Mitochondrial DNA damage analysis in bronchoalveolar lavage cells of preterm infants

In mechanically ventilated preterm infants, the combination of immaturity, volutrauma, oxidative stress, and inflammatory processes can lead to chronic lung injury. Mitochondrial DNA (mtDNA) is more susceptible to oxidative damage than nuclear DNA. We aimed to investigate the level of mtDNA damage (deletions, mutations and changes in copy number) in bronchoalveolar lavage (BAL) cells from 10 preterm infants (27-30 weeks). A first BAL (BAL1) was done within 24 h of endotracheal intubation and BAL2 was performed 30-103 h thereafter. Deletions were analyzed by long range PCR, point mutations by heteroduplex analysis of the D-loop region, and copy number changes by real-time PCR. Using these methods, no deletions were found in any of the BAL samples. When BAL1 and BAL2 samples were compared no new mutations were found. In contrast, a marked decrease in mtDNA copy number was observed in 5 patients. In conclusion, we found that exposure of preterm infants to short term mechanical ventilation did not lead to detrimental consequences for the mtDNA in the form of mutations or deletions

Is There a Need for Including Spiritual Care in Interdisciplinary Rehabilitation of Chronic Pain Patients?: Investigating an Innovative Strategy

OBJECTIVE: Chronic noncancer pain influences patient's quality of life and their ability to cope. Pain relieving medication and other specific treatments commonly integrated in biopsychosocial rehabilitation demonstrate modest benefits in pain relief and improved functioning of individuals. Spiritual care, covering the fourth dimension provides insight, inspires hope and purpose, and is thought to mediate mental and physical health for patients. This study explores the need for its inclusion in interdisciplinary pain rehabilitation and describes the requirements and test environment for evaluation. METHODS: Outcomes of spiritual care and interdisciplinary pain rehabilitation in follow-up studies of randomized controlled trials contained in systematic reviews were summarized. Pubmed, Cochrane, and PsycINFO were searched, citation tracking was applied, articles of follow-up studies therein were located. Literature was searched for insights pertaining to requirements for an assessment of including this fourth dimension. RESULTS: No systematic reviews for spiritual care were identified. Five systematic reviews of biopsychosocial rehabilitation containing 14 studies describing long-term outcomes were retrieved. The importance of coping in maintaining long-term outcomes was empirically illustrated. The required test environment is provided by a structured multidimensional care pathway separating spirituality from well-being and mental health, with measures of treatment outcome installed enabling a comparison with benchmarks. CONCLUSIONS: Active coping seems beneficial for maintaining positive long-term outcomes of interdisciplinary pain rehabilitation Spiritual care may be conducive to active coping. Further research is warranted to explore the additive value of this spiritual care in the context of a multidimensional care pathway

No Fabry Disease in Patients Presenting with Isolated Small Fiber Neuropathy

Screening for Fabry disease in patients with small fiber neuropathy has been suggested, especially since Fabry disease is potentially treatable. However, the diagnostic yield of testing for Fabry disease in isolated small fiber neuropathy patients has never been systematically investigated. Our aim is to determine the presence of Fabry disease in patients with small fiber neuropathy.Patients referred to our institute, who met the criteria for isolated small fiber neuropathy were tested for Fabry disease by measurement of alpha-Galactosidase A activity in blood, lysosomal globotriaosylsphingosine in urine and analysis on possible GLA gene mutations.725 patients diagnosed with small fiber neuropathy were screened for Fabry disease. No skin abnormalities were seen except for redness of the hands or feet in 30.9% of the patients. Alfa-Galactosidase A activity was tested in all 725 patients and showed diminished activity in eight patients. Lysosomal globotriaosylsphingosine was examined in 509 patients and was normal in all tested individuals. Screening of GLA for mutations was performed for 440 patients, including those with diminished α-Galactosidase A activity. Thirteen patients showed a GLA gene variant. One likely pathogenic variant was found in a female patient. The diagnosis Fabry disease could not be confirmed over time in this patient. Eventually none of the patients were diagnosed with Fabry disease.In patients with isolated small fiber neuropathy, and no other signs compatible with Fabry disease, the diagnostic yield of testing for Fabry disease is extremely low. Testing for Fabry disease should be considered only in cases with additional characteristics, such as childhood onset, cardiovascular disease, renal failure, or typical skin lesions

Diagnostic algorithm for confirming Fabry disease in SFN patients.

<p>(A) Diagnostic algorithm for men. (B) Diagnostic algorithm for women. α-Gal A: α-galactosidase A, FD: Fabry disease, Lyso-GB3: lysosomal globotriaosylceramide. ^a Abnormal findings of the GLA gene include class 3 variants (uncertain to be pathogenic), class 4 variants (likely to be pathogenic), and class 5 variants (certain pathogenic). ^b The diagnosis FD is confirmed in women when the abnormal findings of the GLA gene complemented with abnormal findings in the biochemical assessment (α-Gal A and Lyso-GB3).</p

Small fiber neuropathy patients analyzed for Fabry disease in the Maastricht University Medical Center.

<p>Illustration of the outcome of investigations to confirm the diagnosis of Fabry disease. α-Gal A: α-galactosidase A, Lyso-GB3: lysosomal globotriaosylceramide, FD: Fabry disease, SFN: small fiber neuropathy. ^a Missing data. ^b The measurement of lyso-GB3 excretion in urine was incorporated in our workflow for SFN patients from April 2012. ^c GLA gene sequencing was performed in all women, and in males in case of reduced α-Gal A enzyme activity. ^d These includes the class 2 variants (unlikely to be pathogenic) and the class 3 variants (uncertain to be pathogenic).</p

Differences in Strength and Timing of the mtDNA Bottleneck between Zebrafish Germline and Non-germline Cells

We studied the mtDNA bottleneck in zebrafish to elucidate size, timing, and variation in germline and non-germline cells. Mature zebrafish oocytes contain, on average, 19.0 × 10(6) mtDNA molecules with high variation between oocytes. During embryogenesis, the mtDNA copy number decreases to ∼170 mtDNA molecules per primordial germ cell (PGC), a number similar to that in mammals, and to ∼50 per non-PGC. These occur at the same developmental stage, implying considerable variation in mtDNA copy number in (non-)PGCs of the same female, dictated by variation in the mature oocyte. The presence of oocytes with low mtDNA numbers, if similar in humans, could explain how (de novo) mutations can reach high mutation loads within a single generation. High mtDNA copy numbers in mature oocytes are established by mtDNA replication during oocyte development. Bottleneck differences between germline and non-germline cells, due to early differentiation of PGCs, may account for different distribution patterns of familial mutations

The unfolding clinical spectrum of holoprosencephaly due to mutations in SHH, ZIC2, SIX3 and TGIF genes

Holoprosencephaly is a severe malformation of the brain characterized by abnormal formation and separation of the developing central nervous system. The prevalence is 1:250 during early embryogenesis, the live-born prevalence is 1:16 000. The etiology of HPE is extremely heterogeneous and can be teratogenic or genetic. We screened four known HPE genes in a Dutch cohort of 86 non-syndromic HPE index cases, including 53 family members. We detected 21 mutations (24.4%), 3 in SHH, 9 in ZIC2 and 9 in SIX3. Eight mutations involved amino-acid substitutions, 7 ins/del mutations, 1 frame-shift, 3 identical poly-alanine tract expansions and 2 gene deletions. Pathogenicity of mutations was presumed based on de novo character, predicted non-functionality of mutated proteins, segregation of mutations with affected family-members or combinations of these features. Two mutations were reported previously. SNP array confirmed detected deletions; one spanning the ZIC2/ZIC5 genes (approx. 100 kb) the other a 1.45 Mb deletion including SIX2/SIX3 genes. The mutation percentage (24%) is comparable with previous reports, but we detected significantly less mutations in SHH: 3.5 vs 10.7% (P=0.043) and significantly more in SIX3: 10.5 vs 4.3% (P=0.018). For TGIF1 and ZIC2 mutation the rate was in conformity with earlier reports. About half of the mutations were de novo, one was a germ line mosaic. The familial mutations displayed extensive heterogeneity in clinical manifestation. Of seven familial index patients only two parental carriers showed minor HPE signs, five were completely asymptomatic. Therefore, each novel mutation should be considered as a risk factor for clinically manifest HPE, with the caveat of reduced clinical penetrance