Nutritional status in post SARS-Cov2 rehabilitation patients.

BACKGROUND & AIMS: After prolonged hospitalization, the assessment of nutritional status and the identification of adequate nutritional support is of paramount importance. In this observational study, we aimed at assessing the presence of a malnutrition condition in SARS-Cov2 patients after the acute phase and the effects of a multidisciplinary rehabilitation program on nutritional and functional status. METHODS: We recruited 48 patients (26 males/22 females) admitted to our Rehabilitation Unit after discharge from acute Covid Hospitals in northern Italy with negative swab for SARS-Cov2. We used the Global Leadership Initiative on Malnutrition (GLIM) criteria to identify patients with different degrees of malnutrition. Patients underwent a 3 to 4-week individual multidisciplinary rehabilitation program consisting of nutritional intervention (energy intake 27to30 kcal/die/kg and protein intake 1-1.3 g/die/kg), exercise for total body conditioning and progressive aerobic exercise with cycle- and arm-ergometer (45 min, 5 days/week). At admission and discharge from our Rehabilitation Unit, body composition and phase angle (PhA) (BIA101 Akern), muscle strength (handgrip, HG) and physical performance (Timed-Up-and-Go, TUG) were assessed. RESULTS: At admission in all patients the mean weight loss, as compared to the habitual weight, was -12.1 (7.6)%, mean BMI was 25.9 (7.9) kg/m2, mean Appendicular Skeletal Muscle Index (ASMI) was 6.6 (1.7) kg/m2 for males and 5.4 (1.4) kg/m2 for females, mean phase angle was 2.9 (0.9)°, mean muscle strength (HG) was 21.1 (7.8) kg for males and 16.4 (5.9) kg for females, mean TUG value was 23.7 (19.2) s. Based on GLIM criteria 29 patients (60% of the total) showed a malnutrition condition. 7 out of those 29 patients (24%) presented a mild/moderate grade and 22 patients (76%) a severe grade. After a rehabilitation program of an average duration of 25 days (range 13-46) ASMI increased, with statistically significant differences only in females (p = 0.001) and HG improved only in males (p = 0.0014). In all of the patients, body weight did not change, CRP/albumin (p < 0.05) and TUG (p < 0.001) were reduced and PhA increased (p < 0.01). CONCLUSIONS: We diagnosed a malnutrition condition in 60% of our post SARS-Cov2 patients. An individualized nutritional intervention with adequate energy and protein intake combined with tailored aerobic and strengthening exercise improved nutritional and functional status.

Efecto del proceso de germinación de granos de Cajanuscajan en la composición nutricional, ácidos grasos, antioxidantes y bioaccesibilidad mineral / Effect of the germination process of Cajanus cajan grains on nutritional composition, fatty acids, antioxidants and mineral bioaccessibility

RESUMEN La germinación es un proceso que involucra cambios en la actividad enzimática de los granos y ruptura de macro-moléculas, los cuales disminuyen los factores antinutricionales e incrementan la síntesis de otros constituyentes. Se obtienen granos con características sensoriales mejoradas, mayor valor nutricional y funcional. En esta investigación se germinaron granos de Cajanus cajan, por 96 horas, a 20°C en oscuridad, para estudiar las variaciones en composición proximal, ácidos grasos, capacidad antioxidante y bioaccesibilidad mineral. Los resultados obtenidos indican una disminución de las proteínas y aumento de cenizas, grasas y carbohidratos. Se determinó el perfil de ácidos grasos y se identificó la presencia de seis ácidos grasos, siendo el más abundante el ácido linoléico, la cantidad de los mismos también varió como consecuencia del proceso aplicado. El contenido de polifenoles y la capacidad antioxidante de la semilla aumentaron debido a la germinación. Se obtuvieron valores de bioaccesibilidad mineral, en un rango entre 16 y 31 % para el magnesio y el potasio, respectivamente. Se concluyó que el quinchoncho representa un alimento de elevado valor nutricional y que a través de la germinación se pueden introducir cambios nutricionales positivos y de esta forma promover su consumo.

ABSTRACT Germination is a process that involves changes in the enzymatic activity of grains and macromolecule breakdown, which decreases anti-nutritional factors and increases the synthesis of other constituents. Germinated grains are thus of higher sensory, nutritional and functional value. In this research, pigeon pea (Cajanus cajan) grains were germinated for 96 hours at 20°C in the dark. Variations in proximal composition, fatty acids composition, antioxidant ability and mineral bioaccessibility were determined. The results obtained showed that because of the germination process, protein content decreased, while ash, fat and carbohydrate content increased. Fatty acid profile was determined and six fatty acids were identified, with the greatest amount of linoleic acid. The fatty acid content changed with the applied process. The polyphenolic content and antioxidant ability also increased as a result of the germination process. The values obtained for the mineral bioaccessibility varied from 16 to 31% for magnesium and potassium, respectively. It was concluded that pigeon pea grains represent a food of high nutritional value, which, as a result of germination, can create positive nutritional changes. Thus, consumption of this food should be promoted.

Caspase-3 activation in human melanoma A375 cell line by a novel selective sigma-2 agonist.

Two novel 8-azabicyclo[3.2.1]octan-3-ol derivatives, 11a and 11b, with high affinity for sigma-2 receptors and a very good sigma-1/sigma-2 selectivity ratio were synthesized. In comparison with several well established sigma-2 selective ligands, 11 b showed a very low sigma-1 receptor affinity. Functional assays demonstrated that 11b acts as an agonist and in A-375 human melanoma cell line is able to lower levels of procaspase-3, thus confirming a potential major role for sigma-2 pure agonists in the treatment of rapid proliferating melanoma cells.

Sodium caseinate induces mouse granulopoiesis.

OBJECTIVE AND DESIGN: Sodium caseinate (CasNa) induces differentiation and M-CSF production in mouse band granulocytes in vitro; however, it is not yet known if this molecule can also induce the proliferation and activation of the granulocyte lineage in vivo. In this work we evaluated the induction in vivo of granulopoiesis and the activation of granulocytes in mice treated with CasNa. MATERIAL OR SUBJECTS: BALB/c male mice 8-12 weeks old were used. TREATMENT: The animals were inoculated intraperitoneally with 1 ml of CasNa (10% in PBS p/v) four times (every 48 h). METHODS: Granulocyte proliferation was evaluated by flow cytometry; activation was evaluated by phagocytic indices. The cytokine was measured using an ELISA assay. RESULTS: We show that CasNa increased bone marrow granulopoiesis percentage (38.35 ± 10.88 vs. 64.94 ± 34.14 BrdU+/Gr-1+ cells) and the granulocytes generated presented increased phagocytic indices (0.3 ± 0.1 vs. 0.6 ± 0.11, p < 0.05). We also show that G-CSF (974 ± 411 vs. 3189 ± 350 pg/ml, p < 0.05) and GM-CSF increased in serum, but only G-CSF in bone marrow plasma. CONCLUSIONS: CasNa induces granulopoiesis with functional granulocytes, suggesting that this molecule could be an innate immune system activator.

Effect of UVA-activated riboflavin on dentin bonding.

Recent studies have reported collagen cross-linking after exposure to riboflavin followed by ultraviolet-A (UVA) exposure. This study is the first to investigate the effect of a riboflavin-containing primer on adhesive interface stability and dentinal matrix metalloproteinase activity. Human dentin was etched with 35% phosphoric acid, treated with 0.1% riboflavin, exposed to UVA for 2 min, and bonded with a two-step etch-and-rinse adhesive. Adhesive was applied to control specimens without riboflavin/UVA. Specimens were subjected to microtensile bond strength tests and pulled to failure after storage for 24 hrs, 6 mos, or 1 yr. Interfacial nanoleakage was evaluated by light and transmission electron microscopy. To investigate dentinal matrix metalloproteinase activity, we performed correlative zymographic assays on protein extracts obtained from phosphoric-acid-etched dentin powder with or without riboflavin/UVA treatment and XP Bond. Ultraviolet-activated riboflavin treatment increased the immediate bond strength to dentin at all aging intervals (p < 0.05 vs. control) and decreased interfacial nanoleakage in aged specimens (1 yr; p < 0.05). Zymograms revealed that riboflavin/UVA pre-treatment inhibited dentinal matrix metalloproteinase activity (especially MMP-9). In conclusion, dentinal collagen cross-linking induced by riboflavin/UVA increased immediate bond strength, stabilized the adhesive interface, and inhibited dentin matrix metalloproteinases, thereby increasing the durability of resin-dentin bonds.

Función adrenocortical en pacientes con micosis sistémica / Adreno cortical function in patients with systemic mycoses

Las micosis sistémicas como paracoccidioidomicosis e histoplasmosis, son la principal causa de insuficiencia suprarrenal en los países donde éstas son endémicas. Venezuela es un país que registra una frecuencia elevada de estas micosis. El objetivo de este estudio fue evaluar la función adrenal glucocorticoide en pacientes hospitalizados con paracoccidioidomicosis e histoplasmosis. Se realizó la prueba de estimulación rápida con ACTH sintética a 12 pacientes con diagnóstico de alguna de estas micosis y a 12 sujetos control, y se recogieron datos de interés epidemiológico. La relación hombre:mujer de los pacientes fue de 5:1 y la edad media fue 35,214,4 años, similar al grupo control. El cortisol plasmático basal estuvo dentro del rango normal en todos los pacientes y fue similar a los del grupo control. Posterior a la administración de ACTH sintética se observó un incremento del cortisol plasmático, dentro del rango considerado normal pero significativamente menor que el del grupo control. Estos resultados sugieren una disminución de la reserva funcional de las glándulas suprarrenales en los pacientes con paracoccidioidomicosis e histoplasmosis. En nuestro medio, es importante evaluar la respuesta a la prueba de estimulación rápida con ACTH en los pacientes con micosis sistémica dada la frecuencia de su afectación.

Two different techniques in the rehabilitation treatment of low back pain: a randomized controlled trial.

AIM: The Back School is a widely accepted and effective method for treating low back pain, whereas no scientific evidence exists about the effects of the Pilates CovaTech method. With this study we wanted to evaluate the efficacy of this new method in patients with low back pain. METHODS: Fifty-three patients with at least 3 months of nonspecific low back pain were entered into a Pilates therapy or a Back School treatment group, 43 of which completed the study. Small exercise groups of 7 patients each followed a daily kinesitherapy protocol for 10 days. Evaluations were performed at the start of the study and then at 1, 3 and 6 months after the beginning of treatment. We used the Oswestry Low Back Pain Disability Scale (OLBPDQ) to assess disability and the visual analog scale (VAS) to evaluate pain. RESULTS: Demographic and baseline clinical characteristics were similar for both groups. A significant reduction in pain intensity and disability was observed across the entire sample. The Pilates method group showed better compliance and subjective response to treatment. CONCLUSIONS: The results obtained with the Pilates method were comparable to those achieved with the Back School method, suggesting its use as an alternative approach to the treatment of non specific low back pain.

In vitro and in vivo tetracycline-controlled myogenic conversion of NIH-3T3 cells: evidence of programmed cell death after muscle cell transplantation.

Ex vivo gene therapy of Duchenne muscular dystrophy based on autologous transplantation of genetically modified myoblasts is limited by their premature senescence. MyoD-converted fibroblasts represent an alternative source of myogenic cells. In this study the forced MyoD-dependent conversion of murine NIH-3T3 fibroblasts into myoblasts under the control of an inducible promoter silent in the presence of tetracycline was evaluated. After tetracycline withdrawal this promoter drives the transcription of MyoD in the engineered fibroblasts, inducing their myogenesis and giving rise to beta-galactosidase-positive cells. MyoD-expressing fibroblasts withdrew from the cell cycle, but were unable to fuse in vitro into multinucleated myotubes. Five days following implantation of engineered fibroblasts in muscles of C57BL/10J mice we observed a sevenfold increase of beta-galactosidase-positive regenerating myofibers in animals not treated with antibiotic compared with treated animals. After 1 week the number of positive fibers decreased and several apoptotic myonuclei were detected. Three weeks following implantation of MyoD-converted fibroblasts in recipient mice, no positive "blue" fiber was observed. Our results suggest that transactivation by tetracycline of MyoD may drive an in vivo myogenic conversion of NIH-3T3 fibroblasts and that, in this experimental setting, apoptosis plays a relevant role in limiting the efficacy of engineered fibroblast transplantation. This work opens the question whether apoptotic phenomena also play a general role as limiting factors of cell-mediated gene therapy of inherited muscle disorders.

GST-pi expression in BCR-ABL+ and BCR-ABL- cells from CML patients.

Our laboratory has been involved in the study of glutathione-sulfhydryl-transferase-pi (GST-pi) for several years. We have recently observed that during haematopoiesis in BMSC liquid cultures from CML patients who were candidates for transplant GST-pi was expressed in presumably malignant cells during different stages of cellular maturation. To confirm this finding, in the present work we are detecting GST-pi expression by immunofluorescence in BCR-ABL+ and BCR-ABL- cells done by FISH of PB from 30 CML patients during different clinical status: treatment (T), hematological relapse (R), blastic crisis (BC) or post-allotrasplant (PT). As well as in PB from 30 Blood-Bank donors. The results were %BCR-ABL+ GST-pi+ cells: T = 1-67, R = 33-69, BC = 90-100 and PT = 1-2; %BCR-ABL- GST-pi+ cells: T = 2-31, R = 5-18, BC = 0-10 and PT = 2-5; %BCR-ABL- GST-pi- cells: T = 2-97, R = 13-62, BC = 0 and PT = 93-96; %BCR-ABL+ GST-pi- cells: T = 0, R = 0, BC = 0 and PT = 0. GST-pi was not expressed in donor cells. The results obtained confirm our previous observations and suggest that GST-pi expression might be used for the evaluation of the minimal residual disease in CML patients.

A bilateral model integrating vergence and the vestibulo-ocular reflex.

The majority of previous modelling studies of vergence and the vestibulo-ocular reflex (VOR) have postulated arbitrary structures mainly on the basis of input-output behavioural relationships. Such models were developed following traditional schemes of oculomotor organization, based upon the notion of independence between different oculomotor subsystems. This impedes the simulation of complex binocular interactions and associated central activities. In contrast to preceding studies, the mathematical model for binocular control presented here was developed fully on physiological and anatomical grounds which reflect the organization and functional properties of known vergence and VOR premotor centres. Computer simulations show the model properly simulates the main observed characteristics in the discharge of several premotor and motor nuclei during slow vergence and the VOR in the dark. In particular, the model reproduces the activity profiles of abducens internuclear neurons, secondary vestibular cells, tonic prepositus hypoglossi neurons and ocular motoneurons during vergence and the VOR. It also simulates the activity of mesencephalic neurons whose discharge is modulated by vergence parameters alone. It is shown that given recent neurophysiological and behavioural findings, ocular reflexes cannot be properly modelled as separate independent subsystems whereas a single, unified modelling approach can produce results consistent with observed data. This study also shows how changes in the functional activity of shared pathways in a single two-sided structure produce vergence and conjugate integrators whose function relies on coupled loops across the brainstem: separate, dedicated operators are not necessary to replicate data. This provides evidence that challenges previous studies supporting the existence of separate vergence and conjugate integrators to transform velocity to position signals in the brainstem. A major implication of this study is that it questions the validity of testing conjugate and vergence systems independently, neglecting potential interactions.

Providing distinct vergence and version dynamics in a bilateral oculomotor network.

Given reported interactions between vergence and version dynamics, ocular reflexes cannot be properly modelled as separate independent subsystems. Using a model structure compatible with known anatomy, we show that a single bilateral system can produce results consistent with observed data both at the central and ocular levels. This model provides for both vergence and conjugate integrators in a single controller, and explains the observed modulation on abducens interneurons and mesencephalic vergence cells during vergence responses. Reported interactions between version and vergence would then be a natural consequence of a shared premotor network. Major implications include: the need to record both eyes in a protocol, since cross-talk is always possible; and adaptation to monocular changes could be distributed in all motor projections to both eyes.

Follow-up of GST-P during hepatocarcinogenesis with DEN-2AAF in F344 rats.

Although it is known that doublets of hepatocytes GST-P+ are produced during the first week after initiation with DEN, the activity levels of the enzyme are not known in the initial stages of the process, neither is its behavior through an initiation-promotion scheme. We consider the latter issues important in order to obtain information of the initiation step and its relation with GST-P.DEN was applied as initiator in a single dose on day 0 to F344 rats (200 mg/kg) and as promoter 2-AAF in 20 mg/kg doses on days 7, 8 and 9 of the scheme. Partial hepatectomy was performed on day 10, and daily during the 28 days in which the experiment lasted. The GST-P activity was determined in postmicrosomal supernatants of respective livers (by immunoadsorption) as well as their histological section (by immunohistochemistry). In both cases antibody anti-GST-pi produced in our laboratory was employed. The results obtained show GST-P appearance on day 5 of the scheme in rats treated with carcinogens. The activity of the enzyme increased slowly reaching its maximum on day 18, together with the appearance of GST-P+ preneoplastic nodules. Our results suggest that GST-P could display an additional function to those previously known, in cellular differentiation this could explain the very frequent expression of this enzyme in preneoplastic as well as in neoplastic cells.

Determination of denzimol, a new anticonvulsant agent, and its main metabolite in biological material by gas chromatography and high-performance liquid chromatography.

Two methods, using gas chromatography (GC) and high-performance liquid chromatography (HPLC), were developed in order to investigate the pharmacokinetics of denzimol hydrochloride, N-[beta-[4-(beta-phenylethyl)phenyl]-beta-hydroxyethyl] imidazole hydrochloride, which is a new anticonvulsant drug, and of its main metabolite, N-[beta-[4-(beta-phenyl-beta(alpha)-hydroxyethyl)phenyl] -beta-hydroxyethyl]-imidazole (referred to as M2), in humans. Both methods involve the use of a homologue of denzimol as an internal standard. The GC method is more sensitive (sensitivity limit 2.5 ng/ml for denzimol and 15 ng/ml for M2) and was utilized for the determination of denzimol and M2 in plasma. The GC method is specific, precise (relative standard deviations are 3.26, 2.12 and 1.72% at 10, 100 and 1000 ng/ml for denzimol and 6.45, 4.17 and 3.38% at 50, 500 and 1000 ng/ml for M2) and accurate (mean recovery +/- S.D. is 102.58 +/- 4.10% for denzimol and 99.72 +/- 7.75% for M2). The HPLC method is very simple and quick to perform. This method has a sensitivity limit of 0.5 micrograms/ml for denzimol and 1 microgram/ml for M2, and allows the determination of both compounds in urine with high selectivity, reproducibility (relative standard deviations are 2.05, 3.50 and 1.02% for denzimol and 2.78, 2.80 and 1.73% for M2, at concentrations of 15, 35 and 70 micrograms/ml) and accuracy (mean recovery +/- S.D. is 103.57 +/- 2.97% for denzimol and 95.91 +/- 1.59% for M2). The common anticonvulsants, when present in plasma, do not interfere with the monitoring of denzimol levels.

Is there a physiological intravascular coagulation in obstetrical cases?

The concentrations of plasmatic fibrinogen and its degradation products (FDPs), and the paracoagulation test using serial dilution of protamine sulphate (SDPS) were determined during the third trimester of pregnancy, labor, and puerperium. Singificant increases in the concentrations of fibrinogen and FDPs were observed throughout the process of pregnancy and birth, combined with both positive and negative SDPS tests. We suggest that these findings do not indicate a process of physiological DIC.