Accessible and reliable neurometric testing in humans using a smartphone platform.

Tests of human brain circuit function typically require fixed equipment in lab environments. We have developed a smartphone-based platform for neurometric testing. This platform, which uses AI models like computer vision, is optimized for at-home use and produces reproducible, robust results on a battery of tests, including eyeblink conditioning, prepulse inhibition of acoustic startle response, and startle habituation. This approach provides a scalable, universal resource for quantitative assays of central nervous system function.

TB in the elderly: clinical features and outcomes.

BACKGROUND: TB in low-incidence countries is characterised by changes in age distribution towards larger numbers of cases among the elderly.OBJECTIVES: To investigate clinical features and outcomes of TB treatment in older patients and identify predictors of poor outcome.METHODS: Multicentre retrospective study of new TB cases from 53 hospitals included in the registry of the Integrated Tuberculosis Research Programme of the Spanish Society of Pulmonology and Thoracic Surgery (Sociedad Española de Neumología y Cirugía Torácica) between 2006 and 2020.RESULTS: We identified 731 patients aged ≥75 years from a cohort of 7,505 patients with TB. In the elderly, weight loss, disseminated disease and normal X-rays or infiltrates without cavitation were more common. All-cause mortality was 16% (5% of deaths due to TB). The elderly had higher rates of toxicity (6.7%) and hospital admissions (36%). In the multivariate analysis of predictors of TB mortality in ≥75-year-olds, only weight, age and treatment with non-standard regimens remained significant.CONCLUSIONS: TB in older patients needs more attention and remains a challenge because of a lack of specific clinical and radiological features. Standard treatment is effective, although mortality is higher than in young patients. Low weight, non-standard regimens and age are significant predictors of TB mortality.

Factors Associated with Fatality during the Intensive Phase of Anti-Tuberculosis Treatment.

OBJECTIVE: To determine the case-fatality rate (CFR) at the end of the intensive phase of tuberculosis (TB) treatment, and factors associated with fatality. METHODS: TB patients diagnosed between 2006 and 2013 were followed-up during treatment. We computed the CFR at the end of the intensive phase of TB treatment, and the incidence of death per 100 person-days (pd) of follow-up. We performed survival analysis using the Kaplan-Meier method and Cox regression, and calculate hazard ratios (HR) and 95% confidence intervals (CI). RESULTS: A total of 5,182 patients were included, of whom 180 (3.5%) died; 87 of these deaths (48.3%) occurred during the intensive phase of treatment, with a CFR of 1.7%. The incidence of death was 0.028/100 pd. The following factors were associated with death during the intensive phase: being >50 years (HR = 36.9;CI:4.8-283.4); being retired (HR = 2.4;CI:1.1-5.1); having visited the emergency department (HR = 3.1;CI:1.2-7.7); HIV infection (HR = 3.4;CI:1.6-7.2); initial standard treatment with 3 drugs (HR = 2.0;CI:1.2-3.3) or non-standard treatments (HR = 2.68;CI:1.36-5.25); comprehension difficulties (HR = 2.8;CI:1.3-6.1); and smear-positive sputum (HR = 2.3-CI:1.0-4.8). CONCLUSION: There is a non-negligible CFR during the intensive phase of TB, whose reduction should be prioritised. The CFR could be a useful indicator for evaluating TB programs.

Ameloblast Modulation and Transport of Cl⁻, Na⁺, and K⁺ during Amelogenesis.

Ameloblasts express transmembrane proteins for transport of mineral ions and regulation of pH in the enamel space. Two major transporters recently identified in ameloblasts are the Na(+)K(+)-dependent calcium transporter NCKX4 and the Na(+)-dependent HPO4 (2-) (Pi) cotransporter NaPi-2b. To regulate pH, ameloblasts express anion exchanger 2 (Ae2a,b), chloride channel Cftr, and amelogenins that can bind protons. Exposure to fluoride or null mutation of Cftr, Ae2a,b, or Amelx each results in formation of hypomineralized enamel. We hypothesized that enamel hypomineralization associated with disturbed pH regulation results from reduced ion transport by NCKX4 and NaPi-2b. This was tested by correlation analyses among the levels of Ca, Pi, Cl, Na, and K in forming enamel of mice with null mutation of Cftr, Ae2a,b, and Amelx, according to quantitative x-ray electron probe microanalysis. Immunohistochemistry, polymerase chain reaction analysis, and Western blotting confirmed the presence of apical NaPi-2b and Nckx4 in maturation-stage ameloblasts. In wild-type mice, K levels in enamel were negatively correlated with Ca and Cl but less negatively or even positively in fluorotic enamel. Na did not correlate with P or Ca in enamel of wild-type mice but showed strong positive correlation in fluorotic and nonfluorotic Ae2a,b- and Cftr-null enamel. In hypomineralizing enamel of all models tested, 1) Cl(-) was strongly reduced; 2) K(+) and Na(+) accumulated (Na(+) not in Amelx-null enamel); and 3) modulation was delayed or blocked. These results suggest that a Na(+)K(+)-dependent calcium transporter (likely NCKX4) and a Na(+)-dependent Pi transporter (potentially NaPi-2b) located in ruffle-ended ameloblasts operate in a coordinated way with the pH-regulating machinery to transport Ca(2+), Pi, and bicarbonate into maturation-stage enamel. Acidification and/or associated physicochemical/electrochemical changes in ion levels in enamel fluid near the apical ameloblast membrane may reduce the transport activity of mineral transporters, which results in hypomineralization.

On-water remote monitoring robotic system for estimating the patch coverage of Anabaena sp. filaments in shallow water.

An on-water remote monitoring robotic system was developed for indirectly estimating the relative density of marine cyanobacteria blooms at the subtidal sandy-rocky beach in Balandra Cove, Baja California Sur, Mexico. The system is based on an unmanned surface vehicle to gather underwater videos of the seafloor for avoiding physical damage on Anabaena sp. cyanobacteria colonies, which grow in tufts of filaments weakly attached to rocks, seagrass, and macroalgae. An on-axis image stabilization mechanism was developed to support a camcorder and minimize wave perturbation while recording underwater digital images of the seafloor. Color image processing algorithms were applied to estimate the patch coverage area and density, since Anabaena sp. filaments exhibit a characteristic green tone. Results of field tests showed the feasibility of the robotic system to estimate the relative density, distribution, and coverage area of cyanobacteria blooms, preventing the possible impact of direct observation. The robotic system could also be used in surveys of other benthos in the sublittoral zone.

NBCe1 (SLC4A4) a potential pH regulator in enamel organ cells during enamel development in the mouse.

During the formation of dental enamel, maturation-stage ameloblasts express ion-transporting transmembrane proteins. The SLC4 family of ion-transporters regulates intra- and extracellular pH in eukaryotic cells by cotransporting HCO3 (-) with Na(+). Mutation in SLC4A4 (coding for the sodium-bicarbonate cotransporter NBCe1) induces developmental defects in human and murine enamel. We have hypothesized that NBCe1 in dental epithelium is engaged in neutralizing protons released during crystal formation in the enamel space. We immunolocalized NBCe1 protein in wild-type dental epithelium and examined the effect of the NBCe1-null mutation on enamel formation in mice. Ameloblasts expressed gene transcripts for NBCe1 isoforms B/D/C/E. In wild-type mice, weak to moderate immunostaining for NBCe1 with antibodies that recognized isoforms A/B/D/E and isoform C was seen in ameloblasts at the secretory stage, with no or low staining in the early maturation stage but moderate to high staining in the late maturation stage. The papillary layer showed the opposite pattern being immunostained prominently at the early maturation stage but with gradually less staining at the mid- and late maturation stages. In NBCe1 (-/-) mice, the ameloblasts were disorganized, the enamel being thin and severely hypomineralized. Enamel organs of CFTR (-/-) and AE2a,b (-/-) mice (CFTR and AE2 are believed to be pH regulators in ameloblasts) contained higher levels of NBCe1 protein than wild-type mice. Thus, the expression of NBCe1 in ameloblasts and the papillary layer cell depends on the developmental stage and possibly responds to pH changes.

Barrier formation: potential molecular mechanism of enamel fluorosis.

Enamel fluorosis is an irreversible structural enamel defect following exposure to supraoptimal levels of fluoride during amelogenesis. We hypothesized that fluorosis is associated with excess release of protons during formation of hypermineralized lines in the mineralizing enamel matrix. We tested this concept by analyzing fluorotic enamel defects in wild-type mice and mice deficient in anion exchanger-2a,b (Ae2a,b), a transmembrane protein in maturation ameloblasts that exchanges extracellular Cl(-) for bicarbonate. Defects were more pronounced in fluorotic Ae2a,b (-/-) mice than in fluorotic heterozygous or wild-type mice. Phenotypes included a hypermineralized surface, extensive subsurface hypomineralization, and multiple hypermineralized lines in deeper enamel. Mineral content decreased in all fluoride-exposed and Ae2a,b(-/-) mice and was strongly correlated with Cl(-). Exposure of enamel surfaces underlying maturation-stage ameloblasts to pH indicator dyes suggested the presence of diffusion barriers in fluorotic enamel. These results support the concept that fluoride stimulates hypermineralization at the mineralization front. This causes increased release of protons, which ameloblasts respond to by secreting more bicarbonates at the expense of Cl(-) levels in enamel. The fluoride-induced hypermineralized lines may form barriers that impede diffusion of proteins and mineral ions into the subsurface layers, thereby delaying biomineralization and causing retention of enamel matrix proteins.

Factores relacionados con el seguimiento de los pacientes en programa de deshabituación tabáquica / Follow-up related factors in patients in the anti-smoking program

Objetivo. El objetivo del presente estudio fue evaluar las características de los pacientes que completan un programa de 12 meses de deshabituación tabáquica, en comparación con los que no lo completan, a fin de determinar el perfil de paciente respondedor. Métodos. Estudio observacional, prospectivo, de corte transversal en el que fueron incluidos todos los sujetos que asistieron a nuestra clínica en busca de información sobre los programas de deshabituación entre 2000 y 2005. A todos los pacientes se les realizó una evaluación individual con recogida de los antecedentes personales, incluyendo cuadros médicos y psicológicos, así como diversos aspectos relativos al consumo de tabaco. La relación entre las distintas variables recogidas y la finalización del programa fue analizada en un estudio mutlivariante. Resultados. Durante el período de estudio, fueron atendidos 1.681 sujetos en nuestra unidad de deshabituación. Las variables que resultaron estar relacionadas con la finalización del programa de deshabituación fueron la presencia de insuficiencia cardíaca (OR: 3,50;IC95%: 1,24-9,87), la puntuación del test de Fagerström (OR: 0,93;IC95%: 0,87-0,99), la cantidad de nicotina en el cigarrillo (OR0,19; IC95% 0,05-0,66), el tratamiento con bupropion (OR: 2,59;IC95%: 1,70-3,94), la presencia de trastornos del ánimo (OR: 0,59;IC95% 0,38-0,91) y el número de sesiones de psicoterapia (OR 2,18;IC95% 1,99-2,39).Conclusiones. El presente estudio encuentra una relación entre las variables descritas y la finalización del seguimiento de un programa de deshabituación a los 12 meses. Esta información podría ser de ayuda para identificar a los pacientes que se beneficiarían de una estrategia de seguimiento que mejore la cumplimentación del programa. (AU)

Objective. The objective of this study was to evaluate the characteristics of the patients who complete a 12 month quit smoking program,in comparison with those who do not complete it to determine there sponding patient profile. Methods. This observational, prospective study included all those subjects attending our clinic in search of information about anti-smoking programs between 2000 and 2005. All patients were evaluated individually, including personal background as well as medical and psychological information. Likewise, various aspects relating to the consumption of tobacco were also included. The relationship between the different variables collected and the finalization of the program were analyzed in a multi-variant study. Results. During the study, 1.681 subjects attended our anti-smoking unit. The variables that turned out to be related to the finalization of the anti-smoking program were the presence of cardiac insufficiency(OR: 3.50; IC95%: 1.24-9.87), the scoring on the Fagerström test(OR: 0.93; IC95%: 0.87-0.99), the quantity of nicotine in the cigarette(OR 0.19; IC95% 0.05-0.66), the treatment with bupropion (OR: 2.59; IC95%: 1.70-3.94), the presence of depression (OR:0.59; IC95% 0.38-0.91) and the number of psychotherapy sessions(OR 2.18; IC95% 1.99-2.39).Conclusions. This study found a relationship between the described variables and the finalization of the follow up of anti-smoking program after 12 months. This information could help identify those patients who would benefit from a follow-up strategy that improves their compliance with the program (AU)

The anion exchanger Ae2 is required for enamel maturation in mouse teeth.

One of the mechanisms by which epithelial cells regulate intracellular pH is exchanging bicarbonate for Cl(-). We tested the hypothesis that in ameloblasts the anion exchanger-2 (Ae2) is involved in pH regulation during maturation stage amelogenesis. Quantitative X-ray microprobe mineral content analysis, scanning electron microscopy, histology, micro-computed tomography and Ae2 immuno-localisation analyses were applied to Ae2-deficient and wild-type mouse mandibles. Immuno-localisation of Ae2 in wild-type mouse incisors showed a very strong expression of Ae2 in the basolateral membranes of the maturation stage ameloblasts. Strikingly, zones of contiguous ameloblasts were found within the maturation stage in which Ae2 expression was extremely low as opposed to neighbouring cells. Maturation stage ameloblasts of the Ae2(a,b)(-/-) mice failed to stain for Ae2 and showed progressive disorganisation as enamel development advanced. Maturation stage enamel of the Ae2(a,b)(-/-) mice contained substantially less mineral and more protein than wild-type enamel as determined by quantitative X-ray microanalysis. Incisor enamel was more severely affected than molar enamel. Scanning electron microscopy revealed that the rod-inter-rod structures of the Ae2(a,b)(-/-) mice incisor enamel were absent. Mineral content of dentine and bone of Ae2(a,b)(-/-) mice was not significantly different from wild-type mice. The enamel from knockout mouse teeth wore down much faster than that from wild-type litter mates. Basolateral bicarbonate secretion via the anionic exchanger Ae2 is essential for mineral growth in the maturation stage enamel. The observed zonal expression of Ae2 in the maturation stage ameloblasts is in line with a model for cyclic proton secretion during maturation stage amelogenesis.

A mechanism for savings in the cerebellum.

The phenomenon of savings (the ability to relearn faster than the first time) is a familiar property of many learning systems. The utility of savings makes its underlying mechanisms of special interest. We used a combination of computer simulations and reversible lesions to investigate mechanisms of savings that operate in the cerebellum during eyelid conditioning, a well characterized form of motor learning. The results suggest that a site of plasticity outside the cerebellar cortex (possibly in the cerebellar nucleus) can be protected from the full consequences of extinction and that the residual plasticity that remains can later contribute to the savings seen during relearning.

Computer simulation of cerebellar information processing.

Although many functions have been ascribed to the cerebellum, the uniformity of its synaptic organization suggests that a single, characteristic computation may be common to all. Computer simulations are useful in examining this cerebellar computation, as they inherently address function at the level of information processing. Progress is facilitated by factors that make the cerebellum particularly amenable to such analysis. We review progress from two contrasting approaches. Top-down simulations begin with hypotheses about computational mechanisms and then ask how such mechanisms might operate within the cerebellum. Bottom-up simulations attempt to build a representation of the cerebellum that reflects known cellular and synaptic components as accurately as possible. We describe recent advances from these two approaches that are leading to an understanding of what information the cerebellum processes and how its neurons and synapses accomplish this task.

Molecular cloning and genomic organization of the mouse AE2 anion exchanger gene.

The molecular organization of the AE2 (SLC4A2) gene, a member of the multigene family encoding sodium-independent chloride/bicarbonate anion exchangers, has previously been described in both humans and rats. In these two species, AE2 shows alternate promoter usages and tissue-specific expression of isoforms in a similar, but not identical, fashion. Here we report the molecular cloning and organization of the entire mouse AE2 gene. The gene consists of 23 exons and 22 introns and spans about 17 kb. Moreover, it drives transcription of N-terminal truncated isoforms from alternate promoter sequences in a way analogous to that described for rat and/or human orthologs. Thus, sequences within intron 2 function as overlapping alternate promoters for truncated isoforms AE2b(1) and AE2b(2), and sequences of intron 5 drive transcription of isoforms AE2c(1) and AE2c(2). Each of these variants has a specific alternative first exon, while remaining exons are common to the complete form of the message AE2a, the diversity at 5' leading to different N-termini in corresponding encoded proteins. As expected, mouse AE2 promoter sequences and the patterns of tissue expression of AE2 isoforms resemble rat counterparts more closely than human ones.

Cerebellar function: coordination, learning or timing?

Theories of cerebellar function have largely involved three ideas: movement coordination, motor learning or timing. New evidence indicates these distinctions are not particularly meaningful, as the cerebellum influences movement execution by feedforward use of sensory information via temporally specific learning.

Timing mechanisms in the cerebellum: testing predictions of a large-scale computer simulation.

We used large-scale computer simulations of eyelid conditioning to investigate how the cerebellum generates and makes use of temporal information. In the simulations the adaptive timing displayed by conditioned responses is mediated by two factors: (1) different sets of granule cells are active at different times during the conditioned stimulus (CS), and (2) responding is not only amplified at reinforced times but also suppressed at unreinforced times during the CS. These factors predict an unusual pattern of responding after partial removal of the cerebellar cortex that was confirmed using small, electrolytic lesions of cerebellar cortex. These results are consistent with timing mechanisms in the cerebellum that are similar to Pavlov's "inhibition of delay" hypothesis.

In situ localization of anion exchanger-2 in the human kidney.

Na+-independent anion exchangers (AE) are a family of membrane carriers that mediate the electroneutral exchange of Cl- for HCO3- ions across plasma membranes. They are involved in intracellular pH and cell volume regulation as well as in transepithelial acid-base transport. While anion exchanger-1 (AE1) has been localized previously in the human kidney, thus far there has been no definite report on anion exchanger-2 (AE2) in this human tissue. Accordingly, immunohistochemistry was carried out on surgical specimens of the human kidney (fixed in formalin and embedded in paraffin), using a specific AE2 monoclonal antibody. Strong immunostaining was observed at the basolateral membrane of cells of thick ascending limbs and distal convoluted tubules, colocalizing with the basal membranous labyrinth of cellular interdigitations, typical of these segments. In fact, AE2 staining was attenuated at the macula densa, where basal infoldings are scarce. Additionally, in situ hybridization experiments on formalin-fixed tissue demonstrated the presence of AE2 mRNA in the same segments of the distal nephron. On the other hand, control immunohistochemistry with a monoclonal antibody against AE1 gave the expected immunoreactivity at the basal pole of the type A intercalated cells of connecting tubules and cortical collecting ducts, and in erythrocytes. Our results indicate that, depending on the nephron segment and corresponding cell types, AE1 and AE2 proteins are differentially involved in the Na+-independent exchange of Cl- for HCO3- at the basolateral membrane of polarized kidney epithelial cells.

Tissue-specific N-terminal isoforms from overlapping alternate promoters of the human AE2 anion exchanger gene.

Previously, we isolated the human AE2 (SLC4A2) gene, a member of the sodium-independent anion exchanger family. Rat ortholog of this gene was reported to drive alternative transcription yielding N-terminal variants of the AE2a message. We thus analyzed the human AE2 gene in this regard. Using HepG2 cells, two alternative first exons, each splicing to exon 3 in alternative transcripts, were found to be transcribed from overlapping sequences of intron 2. Exon 1b(1) corresponds to the rat variant "b" and encodes three initial residues (MTQ) in AE2b(1) isoform that replace the first 17 amino acids of AE2a protein, while the novel exon 1b(2) encodes eight initial residues (MDFLLRPQ) in AE2b(2) isoform. The relative abundance of AE2b(1) and AE2b(2) mRNAs was about 10% of AE2a mRNA each. Alternate promoter sequences have multiple potential binding motifs for liver-enriched factors, and dual-luciferase assays indicated that they possess the ability for driving transcription in transiently transfected HepG2 cells. Tissue survey showed that expression of human AE2b(1) and AE2b(2) transcripts is restricted to liver and kidney, while AE2a mRNA was encountered in all examined tissues. Our findings reveal a characteristic tissue-specific expression of two N-terminal variants of human AE2 from overlapping sequences within intron 2, one of which is a novel isoform.

Leukotriene A4 hydrolase: a critical role of glutamic acid-296 for the binding of bestatin.

Leukotriene A(4) hydrolase is a bifunctional Zn(2+)-containing enzyme catalysing the formation of the potent chemotaxin leukotriene B(4). From an analysis of three mutants of Glu-296 we have found that this catalytic residue is critical for the binding of bestatin, a classical aminopeptidase inhibitor. For bestatin, but not for three other tight-binding inhibitors, the IC(50) values for inhibition of the epoxide hydrolase activity decreased in the mutants to 0.7-0.003% of the control. Hence Glu-296 is an important structural determinant for binding of bestatin to leukotriene A(4) hydrolase; this conclusion might also apply to other members of the M1 family of metallopeptidases.

Mechanisms of cerebellar learning suggested by eyelid conditioning.

Classical eyelid conditioning has been used to great advantage in demonstrating that the cerebellum helps to improve movements through experience, and in identifying the underlying mechanisms. Results from recent studies support the hypotheses that learning occurs in both the cerebellar nucleus and cortex, and that these sites make different contributions. Specifically, results indicate that the cerebellar cortex is responsible for temporally specific learning. A combination of experimental and computational studies has been important for arriving at these conclusions, which seem to be applicable to the broad range of movements to which the cerebellum contributes.