, uric acid and cystine. A high water intake is recommended (> 2.5 l / day), varying the type of recommended or unadvisable food depending on the type of calculus. In oxalic lithiasis it is recommended to reduce the consumption of meat, moderate that of spinach, Swiss chard, asparagus, chocolate and avoid excessive sodium intake. The administration of oxalate-degrading probiotics (Lactobacillus) may reduce intestinal absorption, although further studies are necessary to corroborate these results. In calcium phosphate acidifying diet is recommended and limit the consumption of coffee and tea. The prevention of uric calculus is based on hydration with alkalizing drinks and vegetarian diet, decreasing foods rich in purines (liver, kidney, fish eggs, anchovies, sardines and seafood) and in calculus of cystine diet is recommended alkalizing. Since the formation of struvite calculi is due to urinary infections, pharmacological treatment and the consumption of acidifying diets, moderation of the intake of phosphate-rich foods and limiting the contribution of fats and citrus fruits are necessary. The nutritional intervention is an effective measure in the prevention of biliary and renal lithiasis and prevent its recurrence.

INTRODUCCIÓN: La colelitiasis y litiasis renal son patologías frecuentes en países desarrollados. Los cálculos biliares pueden ser pigmentarios, de colesterol (75%) o mixtos. La edad, el sexo femenino, la obesidad, la pérdida rápida de peso, el consumo de azúcares refinados, la grasa saturada, la deficiencia de hierro y de vitamina D y la baja ingesta de fibra y vitamina C son factores asociados a mayor riesgo de colelitiasis. Por otro lado, la ingesta de ácidos grasos ω-3, ácido oleico, calcio, magnesio, fibra, frutas y verduras, lácteos, frutos secos, café, un consumo moderado de alcohol, suplementos de vitamina C, ejercicio físico y una alimentación regular desempeñan un papel protector. Las litiasis renales más frecuentes son de oxalato cálcico, seguidas de las mixtas (oxalato y fosfato cálcico), estruvita, ácido úrico y cistina. Se aconseja una elevada ingesta hídrica (> 2,5 l/día), variando el tipo de alimentos recomendados o desaconsejados en función del tipo de cálculo. En litiasis oxálica se recomienda reducir el consumo de carne, moderar el de espinacas, acelgas, espárragos, chocolate y evitar el aporte excesivo de sodio. La administración de probióticos degradantes de oxalato (Lactobacillus) podría reducir su absorción intestinal, aunque son necesarios más estudios para corroborar estos resultados. En las de fosfato cálcico se aconseja dieta acidificante y limitar el consumo de café y té. La prevención de cálculos de úrico está basada en la hidratación con bebidas alcalinizantes y dieta vegetariana, disminuyendo los alimentos ricos en purinas (hígado, riñón, huevas de pescado, anchoas, sardinas y mariscos), y en cálculos de cistina se aconseja el consumo de dietas alcalinizantes. Dado que la formación de cálculos de estruvita se debe a infecciones urinarias, es necesario un tratamiento farmacológico y el consumo de dietas acidificantes, moderar la ingesta de alimentos ricos en fosfatos y limitar el aporte de grasas y cítricos. La intervención nutricional es una medida eficaz en la prevención de la litiasis biliar y renal y para evitar su recurrencia.

[Controversies and errors related to nutrition and breastfeeding. Guidelines for improvement]. / Controversias y errores en relación con la nutrición y la lactancia materna. Pautas para la mejora.

INTRODUCTION: Although the benefits of breastfeeding are clear and well documented, for both newborns and nursing mothers, breastfeeding rates worldwide are not optimal in many cases. There are multiple myths and errors related to breastfeeding: that certain foods cannot be consumed during breastfeeding, that, however, certain foods must be consumed to increase milk production, that composition of breast milk is not adequate in some cases, that breastfeeding is associated with more risk of suffering caries… Therefore, the objective of this article is to clarify the reality about these aspects of breastfeeding.

INTRODUCCIÓN: Aunque los beneficios de la lactancia materna son claros y están bien documentados tanto para los recién nacidos como para las madres lactantes, las tasas de lactancia materna a nivel mundial en muchos casos no son óptimas. Existen múltiples mitos y errores en relación con la lactancia materna, como que no pueden consumirse determinados alimentos, que deben consumirse determinados alimentos para aumentar la producción láctea, que la composición de la leche materna en algunos casos no es adecuada, que la lactancia materna se asocia a más riesgo de caries… Por todo ello, el objetivo de este artículo es aclarar la realidad acerca de estos aspectos sobre la lactancia materna.

Purification and characterization of hieronymain III. Comparison with other proteases previously isolated from Bromelia hieronymi Mez.

A new proteolytic enzyme, named hieronymain III, has been purified by ion-exchange chromatography from unripe fruits of Bromelia hieronymi Mez. The new peptidase belongs to the cysteine catalytic type, as well as hieronymain I and II, the other two peptidases previously isolated from this species. Hieronymain III showed optimum alkaline pH range (8.6-9.3) and the molecular mass (MALDI-TOF) was 23713 Da. The N-terminal sequence (AVPQSIDWRRYGAVTTSRNQG) exhibited a higher percentage identity with hieronymain II (93%) than with hieronymain I (71%). The three peptidases showed notable differences on synthetic substrates degradation: whereas hieronymain III was the only one able to hidrolyze Z-Arg-Arg-p-nitroanilide, hieronymain I and II could degrade Z-Phe-Arg-p-nitroanilide; on the other hand, PFLNA was only split by hieronymain I. Finally, the three proteases showed different preferences on N-alpha-CBZ-p-nitrophenyl aminoacid ester substrates. From a biotechnological point of view, cleavage specificity differences are significant enough to use these enzymes as potential tools in that area.

Granulosain I, a cysteine protease isolated from ripe fruits of Solanum granuloso-leprosum (Solanaceae).

A new cysteine peptidase (Granulosain I) was isolated from ripe fruits of Solanum granuloso-leprosum Dunal (Solanaceae) by means of precipitation with organic solvent and cation exchange chromatography. The enzyme showed a single band by SDS-PAGE, its molecular mass was 24,746 Da (MALDI-TOF/MS) and its isoelectric point was higher than 9.3. It showed maximum activity (more than 90%) in the pH range 7-8.6. Granulosain I was completely inhibited by E-64 and activated by the addition of cysteine or 2-mercaptoethanol, confirming its cysteinic nature. The kinetic studies carried out with PFLNA as substrate, showed an affinity (Km 0.6 mM) slightly lower than those of other known plant cysteine proteases (papain and bromelain). The N-terminal sequence of granulosain I (DRLPASVDWRGKGVLVLVKNQGQC) exhibited a close homology with other cysteine proteases belonging to the C1A family.