[Development of analytics in newborn screening-from the Guthrie card to genetics]. / Entwicklung der Analytik im Neugeborenen-Screening ­ Von der Guthrie-Karte zur Genetik.

For more than five decades, all newborns in Germany have been offered a screening examination for the early detection of congenital treatable diseases. Since its inception, about 35 million children have been screened in this way.Originally, screening exams only included early detection of phenylketonuria, which, without timely treatment, would lead to mental retardation that could no longer be corrected. The bacteriological Guthrie test allowed the detection of elevated concentrations of phenylalanine. The methods used today are the result of decades of development. They have been expanded to include tests to determine enzyme activities, immunoassays for the early detection of important hormonal disorders such as congenital hypothyroidism, and high-pressure liquid chromatography for the diagnosis of pathologic hemoglobins. The very sophisticated tandem mass spectrometry enables the simultaneous detection of amino acid and fatty acid compounds. Steroids can also be identified. The specificity can be further increased by combining tandem mass spectrometry with chromatographic pre-separation. In recent years, chemical-analytical analyses have been supplemented by genetic diagnostic methods such as quantitative or qualitative polymerase chain reaction (PCR).The current state of laboratory technology is by no means final. Both classical analytics and especially genetic methods are facing further rapid development. Although the expansion of screening is also a consequence of technical development, the inclusion of further congenital diseases is fundamentally dependent on the given therapy. But it is precisely here that many innovations are currently being investigated. Gene therapy is at the forefront of interest.

How "Universal" is Adhesion? Shear Bond Strength of Multi-mode Adhesives to Enamel and Dentin.

PURPOSE: The purpose of this in vitro study was to compare the shear bond strength of different universal adhesives to enamel and dentin with and without additional phosphoric acid etching before and after thermocycling (TC). MATERIALS AND METHODS: The mesial, distal, lingual, and buccal surfaces of the teeth were ground flat to expose two enamel areas and two dentin areas. Specimens were randomly divided into 36 groups (n = 10). Four adhesives were tested: Scotchbond Universal (SBU), Prime & Bond Elect (PBE), All-Bond Universal (ABU), and iBond Universal (IBU). The application was performed in the self-etch mode (SE) or in the etch-and-rinse mode (ER). Optibond FL (OFL) was used as an etch-and-rinse control. After composite buildups were made, half of the groups were tested before and the other half after TC. The failure mode was evaluated using a light microscope. RESULTS: In enamel, mean bond strengths ranged from 13.4 to 21.9 MPa in the SE mode before TC. When used as an ER adhesive, mean bond strengths exceeded 30 MPa. TC did not influence the mean bond strengths in either application mode. Regarding dentin, significant differences in the SE mode were adhesive dependent. Differences were only detected between SBU and IBU before TC and were not present after TC. When SBU, PBE, and ABU were used as ER adhesives, no differences were detected before TC. TC only affected IBU when compared to the other adhesives. CONCLUSION: Universal adhesives benefit from phosphoric acid etching, as bond strengths increased especially at enamel surfaces.

Hypoglycemic Toxins and Enteroviruses as Causes of Outbreaks of Acute Encephalitis-Like Syndrome in Children, Bac Giang Province, Northern Vietnam.

We investigated the cause of seasonal outbreaks of pediatric acute encephalitis-like syndrome associated with litchi harvests (May-July) in northern Vietnam since 2008. Nineteen cerebrospinal fluid samples were positive for human enterovirus B, and 8 blood samples were positive for hypoglycemic toxins present in litchi fruits. Patients who were positive for hypoglycemic toxins had shorter median times between disease onset and admission, more reports of seizures, more reports of hypoglycemia (glucose level <3 mmol/L), lower median numbers of leukocytes in cerebrospinal fluid, and higher median serum levels of alanine aminotransferase and aspartate transaminase than did patients who were positive for enteroviruses. We suggest that children with rapidly progressing acute encephalitis-like syndrome at the time of the litchi harvest have intoxication caused by hypoglycemic toxins, rather than viral encephalitis, as previously suspected. These children should be urgently treated for life-threatening hypoglycemia.

Tyrosinemia type 1 and irreversible neurologic crisis after one month discontinuation of nitisone.

Tyrosinemia type I is an autosomal recessively inherited metabolic disease of tyrosine metabolism due to the deficiency of fumarylacetoacetate hydrolase. Clinical manifestations include hepatic failure, cirrhosis, hepatocellular carcinoma, renal fanconi syndrome, and neurologic crisis. With the introduction of 2-(2-nitro-4-trifluoro-methylbenzyol)-1,3 cyclohexanedione treatment the prognosis improved with reduced rate of complications. "Neurologic crisis" of tyrosinemia type I is a rare complication seen after discontinuation of treatment characterized with anorexia, vomiting, and hyponatremia in the initial phase continuing with paresthesia and paralysis of the extremities and the diaphragm. Here, we report a tyrosinemia type I patient who admitted to the hospital with nonspecific symptoms such as vomiting, anorexia, weakness, and restlessness only after one month discontinuation of nitisone and diagnosed as neurological crisis.

The Application of Cohen's Stress-Buffering Model for Weight Bias Internalization in Prebariatric Patients.

INTRODUCTION: Weight bias internalization (WBI) is associated with reduced psychological well-being in individuals with obesity. The aim of this study was to investigate the application of Cohen's stress-buffering model of social support for WBI on well-being in patients presenting for bariatric surgery. METHODS: In N = 804 adult prebariatric patients, WBI, social support, depression severity, health-related quality of life (HRQOL), and self-esteem were assessed by self-report questionnaires. Structural Equation Modeling was applied to test for direct associations between social support and well-being and for a buffering effect of social support on the relationship between WBI and well-being. RESULTS: After controlling for age, sex, and body mass index, greater social support was directly associated with reduced depression severity and increased self-esteem, but not with increased HRQOL. Contrary to Cohen's stress-buffering model, social support showed no moderating effects on the association between WBI and depression severity, HRQOL, and self-esteem. CONCLUSION: These cross-sectional results may indicate that greater social support is associated with improved well-being, supporting it as a potential coping resource in bariatric surgery. Given the absence of supporting evidence for the buffering effect in the present study, future prospective research may reevaluate the existence of a moderating effect of social support and investigate whether support-focused interventions improve psychological well-being.

Psychometric Properties of the WBIS/M in a Representative Prebariatric Sample: Evidence for an Improved 10-Item Version.

INTRODUCTION: The Weight Bias Internalization Scale and the Modified Weight Bias Internalization Scale are well-established self-report questionnaires for assessing weight bias internalization, which is widespread among bariatric patients. However, among this group, psychometric properties of the Weight Bias Internalization Scale have only been examined in small samples showing unsatisfactory model fit and have not been explored for the modified questionnaire. METHODS: This study psychometrically evaluated and compared the Weight Bias Internalization Scale and Modified Weight Bias Internalization Scale in a large sample of prebariatric patients (N = 825, mean age = 46.75 years, SD = 11.55) regarding item characteristics, model fit to unidimensionality, reliability, construct validity, and measurement invariance. RESULTS: Item 4 of both questionnaires showed low corrected item-total correlations (<0.40) and was therefore removed from the scales. The new 10-item versions showed improved item characteristics, internal consistency, model fit to unidimensionality, and convergent and divergent validity when compared to the 11-item versions. The best psychometric properties were found for the 10-item version of the Modified Weight Bias Internalization Scale. CONCLUSION: The 10-item version of the Modified Weight Bias Internalization Scale surpasses the other versions studied in all psychometric properties. Therefore, it should be used in prebariatric patients to detect weight bias internalization and provide them with psychological interventions that could improve bariatric surgery outcomes.

Depressive Symptoms among Bariatric Surgery Candidates: Associations with Stigmatization and Weight and Shape Concern.

Bariatric surgery candidates (BSC) are a highly vulnerable group for mental health impairments. According to the theoretical model of weight stigma, weight-related experienced stigmatization (ES) negatively influences mental health through weight bias internalization (WBI). This study tested this model among BSC and investigated whether this association depends on a negative body image in terms of weight and shape concern as a potential moderator. As part of a German multicenter study, ES, WBI, weight and shape concern, and depressive symptoms were assessed via self-report questionnaires among n = 854 BSC. Simple and moderated mediation analyses were applied to analyze whether WBI influences the relationship between ES and depressive symptoms, and whether this influence depends on weight and shape concern. WBI significantly mediated the relationship between ES and depressive symptoms by partially reducing the association of ES with depressive symptoms. Weight and shape concern emerged as significant moderators in the overall model and specifically for associations between WBI and depressive symptoms. The results suggest that the association between ES and depressive symptoms among BSC is stronger in those with high WBI. This association is strengthened by weight and shape concern, especially at low and mean levels. Studies evaluating longitudinal associations between weight-related stigmatization and mental health are indicated, as well as intervention studies targeting WBI in order to reduce adverse effects of ES on mental health in BSC.

Tissue Specific Distribution and Activation of Sapindaceae Toxins in Horses Suffering from Atypical Myopathy.

Equine atypical myopathy is caused by hypoglycin A (HGA) and methylenecyclopropylglycine (MCPrG), the known protoxins of sycamore maple (Acer pseudoplatanus). Various tissues from five atypical myopathy cases were analyzed but only HGA was found. Whether deamination of MCPrG has already occurred in the intestine as the first stage of metabolization has not been investigated. Activation of the protoxins to methylenecyclopropylacetyl (MCPA)-CoA and methylenecyclopropylformyl (MCPF)-CoA, respectively, occurred mainly in the skeletal muscles, as evidenced by very high concentrations of MCPA-carnitine and MCPF-carnitine in this tissue. Inhibition of the acyl-CoA dehydrogenases of short- and medium-chain as well as branched-chain fatty acids by the toxins led to a strong increase in the corresponding acylcarnitines, again preferentially in skeletal muscles. An accumulation of the long-chain acylcarnitines beyond the level of the control samples could not be detected in the tissues. As a high amount of HGA was always found unmetabolized in the organs, we speculate that targeting the interruption of further metabolization might be a way to stop the progression of intoxication. Inhibition of the mitochondrial branched-chain amino acid aminotransferase, i.e., the first enzyme responsible for the activation of sycamore maple protoxins, could be a therapeutic approach.

Nonnormative Eating Behaviors and Eating Disorders and Their Associations With Weight Loss and Quality of Life During 6 Years Following Obesity Surgery.

Importance: Individuals with severe obesity presenting for obesity surgery (OS) frequently show nonnormative eating behaviors (NEBs) and eating disorders (EDs), but the long-term course and prospective associations with weight loss and health-related quality of life (HRQOL) remain unclear. Objective: To examine the prevalence and prospective relevance of presurgical and postsurgical NEBs and EDs according to the Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition, diagnosed through clinical interview, for weight loss and HRQOL up to 6 years following OS. Design, Setting, and Participants: In the prospective, multicenter Psychosocial Registry for Obesity Surgery cohort study, patients seeking OS were recruited at 6 OS centers in Germany and assessed at baseline before surgery and at 6 months and 1 to 6 years after surgery. From a consecutive sample of 1040 volunteers with planned OS from March 1, 2012, to December 31, 2020, a total of 748 (71.92%) were included in this study. Across follow-up, 93 of the 748 patients (12.43%) dropped out. Data were analyzed from April to November 2021. Interventions: Laparoscopic Roux-en-Y gastric bypass or sleeve gastrectomy. Main Outcomes and Measures: Both NEBs and EDs were identified using the Eating Disorder Examination interview. Main outcomes were the percentage of total body weight loss (%TBWL) and HRQOL (Impact of Weight on Quality of Life-Lite; range, 0-100, with 0 indicating worst and 100 indicating best). Results: In 748 patients undergoing OS (mean [SD] age, 46.26 [11.44] years; mean [SD] body mass index [calculated as weight in kilograms divided by height in meters squared], 48.38 [8.09]; 513 [68.58%] female), the mean (SD) %TBWL was 26.70% (9.61%), and the mean (SD) HRQOL improvement was 35.41 (20.63) percentage points across follow-up. Both NEBs and EDs were common before surgery, with postsurgical improvements of varying degrees. Whereas NEBs and EDs did not reveal significant prospective associations with %TBWL, loss-of-control eating at follow-up was concurrently associated with lower %TBWL (estimate, -0.09; 95% CI, -0.14 to -0.04). Loss-of-control eating (estimate, -0.10; 95% CI, -0.17 to -0.03 percentage points) and binge-eating disorder of low frequency and/or limited duration (estimate, -6.51; 95% CI, -12.69 to -0.34 percentage points) at follow-up showed significant prospective associations with lower HRQOL. Conclusions and Relevance: This cohort study found prospective relevance of loss-of-control eating and binge-eating disorder of low frequency and/or limited duration for reduced long-term HRQOL following OS. These findings underline the importance of monitoring both NEBs, especially loss-of-control eating, and EDs in the long term postsurgically to identify patients in need of targeted prevention or psychotherapy.

Cyclosporine A: impact on mitochondrial function in endothelial cells.

INTRODUCTION: Although cyclosporine A (CSA) is considered to be an efficient immunosuppressive compound in transplantation, vascular side effects like arterial hypertension, neurologic complications and other adverse reactions occur. Interference of CSA with mitochondrial function may be responsible for these side effects. METHODS: We evaluated the effect of CSA on mitochondrial and glycolytic function by measuring fatty acid oxidation (FAO), activities of respiratory chain complexes (RC) and citratesynthase (CS), lactate/pyruvate-ratios, energy-rich phosphates as well as activities of some glycolytic enzymes in human umbilical vein endothelial cells. RESULTS: After 48 h of CSA incubation, global FAO, RC-complexes 1 + 3; 4 and 5 as well as CS were compromised while energy charges were not reduced. Lactate/pyruvate-ratios increased; cellular lactate dehydrogenase (LDH)-, hexokinase- and phosphofructokinase-activities were not impaired by CSA. Moderate cellular toxicity, assessed by LDH leakage, appeared only at the highest CSA concentration. CONCLUSION: Part of CSA toxicity may arise from alterations in mitochondrial function as judged by impaired FAO and respiratory chain enzymes. To some extent, energy balance seems to be maintained by cytosolic energy production. Although only demonstrated for endothelial cells, it is conceivable that such effects will alter energy metabolism of different organs with high oxidative energy demands.

Severe Inhibition of Long-Chain Acyl-CoA Enoylhydratase (EC 4.2.1.74) in a Newborn Foal Suffering From Atypical Myopathy.

In horses, congenital defects of energy production from long-chain fatty acids have not been described so far. In contrast, inhibition of fatty acid degradation caused by the toxins hypoglycin A and methylenecyclopropylglycine from various maple species are observed frequently. These non-proteinogenic aminoacids are passed on placentally to fetuses or with collostrum or milk to newborn foals. Nevertheless, newborn foals become very rarely symptomatic. Vertical transmission apparently is not sufficient to induce clinical disease without a particular genetic constellation being present. One of these rare cases was investigated here using samples from a mare and her foal. Intoxication by hypoglycin A and methylenecyclopropylglycine is also of interest to human pathology, because these toxins have caused fatal poisonings after consumption of certain fruits many times, especially in children. Maple toxins, their metabolites and some short-chain acyl compounds were quantified by ultrahigh-pressure liquid chromatography/tandem mass spectrometry. An comprehensive spectrum of long-chain acylcarnitines was prepared using electrospray ionization tandem mass spectrometry. Organic acids and acylglycines were determined by gas chromatography mass spectrometry. For evaluation, results of other horses poisoned by maple material as well as unaffected control animals were used. In the serum of the foal, hypoglycin A was detected at a low concentration only. Toxin metabolites reached <3.5% of the mean of a comparison group of horses suffering from atypical myopathy. The spectrum of acylcarnitines indicated enzyme inhibition in short-chain and medium-chain regions typical of acer poisoning, but the measured concentrations did not exceed those previously found in clinically healthy animals after maple consumption. The values were not sufficient to explain the clinical symptoms. In contrast, a remarkably strong enrichment of tetradecenoylcarnitine and hexadecenoylcarnitine was observed. This proves a blockade of the long-chain enoyl-CoA hydratase (EC 4.2.1.74). Vertical transfer of maple toxins to a newborn foal is sufficient for induction of clinical disease only if there is an additional specific reactivity to the active toxins. This was found here in an inhibition of long-chain enoyl-CoA hydratase. Isolated dysfunction of this enzyme has not yet been reported in any species. Further studies are necessary to prove a specific genetic defect.

Detection of maple toxins in mare's milk.

BACKGROUND: Plants from the Sapindaceae family that are consumed by horses (maple) and humans (ackee and litchi) are known to contain the toxins hypoglycin A and methylenecyclopropylglycine which cause seasonally occurring myopathy in horses and entero-encephalopathic sickness in humans. Vertical transmission of these toxins from a mare to her foal has been described once. However the mare's milk was not available for analysis in this case. We investigated mare's milk in a similar case. OBJECTIVE: We hypothesized that hypoglycin A and methylenecyclopropylglycine, like other amino acids' are secreted into the milk. ANIMALS: Mare with atypical myopathy. METHODS: A sample of the mare's milk and 6 commercial horse milk samples were extracted with a methanolic standard solution and analyzed for hypoglycin A, methylenecyclopropylglycine, and metabolites using tandem mass spectrometry after column chromatographic separation. RESULTS: There were hypoglycin A (0.4 µg/L) and the associated metabolites methylenecyclopropylacetyl glycine and carnitine (18.5 and 24.6 µg/L) plus increased concentrations of several acylcarnitines in the milk. The milk also contained methylenecyclopropylformyl glycine and carnitine (0.8 and 60 µg/L). The latter substances were also detected in 1 of 6 commercial horse milk samples. CONCLUSIONS AND CLINICAL IMPORTANCE: Transmission of the maple toxins can occur through mare's milk. Vertical transmission of Sapindacea toxins might also have importance for human medicine, for example, after consumption of ackee or litchi.