MSTO1 mutations cause mtDNA depletion, manifesting as muscular dystrophy with cerebellar involvement.

MSTO1 encodes a cytosolic mitochondrial fusion protein, misato homolog 1 or MSTO1. While the full genotype-phenotype spectrum remains to be explored, pathogenic variants in MSTO1 have recently been reported in a small number of patients presenting with a phenotype of cerebellar ataxia, congenital muscle involvement with histologic findings ranging from myopathic to dystrophic and pigmentary retinopathy. The proposed underlying pathogenic mechanism of MSTO1-related disease is suggestive of impaired mitochondrial fusion secondary to a loss of function of MSTO1. Disorders of mitochondrial fusion and fission have been shown to also lead to mitochondrial DNA (mtDNA) depletion, linking them to the mtDNA depletion syndromes, a clinically and genetically diverse class of mitochondrial diseases characterized by a reduction of cellular mtDNA content. However, the consequences of pathogenic variants in MSTO1 on mtDNA maintenance remain poorly understood. We present extensive phenotypic and genetic data from 12 independent families, including 15 new patients harbouring a broad array of bi-allelic MSTO1 pathogenic variants, and we provide functional characterization from seven MSTO1-related disease patient fibroblasts. Bi-allelic loss-of-function variants in MSTO1 manifest clinically with a remarkably consistent phenotype of childhood-onset muscular dystrophy, corticospinal tract dysfunction and early-onset non-progressive cerebellar atrophy. MSTO1 protein was not detectable in the cultured fibroblasts of all seven patients evaluated, suggesting that pathogenic variants result in a loss of protein expression and/or affect protein stability. Consistent with impaired mitochondrial fusion, mitochondrial networks in fibroblasts were found to be fragmented. Furthermore, all fibroblasts were found to have depletion of mtDNA ranging from 30 to 70% along with alterations to mtDNA nucleoids. Our data corroborate the role of MSTO1 as a mitochondrial fusion protein and highlight a previously unrecognized link to mtDNA regulation. As impaired mitochondrial fusion is a recognized cause of mtDNA depletion syndromes, this novel link to mtDNA depletion in patient fibroblasts suggests that MSTO1-deficiency should also be considered a mtDNA depletion syndrome. Thus, we provide mechanistic insight into the disease pathogenesis associated with MSTO1 mutations and further define the clinical spectrum and the natural history of MSTO1-related disease.

Smoking behaviours of current cancer patients in Canada.

Evidence shows that continued smoking by cancer patients leads to adverse treatment outcomes and affects survival. Smoking diminishes treatment effectiveness, exacerbates side effects, and increases the risk of developing additional complications. Patients who continue to smoke also have a higher risk of developing a second primary cancer or experiencing a cancer recurrence, both of which ultimately contribute to poorer quality of life and poorer survival. Here, we present a snapshot of smoking behaviours of current cancer patients compared with the non-cancer patient population in Canada. Minimal differences in smoking behaviours were noted between current cancer patients and the rest of the population. Based on 2011-2014 data from the Canadian Community Health Survey, 1 in 5 current cancer patients (20.1%) reported daily or occasional smoking. That estimate is comparable to findings in the surveyed non-cancer patient population, of whom 19.3% reported smoking daily or occasionally. Slightly more male cancer patients than female cancer patients identified as current smokers. A similar distribution was observed in the non-cancer patient population. There is an urgent need across Canada to better support cancer patients in quitting smoking. As a result, the quality of patient care will improve, as will cancer treatment and survival outcomes, and quality of life for these patients.

Coffin-Siris syndrome is a SWI/SNF complex disorder.

Coffin-Siris syndrome (CSS) is a congenital disorder characterized by intellectual disability, growth deficiency, microcephaly, coarse facial features, and hypoplastic or absent fifth fingernails and/or toenails. We previously reported that five genes are mutated in CSS, all of which encode subunits of the switch/sucrose non-fermenting (SWI/SNF) ATP-dependent chromatin-remodeling complex: SMARCB1, SMARCA4, SMARCE1, ARID1A, and ARID1B. In this study, we examined 49 newly recruited CSS-suspected patients, and re-examined three patients who did not show any mutations (using high-resolution melting analysis) in the previous study, by whole-exome sequencing or targeted resequencing. We found that SMARCB1, SMARCA4, or ARID1B were mutated in 20 patients. By examining available parental samples, we ascertained that 17 occurred de novo. All mutations in SMARCB1 and SMARCA4 were non-truncating (missense or in-frame deletion) whereas those in ARID1B were all truncating (nonsense or frameshift deletion/insertion) in this study as in our previous study. Our data further support that CSS is a SWI/SNF complex disorder.

Genotype and clinical care correlations in craniosynostosis: findings from a cohort of 630 Australian and New Zealand patients.

Craniosynostosis is one of the most common craniofacial disorders encountered in clinical genetics practice, with an overall incidence of 1 in 2,500. Between 30% and 70% of syndromic craniosynostoses are caused by mutations in hotspots in the fibroblast growth factor receptor (FGFR) genes or in the TWIST1 gene with the difference in detection rates likely to be related to different study populations within craniofacial centers. Here we present results from molecular testing of an Australia and New Zealand cohort of 630 individuals with a diagnosis of craniosynostosis. Data were obtained by Sanger sequencing of FGFR1, FGFR2, and FGFR3 hotspot exons and the TWIST1 gene, as well as copy number detection of TWIST1. Of the 630 probands, there were 231 who had one of 80 distinct mutations (36%). Among the 80 mutations, 17 novel sequence variants were detected in three of the four genes screened. In addition to the proband cohort there were 96 individuals who underwent predictive or prenatal testing as part of family studies. Dysmorphic features consistent with the known FGFR1-3/TWIST1-associated syndromes were predictive for mutation detection. We also show a statistically significant association between splice site mutations in FGFR2 and a clinical diagnosis of Pfeiffer syndrome, more severe clinical phenotypes associated with FGFR2 exon 10 versus exon 8 mutations, and more frequent surgical procedures in the presence of a pathogenic mutation. Targeting gene hot spot areas for mutation analysis is a useful strategy to maximize the success of molecular diagnosis for individuals with craniosynostosis.

Russell-Silver syndrome due to paternal H19/IGF2 hypomethylation in a patient conceived using intracytoplasmic sperm injection.

Epigenetic alterations at several maternal loci have been associated with imprinting disorders in children conceived using assisted reproductive technologies. To date, epimutations at paternal loci have been observed in the spermatozoa of infertile men, but there is little evidence of paternal epimutations in babies conceived using assisted reproductive treatment. This is a report of a female infant with classic Russell-Silver Syndrome (RSS) who was conceived using intracytoplasmic injection of spermatozoa obtained from testicular aspiration. Methylation studies revealed hypomethylation of the paternally derived H19/IGF2 locus. As far as is known, this is the second assisted reproduction treatment-conceived patient with classic RSS and this epigenotype. This case provides further evidence that epimutations affecting paternal alleles might be associated with assisted reproductive treatment.

Mowat-Wilson syndrome.

MWS is a multiple congenital anomaly syndrome, first clinically delineated by Mowat et al in 1998. Over 45 cases have now been reported. All patients have typical dysmorphic features in association with severe intellectual disability, and nearly all have microcephaly and seizures. Congenital anomalies, including Hirschsprung disease (HSCR), congenital heart disease, hypospadias, genitourinary anomalies, agenesis of the corpus callosum, and short stature are common. The syndrome is the result of heterozygous deletions or truncating mutations of the ZFHX1B (SIP1) gene on chromosome 2q22.

Aortic blood velocity measurement in healthy adults using a simple ultrasound technique.

The normal range of aortic blood velocity has been established in 140 healthy adults using the non-invasive technique of transcutaneous aortovelography (TAV). Velocity is independent of sex, body surface area and blood pressure but declines progressively with age so that at age 70 the mean value of peak velocity is 55% of that at age 20. Integration of the area under the velocity time curve gives an indication of stroke volume and cardiac output. These indices also decrease with age as does acceleration which may reflect left ventricular function. Measurement of aortic blood velocity and its derivatives is a safe, simple and physiologically meaningful way of assessing cardiac output and function.

Deficiency of the syntrophins and alpha-dystrobrevin in patients with inherited myopathy.

The syntrophins and dystrobrevins are members of the dystrophin-associated protein complex, and are thought to function as modular adaptors for signalling proteins recruited to the sarcolemmal membrane. We have characterised the expression of the syntrophins (alpha-, beta1-, and beta2-) and alpha-dystrobrevin by immunohistochemistry in normal human muscle and in biopsies from 162 patients with myopathies of unknown aetiology (with normal staining for dystrophin and other dystrophin-associated proteins). Unlike mice, beta2-syntrophin is expressed at the sarcolemma in post-natal human skeletal muscle. Deficiency of alpha-dystrobrevin +/- beta2-syntrophin was present in 16/162 (10%) patients, compared to age-matched controls. All patients presented with congenital-onset hypotonia and weakness, although there was variability in clinical severity. Two major clinical patterns emerged: patients with deficiency of beta2-syntrophin and alpha-dystrobrevin presented with severe congenital weakness and died in the first year of life, and two patients with deficiency of alpha-dystrobrevin had congenital muscular dystrophy with complete external ophthalmoplegia. We have sequenced the coding regions of alpha-dystrobrevin and beta2-syntrophin in these patients, and identified a new isoform of dystrobrevin, but have not identified any mutations. This suggests that disease causing mutations occur outside the coding region of these genes, in gene(s) encoding other components of the syntrophin-dystrobrevin subcomplex, or in gene(s) responsible for their post-translational modification and normal localisation.

Duplication within chromosome 5q characterized by fluorescence in situ hybridization.

We present a 16-month-old boy with developmental delay, minor anomalies, small penis, and lymphedema of the upper limbs. Routine cytogenetic analysis suspected a duplication of 5q. Fluorescent in situ hybridization (FISH) with a cosmid probe (MCC at the 5q22 APC region) showed tandemly duplicated fluorescent signals on one of chromosomes 5, whereas FISH with three YAC probes (TYAC12 at 5q35, HTY3182 at 5q34, and TYAC139 at 5q31) did not give duplicated signals. These findings indicate a duplication of 5q22 band in one chromosome 5. The boy we describe here is the first case of a pure partial duplication of 5q to be proven by FISH techniques. A review of previously reported cases of putative partial 5q duplication showed no consistent phenotype.

Genes on the X chromosome are important in undiagnosed mental retardation.

The clinical genetic diagnosis was reviewed in 429 subjects with intellectual disability in the Australian Child and Adolescent Development (ACAD) study of behavioural problems. With minor differences, the overall "general distribution by causation" was similar to that to that found by the Consensus Conference of the American College of Medical Genetics in 1995. There was a significant male excess in the whole series which was shown to reside in those with "autism," those with undiagnosed nonsyndromic mental retardation (NSMR) and those with X-linked monogenic disorders. It is argued that a substantial proportion of undiagnosed NSMR is caused by genes on the X chromosome. Some of the practical problems of assigning individuals to diagnostic groups are discussed.

Effects of chromium on health status, blood neutrophil phagocytosis and in vitro lymphocyte blastogenesis of dairy cows.

The objective of this study was to determine the effects of supplemental dietary chromium (Cr) on health status and mastitis-related parameters, as well as neutrophil phagocytic activity of dairy cows during late pregnancy and early lactation. In addition, possible interactions and involvements of Cr with insulin, cortisol, somatotropin (rBST) and insulin-like growth factor-1 (IGF-1) were directly investigated in vitro based on blastogenic responses using peripheral blood lymphocytes (PBL) of eight cows fed the control diet. Forty pregnant Holsteins, 18 primiparous and 22 multiparous, at week (wk) 6 before the expected calving dates were randomly assigned to treatments: control and supplemental chelated Cr (0.5 ppm) in the diet. All cows were managed in a normal production operation and health status was assessed by recording the incidence of health problems during the experimental period of wks 6 before and 16 after calving (-6 to 16 wks). Mastitis-related parameters included somatic cell counts (SCC), bacterial colony counts of milk samples from each mammary gland quarter of all cows during wk 1 to wk 8 postpartum. Peripheral blood neutrophil phagocytic function of eight cows fed either control or supplemental Cr diet was determined by the ability to take up uniform fluorescent beads measured by flow cytometry. Supplemental Cr had no effect (P > 0.10) on health status of cows during late pregnancy and early lactation, or on SCC and bacterial colonies of quarter milk samples from early lactation (wks 1 to 8). Supplemental Cr also did not affect neutrophil phagocytic function of cows from 6 wks prepartum to 6 wks postpartum. However, in the in vitro study of PBL (from control animals, not fed Cr diet) blastogenesis with addition of insulin or cortisol at two levels (0.05 and 0.5 ng ml-1), insulin and cortisol enhanced or had no effect on PBL proliferations with or without concanavalin A (con A) simulation. However, further Cr addition in the culture medium containing supplemental insulin or cortisol, particularly CrCl3, additively increased (P < 0.05) BPL blastogenic activities with or without con a stimulation. Conversely, addition of rBST or IFG-1 (0.5 and 5.0 ng ml-1) in the culture medium enhanced PBL proliferation, but addition of Cr gave no additional effect. These results indicated that supplemental Cr had no beneficial effect on health status, mastitis-related parameters or neutrophil phagocytic activity of dairy cows. However, in vitro study confirmed and extended our previous observations that Cr has an effect on lymphocyte proliferation and this may associate with insulin or cortisol actions.

Immunomodulatory activity of blood serum from chromium-supplemented periparturient dairy cows.

Our previous research showed enhanced immune responses, including mitogen-induced blastogenesis of peripheral blood mononuclear cells from feedlot calves and periparturient dairy cows supplemented with dietary chromium (Cr). The objective of the present study were to test whether blood sera from Cr-supplemented periparturient cows contained immunomodulatory activity for mitogen-stimulated peripheral blood mononuclear cells and, if so, to determine if this activity was explicable by differences in blood profiles of some glucose-regulating hormones (insulin, cortisol, growth hormone, insulin-like growth factor I, and tumor necrosis factor-alpha) between Cr-supplemented and unsupplemented (control) animals. Blood sera from ten unsupplemented cows and nine Cr-supplemented cows (0.5 ppm day-1) were collected weekly from 2 weeks before to 6 weeks after parturition, and were used to supplement (1, 10, and 20% vol/vol) culture medium supporting concanavalin A (Con A) stimulated mononuclear cells enriched from blood of four nulliparous donor cows. Hormone concentrations were determined using radioimmunoassays. Con A-induced blastogenesis was enhanced when 1, 10, and 20% sera from Cr-supplemented cows was added to the mononuclear cell cultures, and this was particularly evident around parturition. Conversely, peripartum sera from unsupplemented cows depressed Con A-induced blastogenesis. Except for a marginal rise in blood cortisol 2-4 weeks after parturition, no significant effects of Cr supplementation on other hormones (insulin, growth hormone, insulin-like growth factor I, tumor necrosis factor-alpha) were observed. These observations suggest that factors in peripheral blood serum from Cr-supplemented cows, other than absolute concentrations of the glucose-regulating hormones studied, modulate Con A-induced blastogenesis of mononuclear leukocytes.

Effect of supplemental chromium on whole-body kinetics of glucose, lactate, and propionate in rams fed a high grain diet.

The isotope dilution technique of [6-3H]glucose, [U-14C]lactate and [l-14C]propionate was used to evaluate the effect of dietary chromium (Cr) supplementation on whole-body kinetics of glucose, lactate, and propionate in rams. Rams were fed a high grain diet at 2% of body weight with or without 0.5 ppm of supplemental Cr from chelated Cr for the initial 14 days, and then intake was increased to 2.5% at body weight for the last 9 days. Weight gain was enhanced (P < 0.01) with Cr supplementation. Plasma concentrations of glucose, lactate, and propionate were not influenced by Cr supplementation. Turnover rates of glucose and lactate, and their interconversion were also not influenced. Propionate turnover rate tended to increase (P = 0.11) and the conversion of propionate to glucose increased (P < 0.05) with Cr supplementation, leading the increased proportional contribution of propionate to glucose turnover rate (P < 0.05). Chromium supplementation may influence the contribution of each glucogenic substrate for glucose production in rams fed a high grain diet.

Tobacco access to youth: beliefs and attitudes of retailers.

Results of a telephone survey provide insights into the knowledge, attitudes and beliefs of tobacco merchants from two local health units. More than 90% of the retailers said they should not be able to sell cigarettes to minors. They are aware of laws prohibiting such sales but are sceptical about the impact on young people. The majority report being motivated to help protect the health of youth, however, they advise that legislation provides the main reason for not selling cigarettes to minors. Other responses and behaviours of the merchants help provide a profile of an important group that is being asked to stop selling tobacco to young people. The authors classify the retailers into three groups according to the potential influence on their behaviour of messages about health and threats of enforcement. One of the health units had implemented a local intervention, therefore we also compare responses between the two health units. This type of information can be used when selecting strategies to strengthen health policies. Such policies and strategies should include input and feedback from retailers of tobacco.

Changes among retailers selling cigarettes to minors.

This study analyzes changes over a three-year period among Ontario retailers selling cigarettes to minors. Under supervision, 13 and 14-year-old minors were sent into stores to attempt to buy cigarettes. These minor-purchase-events (MPEs) were carried out in a local health unit that had implemented a community-based intervention and in an adjoining comparison health unit. After the local program we observed a large reduction (from 46% to 6%) in merchants willing to sell tobacco to minors. In the neighbouring health unit, a high rate of selling continued until a federal program using a similar intervention was implemented, after which a large reduction (from 47% to 2%) was observed. This magnitude of change has been unprecedented, except when active enforcement was implemented by police officers. Thus, from a public health perspective, it is important to understand what is influencing the store operators.

Effect of level of supplemental chromium on performance, serum constituents, and immune status of stressed feeder calves.

The effects of level of supplemental Cr from high-Cr yeast on performance, blood chemistry profile, morbidity, and immune status were investigated using 84 Charolais-crossed steer calves in a completely randomized design. Calves of 236-kg average weight, after transportation from Saskatchewan to Ontario, were randomly assigned to four treatments; 0, .2, .5, and 1 ppm of supplemental Cr, incorporated into a corn-silage diet. Blood was collected via jugular venipuncture at d 0, 7, 14, 21, and 28 and analyzed for metabolites, minerals, immunoglobulins, hematocrit (Hct), and leucocyte counts. Hemagglutinating antibody titers to human red blood cells (HRBC) were quantified after immunizations on d 0 and 14. Contact sensitivity after sensitization and challenge with dinitrochlorobenzene was also measured. A 27% increase (P < .05) in ADG was observed at d 30 for calves that were fed .2 and 1 ppm of supplemental Cr. Dry matter intake also increased (P < .05) for the .2- and 1-ppm Cr treatments. A linear decrease (P < .05) in serum cortisol with increasing Cr level was observed at d 28. Chromium supplementation decreased (P < .05) morbidity, as well as rectal temperatures at d 2 and 5. Peak primary antibody titers to HRBC (P < .05) and immunoglobulin G1 concentrations (P = .06) at d 14 were higher for steers that received the Cr supplementation. However, Cr treatment had no effect on expression of contact sensitivity. Chromium supplementation increased (P < .05) Hct on d 14 and 21 and serum Ca and Mg on d 7.(ABSTRACT TRUNCATED AT 250 WORDS)

Supplemental chromium for stressed and growing feeder calves.

The effects of supplemental chromium (Cr) from high-Cr yeast were investigated with steer calves fed corn silage diets. One hundred eight Charolais-crossed calves, weighing 245 kg after marketing and transport, were allotted to one of four treatments during the initial 28-d stress period: control, .4 ppm of Cr in the diet, long-acting injectable oxytetracycline (LAOTC), and Cr + LAOTC. Those fed Cr received 4 mg of Cr/d for the first 3 d sprinkled onto a small amount of hay over the silage. Chromium without LAOTC increased (P less than .05) ADG by 30% (.61 vs .79 kg/d) and ADG/DMI by 27% (.123 vs .156). Oxytetracycline alone increased (P less than .05) ADG by 30% and DMI by 15%. Chromium had no effect on morbidity. However, LAOTC tended (P less than .14) to reduce morbidity (26.0 vs 14.0%) after its administration. After d 28, steers were processed. Two weeks later, they were rerandomized within Cr groups to urea-corn vs soybean meal supplementation of corn silage during a 70-d growing period. Level of Cr was reduced to .2 ppm. Jugular blood was collected from eight steers on each treatment on two occasions. Chromium had no effect on ADG or ADG/DMI. However, Cr decreased (P less than .05) serum cortisol (75.0 vs 55.6 nmol/L). Furthermore, Cr increased (P less than .05) serum immunoglobulin M and total immunoglobulins in calves fed diets with soybean meal but had no effect in calves with urea-corn supplementation.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of supplemental chromium on immune responses of periparturient and early lactation dairy cows.

The objective of the present study was to determine the effects of supplemental dietary chromium on immune responses of dairy cows subjected to physical and metabolic stresses associated with late pregnancy, calving, early lactation, and peak milk yield. Nine periparturient dairy cows were supplemented with chelated Cr (.5 ppm/d) from 6 wk prepartum (wk -6) through 16 wk postpartum (wk 16), and 10 cows were unsupplemented controls. To assess humoral immune responses, all cows were immunized with ovalbumin (OVA; s.c.) and human erythrocytes (HRBC; i.v.) on wk -2 and 2, and sera from weekly blood samples were assayed for content of antigen-specific antibody. Cell-mediated immunity was assessed in vitro using antigen (OVA)- and mitogen-stimulated peripheral blood mononuclear cell (PBMC) blastogenesis of cells collected biweekly from wk -2 and 6. Supplemental Cr caused anti-OVA antibody responses (P < .01) and mitogen-stimulated blastogenic responses of PBMC (P = .05) to be elevated, was associated with lowered OVA-stimulated blastogenic responses of PBMC (P < .01), and had no overall effect on antibody responses to HRBC (P > .10) relative to responses of control cows. These results confirmed and extended our previous observations that supplemental Cr can alter specific immune responses of stressed cattle.

The influence of supplemental chromium and vaccines on the acute phase response of newly arrived feeder calves.

The acute phase response as indicated by serum haptoglobin and total haemolytic complement activity (CH50) was measured in 72 cross-bred steer calves purchased at sales in Ontario. During the 28 day (d) trial, 18 steers were randomly assigned to each of the following groups: 1) control; 2) vaccinated (Infectious Bovine Rhinotracheitis, Parainfluenza-3, Bovine Viral Diarrhea, Bovine Respiratory Synctial Virus vaccine plus Pasteurella haemolytica vaccine); 3) supplemental chelated Cr (0.14 mg/kg); and 4) Cr plus vaccines. Haptoglobin concentrations were low at arrival, increased (P < 0.05) on day 7, and returned to near initial levels (P > 0.05) by day 14. Supplemental Cr reduced (P < 0.05) haptoglobin on day 7 when morbidity was highest. Following antibiotic treatment for respiratory disease haptoglobin was lower (P < 0.05) than during morbidity; however, during morbidity, haptoglobin concentrations were not greater in sick calves (P > 0.05) than in healthy calves. Complement activity was lowest on day 7 (P < 0.05) and peaked on day 14 (P < 0.05). Complement activity tended to be lower on day 7 for vaccine, Cr, and Cr+ vaccine groups; however, the difference from controls was not significant (P > 0.10). Complement activity did not increase on day 14 (P > 0.05) with Cr supplementation as in other treatments. Morbid calves had lower (P < 0.05) CH50 activity than healthy calves on day 14. Following antibiotic treatment, the Cr-supplemented group had higher (P < 0.05) CH50 than during morbidity. In general, chromium supplementation reduced the acute phase response in newly arrived feeder calves.