Genome sequencing reveals underdiagnosis of primary ciliary dyskinesia in bronchiectasis.

BACKGROUND: Bronchiectasis can result from infectious, genetic, immunological and allergic causes. 60-80% of cases are idiopathic, but a well-recognised genetic cause is the motile ciliopathy, primary ciliary dyskinesia (PCD). Diagnosis of PCD has management implications including addressing comorbidities, implementing genetic and fertility counselling and future access to PCD-specific treatments. Diagnostic testing can be complex; however, PCD genetic testing is moving rapidly from research into clinical diagnostics and would confirm the cause of bronchiectasis. METHODS: This observational study used genetic data from severe bronchiectasis patients recruited to the UK 100,000 Genomes Project and patients referred for gene panel testing within a tertiary respiratory hospital. Patients referred for genetic testing due to clinical suspicion of PCD were excluded from both analyses. Data were accessed from the British Thoracic Society audit, to investigate whether motile ciliopathies are underdiagnosed in people with bronchiectasis in the UK. RESULTS: Pathogenic or likely pathogenic variants were identified in motile ciliopathy genes in 17 (12%) out of 142 individuals by whole-genome sequencing. Similarly, in a single centre with access to pathological diagnostic facilities, 5-10% of patients received a PCD diagnosis by gene panel, often linked to normal/inconclusive nasal nitric oxide and cilia functional test results. In 4898 audited patients with bronchiectasis, <2% were tested for PCD and <1% received genetic testing. CONCLUSIONS: PCD is underdiagnosed as a cause of bronchiectasis. Increased uptake of genetic testing may help to identify bronchiectasis due to motile ciliopathies and ensure appropriate management.

Double layer SiO2-TiO2 sol-gel thin films on glass for antireflection, antifogging, and UV recoverable self-cleaning.

Double layer thin films, mechanically stable and adhering to glass, were produced through the sol-gel process, using tetraethyl orthosilicate and titanium butoxide as precursors. The refractive index of the titania and silica-titania composite layers were typically 2.1 and 1.7, and their physical thicknesses were approximately 65 nm and 81 nm, respectively, as determined by ellipsometry. These optical constants allowed attainment of quarter-wave optical thicknesses at the center of the visible spectrum (550 nm) as designed, with an increase of 3.4% in transmittance. The nanometric surface roughness, measured by optical profilometry, was effective to decrease light scattering and water contact angles to below 10°. As novelty in dip-coated sol-gel films, superhydrophilicity for self-cleaning, antifogging, and antireflection in the mid-visible spectrum were simultaneously attained with durability of 9 weeks in the dark. Further application of UV light allowed regeneration of contact angles for self-cleaning.

Spectrally extended, near-surface refractive index determination: atomic layer deposited TiO2 on Si.

The optical performance of very thin films has always been difficult to measure, and it has been a gray area due to interface and coating imperfections. Atomic layer deposition has produced pinhole-free, continuous, and extremely pure Titania films, with precise control at monolayer level, on native oxide/Si substrates, whose atomic force microscopy imaged nano-topology is approximately followed. For interface probing, the Abelès method is extended to the visible spectrum, and Cauchy dispersion curves are fitted to the refractive index experimental data, showing excellent compatibility with ellipsometric analysis. This is possible with decreasing optical thickness until ∼1/10 quarterwave is reached, which sets a limit for the usual optical film behavior.

Potential of coffee straw biochal as a substrate conditioner in seed lettuce and sorghum germination and vigority.

The use of residues from coffee production to obtain biochar is a sustainable approach, which aims to minimize the environmental impact of these materials. In this study, the effect of adding coffee straw biochar on the physiological quality of lettuce and sorghum seeds was investigated. Thus, the objective of this work was to study the effect of adding different concentrations of coffee biochar in the substrate composition on the physiological quality of lettuce (Lactuca sativa) and sorghum (Sorghum bicolor) seeds. The experimental design used was completely randomized, with five concentrations of biochar (0; 7.5; 15; 30 and 60%), conducted with four replications of 25 seeds. The use of biochar in the concentrations studied does not provide an increase in the average germination percentage and vigor of lettuce and sorghum seeds. The increase in the concentration of biochar caused less seed vigor, suggesting a toxic effect. For seed germination, there was no significant difference between lettuce and sorghum species, regardless of treatment. For the germination speed index, sorghum seeds have higher means, except for the treatment with the addition of 15% coffee straw biochar. Lettuce seeds have higher shoot length averages, except for treatment with 100% commercial substrate. The sorghum seeds have higher mean root length and dry mass than lettuce, regardless of the treatment.

Heavy metal tolerance in the psychrotolerant Cryptococcus sp. isolated from deep-sea sediments of the Central Indian Basin.

A deep-sea isolate of the psychrotolerant yeast Cryptococcus sp. (NIOCC#PY13) obtained from polymetallic nodule-bearing sediments of the Central Indian Basin was examined for its capacity to grow in the presence of various concentrations of the heavy metal salts i.e., ZnSO4 , CuSO4 , Pb(CH3 COO)2 and CdCl2 . It demonstrated considerable growth in the presence of 100 mg/l concentrations of the above-mentioned four heavy metal salts both at 30°C and 15°C. This strain tolerated comparatively higher levels of these four metal salts than other deep-sea and terrestrial yeast isolates belonging to Cryptococcus, Rhodotorula, Rhodosporidium and Sporidiobolus spp. Optimum pH for growth of this isolate was in the range of 6-8 in the presence of heavy metal salts at these two temperatures. Scanning electron microscopic (SEM) studies exhibited altered cell surface morphology of the cells under the influence of heavy metals compared to that with control. The adsorption of heavy metals to the cells was demonstrated by FTIR and EDAX analysis. As evidenced by atomic absorption spectrophotometric (AAS) analysis, about 30-90% of the heavy metals were removed from the culture supernatant after 4 days of growth at 30°C. This deep-sea yeast isolate appears to be a potential candidate for bioremediation of metal-contaminated sites. Moreover, its metal tolerance properties provide a significant insight into its ecological role and adaptations to growth in such extreme conditions.

Focal adhesion kinase governs cardiac concentric hypertrophic growth by activating the AKT and mTOR pathways.

The heart responds to sustained overload by hypertrophic growth in which the myocytes distinctly thicken or elongate on increases in systolic or diastolic stress. Though potentially adaptive, hypertrophy itself may predispose to cardiac dysfunction in pathological settings. The mechanisms underlying the diverse morphology and outcomes of hypertrophy are uncertain. Here we used a focal adhesion kinase (FAK) cardiac-specific transgenic mice model (FAK-Tg) to explore the function of this non-receptor tyrosine kinase on the regulation of myocyte growth. FAK-Tg mice displayed a phenocopy of concentric cardiac hypertrophy, reflecting the relative thickening of the individual myocytes. Moreover, FAK-Tg mice showed structural, functional and molecular features of a compensated hypertrophic growth, and preserved responses to chronic pressure overload. Mechanistically, FAK overexpression resulted in enhanced myocardial FAK activity, which was proven by treatment with a selective FAK inhibitor to be required for the cardiac hypertrophy in this model. Our results indicate that upregulation of FAK does not affect the activity of Src/ERK1/2 pathway, but stimulated signaling by a cascade that encompasses PI3K, AKT, mTOR, S6K and rpS6. Moreover, inhibition of the mTOR complex by rapamycin extinguished the cardiac hypertrophy of the transgenic FAK mice. These findings uncover a unique role for FAK in regulating the signaling mechanisms that governs the selective myocyte growth in width, likely controlling the activity of PI3K/AKT/mTOR pathway, and suggest that FAK activation could be important for the adaptive response to increases in cardiac afterload. This article is part of a Special Issue entitled "Local Signaling in Myocytes".

Characterization of films of weak polyelectrolytes incorporated with poly(vinyl-pyrrolidone)-stabilized gold nanoparticles.

The incorporation process of gold nanoparticles (Au NPs) into self-assembled films was obtained using the layer-by-layer (LbL) technique where two weak polyelectrolytes, i.e., poly(allylamine hydrochloride) (PAH) as polycation and poly(acrylic acid) (PAA) as polyanion, were sequentially adsorbed. To evaluate this process, the films of PAH and PAA were immersed in gold solutions containing different sizes of Au NPs (6 nm and 10 nm) stabilized with poly(vinyl-pyrrolidone) (PVP) before and after treatment with a 0.1 mol L(-1) solution of hydrochloric acid. The systems were analyzed using a multi-method approach involving UV-Visible spectroscopy (UV-Vis), X-ray diffraction (XRD), atomic force (AFM) and transmission electron microscopy (TEM). The results showed that the self-assembled polyelectrolyte films that were not treated by acid showed greater incorporation and better distribution of Au NPs.

Amino hydroxyapatite/chitosan hybrids reticulated with glutaraldehyde at different pH values and their use for diclofenac removal.

Diclofenac sodium (DS) is an emergent pollutant, and among the methods investigated for its removal, adsorption is the most widely utilized technique. Hydroxyapatite and chitosan are biomaterials often used for adsorption. However, both biomaterials are limited due to their low chemical stability in an acidic medium; furthermore, pure hydroxyapatite interacts poorly with diclofenac. In this work, hydroxyapatite was organofunctionalized with 3-aminopropyltrimethoxysilane and further used to obtain amino hydroxyapatite /chitosan hybrids by crosslinking with glutaraldehyde at pH 3, 4, 5, and 6. X-ray diffraction patterns indicated the preservation of the hydroxyapatite phase under all pH conditions. Based on the control reaction of the amino hydroxyapatite with glutaraldehyde and its further reduction in sodium borohydride, the formation of CN moieties was highlighted as the main interaction mechanism between the aldehyde and amino groups. Therefore, crosslinking with glutaraldehyde was evaluated by infrared, Raman spectroscopy, and 13C NMR techniques; the results suggested contributions of imine formation and hydrogen bonding. The hybrid obtained at pH 3 exhibited an enhanced adsorption capacity of 125 mg g-1 at 15 min. The synergy between amino hydroxyapatite and chitosan crosslinked by glutaraldehyde was demonstrated.

Study pharmacologic of the GABAergic and glutamatergic drugs on seizures and status epilepticus induced by pilocarpine in adult Wistar rats.

This work was designed to study the influence of drugs during seizures and status epilepticus (SE) induced by pilocarpine and mortality in adult rats. Glutamate (10 and 20 mg/kg), N-methyl-d-aspartate (NMDA, 5 and 10 mg/kg), ketamine (1.5 and 2.0 mg/kg), gabapentin (200 and 250 mg/kg), phenobarbital (50 and 100 mg/kg) and vigabatrin (250 and 500 mg/kg) were administered intraperitoneally, 30 min prior to pilocarpine (400 mg/kg, i.p.). The animals were observed (24 h) to determine: number of peripheral cholinergic signs, tremors, stereotyped movements, seizures, SE, latency to first seizure and number of deaths after pilocarpine treatment. NMDA and glutamate had pro-convulsive effects in both doses tested. Smaller and higher doses of these drugs no protected and increased pilocarpine-induced seizures and/or mortality. Gabapentin, vigabatrin, phenobarbital and ketamine protected against seizures and increased the latency to first seizure. Thus, these results suggest that caution should be taken in the selection of pharmacotherapy and dosages for patients with seizures and SE because of the possibility of facility the convulsive process toxicity, SE and the mortality of adult animals in this seizures model that is similar temporal lobo epilepsy in humans.

A novel procedure for the identification of chaos in complex biological systems.

We demonstrate the presence of chaos in stochastic simulations that are widely used to study biodiversity in nature. The investigation deals with a set of three distinct species that evolve according to the standard rules of mobility, reproduction and predation, with predation following the cyclic rules of the popular rock, paper and scissors game. The study uncovers the possibility to distinguish between time evolutions that start from slightly different initial states, guided by the Hamming distance which heuristically unveils the chaotic behavior. The finding opens up a quantitative approach that relates the correlation length to the average density of maxima of a typical species, and an ensemble of stochastic simulations is implemented to support the procedure. The main result of the work shows how a single and simple experimental realization that counts the density of maxima associated with the chaotic evolution of the species serves to infer its correlation length. We use the result to investigate others distinct complex systems, one dealing with a set of differential equations that can be used to model a diversity of natural and artificial chaotic systems, and another one, focusing on the ocean water level.

Effects of physical activity on weight reduction in obese middle-aged women.

Increased physical activity consisting of jog-walking 2.5 miles and 1 hr of calisthenics/week was the primary focus of a 17-week weight reduction program in 22 obese (X = 40% body fat) women ages 30 to 52, many of whom had failed at previous attempts to lose weight by dieting alone. Regular exercise was also increased substantially on an individual basis. Caloric restriction was self-determined and was generally moderate, accounting for about 60% of the total mean energy deficit. Initial and final evaluations included body composition by hydrostatic weighing, progressive multistage exercise testing, and plasma lipid and lipoprotein analyses. A mean relative body fat reduction of 5% (P less than or equal to 0.001) was achieved by a drop in fat body weight (X = 5.4 kg; P less than or equal to 0.001) which closely paralleled that of total body weight (X = 4.2 kg; P less than or equal to 0.001). Mean heart rates and systolic blood pressures at identical submaximal exercise intensities were significantly lower at the time of reevaluation. Mean plasma triglyceride and total cholesterol concentrations did not change significantly (P greater than 0.05). However, the high-density lipoprotein cholesterol/low-density lipoprotein cholesterol ratio increased significantly (P less than or equal to 0.05). Increased physical activity combined with moderate dieting is a feasible approach to weight reduction in middle-aged women.

The HLA-class II-associated chondroitin sulfate proteoglycan expressed by class II positive T and monocyte-like cell lines is larger than that expressed by EBV-transformed B-lymphoblastoid cell lines.

The human class II-associated chondroitin sulfate proteoglycan (CSPG) was originally detected as an approximately 40-70 kd species from normal human tonsil cells and Epstein-Barr virus (EBV) transformed B-lymphoblastoid cell lines. The identification of the invariant chain as the core protein of the CSPG allowed us to directly assay for the CSPG on both class II positive and negative immunocompetent cells of other lineages. Our results indicate that the CSPG is present on the class II-positive monocyte-like cell line U937 and T-cell line HUT-102, but not on the class-II negative T-cell line CCRF/CEM. No class II positive cells were found that did not also express the CSPG. The expression of the CSPG on U937 cells is increased after stimulation with gamma-interferon and PMA, paralleling the previously described increase in class II and invariant chain expression. In addition, the CSPGs from U937, HUT-102, and Raji, all cell lines derived from human malignancies, migrate as an approximately 55-90 kd species, larger than the CSPG previously characterized. However, the core proteins of the CSPG from all cells studied appear as two bands of 38 and 28 kd, indicating the size difference in the CSPG is attributable to differences in the glycosaminoglycan chains.

Phrenic nerve injury associated with venipuncture of the internal jugular vein.

Coincident with repeated, unsuccessful efforts to place a transvenous pacemaker through the right internal jugular vein, and 81-year-old man developed paralysis of the right hemidiaphragm. The patient had sudden shortness of breath and hypoxemia that rapidly subsided, although the loss of diaphragmatic function appears to be permanent.

Immunohistochemical study of the expression of MUC5AC and MUC6 in breast carcinomas and adjacent breast tissues.

AIM: To study the protein expression patterns of MUC5AC and MUC6 in normal and diseased breast tissues and to compare their expression with that of a mucin (MUC1) normally expressed in mammary tissues. METHODS: Formalin fixed, paraffin wax embedded tissue from 69 cases of invasive breast carcinoma and surrounding breast tissue was studied immunohistochemically with monoclonal antibodies against MUC1 (SM3), MUCSAC (CLH2), and MUC6 (CLH6), using the avidin-biotin-peroxidase method. RESULTS: MUC5AC was detected in five of 68 cases of invasive carcinoma including one of three cases of pure colloid carcinoma. MUCSAC expression in the adjacent normal breast epithelium was present in one of 29 cases and in one of two cases of ductal carcinoma in situ. None of 15 cases of ductal hyperplasia without atypia was positive for MUCSAC. MUC6 was present in 15 of 65 cases of invasive carcinoma, in four of 29 cases of normal adjacent epithelium, two of 15 cases of ductal hyperplasia without atypia, and one of two cases of ductal carcinoma in situ. MUC1 immunoreactivity detected by the SM3 antibody was present in 50 of the 67 cases of invasive carcinoma, but expression was also detected in benign epithelium. All invasive carcinomas expressing MUCSAC were positive for MUC1 and four were positive for MUC6. No significant association was found between the expression of these mucins and tumour size, histological grade, node status, oestrogen receptor status, p53 positivity, or c-ErbB-2 overexpression. CONCLUSIONS: This study documents the expression of two different mucins (MUCSAC and MUC6) not described as being expressed by normal breast tissues in a minority of breast carcinomas, as well as in normal and hyperplastic epithelium. Although the role of mucins in malignant transformation and the progression of breast cancer is not well understood, in some cases, there is probably an upregulation of several genes that encode distinct mucin proteins.

[Susacs syndrome: case report]. / Síndrome de Susac: relato de caso.

We present the case of a 34-year-old woman with clinical picture suggestive of Susacs syndrome, or retinocochlear vasculopathy. This syndrome, which was described for the first time in 1979, is characterized by a clinical triad of encephalopathy, neurosensorial deafness and visual deficit. Its pathogenesis and treatment are still disputed. We have called special attention to differential diagnosis, since this entity has not yet been described in Brazil, and is probably underdiagnosed.

Detection of metabolic disorders in high-risk patients. A pilot study in Salvador, Bahia.

The purpose of this pilot-study was to evaluate the applicability of a screening protocol for the detection of inborn errors of metabolism (IEM) in high-risk patients. The protocol was applied in 65 patients referred to the Medical Genetics Laboratory of the University Hospital Professor Edgard Santos due to the suspicion of an IEM. Eight of these patients (12.3%) displayed an abnormal result in the screening protocol. These patients, along with 22 who displayed normal results in the screening protocol but who presented clinical symptoms or signs suggestive of an IEM not detectable by the tests applied, were selected for a further diagnostic investigation. In 5 of these 30 patients (7.7% of the total sample) it was possible to establish the diagnosis of an specific IEM. The results indicate that the designed screening protocol was successfully applied, allowing the detection of affected patients in a frequency comparable to that observed in larger studies performed elsewhere. The continuation of this study and the enlargement of the sample will help to delineate the profile of IEM in northeast of Brazil and will allow the identification of a significative number of patients and families, who could benefit from the therapeutic and preventive measures available for these diseases.

Comparison of screening methods for conductive hearing loss identification in children: low-cost proposal.

OBJECTIVE: To evaluate the effectiveness of two screening methods (imitanciometry screening and questionnaire) to identify children at risk for conductive hearing loss, comparing this data with complete audiologic evaluation. METHODS: Of 507 children aged between three and six, 111 completed all procedures. The observational methods used were: imitanciometry screening, a questionnaire to identify risk factors for hearing loss and complete audiologic evaluation. Results obtained in the first two instruments were compared with results from complete audiologic evaluation (gold standard). From these comparisons, sensitivity and specificity, accuracy, positive and negative predictive values, and odds ratio were determined for the two screening methods and for the combination of both methods. RESULTS: The two methods applied in series (questionnaire and after imitanciometry screening) showed a greater odds ratio and better correlation between sensitivity and proportion of false-positives (ROC curve). CONCLUSION: Combining the two tests in series improved screening accuracy. This combination was the best tool for identifying children at risk for conductive hearing loss.

Efeito da endogamia sobre a sobrevivência e a longevidade no escargot da espécie helix aspersa / Effect of inbreeding on viability and longevity of edible snail helix aspersa

Estudou-se o efeito da endogamia sobre as características sobrevivência (intervalos de 0-30; 30-60; 60-90 e 90-120 dias) e longevidade do escargot da espécie Helix aspersa em três gerações consecutivas de acasalamentos entre irmãos perfeitos. Os animais foram criados em laboratório com temperatura entre 20 e 25ºC e umidade relativa entre 70 a 90 por cento. O menor percentual de sobrevivência ocorreu no primeiro intervalo, mas, com o aumento da endogamia, observou-se diminuição na sobrevivência nos quatro intervalos testados. A longevidade média foi de 314 e 306 dias para as gerações um e três, respectivamente, não tendo sido observados animais em hibernação. O estudo de simulação para o caráter sobrevivência, em função do coeficiente de endogamia, indicou que se deve iniciar uma criação com, pelo menos, 100 reprodutores não aparentados.

The effect of inbreeding on longevity and viability (intervals of 0-30, 30-60, 60-90, and 90-120 days of birth) in three consecutive generations of full sibs of escargot Helix aspersa raised under laboratory conditions (20-25ºC temperature; 70-90 percent relative humidity) was studied. Inbreeding effects decreased the viability for all the studied intervals. The longevity means were 314 and 306 days, respectively to the first and third generations. Hibernating animals were not observed. Population size of at least 100 non-related mating escargots replaced after 10 generations to avoid inbreeding effect is recommended.