A novel, blocking, Fc-silent anti-CD40 monoclonal antibody prolongs nonhuman primate renal allograft survival in the absence of B cell depletion.

CD40-CD154 pathway blockade prolongs renal allograft survival in nonhuman primates (NHPs). However, antibodies targeting CD154 were associated with an increased incidence of thromboembolic complications. Antibodies targeting CD40 prolong renal allograft survival in NHPs without thromboembolic events but with accompanying B cell depletion, raising the question of the relative contribution of B cell depletion to the efficacy of anti-CD40 blockade. Here, we investigated whether fully silencing Fc effector functions of an anti-CD40 antibody can still promote graft survival. The parent anti-CD40 monoclonal antibody HCD122 prolonged allograft survival in MHC-mismatched cynomolgus monkey renal allograft transplantation (52, 22, and 24 days) with accompanying B cell depletion. Fc-silencing yielded CFZ533, an antibody incapable of B cell depletion but still able to potently inhibit CD40 pathway activation. CFZ533 prolonged allograft survival and function up to a defined protocol endpoint of 98-100 days (100, 100, 100, 98, and 76 days) in the absence of B cell depletion and preservation of good histological graft morphology. CFZ533 was well-tolerated, with no evidence of thromboembolic events or CD40 pathway activation and suppressed a gene signature associated with acute rejection. Thus, use of the Fc-silent anti-CD40 antibody CFZ533 appears to be an attractive approach for preventing solid organ transplant rejection.

[Acute symptomatic epileptic seizures. A clinical-electroencephalographic etiological description and prognosis of an oncopediatric series]. / Crisis epilépticas sintomáticas agudas. Descripción clinicoelectroencefalográfica etiológica y pronóstico de una serie oncopediátrica.

AIM: To determine clinical, electroencephalographic, therapeutic and evolutive characteristics of a series of oncopediatric patients with acute symptomatic seizures. PATIENTS AND METHODS: We performed a retrospective and prospective descriptive analysis of clinical records of oncopediatric children evaluated by neurology at the comprehensive outpatient Center for Hemato-Oncological Patients during 2017-2021. We included children aged one month to 17 years with intracranial and extracranial tumors who presented with acute symptomatic seizure (ASC). We defined acute symptomatic seizure according to the 2010 International League Against Epilepsy. We classified seizures according to 2017 International League Against Epilepsy classification. We excluded any patient with a diagnosis of previous epilepsy and non-epileptic paroxysmal episodes. RESULTS: We analyzed 44 cases with a median of 4 years (range: 1 month-17 years) and mean of 5.75 months (range: 1 month-11 months) and 8.33 years (2-17 years). The main etiologies were neurotoxicity and post-surgical context. Four patients presented dysnatremias and two associated with endocranial hypertension. Forty-one electroencephalograms were performed with intercritical results with abnormalities in the baseline rhythm, but without foci or paroxysms. There were no critical recordings. Focal seizures were 25 (56.8%) and generalized seizures 19 (43.18%). Levetiracetam was the most commonly used drug for acute management. CONCLUSIONS: Our cohort shows that ASC, in this population, do not show considerable differences between focal motor and generalized seizures and occur mostly in neurotoxic and post-surgical contexts. Dysnatremias and endocranial hypertension associated with ASC were also recorded. Postcrisis electroencephalograms were without foci or paroxysms and good seizure evolution.

TITLE: Crisis epilépticas sintomáticas agudas. Descripción clinicoelectroencefalográfica etiológica y pronóstico de una serie oncopediátrica.Objetivo. Determinar las características clínicas, electroencefalográficas, terapéuticas y evolutivas de una serie de pacientes oncopediátricos con convulsiones sintomáticas agudas. Pacientes y métodos. Efectuamos un análisis descriptivo retrospectivo y prospectivo de registros clínicos de niños oncopediátricos evaluados por neurología en el Centro Ambulatorio Integral de Pacientes Hematooncológicos durante 2017-2021. Incluimos a niños de 1 mes a 17 años con tumores intracraneales y extracraneales que presentaron convulsiones sintomáticas agudas (CSA). Definimos convulsión sintomática aguda según la clasificación de la Liga Internacional contra la Epilepsia de 2010. Clasificamos las crisis epilépticas según la clasificación de la Liga Internacional contra la Epilepsia de 2017. Excluimos a todo paciente con diagnóstico de epilepsia previa y de episodios paroxísticos no epilépticos. Resultados. Analizamos 44 casos, con una mediana de 4 años (rango: 1 mes-17 años) y una media de 5,75 meses (rango: 1 mes-11 meses) y 8,33 años (2-17 años). Registramos como principales etiologías la neurotoxicidad y el contexto posquirúrgico, con cuatro pacientes asociados a disnatremias y dos a hipertensión endocraneana. Se realizaron 41 electroencefalogramas, con resultados intercríticos con anormalidades en el ritmo de base, pero sin focos ni paroxismos. No hubo registros críticos. Las convulsiones focales fueron 25 (56,8%), y las generalizadas, 19 (43,18%). El levetiracetam fue el fármaco más utilizado para el tratamiento agudo. Conclusiones. Nuestra cohorte muestra que las CSA, en esta población, no evidencian diferencias considerables entre convulsiones focales motoras y generalizadas, y ocurren mayormente en un contexto neurotóxico y posquirúrgico. También se registraron disnatremias e hipertensión endocraneana asociadas a CSA. Los electroencefalogramas poscrisis fueron sin focos o paroxismos y con evolución de las crisis.

High NaCl concentrations induce the nod genes of Rhizobium tropici CIAT899 in the absence of flavonoid inducers.

The nodulation (nod) genes of Rhizobium tropici CIAT899 can be induced by very low concentrations (micromolar to nanomolar range) of several flavonoid molecules secreted by the roots of leguminous plants under a number of different conditions. Some of these conditions have been investigated and appear to have a great influence on the concentration and the number of different Nod factors, which can induce root nodule primordia and pseudonodules in several leguminous plant roots. In one such condition, we added up to 300 mM NaCl to the induction medium of R. tropici CIAT899 containing the nod gene inducer apigenin. At the higher concentrations of NaCl, larger amounts and more different Nod factors were produced than in the absence of extra NaCl. To our surprise, under control conditions (300 mM NaCl without apigenin), some Nod-factor-like spots were also observed on the thin-layer plates used to detect incorporation of radiolabeled glucosamine into newly synthesized Nod factors. This phenomenon was further investigated with thin-layer plates, fusions of nod genes to the lacZ gene, high-performance liquid chromatography, mass spectrometry, and the formation of pseudonodules on bean roots. Here, we report that, in the absence of flavonoid inducers, high concentrations of NaCl induced nod genes and the production of Nod factors.

Total suspended particulate (TSP), polychlorinated dibenzodioxin (PCDD) and polychlorinated dibenzofuran (PCDF) emissions from medical waste incinerators in Antioquia, Colombia.

Results of a preliminary survey of particulate and dioxin emissions in combustion gases from hospital waste incinerators in Antioquia-Colombia are presented. Base line data of total suspended particulate (TSP) and polychlorinated dibenzodioxin and dibenzofuran (PCDD/PCDF) emissions in incinerators from health care institutions in Antioquia-Colombia will be used to improve the management of medical waste in the local context. All monitored incinerators are batch operated. TSP exceeds 80 mg Nm(-3) in 8 out of 12 incinerators. Dioxin emissions are in the range from about 7 to 700 I-TEQ (ng Nm(-3)). Such a significant amount of dioxin emissions did correlate with entrained particulate matter, mainly as a consequence of poor control of operation parameters. Several suggestions are made to improve medical waste management practices in Colombia.

Circulating tumor cells (CTCs) and cytokeratin 19 (CK19) mRNA as prognostic factors in heavily pretreated patients with metastatic breast cancer.

INTRODUCTION: Circulating tumor cell (CTC) count and cytokeratin 19 (CK19) mRNA expression have a prognostic value for patients with metastatic breast cancer (MBC), but their clinical utility remains controversial. We studied CTC count and CK19 mRNA expression in the peripheral blood samples from heavily pretreated patients with MBC and their correlations with prognosis and response to the subsequent line of therapy. METHODS: This prospective observational study included 67 consecutive patients with MBC who were on progression to systemic therapy, and criteria for a new line of systemic treatment were proposed outside a clinical trial. CTC counts and CK19 mRNA expression were measured by the CellSearch® and RT-PCR methods, respectively, before and after the first cycle of treatment. Progression-free survival (PFS) was defined as the time elapsed between the initiation of the treatment and either the date of clinical or radiological tumor progression or death from any cause or the last follow-up. Cox proportional hazards regression model was used to assess the univariate prognostic value of CTC and CK19 mRNA expression on PFS and Kaplan-Meier estimates. A multivariate Cox model was also used to additionally account for phenotype and visceral disease. RESULTS: The mean age was 60 (range 35-86) years, and the average number of previous treatments was 3 (range 1-10); 42 patients (62.6%) were ER+ and 38 patients (56.7%) had visceral disease. The median PFS rate was 8 months (95% CI: 3.7-8.2). Univariate analyses showed a significant effect of the initial value of CK19 mRNA expression (HR = 2.00; 95% CI: 1.05-3.8; p = 0.03) and for the second value of CTC (HR = 2.18; 95% CI: 1.22-3.9; p = 0.009) but did not reach statistical significance for the initial value of CTC and the second value of CK19 mRNA expression. The estimated PFS rates at 6 and 12 months were 75% and 31% for patients with a low initial value of CK19 mRNA expression and 36% and 10% for those with a high initial value of CK19 mRNA expression, respectively (p: 0.022). Further, the estimated PFS rates at 6 and 12 months were 86% and 65% for patients with a low second value of CTC and 76% and 47% for those with a high second value of CTC, respectively (p: 0.004). In the multivariate analysis adjusted for phenotype, visceral disease, and the last treatment performed, only the effect of the second value of CTC remained significant (HR = 2.7, p = 0.004). CONCLUSIONS: CK19 mRNA expression and CTC count appeared clinically meaningful in pretreated patients with MBC, even when adjusted for phenotype and visceral disease involvement. These results support the use of CK19 and CTC as relevant biomarkers for predicting clinical response in MBC.

Purification and characterization of a galactose-specific lectin from corn (Zea mays) coleoptile.

We purified and characterized a lectin from the corn coleoptyle (Zea mays). The lectin (CCL) was purified by affinity chromatography on a Lactosyl-Sepharose 4B column. It is a glycoprotein of 88.7 kDa, composed mainly by glutamic, aspartic, glycine, and Ser residues; in a minor proportion, it contained methionine and cysteine residues. Carbohydrates that constituted 12% of the total weight comprised galactose, mannose, and N-acetyl-D-glucosamine. The lectin contained the blocked amino-terminus. Analysis of the lectin, determined from peptides obtained after trypsin digestion by MALDI-TOF (matrix-assisted laser desorption ionization-time of flight), indicated that CCL has 18% homology with a putative calcium-dependent Ser/Thr protein kinase, from Arabidopsis thaliana, and 39% homology with a NADPH-dependent reductase from Z. mays. The lectin showed hemagglutinating activity toward several erythrocytes, including human A, B, and O. Hapten inhibition assays indicated that the lectin interacts specifically with the OH on C4 from galactose residues. OH- on C1 plays a relevant role in the interaction with CCL, since beta-galactose residues are better recognized than those from the anomeric alpha-galactose. Lack of lectin activity was observed in corn extracts; the highest specific activity was obtained from coleoptyle obtained at the 7th day after seeding.

Regulation of nitrite uptake and nitrite reductase expression in Chlamydomonas reinhardtii.

Expression of nitrite uptake and nitrite reductase activities has been studied in Chlamydomonas reinhardtii under different nutritional conditions. Both activities were expressed at a low level in derepressed cells (with no nitrogen source) and at a high level in induced cells (with nitrate or nitrite). Nitrate was required for both activities to be maximally expressed. Ammonium-grown cells did not show nitrite uptake capability and had a basal nitrite reductase activity. Nitrite uptake but not nitrite reductase levels decreased very significantly in nitrate-induced cells subject to cycloheximide treatment, which suggests that protein(s) involved in the uptake are under a rapid turnover. Nitrite uptake expression was strongly inhibited by the presence of the glutamine synthetase inhibitor L-methionine-D,L-sulfoximine under either derepression or induction conditions, whereas that of nitrite reductase was not affected under the same conditions. Our results indicate that nitrite uptake expression is regulated primarily by ammonium, and that of nitrite reductase by both ammonium and ammonium derivative(s).

Purification and Characterization of Two Distinct NAD(P)H Dehydrogenases from Onion (Allium cepa L.) Root Plasma Membrane.

Highly purified plasma membrane fractions were obtained from onion (Allium cepa L.) roots and used as a source for purification of redox proteins. Plasma membranes solubilized with Triton X-100 contained two distinct polypeptides showing NAD(P)H-dependent dehydrogenase activities. Dehydrogenase I was purified by gel filtration in Sephacryl S-300 HR, ion-exchange chromatography in DEAE-Sepharose CL-6B, and dye-ligand affinity chromatography in Blue-Sepharose CL-6B after biospecific elution with NADH. Dehydrogenase I consisted of a single polypeptide of about 27 kD and an isoelectric point of about 6. Dehydrogenase II was purified from the DEAE-unbound fraction by chromatography in Blue-Sepharose CL-6B and affinity elution with NADH. Dehydrogenase II consisted of a single polypeptide of about 31 kD and an isoelectric point of about 8. Purified dehydrogenase I oxidized both NADPH and NADH, although higher rates of electron transfer were obtained with NADPH. Maximal activity was achieved with NADPH as donor and juglone or coenzyme Q as acceptor. Dehydrogenase II was specific for NADH and exhibited maximal activity with ferricyanide. Optimal pH for both dehydrogenases was about 6. Dehydrogenase I was moderately inhibited by dicumarol, thenoyltrifluoroacetone, and the thiol reagent N-ethyl-maleimide. A strong inhibition of dehydrogenase II was obtained with dicumarol, thenoyltrifluoroacetone, and the thiol reagent p-hydroxymercuribenzoate.

Role of Apoplastic and Cell-Wall Peroxidases on the Stimulation of Root Elongation by Ascorbate.

Elongation of onion (Allium cepa L.) roots was highly stimulated by ascorbate (ASC) and its natural precursor I-galactone-[gamma]-lactone (GL). When incubation media were supplemented with lycorine (Lyc), an inhibitor of the ASC biosynthesis, root growth was negligible even in the presence of ASC or GL. ASC completely inhibited in vitro guaiacol peroxidase activities that were isolated from both the apoplast and the cell wall. However, ferulic-acid-dependent peroxidase from the cell wall was partially inhibited by ASC, whereas ferulic acid peroxidase activity from the apoplastic fluid was completely inhibited by ASC as long as ASC was present in the assay medium. ASC content in cells was increased by preincubations with ASC or GL, whereas Lyc reduced it. On the other hand, ASC or GL treatments decreased both apoplast and cell-wall-bound peroxidase activities, whereas Lyc had a slight stimulating effect. These results are discussed on the basis of a possible control of root elongation by ASC via its action on peroxidases that are involved in the regulation of cell-wall extensibility.

[Colombian validation of the Pittsburgh Sleep Quality Index]. / Validación colombiana del índice de calidad de sueño de Pittsburgh.

INTRODUCTION: Many studies establish an elevated frequency of sleep disorders. The prevalence during life is of 35%. This situation has lead to develop a great number of instruments with the intention to evaluate better these alterations. Nevertheless, the majority lacks a suitable and national validation. Buysse et al designed the Pittsburgh Sleep Quality Index (PSQI) considered the most able instrument to establish the quality of the sleep. This has been used and validated widely. Nor it exists a version validated to the Castilian for Colombia, nor similar instruments that allow making studies in sleep. AIM: Validation of the Pittsburg Sleep Quality Index in Colombia (ICSP-VC). SUBJECTS AND METHODS: Experts consensus for face validity, pilots studies for evaluating understanding of the questionnaire, with double interview method. Evaluation of internal consistency. RESULTS. Internal consistency was Cronbach's alpha = 0.78. There were different scores between subjects that mentioned some subjective sleep disturbance (U Mann Whitney z = -5.635, p = 0.000), between them that qualify as 'so bad' or not their sleep quality (U Mann Whitney z = -2.90, p = 0.0037), between they that were consuming or not, habitually hypnotics (U Mann Whitney z = -4.289, p = 0.0000) and between they that referred conciliation insomnia or not (U Mann Whitney z = -4.769, p = 0.0000). There were clinical differences in ICSP-VC scores between adult older and younger (U Mann Whitney z = -1.926, p = 0.0542). There were not differences by gender. Subjects with extreme values scores had clinical differences with others. CONCLUSIONS: ICSP-VC is a valid version of PQSI useful for studying sleep disorders.

Effects of concanavalin A on protein-C activity.

Concanavalin A interacts specifically with the oligosaccharides from protein-C and modifies its anticoagulant activity. The lectin activates the protein-C activity in a dose dependent manner as demonstrated by in vitro and in vivo assays. Concanavalin A at low concentration (0.1 to 2 microg/mL) induces an increase on the catalytic activity of protein-C; at higher concentrations (5 to 20 microg/mL), the catalytic activity returns to the baseline. The effect of concanavalin A was prevented by incubating the protein-C with alpha-methyl-mannoside or by treating the purified protein-C with alpha-mannosidase; furthermore, cleavage of mannosidic residues diminishes its catalytic activity. Our results indicate that the oligomannosidic portion of protein-C participates in the regulation of the catalytic activity of this protein.

Modification of the Smith and Naylor technique for the identification of ferruginous bodies.

Smith and Naylor's technique has been successfully used by many authors around the world to recover inorganic particles from the lungs. In this short report we compare the results of counting inorganic particles recovered from human lung tissue by two different methods: Smith and Naylor's technique and a modification to it. We used ferruginous bodies (FB) as markers for inorganic particles and we compared the results of the FB counts in both methods. In the traditional technique the interface formed in the 1:1 ethanol-chloroform mixture is discharged and FB are counted only in the formed pellet. Our modification also quantified FB in the interface fraction. Post-mortem samples from each of the lung lobes were taken from 22 individuals, totalling 198 samples. Each sample was digested in liquid commercial bleach and processed by both techniques. Our results showed that the modified technique was more sensitive in the detection of FB than the traditional method, with 16 out of 22 individuals showing positive identification against 14 out of 22, respectively. Furthermore, the modified technique proved not only to be more sensitive, but also almost twice as accurate: 49 FB/g vs. 26.7 FB/g of dry tissue.

Changes in neurotransmitter amino acids and protein in CNS areas of mice subjected to differential housing conditions.

The amino acid and protein content of mice exposed to enriched, restricted and impoverished environments have been studied in six discrete CNS areas. Differences between enriched and either restricted or impoverished groups were found whereas no difference was observed between restricted and impoverished ones. In the first case, a significant increase for aspartate was found in spinal cord, whereas glutamate significantly decreased in colliculi and cerebral cortex. Similarly, glycine increased in cerebral cortex and decreased in colliculi and pons-medulla, and gamma-aminobutyrate (GABA) increased in spinal cord, pons-medulla and cerebellum and decreased in thalamus-hypothalamus. No changes in concentrations of five non-transmitter amino acids (serine, threonine, alanine, isoleucine, leucine) were observed. Significant increases of the protein concentration in cerebellum and spinal cord were found. The changes were due to enrichment, not to aggregation conditions. The results corroborate the proposed plasticity of the aminoacidergic system.

Differential distribution of neurotransmitter amino acids from the limbic system of aggressive and non-aggressive bull strains.

The amino acid content of crude synaptosomal fractions from the limbic system and related CNS regions showed significant differences between the aggressive Spanish fighting-bull and the non-aggressive Friesan bull breeds. Neurotransmitter amino acids (glutamate, aspartate, GABA and glycine) were the most unequally distributed. A higher ratio of excitatory to inhibitory neurotransmitter amino acids was always found in all the CNS regions studied in the aggressive breed. The concentrations of five non-transmitter amino acids (threonine, alanine, serine, leucine and isoleucine) showed minor variations between both studied bull strains and cannot be ascribed to differences in central energy metabolism. The results are explained in terms of a possible relationship between the amino acid neurotransmitter levels and the innate aggressiveness of the Spanish fighting-bull.

[Assessing the use of low-molecular-weight heparins fo venous thromboembolism prophylaxis in internal medicine patients]. / Evaluación de la utilización de heparinas de bajo peso molecular como profilaxis de tromboembolismo venoso en pacientes de medicina interna.

BACKGROUND: Sufficient evidence exists to recommend routine thromboembolic disease prophylaxis for medical inpatients with risk factors --with low-molecular-weight-- heparins being (LMWH) the most suitable treatment option. The objective is to determine the thromboembolic risk level of Internal Medicine patients with LMWH prophylaxis, prescription habits and their adequacy to hospital's standards, as well as prevalence of non-treated patients at risk. MATERIAL AND METHODS: Descriptive and prospective study of internal medicine patients for 2 months. Patients with prophylactic LMWH prescription were chosen, and their thromboembolic risk level and suitable LMWH dose was determined according to the hospital's "thromboembolic disease prevention standards". On the other hand, patients with no LMWH prophylaxis were analysed in order to judge their candidacy. RESULTS: 30% of patients had a prophylactical LMWH prescription, with 43.5% of these prescriptions being adequate to the risk level. The main risk factors were: age, bed-stay, hypertension, cardiopathy with risk factors, diabetes mellitus, dislipemias and COPD. Chi2(0.05) test between risk level and prescribed LMWH revealed no association. 72% of patients without LMWH prescription had a moderate or high risk level. CONCLUSIONS: 1. A high proportion of the patients studied have a considerable thromboembolic risk level. 2. There is not a statistical relationship between thromboembolic risk level and LMWH prescription. 3. There is a high percentage of patients with no LMWH prophylaxis which could be eligible for it. 4. A pharmaceutical intervention would be useful to approach pharmacological prophylaxis to each patient's risk.