Exploring the utility of whole‐exome sequencing as a diagnostic tool in a child with atypical episodic muscle weakness

The advent of whole‐exome next‐generation sequencing (WES) has been pivotal for the molecular characterization of Mendelian disease; however, the clinical applicability of WES has remained relatively unexplored. We describe our exploration of WES as a diagnostic tool in a 3½‐year old female patient with a 2‐year history of episodic muscle weakness and paroxysmal dystonia who presented following a previous extensive but unrevealing diagnostic work‐up. WES was performed on the proband and her two parents. Parental exome data was used to filter potential de novo genomic events in the proband and suspected variants were confirmed using di‐deoxy sequencing. WES revealed a de novo non‐synonymous mutation in exon 21 of the calcium channel gene CACNA1S that has been previously reported in a single patient as a rare cause of atypical hypokalemic periodic paralysis. This was unexpected, as the proband's original differential diagnosis had included hypokalemic periodic paralysis, but clinical and laboratory features were equivocal, and standard clinical molecular testing for hypokalemic periodic paralysis and related disorders was negative. This report highlights the potential diagnostic utility of WES in clinical practice, with implications for the approach to similar diagnostic dilemmas in the future.     

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Background: Addressing the long-term consequences of stroke is a top 10 research priority, and understanding the views of different stakeholders is essential in designing appropriate patient-centred multidisciplinary rehabilitation.

Aims: This study reports on the perspectives of 38 individuals who live with or work with aphasia: people with aphasia (PWA), family members (FM) and friends, and speech and language therapists (SLTs), about its consequences in the daily lives of PWA.

Method &; Resources: In total, 14 PWA, 14 FM and 10 SLTs participated in the study. All participants were interviewed about the consequences of stroke and aphasia. Small focus groups were used with some participants. In-depth semi-structured interviews were used with other participants. Interviews were carried out in groups or individually in order to accommodate severe aphasic impairments or respondents with fewer years of education. Interviews were video or audio-recorded. Verbal and non-verbal communication was transcribed and analysed qualitatively using content analysis. Data was coded and mapped onto the ICF framework.

Outcomes &; Results: All three groups reported consequences in Body Functions and Body Structures, primarily in Mental Functions (Specific Mental Functions and Global Mental Functions) and Neuromusculoskeletal and Movement-Related Functions. Within Specific Mental Functions, PWA emphasised Expression of Oral Language, whereas FM and SLTs emphasised Emotional Functions. Within Global Mental Functions, PWA and FM emphasised Energy and Drive Functions, whereas SLTs emphasised Temperament and Personality Functions. Some consequences were shared between PWA and SLTs (Sensory Functions and Pain, and Functions of the Digestive, Metabolic and Endocrine Systems), whilst others were specific to SLTs alone (Voice and Speech Functions, and Cardiovascular, Haematological, Immunological and Respiratory Systems). All three groups reported consequences in Activities and Participation in Major Life Areas; Community, Social and Civic Life; Interpersonal Interactions and Relationships; Communication; and Mobility. Some consequences were shared by PWA and FM (Domestic Life), and others by FM and SLT (Self Care). Consequences in General Tasks and Demands were only described by SLTs and in Learning and Applying Knowledge by PWA. Both PWA and FM reported a general Loss of Autonomy.

Conclusions: PWA, FM and SLTs reported a considerable range of stroke and aphasia consequences in the daily life of those who live with it. They were concerned to different degrees with language, physical and emotional changes, and impact on communication, mobility, self-care, relationships, leisure and work. Professionals’ practices in Portugal need to be changed in order to consider these findings.     

Renal thorium deposition associated with transitional cell carcinoma: radiologic demonstration in two patients

Abdominal radiography, excretory urography, retrograde pyelography, and computed tomography were performed in two patients who had undergone retrograde pyelography with thorium dioxide (Thorotrast) approximately 40 years ago. Both patients developed a transitional cell carcinoma due to suburothelial thorium deposition. Typical thorium densities were demonstrated at CT in the peripelvicalyceal area as well as in retroperitoneal lymph nodes. Elderly patients in whom radiographic examination reveals retained Thorotrast in the kidney should be followed up because of the high risk of renal carcinoma.     

Total parenteral nutrition (TPN) and gallbladder diseases in neonates. Sonographic assessment

During a 1-year period, a prospective ultrasound (US) study of the biliary tract was performed in 41 neonates receiving total parenteral nutrition (TPN). Gallbladder sludge appeared in 18 neonates (44%) after a mean period of 10 days of TPN. The occurrence of this abnormality seems related to three main factors: prematurity with immaturity of the enterohepatic circulation of bile acids, duration of TPN, and lack of enteral nutrition. In five neonates (12%), an evolution of sludge to "sludge balls" was observed. Two neonates (5%) went on to develop uncomplicated gallstones and a spontaneous resolution occurred in one of them 6 months after the examination. This finding suggests conservative management in such cases. However, since cholecystitis or biliary tract obstruction are classical complications in older children, a US survey of neonates receiving TPN is useful to follow the abnormal gallbladder content, and rapid introduction of enteral feeding is advisable.     

Influence of exposure time to saliva and antioxidant treatment on bond strength to enamel after tooth bleaching: an in situ study

Objectives

This study evaluated the influence of different exposure times to saliva in situ in comparison with an antioxidant treatment on composite resin bond strength to human enamel restored after tooth bleaching.

Material and Methods

Forty human teeth specimens measuring 5x5 mm were prepared and randomly allocated into 5 groups with 8 specimens each: Gct (control group, restored on unbleached enamel); Gbl (restored immediately after bleaching); Gsa (bleached, treated with 10% sodium ascorbate gel for 60 min and restored); G7d (bleached, exposed to saliva in situ for 7 days and restored); and G14d (bleached, exposed to saliva in situ for 14 days and restored). Restored samples were cut into 0.8 mm² sticks that were tested in microtensile. Specimens were microscopically analyzed and failure modes were classified as adhesive, cohesive, or mixed. Pretest and cohesive failures were not considered in the statistical analysis, which was performed with one-way ANOVA and Tukey''s post-hoc test (α=0.05), with the dental specimen considered as the experimental unit.

Results

Mean bond strength results found for Gbl in comparison with Gct indicated that bleaching significantly reduced enamel adhesiveness (P<0.01). However, no statistically significant differences were found between Gct, Gsa and G7d (P>0.05). Bond strength found for G14d was significantly higher than for Gsa (P<0.01). Fractures modes were predominantly of a mixed type.

Conclusions

Bonding strength to bleached enamel was immediately restored with the application of sodium ascorbate and exposure to human saliva in situ for at least 7 days. Best results were obtained with exposure to human saliva in situ for 14 days. Treatment with sodium ascorbate gel for 60 min may be recommended in cases patients cannot wait for at least 7 days for adhesive techniques to be performed.     

Gamma-carboxylated isoforms of recombinant human protein S with different biologic properties

Human protein S (HPS), a regulator of hemostasis, is a vitamin K- dependent plasma protein with potential clinical utility. We have obtained high-level expression of the cDNA for HPS in two mammalian cell lines. Both cell lines secreted single chain recombinant HPS (rHPS) in serum-free medium as determined by Western blot analysis. The ability of the rHPS from both cell lines to act as a cofactor for human protein C (HPC) was determined; the rHPS secreted from the human 293 cell line had an activity six times that of the rHPS from the AV12-664 Syrian hamster cell line. Furthermore, the relative specific cofactor activity of rHPS from the 293 cell line was actually 2.5-fold higher than that of single-chain human plasma-derived HPS. Essentially all of the rHPS secreted from the 293 cell line exhibited a calcium-dependent elution profile on anion exchange chromatography, whereas only 25% to 35% of the hamster cell-derived rHPS exhibited this profile. However, the calcium-eluted rHPS from the AV12 cell line had a high specific cofactor activity, equivalent to that of the 293-derived rHPS. A NaCl- elutable rHPS fraction (calcium nondependent) was isolated from the recombinant AV12-664 cell line, further purified, and found to have reduced activity, only 40% that of the calcium-dependent rHPS. The only observable difference in the calcium-dependent and nondependent rHPS molecules was in the content of gamma-carboxyglutamic acid (Gla); the calcium-dependent material contained approximately 10 mol Gla/mol protein whereas the calcium-nondependent material contained only approximately 8 mol Gla/mol of protein. In addition, the calcium- nondependent rHPS had reduced ability to interact with phospholipid vesicles as evidenced by an eightfold increase in the apparent kd. Our data demonstrate the isolation of rHPS with high specific activity, and show that a reduction in as few as two Gla residues dramatically decreases its functional cofactor activity for HPC, due to a reduction in ability to interact with the phospholipid bilayer.     

Ausência de associação entre os polimorfismos do gene interleucina-18 e artrite reumatoide

ObjectiveTo assess the association of the polymorphisms of the interleukin-18 (IL-18) gene with rheumatoid arthritis (RA) and with risk factors for cardiovascular diseases (CVD).MethodsThis sample comprised 97 patients with RA and 151 healthy controls. In the patients, risk factors for CVD were analyzed, such as cholesterol levels, arterial hypertension, smoking habit, C-reactive protein (CRP) level, and rheumatoid factor. DNA was extracted and the single nucleotide polymorphisms (SNP) at the ?607C/A and ?137G/C positions of the IL-18 gene were assessed in both groups. The Hardy-Weinberg equilibrium (HWE) was calculated and the odds ratio (OR) test performed, considering a 95% CI and P < 0.05.ResultsThe frequencies of the ?607A allele in patients with RA and in controls were 0,443 and 0.424, respectively, and of the ?137C allele, 0.304 and 0.291, respectively. The genotype frequencies were in HWE, except for controls in the ?137 locus (P = 0.006). Association of the polymorphisms of the IL-18 gene was found with neither RA nor risk factors for CVD, including cholesterol level and CRP (P > 0.05). In addition, more smokers were found among patients with RA as compared with controls (OR = 1.691; P = 0.088), and the CRP levels were slightly higher in patients who smoked than in patients who did not (OR = 2.673; P = 0.061).ConclusionsIn this sample of patients with RA in the South of Brazil, association of the polymorphisms of the IL-18 gene was observed with neither RA nor risk factors for CVD.     

Traumatic brain injury biomarkers in pediatric patients: a systematic review

Neurosurgical Review - Traumatic brain injury (TBI) is the main cause of pediatric trauma death and disability worldwide. Recent studies have sought to identify biomarkers of TBI for the purpose of...