Macromolecular crowding sensing during osmotic stress in plants.

Osmotic stress conditions occur at multiple stages of plant life. Changes in water availability caused by osmotic stress induce alterations in the mechanical properties of the plasma membrane, its interaction with the cell wall, and the concentration of macromolecules in the cytoplasm. We summarize the reported players involved in the sensing mechanisms of osmotic stress in plants. We discuss how changes in macromolecular crowding are perceived intracellularly by intrinsically disordered regions (IDRs) in proteins. Finally, we review methods for dynamically monitoring macromolecular crowding in living cells and discuss why their implementation is required for the discovery of new plant osmosensors. Elucidating the osmosensing mechanisms will be essential for designing strategies to improve plant productivity in the face of climate change.

Using NIR irradiation and magnetic bismuth ferrite microparticles to accelerate the removal of polystyrene microparticles from the drinking water.

Magnetic bismuth ferrite (BiFO) microparticles were employed for the first time for the removal of polystyrene (PS) nano/microplastics from the drinking water. BiFO is formed by porous agglomerates with sizes of 5-11 µm, while the PS nano/microparticles have sizes in the range of 70-11000 nm. X-ray diffraction studies demonstrated that the BiFO microparticles are composed of BiFeO3/Bi25FeO40 (the content of Bi25FeO40 is ≈ 8.6%). Drinking water was contaminated with PS nano/microparticles (1 g L-1) and BiFO microparticles were also added to the contaminated water. Later, the mixture of PS-particles + BiFO was irradiated with NIR light (980 nm). Consequently, PS nano/microparticles melted on the BiFO microparticles due to the excessive heating on their surface. At the same time, the NIR (near infrared) light generated oxidizing agents (∙OH and h+), which degraded the by-products formed during the photocatalytic degradation of PS nano/microparticles. Subsequently, the NIR irradiation was stopped, and a Neodymium magnet was utilized to separate the BiFO microparticles from the water. This last procedure also permitted the removal of PS nano/microparticles by physical adsorption. Zeta potential measurements demonstrated that the BiFO surface was positively charged, allowing the removal of the negatively charged PS nano/microparticles by electrostatic attraction. The combination of the photocatalytic process and the physical adsorption permitted a complete removal of PS nano/microparticles after only 90 min as well as a high mineralization of by-products (≈95.5% as confirmed by the total organic carbon measurements). We estimate that ≈23.6% of the PS nano/microparticles were eliminated by photocatalysis and the rest of PS particles (≈76.4%) by physical adsorption. An outstanding adsorption capacity of 195.5 mg g-1 was obtained after the magnetic separation of the BiFO microparticles from the water. Hence, the results of this research demonstrated that using photocatalysis + physical-adsorption is a feasible strategy to quickly remove microplastic contaminants from the water.

14-3-3ε protein-immobilized PCL-HA electrospun scaffolds with enhanced osteogenicity.

Adipose-derived mesenchymal stem cells (ASCs) accelerate the osteointegration of bone grafts and improve the efficiency in the formation of uniform bone tissue, providing a practical and clinically attractive approach in bone tissue regeneration. In this work, the effect of nanofibrous biomimetic matrices composed of poly(ε-caprolactone) (PCL), nanometric hydroxyapatite (nHA) particles and 14-3-3 protein isoform epsilon on the initial stages of human ASCs (hASCs) osteogenic differentiation was investigated. The cells were characterized by flow cytometry and induction to differentiation to adipogenic and osteogenic lineages. The isolated hASCs were induced to differentiate to osteoblasts over all scaffolds, and adhesion and viability of the hASCs were found to be similar. However, the activity of alkaline phosphatase (ALP) as early osteogenic marker in the PCL-nHA/protein scaffold was four times higher than in PCL-nHA and more than five times than the measured in neat PCL.

Plasticity in sunflower leaf and cell growth under high salinity.

A group of sunflower lines that exhibit a range of leaf Na(+) concentrations under high salinity was used to explore whether the responses to the osmotic and ionic components of salinity can be distinguished in leaf expansion kinetics analysis. It was expected that at the initial stages of the salt treatment, leaf expansion kinetics changes would be dominated by responses to the osmotic component of salinity, and that later on, ion inclusion would impose further kinetics changes. It was also expected that differential leaf Na(+) accumulation would be reflected in specific changes in cell division and expansion rates. Plants of four sunflower lines were gradually treated with a relatively high (130 mm NaCl) salt treatment. Leaf expansion kinetics curves were compared in leaves that were formed before, during and after the initiation of the salt treatment. Leaf areas were smaller in salt-treated plants, but the analysis of growth curves did not reveal differences that could be attributed to differential Na(+) accumulation, since similar changes in leaf expansion kinetics were observed in lines with different magnitudes of salt accumulation. Nevertheless, in a high leaf Na(+) -including line, cell divisions were affected earlier, resulting in leaves with proportionally fewer cells than in a Na(+) -excluding line. A distinct change in leaf epidermal pavement shape caused by salinity is reported for the first time. Mature pavement cells in leaves of control plants exhibited typical lobed, jigsaw-puzzle shape, whereas in treated plants, they tended to retain closer-to-circular shapes and a lower number of lobes.

Human papillomavirus infection in cyclosporin-induced gingival overgrowth in renal allograft recipients.

BACKGROUND: Host immunity plays an important role in the development of human papillomavirus (HPV)-associated disease. The HPV infection in oral cyclosporin-induced gingival overgrowth in renal transplant recipients has not been investigated previously. The aim of this study was to establish the HPV infection of cyclosporin-induced gingival hyperplasia in renal transplant recipients through morphological changes and use of the in situ hybridization technique. METHODS: We examined 13 renal transplant recipient biopsies with gingival overgrowth lesions and 4 healthy mucosa samples of these patients. The histopathological diagnoses were established on the basis of widely accepted criteria, and the pathologist was not aware of the HPV result. An in situ molecular hybridization was carried out under low stringent conditions to detect HPV species with mixed biotin-labeled probes of HPV 6 and HPV 11, and under high stringent conditions with HPV 6, HPV 11, HPV 16, and HPV 18 probes for HPV typing. RESULTS: The HPV prevalence among the 13 samples studied was 92.31% (12/13), of which 4 tested positive for HPV 6-11 and 1 for HPV 16. The 4 biopsies of normal mucosa from gingival overgrowth patients were also reactive for HPV DNA. In 11/12 (91.7%) HPV-positive cases, koilocytotic atypia was found. CONCLUSIONS: The suppression of T-cell function by cyclosporin therapy can result in an increase of HPV infection, adding to the proliferative activity of cyclosporin in the oral mucosa.

Loss of maternally derived human herpesvirus-6 immunity and natural infection in Argentinian infants.

BACKGROUND: Human herpesvirus-6 (HHV-6) infection is widespread throughout the world. No data are available in Argentina about the loss of maternally derived HHV-6 immunity and natural infection in infants. METHODS: A population of 100 pregnant women and 407 children between 1 and 15 months of age were assayed by indirect immunofluorescence to detect and quantify specific IgG anti-human herpesvirus-6 (anti-HHV-6) antibodies in Córdoba City, Argentina. RESULTS: There was no significant difference in the positive rate between infants aged 1 to 9 months (range, 43.6 35.5%) and pregnant women (37%). Seropositive ratio dropped in the 10-month group (23.33% seropositive) and rose sharply in the 11-month group (38.89%), 12-month (60.61%), and 13- to 15-month group (63.46%). The geometric mean titer (GMT) for infants in the 12 to 15 months age group (23.4 41.64) was significantly higher than the GMT for infants 10 months of age (11.04) (P < 0.05 with the Tukey-HSD test). CONCLUSIONS: This study shows a significant association between loss of passive HHV-6 antibody and age among infants. The results support evidence that HHV-6 enters the susceptible population at 11 months, leading to a high prevalence of antibodies in children between 13 and 15 months of age.

Antibacterial activity of some medicinal plants from San Juan, Argentina.

The antibacterial activities and preliminary phytochemical screening of 13 plants used as folk medicine in San Juan, Argentina, are reported.

Quantification of fecal neutrophils by MPO determination (myeloperoxidase) in patients with invasive diarrhea. Cuantificación de neutrofilos fecales mediante la determinación de MPO (Mieloperoxidasa) en pacientes con diarrea invasiva.

Myeloperoxidase (MPO), a specific polymorphonuclear leukocyte enzyme, has been used previously to quantify the number of neutrophils in tissue. MPO activity was found to be linearly related to the number of neutrophil cells. In an attempt to use this method in leukocytes measuring in stool, fecal MPO was solubilized with hexadecyltrimethylammonium bromide and the MPO activity was measured by a dianisidine-H2O2 assay. Stools from 10 normal subjects and 39 patients with diarrhea produced by enteropathogenic bacteria were examined for leukocytes by MPO activity as well as microscopically using methylene blue stain, MPO activity was positive in 36 patients (92%) and leukocytes were present by microscopic observation in 30 (77%). Fecal leukocytes were not found in healthy controls and the MPO activity was undectable. Stool MPO activity had a range of from 1.6 to 2,830.0 x 10(3) UMPO per gram of feces (median 460.0). The number of neutrophils obtained through MPO activity had a range of 6.0 to 13,216.0/ mm3 (median 1,261.0). Fecal MPO activity is a simple biochemical assay for the detection and quantification of fecal leukocytes.

Conversion from azathioprine [correction of azathioprina] to mycophenolate mofetil in pediatric renal transplant recipients with chronic rejection.

BACKGROUND: Chronic rejection is the leading cause of graft failure. Both nonimmunological and immunological mechanisms contribute to this pathology. METHODS: We studied changes in kidney function, mixed lymphocyte culture, cell-mediated lympholysis, serum HLA class I antigens, cytotoxic antibodies, and lymphocyte population before and after 6 months of follow-up in 22 pediatric renal transplanted patients. The immunosuppressive protocol used was: cyclosporine, azathioprine, and corticosteroids. Eight patients demonstrated chronic graft rejection (by biopsy), group I; and eight patients had no clinical evidence of chronic and/or acute rejection, group II. Substitution of mycophenolate mofetil (MMF) (600 mg/m2 bid for azathioprine was done in patients of groups I and II. Another six patients with chronic rejection, did not receive MMF, group III. RESULTS: Creatinine clearance increased in group I (44+/-5 vs. 51.1+/- ml/min/1.73 m2, P<0.03) but it decreased in group III (30+/-3 vs. 25+/-2, P<0.01). Urinary protein excretion decreased only in group I (0.3+/-0.03 to 0.06+/-0.03 g/24 hr, P<0.03). During MMF therapy antidonor mixed lymphocyte culture decreased 62 and 70% (P<0.05) in group I and II. Cell-mediated lympholysis against lymphocyte of the donor decreased 65% (P<0.05) in group I. Cell-mediated lympholysis toward control cells decreased 54% (P<0.01) in group II. Serum HLA class I antigens, only decreased from 0.7+/-0.1 to 0.5+/-0.1 microl/ml, P<0.05, in group I. CD19+ decreased from 7.9+/-1.1 to 5.6+/-0.8%, P<0.05, and 7.8+/-1.2 to 5.5+/-0.9%, P<0.05, in groups I and II, respectively. CD16+ increased from 5.7+/-1.1 to 8.6+/-1.3 (P<0.05) only in group I. CONCLUSIONS: Our data suggest that substituting MMF for azathioprine therapy leads to an improvement in the immunosuppression and renal function in children with on-going chronic rejection.

Biologically active alkaloids and a free radical scavenger from Prosopis species.

The biological activity of extracts from the aerial parts of five Argentinian Prosopis species and the exudate of P. flexuosa were assessed for DNA binding, beta-glucosidase inhibition and free radical scavenging effect using the DPPH decoloration assay. DNA binding effect was found mainly in the basic fraction. The alkaloids tryptamine as well as piperidine and phenethylamine derivatives were isolated from the basic extracts. At 0.50 mg/ml, DNA binding activities ranged from 28% for tryptamine to 0-27% for the phenethylamine and 47-54% for the piperidine derivatives. Tryptamine and 2-beta-methyl-3-beta-hydroxy-6-beta-piperidinedodecanol showed a moderate inhibition (27-32%) of the enzyme beta-glucosidase at 100 microg/ml. The exudate of P. flexuosa displayed a strong free radical scavenger effect in the DPPH decoloration assay. The main active constituent was identified as catechin.

Cuantificación de neutrófilos fecales mediante la determinacián de MPO (Mieloperoxidasa) en pacientes con diarrea invasiva / Quantify of fecal neutrophils by MPO (myeloperoxidase) determination in patients with invasive diarrhea

La mieloperoxidasa (MPO) es una enzima especifíca de los polimorfonucleares neutrófilos, la cual ha sido previamente usada para cuantificar elmnúmero de neutrófilos en tejidos, desde que su actividad se correlaciona linealmente con el número de neutrófilos. Con el objetivo de demostrar la presencia y realizar la cuantificación de leucocitos en materia fecal la MPO se disolvió en Bromuro de hexadeciltrimetilamonio y la actividad fue medida usando un ensayo de H2O2O-dianosid-ina. Se midió la actividad de MPO en 39 pacientes con diarrea producida por bacterias enteropatógenas y en 10 sujetos control. La presencia de leucocitos fue también determinada mediante la observación microscópica usando azul de metileno. La actividad de MPO fue positiva en 36 (92 por ciento) de los pacientes y la observació microscópica resultó positiva en 30 (77 por ciento). En los sujetos control la actividad de MPO fue indetectable y no se encontraron leucocitos en material fecal. En los pacientes la actividad de MPO en materia fecal tuvo un recuento de 1.6 a 2830 x 10(3) UMPO por gramo de heces (mediana: 46.0). El número de neutrófilos obtenido a través de la actividad de MPO tuvo un recuento de 6 a 13216.0 x mm(3) (mediana: 1261.0), la actividad fecal de MPO es una determinación bioquímica simple para la detección y cuantificación de leucocitos en materia fecal.

Quantification of fecal neutrophils by MPO determination (myeloperoxidase) in patients with invasive diarrhea. Cuantificación de neutrofilos fecales mediante la determinación de MPO (Mieloperoxidasa) en pacientes con diarrea invasiva

Myeloperoxidase (MPO), a specific polymorphonuclear leukocyte enzyme, has been used previously to quantify the number of neutrophils in tissue. MPO activity was found to be linearly related to the number of neutrophil cells. In an attempt to use this method in leukocytes measuring in stool, fecal MPO was solubilized with hexadecyltrimethylammonium bromide and the MPO activity was measured by a dianisidine-H2O2 assay. Stools from 10 normal subjects and 39 patients with diarrhea produced by enteropathogenic bacteria were examined for leukocytes by MPO activity as well as microscopically using methylene blue stain, MPO activity was positive in 36 patients (92

) and leukocytes were present by microscopic observation in 30 (77

). Fecal leukocytes were not found in healthy controls and the MPO activity was undectable. Stool MPO activity had a range of from 1.6 to 2,830.0 x 10(3) UMPO per gram of feces (median 460.0). The number of neutrophils obtained through MPO activity had a range of 6.0 to 13,216.0/ mm3 (median 1,261.0). Fecal MPO activity is a simple biochemical assay for the detection and quantification of fecal leukocytes.

Cuantificación de neutrófilos fecales mediante la determinacián de MPO (Mieloperoxidasa) en pacientes con diarrea invasiva / Quantify of fecal neutrophils by MPO (myeloperoxidase) determination in patients with invasive diarrhea

La mieloperoxidasa (MPO) es una enzima especifíca de los polimorfonucleares neutrófilos, la cual ha sido previamente usada para cuantificar elmnúmero de neutrófilos en tejidos, desde que su actividad se correlaciona linealmente con el número de neutrófilos. Con el objetivo de demostrar la presencia y realizar la cuantificación de leucocitos en materia fecal la MPO se disolvió en Bromuro de hexadeciltrimetilamonio y la actividad fue medida usando un ensayo de H2O2O-dianosid-ina. Se midió la actividad de MPO en 39 pacientes con diarrea producida por bacterias enteropatógenas y en 10 sujetos control. La presencia de leucocitos fue también determinada mediante la observación microscópica usando azul de metileno. La actividad de MPO fue positiva en 36 (92 por ciento) de los pacientes y la observació microscópica resultó positiva en 30 (77 por ciento). En los sujetos control la actividad de MPO fue indetectable y no se encontraron leucocitos en material fecal. En los pacientes la actividad de MPO en materia fecal tuvo un recuento de 1.6 a 2830 x 10(3) UMPO por gramo de heces (mediana: 46.0). El número de neutrófilos obtenido a través de la actividad de MPO tuvo un recuento de 6 a 13216.0 x mm(3) (mediana: 1261.0), la actividad fecal de MPO es una determinación bioquímica simple para la detección y cuantificación de leucocitos en materia fecal. (Au)

Quantification of fecal neutrophils by MPO determination (myeloperoxidase) in patients with invasive diarrhea. Cuantificación de neutrofilos fecales mediante la determinación de MPO (Mieloperoxidasa) en pacientes con diarrea invasiva.

Myeloperoxidase (MPO), a specific polymorphonuclear leukocyte enzyme, has been used previously to quantify the number of neutrophils in tissue. MPO activity was found to be linearly related to the number of neutrophil cells. In an attempt to use this method in leukocytes measuring in stool, fecal MPO was solubilized with hexadecyltrimethylammonium bromide and the MPO activity was measured by a dianisidine-H2O2 assay. Stools from 10 normal subjects and 39 patients with diarrhea produced by enteropathogenic bacteria were examined for leukocytes by MPO activity as well as microscopically using methylene blue stain, MPO activity was positive in 36 patients (92

) and leukocytes were present by microscopic observation in 30 (77

). Fecal leukocytes were not found in healthy controls and the MPO activity was undectable. Stool MPO activity had a range of from 1.6 to 2,830.0 x 10(3) UMPO per gram of feces (median 460.0). The number of neutrophils obtained through MPO activity had a range of 6.0 to 13,216.0/ mm3 (median 1,261.0). Fecal MPO activity is a simple biochemical assay for the detection and quantification of fecal leukocytes.

[Human papillomavirus detection in oral cancer lesions in the city of Córdoba]. / Detección del virus papiloma humano en lesiones cancerosas orales en la ciudad de Córdoba.

Oral cancer is a process that involves different etiological factors and mechanisms in the light of current view of viral cocarcinogenesis. Evidence from histology and DNA hybridization studies suggests that HPV is engaged in oral carcinogenesis. The Pathology Laboratory of the Dentistry School, National University of Córdoba, admits approximately 20% of all patients with cancerous lesions in this city. In the January 1992-December 1997 lapse, we examined 1950 biopsies with oral lesions, 4.77% (93/1950) of which were malignant neoplasms, 79.57% (74/93) were oral carcinomas. Thirty-three oral carcinomas (44.6%; 33/74) were selected at random and included in this study, 33 cells smears of normal oral mucosa of controls individuals were included. They were analyzed by conventional light microscopy and an in situ hybridization technique for the detection of HPV. Data were analyzed with chi square test. The prevalence of HPV among the 33 cancer samples studied was 27.27%, 9/33 tested positive for HPV in low stringent conditions. Only one was positive in high stringent condition for HPV16, a verrugous carcinoma. No HPV-DNA was detected in cells smears of controls. Among the HPV positive, 3/9 (33.33%) were squamous carcinomas and 5/9 (55.56%) were verrugous carcinomas. Only one was a melanoma. Verrugous carcinoma was the carcinoma most associated with the HPV infection (x2 = 20.5; 95% level of confidence). This would indicate a major role of HPV in the pathogenesis of verrucous carcinomas. The viral prevalence found in cancerous lesions reinforces the concept of heterogenic natures of oral cancer. HPV is a circumstance that increase the probability of malignancy, and when reducing, diminish the frequency of cancer.

Colonic proteinases: increased activity in patients with ulcerative colitis.

In order to study the colonic intraluminal proteinase-antiproteinase imbalance under inflammatory conditions, we determined proteolytic activity (PA), alpha-1-antitrypsin and the activities of trypsin, chymotrypsin and neutrophil elastase in feces from patients with ulcerative colitis (UC) comparing the results with a control group. A fecal sample was obtained from each 25 patients with ulcerative colitis and 10 control subjects were studied. The severity of the disease was assessed by the Truelove index. Proteolytic activity was measured using azocasein as proteolytic substrate. The fecal concentration of alpha-1-antitrypsin was measured by radial immunodiffusion and the activities of the enzymes were measured using specific substrates. We found an increase in fecal PA, alpha-1-antitrypsin and neutrophil elastase in patients with UC and the correlation between the severity of the disease and the PA was statistically significant (r = 0.62, P < 0.05). We conclude that elevated colonic proteinase activity could contribute to the pathophysiology of ulcerative colitis.

Las formas elementales de la enfermedad: la sistematización de Laplantine / Elemental forms of the illness: the Laplantine systematization

En este artículo, se analiza el importante aporte de Francois Laplantine en el estudio de la antropología de la enfermedad, vía de acceso a la antropología de la salud en un sentido amplio. La importancia de este trabajo es que va más allá del análisis de los sistemas médicos tradicionales, sistematizando los discursos actuales válidos para la cultura occidental sobre el proceso salud-enfermedad, revelando la influencia de los factores culturales sobre el comportamiento y creencias de los enfermos, fundamental para mejorar la eficacia de los programas médicos en nuestros países latinoamericanos, cuya cultura básica es sincrética y oral. Al proponerse el concepto "Antropología de la enfermedad", se permite haciendo de la enfermedad un objeto de análisis ampliar el horizonte de custionamiento antropológico sobre la naturaleza del hombre y la cultura, antecedente básico para el desarrollo de una reflexión bioética en nuestro continente que repose en el "hombre total", no desinserto de su cultura y de su historia

Hydroxylation of dehydroabietic acid by Fusarium species.

A novel compound, 1 beta-hydroxydehydroabietic acid has been obtained by the microbial transformation of dehydroabietic acid, using cultures of Fusarium oxysporum and F. moniliforme. Its antibacterial activity was also tested.