La telerehabilitación durante la pandemia COVID-19: experiencia de las familias de niños con condiciones crónicas complejas / Telerehabilitation during the COVID-19 pandemic: experience of families of children with complex chronic conditions

La telerehabilitación ha sido una solución rápida y efectiva para la atención de pacientes durante la pandemia COVID-19. Nuestro objetivo ha sido describir la experiencia de las familias de niños con condiciones crónicas complejas (CCC) asistidos con la modalidad de telerehabilitación durante la pandemia. Materiales y métodos: Se ha realizado un estudio observacional, descriptivo y transversal de tipo encuesta online. Se encuestaron familias de niños (1 mes a 5 años de edad) con al menos una CCC que recibieron seguimiento interdisciplinario a distancia durante la pandemia de 2 o más áreas de rehabilitación (kinesiología, fonoaudiología y terapia ocupacional) pertenecientes al Servicio de Kinesiología del Hospital de Pediatría Juan P Garrahan, durante marzo a octubre de 2020. Resultados: El 88,3% de los participantes (n=43) se expresó satisfecho con la modalidad de telerehabilitación. Entre los facilitadores seleccionados por los participantes se mencionan la reducción de tiempos, mayor frecuencia de consultas, disminución de gastos por traslados y mayor comodidad, mientras que entre las barreras se destacaron: el no vínculo con profesionales, alargue en tiempos de tratamiento, problemas de conectividad. El análisis estadístico no indicó diferencias significativas entre quienes se reportaron como más satisfechos según su estrato social (alto o bajo), la disponibilidad de wifi propio, o respecto a la edad del niño. Se reportaron diferencias significativas en cuanto a la distancia al hospital (p=0.034). Conclusión: Esta forma de intervención ofreció nuevas posibilidades de atención que podrían considerarse a futuro en el seguimiento de nuestros pacientes. (AU)

Telerehabilitation has been a fast and effective solution in patient care during the COVID-19 pandemic. Our aim was to describe the experience of families of children with complex chronic conditions (CCC) treated through telerehabilitation during the pandemic. Materials and methods: An observational, descriptive, cross-sectional, online survey study was conducted. Families of children (1 month to 5 years of age) with at least one CCC who received interdisciplinary remote follow-up during the pandemic from two or more rehabilitation areas (physical therapy, speech therapy, and occupational therapy) belonging to the Department of Physical Therapy of Hospital de Pediatría Juan P Garrahan, from March to October 2020, were surveyed. Results: 88.3% of the participants (n=43) expressed satisfaction with the telerehabilitation modality. Among the facilitators selected by the participants, the following were mentioned: time saving, greater frequency of consultations, reduction of travel expenses and greater comfort, while among the barriers, the following stood out: no connection with the professionals, longer treatment times, connectivity problems. Statistical analysis did not show significant differences between those who reported being more satisfied according to socioeconomic level (high or low), availability of their own wifi connection, or age of the child. A significant difference was found for distance to the hospital (p=0.034). Conclusion: This type of intervention provided new possibilities of care that could be considered in the future follow-up of our patients (AU)

Disfagia pediátrica: rol del fonoaudiólogo como parte de un equipo interdisciplinario en el Hospital Juan P. Garrahan / Dysphagia in children: The role of the speech therapist in the interdisciplinary team of Hospital Juan P. Garrahan

Introducción: El 80% de los niños con condiciones médicas crónicas complejas presentan alteración de la conducta alimentaria. Dada la heterogeneidad de los diagnósticos y la complejidad en el manejo de la disfagia pediátrica, es muy importante la intervención de equipos especializados. Objetivo: Evaluación de la evolución de los niños con trastornos de la deglución y/o conducta alimentaria atendidos durante el año 2017 por el equipo interdisciplinario de deglución y conducta alimentaria, del Hospital de Pediatría Juan P. Garrahan. Material y métodos: Estudio analítico, prospectivo y longitudinal con la intervención de un equipo interdisciplinario. Se incluyeron todos los pacientes evaluados durante el año 2017. Se realizó evaluación clínica de la deglución que permitió detectar dificultades durante el momento de la alimentación. Se dieron pautas de estimulación y modificación de consistencias y se derivó a tratamiento oportuno. Se midió porcentaje de destete de soporte nutricional (SN) y/o aumento del aporte por vía oral. Resultados: evaluamos 153 pacientes, 39% mujeres, 90% menores de 3 años de edad y el 75% en apoyo nutricional. El 72% presentó trastorno de la deglución exclusivamente o asociado a trastorno de la conducta alimentaria. El 68% fue seguido en más de una oportunidad. El 36% de los pacientes que ingresaron con requerimiento de SN lograron el destete (sin diferencia significativa entre los que tenían o no trastorno deglutorio p=0.85 y los que tenían o no traqueostomía p=0,88) y 40% aumentó el aporte por vía oral dentro del grupo que no logró el destete. Se observó una diferencia estadísticamente significativa en el destete de los pacientes que concurrieron al espacio de la clínica con respecto a los que no(p=0,016) y mayor tiempo de intervención entre quienes lograron el destete y quienes no, 5,2 ± 3,1 vs 3,45 ± 3,1 meses (p=0,0099). Conclusión: Es esencial el trabajo interdisciplinario y especializado en niños con trastornos de la deglución. La intervención del fonoaudiólogo como parte del equipo es fundamental para una detección precoz y correcto abordaje de la disfagia (AU)

Of all children with chronic complex medical conditions, 80% have eating behavior disorders. Given the heterogeneity of the diagnoses and the complexity of the management of dysphagia in children, intervention of a specialized medical team is essential. Objective: Evaluation of the outcome in children with swallowing and/or eating behavior disorders seen during 2017 by the interdisciplinary group of swallowing and eating behavior disorders at Hospital de Pediatría Juan P. Garrahan. Material and methods: An analytical, prospective and longitudinal study with intervention by an interdisciplinary team. All patients evaluated during 2017 were included. Swallowing was clinically assessed to identify eating disorders. Indications were given for stimulation and food consistency and patients were referred for adequate treatment. The rates of weaning from nutritional support (NS) and/or increase of oral food intake were measured. Results: 153 patients were evaluated, 39% were female, 90% younger than 3 years of age, and 75% AN. Overall, 72% had swallowing difficulties only or associated with an eating behavior disorder; 68% was followed on more than one occasion. Thirty-six percent of the patients who were admitted with NS requirement could be weaned (without a significant difference between those who had a swallowing disorder and those who did not p=0.85 and those that did and did not have a tracheostomy p=0.88) and oral food intake increased in 40% of the patients in the group that could not be weaned. A statistically significant difference was found in the weaning of patients who attended the clinic and those who did not (p=0.016) and longer intervention time between those who could be weaned and those who could not, 5.2 ± 3.1 vs 3.45 ± 3.1 months (p=0.0099). Conclusion: Interdisciplinary and specialized care is necessary for children with swallowing disorders. Intervention of a speech therapist as part of the team is fundamental for the early detection and adequate management of dysphagia (AU)

[The role of endoscopy in gastroenteropancreatic neuroendocrine tumors]. / Il ruolo dell'endoscopia nei tumori neuroendocrini gastroenteropancreatici.

Versione italiana Riassunto: Il ruolo dell'endoscopia nei tumori neuroendocrini gastroenteropancreatici. L. Magno, L. Sivero, V. Napolitano, S. Ruggiero, G. Fontanarosa, S. Massa I tumori neuroendocrini (NET) gastro-entero-pancreatici (GEP) sono neoplasie rare che originano dalle cellule neuroendocrine del tubo digerente e del pancreas. L'endoscopia digestiva e l'ecoendoscopia rivestono un ruolo importante nella diagnosi, stadiazione e sorveglianza dei pazienti con NET. Inoltre, in casi selezionati, le tecniche endoscopiche operative consentono il trattamento di queste neoplasie in fase precoce. English version Summary: The role of endoscopy in gastroenteropancreatic neuroendocrine tumors. L. Magno, L. Sivero, V. Napolitano, S. Ruggiero, G. Fontanarosa, S. Massa Gastroenteropancreatic (GEP) neuroendocrine tumors (NET) are rare neoplasia arisen from neuroendocrine cells present in the gut mucosa and pancreas. Digestive endoscopy and endoscopic ultrasonography play a relevant role in NET diagnosis, stadiation and surveillance. Moreover, in selected patients, surgical endoscopy allows the tratment of these cancers at an early stage.

Occurrence of Giardia and Cryptosporidium in Italian water supplies.

A total of 21 samples: raw water (RW) samples; water samples after coagulation with aluminium sulfate (clarified water: CW); and water after chlorination (treated water: TW) from a water purification plant that treats river surface water from the neighbourhood of Foggia (Italy), were analysed for the presence of Giardia cysts and Cryptosporidium oocysts. Bacteriological indicator of faecal contamination (total and faecal coliforms, faecal streptococci,), total bacterial count at 22 and 36 degrees C and physicochemical parameters (turbidity, temperature, pH) were evaluated. Cryptosporidium oocysts were not found in any samples examined, while Giardia cysts were found only in RW samples, with the maximal concentration of 8 cysts/100 l. A positive correlation was found between the Giardia densities and quality parameters such as TC, FC and TBC at 22 degrees C. Giardia levels in raw water samples correlated (p < 0.05) with TC, FC and with temperature. No other water quality parameters was consistently correlated with cysts level.

Immunomodulatory activity of a plant extract containing human papillomavirus 16-E7 protein in human monocyte-derived dendritic cells.

This study reports the immunomodulatory activity on human monocyte derived dendritic cells (MDDCs) of a vaccine preparation shown to be effective against an HPV16-related tumour in an animal model. The vaccine is composed of extract from Nicotiana benthamiana leaves containing HPV16 E7 protein expressed by a potato virus X-derived vector (NbPVX-E7). The effect of the extract was evaluated on MDDC differentiation and maturation by monitoring the phenotypic expression of specific markers. The results show that NbPVX-E7 does not induce monocyte differentiation to dendritic cells, but does induce MDDC maturation. Plant extract does not influence MDDC-uptake of E7-FITC while it significantly improves the Ovalbumin-FITC uptake, considered as a model antigen. Importantly, NbPVX-E7-pulsed MDDCs/PBMCs are able to prime human blood-derived lymphocytes from healthy individuals to induce HPV16 E7-specific cytotoxic activity. This is a propaedeutic study for a possible use of E7-containing plant extract in human immunotherapy of HPV-related lesions.

Exploiting the plant secretory pathway to improve the anticancer activity of a plant-derived HPV16 E7 vaccine.

The human papillomavirus 16 (HPV16) E7 oncoprotein can be considered a "tumor-specific antigen", and therefore it represents a promising target for a therapeutic vaccine against HPV-associated tumors. Efficient production of E7 protein with a plant-based transient expression system has been already described and it was demonstrated that E7-containing crude plant extracts confer partial protection against tumor challenge in a mouse model system. Before adopting the plant-based system as a cost-effective method for the production of an E7-based anti-cancer vaccine, some aspects, such as the oncoprotein yield, need further investigation. In the present study, we report the transient expression, mediated by a potato virus X (PVX)-derived vector, of the E7 protein targeted to the secretory system of Nicotiana benthamiana plants by using a plant-derived signal sequence. Targeting the antigen to the secretory pathway enhanced the E7 protein expression levels about five-fold. Mice immunized by s.c. administration with crude foliar extracts containing E7 showed strong stimulation of cell-mediated immune response after five boosters, as detected by ELISPOT. After challenging with the E7-expressing C3 tumor cells, tumor growth was completely inhibited in 80% of the vaccinated animals and a drastic reduction of tumor burden was observed in the remaining tumor-affected mice. These data demonstrate that, by enhancing E7 yield, it is possible to improve the anti-cancer activity of the plant-based experimental vaccine and open the way for a large-scale production of the E7 protein which could be purified or used as in planta formulation, also suitable for oral therapeutic vaccination.

Leukocyte common-antigen-related tyrosine phosphatase receptor: altered expression of mRNA and protein in the New England Deaconess Hospital rat line exhibiting spontaneous pheochromocytoma.

Regulation of cell proliferation by protein tyrosine phosphatases (PTPs) suggests that PTPs are important tumor suppressor genes. The gene encoding the leukocyte common-antigen-related (LAR) PTP receptor maps to chromosome 1p32-33, a region in which loss of heterozygosity is associated with human pheochromocytoma and other neuroectodermal tumors. The rat pheochromocytoma PC12 cell line was originally derived from the transplantable P259 tumor originating from the New England Deaconess Hospital (NEDH) line of Wistar inbred rats. Compared with their Wistar counterparts, 1-2-year-old NEDH rats exhibit a high incidence of spontaneous pheochromocytomas. This study investigates whether levels of LAR transcripts and protein are altered in NEDH adrenal tissue prior to tumor onset. In addition, alternative splicing of an LAR extracellular domain [LAR alternatively spliced element-c (LASE-c)], regulating LAR interaction with extracellular matrix components, was examined. These changes in LAR expression and alternative splicing were hypothesized to be more pronounced in tumor tissue and PC12 cells. Northern blot analysis demonstrated the presence of the approximately 5 kb LAR transcript in all cell lines examined, except PC12. In adrenal medulla tissue harvested from 2-3-month-old rats, LAR approximately 8 and approximately 5 kb transcript expression was decreased in NEDH compared with Wistar samples. RT-PCR demonstrated increased splicing of the LASE-c 27 bp alternatively spliced insert in the LAR extracellular domain in NEDH adrenal medulla tissue. Even greater LASE-c splicing was detected in adrenal medulla tumor tissue derived from 12-month-old NEDH rats and in PC12 cells. Western blot analysis demonstrated decreased levels of LAR protein and increased levels of LASE-c containing LAR protein isoforms in NEDH adrenal medulla tissue. These studies demonstrate that patterns of altered LAR expression present in PC12 cells and in pheochromocytoma tumor tissue are also present in adrenal tissue predisposed to a high incidence of spontaneous pheochromocytoma.

Leukocyte common antigen-related tyrosine phosphatase receptor: increased expression and neuronal-type splicing in breast cancer cells and tissue.

The findings that protein tyrosine phosphatases (PTPs) regulate cell proliferation, response to growth factors, and cellular adhesion and the discovery that mutations in PTP genes are associated with breast cancer suggest that altered expression of PTPs contributes to the breast cancer cell phenotype. The leukocyte common antigen-related (LAR) PTP receptor is a prototype member of the class of PTP receptors containing cell adhesion domains. Full-length constitutively spliced LAR transcripts are expressed in breast and other tissues, whereas alternatively spliced isoforms are preferentially expressed in the nervous system. As a first step in evaluating the hypothesis that LAR-type PTPs influence breast cancer cell behavior, LAR expression and neuronal-type alternative splicing were examined in normal and breast cancer cell lines and tissues. Northern blot analysis demonstrated markedly increased LAR mRNA levels in breast cancer cell lines and tissues. Western blot analysis showed a greater than tenfold increase in LAR protein levels in breast cancer tissues. Reverse transcription-polymerase chain reaction was used to assess alternative splicing of extracellular and proximal membrane exons. Differential patterns of extracellular alternative splicing were found in normal versus carcinoma cell lines and tissues. Western blot analysis demonstrated increased levels of LAR protein isoforms encoded by alternatively spliced transcripts in breast cancer cell lines. This study is the first demonstration of increased LAR mRNA and LAR protein expression in breast cancer tissue and nontransformed cell lines and helps to elucidate the role of LAR in human breast cancer. The differential patterns of alternative splicing of LAR transcripts introduce LAR isoforms as candidate markers for future studies correlating differential gene expression and tumor behavior.

Geometrically and conformationally restrained cinnamoyl compounds as inhibitors of HIV-1 integrase: synthesis, biological evaluation, and molecular modeling.

Various cinnammoyl-based structures were synthesized and tested in enzyme assays as inhibitors of the HIV-1 integrase (IN). The majority of compounds were designed as geometrically or conformationally constrained analogues of caffeic acid phenethyl ester (CAPE) and were characterized by a syn disposition of the carbonyl group with respect to the vinylic double bond. Since the cinnamoyl moiety present in flavones such as quercetin (inactive on HIV-1-infected cells) is frozen in an anti arrangement, it was hoped that fixing our compounds in a syn disposition could favor anti-HIV-1 activity in cell-based assays. Geometrical and conformational properties of the designed compounds were taken into account through analysis of X-ray structures available from the Cambridge Structural Database. The polyhydroxylated analogues were prepared by reacting 3,4-bis(tetrahydropyran-2-yloxy)benzaldehyde with various compounds having active methylene groups such as 2-propanone, cyclopentanone, cyclohexanone, 1,3-diacetylbenzene, 2, 4-dihydroxyacetophenone, 2,3-dihydro-1-indanone, 2,3-dihydro-1, 3-indandione, and others. While active against both 3'-processing and strand-transfer reactions, the new compounds, curcumin included, failed to inhibit the HIV-1 multiplication in acutely infected MT-4 cells. Nevertheless, they specifically inhibited the enzymatic reactions associated with IN, being totally inactive against other viral (HIV-1 reverse transcriptase) and cellular (RNA polymerase II) nucleic acid-processing enzymes. On the other hand, title compounds were endowed with remarkable antiproliferative activity, whose potency correlated neither with the presence of catechols (possible source of reactive quinones) nor with inhibition of topoisomerases. The SARs developed for our compounds led to novel findings concerning the molecular determinants of IN inhibitory activity within the class of cinnamoyl-based structures. We hypothesize that these compounds bind to IN featuring the cinnamoyl residue C=C-C=O in a syn disposition, differently from flavone derivatives characterized by an anti arrangement about the same fragment. Certain inhibitors, lacking one of the two pharmacophoric catechol hydroxyls, retain moderate potency thanks to nonpharmacophoric fragments (i.e., a m-methoxy group in curcumin) which favorably interact with an "accessory" region of IN. This region is supposed to be located adjacent to the binding site accommodating the pharmacophoric dihydroxycinnamoyl moiety. Disruption of coplanarity in the inhibitor structure abolishes activity owing to poor shape complementarity with the target or an exceedingly high strain energy of the coplanar conformation.

Effect of genetic modification of acute inflammatory responsiveness on tumorigenesis in the mouse.

Two distinct bidirectional selective breedings for quantitative traits were initiated from identical genetically heterogeneous mouse populations. The resulting lines are characterized by maximal or minimal acute inflammatory responsiveness (AIR): AIRmax and AIRmin lines, respectively, and by resistance or susceptibility to chemical skin tumorigenesis: Car-R and Car-S lines, respectively. The AIR response to s.c. injection of polyacrylamide microbeads, measured by cell content in the local exudate, was 10 times higher in AIRmax than in AIRmin mice. The response to selection was asymmetrical: the realized heritability was 0.26 in AIRmax and 0.008 in AIRmin, and resulted from the additive effect of 7-11 quantitative trait loci (QTL). Low responsiveness was globally dominant in F1 and 48% of F2 segregant variance was found to be due to genetic factors. These findings are the first demonstration of innate regulation of AIR by germ line genes. Susceptibility to skin tumorigenesis induced by a two-stage initiation (DMBA)-promotion (TPA) protocol was lower in AIRmax mice than in AIRmin mice, a 6-fold difference in tumor induction rate. Intense AIR was found to be associated with resistance, and low AIR with susceptibility to tumorigenesis, in F2 segregants chosen for extreme AIR phenotypes. At least some of the AIR QTLs therefore contain genes controlling tumorigenesis. Tumor phenotypes differed more in Car-R and Car-S than in AIRmax and AIRmin lines, indicating that QTLs unrelated to AIR, contribute to the host response to tumorigenesis. The extreme phenotypes/genotypes of the four selected lines and the known genetic constitution of their foundation population, offer new possibilities to discriminate the genes/mechanisms controlling two important traits: AIR and response to chemical tumorigenesis. Collaborative projects will be favorably considered. The description of tumor resistance genes in AIRmax and Car-R mice may be helpful for epidemiology and therapy of human cancer.

Sulfone derivatives with anti-HIV activity.

Following the discovery of anti-HIV properties of suramin great efforts were devoted to design novel NNRT agents with the aims to find novel drugs for the clinical therapeutic management of AIDS. Sulfone and sulfonamide derivatives were studied by NCI at Bethesda as potential anti-HIV-1 agents and nitrophenyl phenyl sulfone (NPPS) was selected as lead compound for further investigations. At the same time Merck Laboratories discovered L-737,126, a potent indolyl aryl sulfone with inhibitory activity against reverse transcriptase. These studies stimulated novel search in the sulfone series and both diarylsulfones and cyclic sulfone derivatives were investigated. Our decennial interest in chemotherapeutic agents containing a pyrrole ring pulsed us to synthesize and test as anti-HIV-1 agents a number of pyrryl aryl sulfones (PASs), pyrrolobenzothiadiazepine (PBTDs) and pyrrolobenzothiazepine related sulfones. The new sulfone derivatives inhibit selectively HIV-1 and were inactive against HIV-2. Most of them were as active as, if not more active than, nevirapine.

Global ischemia induces apoptosis-associated genes in hippocampus.

Using in situ hybridization, Northern blotting and RT-PCR we studied the post-ischemic expression of bcl-2, bcl-x, bax and ICE. One day following 5 min or 10 min of global ischemia bcl-2 and bcl-x mRNAs were induced in CA1 hippocampal pyramidal neurons while bax was unchanged. By 72 h after ischemia the expression of bcl-2, bcl-x and bax mRNAs decreased in CA1. The large isoform of bcl-x (bcl-xL), detected using RT-PCR, decreased in whole hippocampus by 24-72 h after ischemia relative to the putative short (bcl-xS) and transmembrane deleted (bcl-x delta TM) forms. Oligonucleotides to interleukin-1 beta convertase (ICE), which detected the expected 2-kb transcript and two lesser 1.5- and 3-kb hybridizing species, demonstrated slight mRNA induction in the CA1 region at 72 h following ischemia. DNA nick end-labeling at 3 days following ischemia showed DNA fragmentation in neurons limited to the CA1 region of hippocampus following 5 min ischemia, while DNA fragmentation was detected in CA1, CA3, dentate gyrus and cortical neurons following 10 min ischemia. The data support the view that hippocampal neurons might undergo an apoptosis-like death after global ischemia. Since global ischemia decreases total protein synthesis especially in the CA1 region, the increases in bcl-2 mRNA levels may not necessarily lead to increased Bcl-2 protein levels. This may explain why the CA1 neurons die despite the prominent induction of the protective bcl-2 gene. The observed decrease by 24 h in the bcl-xL/bcl-xS ratio which preceded DNA fragmentation may participate in the cell death produced by ischemia. However, because of the ischemia-induced decrease in total protein synthesis, the decreased bcl-xL/bcl-xS ratio does not necessarily lead to a changed ratio in the amount of the appropriate proteins. Since ICE-like mRNA was induced at 72 h when the CA1 neurons were dead, the significance of this ICE-like mRNA induction remains unclear.

Synthesis and anti-HIV activity of 10,11-dihydropyrrolo [1,2-b][1,2,5]benzothiadiazepine-11-acetic acid 5,5-dioxide derivatives and related compounds.

The synthesis and the in vitro anti-HIV-1 activity of novel pyrrolo annulated benzothiadiazepine acetic acids and some related derivatives are reported. The new compounds share chemical features with pyrrolo[1,2-d][1,4]benzodiazepin-6-one 1 and Ro 5-3335 pyrrylbenzodiazepinone 4, two inhibitors of HIV-replication at the level of reverse transcriptase (RT) and transcriptional transactivation by Tat, respectively. Two derivatives, namely methyl 10,11-dihydropyrrolo[1,2-b][1,2,5]benzothiadiazepine-11-acetic-5,5 -dioxide (5a) and 1,12b-dihydro-2H-azeto[2,1-d]pyrrolo[1,2-b][1,2,5]benzoth iadiazepin-2-one 8,8-dioxide (7a), were found to exhibit a significant, although not very potent, activity against human immunodeficiency virus Type 1 (HIV-1).

Heme oxygenase-1 (HO-1) protein induction in rat brain following focal ischemia.

The induction of the heme oxygenase-1 (HO-1) protein, also called HSP32, was compared to HSP70 heat shock protein induction following focal ischemia. Adult Sprague-Dawley male rats (n = 14) were subjected to either 30 min or 2 h of focal cerebral ischemia using the suture, middle-cerebral-artery (MCA) occlusion model. Controls (n = 4) had sham surgery. Following 24 h of reperfusion, subjects were killed and their brains stained immunocytochemically for HO-1 and the HSP70 heat shock proteins. One day following 30 min of ischemia, HO-1 and HSP70 staining in striatum occurred mainly in endothelial cells in infarcts and in glial cells surrounding the areas of infarction. Following the 30 min ischemia HO-1 was not induced in cortex whereas HSP70 was induced in cortical neurons in the MCA distribution. One day following 2 h of MCA ischemia, both HO-1 and HSP70 were induced in neurons in cortex in the MCA distribution. HO-1, however, was induced in glial cells throughout ipsilateral cortex, inside as well as outside the MCA distribution. This suggests that translation and/or transcription of the HO-1 and HSP70 genes are blocked in neurons and glia destined to die within infarcts, whereas translation of these stress genes continues in the endothelial cells. The duration of ischemia required to induce HSP70 in cortical neurons appears to be less than that required to induce HO-1 in cortical glia. Prolonged spreading depression and/or diffuse hemispheric ischemia may induce HO-1 in glia throughout the ipsilateral cortex via immediate early gene activation of the AP-1 site in the HO-1 promoter. Since HO-1 degrades heme, a pro-oxidant, to antioxidant molecules, the induction of HO-1 may augment oxidative defense mechanisms compromised by cerebral ischemia.

L-29, an endogenous lectin, binds to glycoconjugate ligands with positive cooperativity.

The soluble mammalian lactose-binding lectins L-14-I and L-29 are both secreted and bind to oligosaccharides on laminin, a large extracellular matrix glycoprotein containing polylactosamine chains. Because of the potential functional significance of these lectin-laminin interactions, we compared quantitative aspects of L-14-I and L-29 binding to immobilized laminin using recombinant lectins labeled with 125I. We report that the concentration-dependent binding of L-29 exhibits positive cooperativity whereas binding of L-14-I does not. Cooperative binding of L-29 can also occur on glycoconjugate substrates other than laminin and is not dependent on cystine bond formation or aggregation in solution. L-29 contains repetitive sequences within the N-terminal domain not present in L-14-I. This domain is not required for binding activity, but is required for positive cooperativity. Though the precise mechanism of interaction of L-29 with laminin remains to be determined, it apparently results in assembly of a lectin aggregate on the substrate surface, which could have important functional consequences.

Isolation and expression of a gene encoding L-14-II, a new human soluble lactose-binding lectin.

In the course of screening a human hepatoma cDNA library with antibody raised against a mammalian lectin with subunit molecular weight of about 14,000, we detected a partial cDNA encoding a related but distinct protein that was possibly a homologous lectin (Gitt and Barondes, 1986). We here report the isolation and sequencing of a full-length cDNA for this protein from a HepG2 cDNA library. The cDNA encodes a protein with subunit molecular weight of 14,650. Expression of the coding sequence in Escherichia coli yields a product that binds to a lactose affinity column and is specifically eluted with lactose, confirming that this new protein is a lectin. Like its well studied relative, here called L-14-I, the new lectin, L-14-II, exists as a homodimer in solution. The two related human lectins have 43% amino acid sequence identity. The genomic DNA encoding L-14-II (LGALS2) contains four exons with similar intron placement to L-14-I (LGALS1); but the genomic upstream region, which contains several sequences characteristic of regulatory elements, differs significantly from L-14-I.

Preliminary results corroborating the polygenic control of immunological tolerance

Tolerance-susceptible (TS) and-resistant (TR) lines of mice are in the process of bidirectional genetic selection starting from a genetically hetrogeneous population achieved by the equilibrated intercrossing of eight inbred lines. Mice are intragastrically pretreated and then immunized with hen ovalbumin or bovine serum albunin and the extreme phenotypes are selected for assortative mating. The normal distribution of agglutinin titers in the F0 population and the significant interline difference already observed in the F2 and F3 generations indicate that oral tolerance is a character controlled by the additive effect of several independet loci. The mean heritability (h2) obtained thus far is 11% for the TS line and 19% for the TR line

Salmonella typhimurium infection in high and low antibody responder mice: inverse correlation between antibody responsiveness and resistance to infection.

Susceptibility to Salmonella typhimurium infection was compared in H (high Ab responder) and L (low Ab responder) mice obtained by several selective breeding experiments (Selections I, II, III, IV and IV A). H mice were always much more susceptible to infection than their L mice counterparts within a continuous LD 50 variation range. In three of the selections (I, II and IV A) the low responsiveness character is known to result mainly from rapid Ag degradation in L mice macrophages. It was hypothesized that resistance to multiplication of intracellular pathogens could be related to an increased catabolic activity towards Ag. This was actually demonstrated, in F2 segregant hybrids of selection IV A, by the significant inverse correlation between capacity for Ab production and resistance to infection.

[Anesthesia with isoflurane in surgery of the anterior segment of the eye]. / Anestesia con isoflurano nella chirurgia del segmento anteriore dell'occhio.

The effects of isoflurane on intraocular pressure (IOP) were studied in 46 patients undergoing cataract surgery. The IOP was measured 30 minutes after premedication, 10 minutes after induction of anesthesia and 10 minutes after administration of isoflurane. Since a significant decrease of IOP was found to occur after the administration of volatile agent, and in account of absence of complications or side effects, the Authors conclude that isoflurane can be considered a suitable anesthetic agent in ophthalmic surgery.

Potential antitumor agents. II. Lappin intramolecular cyclization of diethyl 3-carbazolylaminomethylenemalonate: a structure assignment.

Diethyl 3-carbazolylaminomethylenemalonate undergoes Lappin cyclization giving 2-carbethoxy-1-oxo-1,4-dihydro-7H-pyrido[2,3-c]carbazole. Formation of an angular pyridocarbazole derivative is consistent with the Markwald rule. The synthesis of 4-ethyl derivatives of the above pyridocarbazole and its 8,9,10,11-tetrahydro derivative is also reported as a goal to potential indoloquinolone antitumoral agents.