Therapeutic Role of Nusinersen on Respiratory Progression in Pediatric Patients With Spinal Muscular Atrophy Type 2 and Nonambulant Type 3.

Background and Objectives: Nusinersen has shown significant functional motor benefit in the milder types of spinal muscular atrophy (SMA). Less is known on the respiratory outcomes in patients with nusinersen-treated SMA. The aim of this study was to describe changes in respiratory function in pediatric patients with SMA type 2 and 3 on regular treatment with nusinersen within the iSMAc international cohort and to compare their trajectory with the natural history (NH) data published by the consortium in 2020. Methods: This is a 5-year retrospective observational study of pediatric SMA type 2 and nonambulant type 3 (age ≤18 years) treated with nusinersen. The primary objective was to compare the slopes of decline in forced vital capacity % predicted (FVC% pred.), FVC, and age when FVC dropped below 60% between the treated patients and a control group from the natural history cohort. Data on peak cough flow and the use of noninvasive ventilation (NIV) and cough assist were collected. Results: Data were available for 69 treated patients, 53 were SMA type 2 and 16 type 3. The mean (SD) age at first injection was 8.5 (3.2) and 9.7 (3.7) years, respectively. The median (interquartile range) treatment duration was 1 (0.7; 1.9) and 1.2 (0.9; 1.9) years, respectively. At the time of the first nusinersen injection, 24 of 52 (46%) patients with SMA type 2 and 2 of 16 (13%) patients with SMA type 3 were on NIV. Forty-three of 53 (81%) and 4 of 16 (25%) patients used cough device. FVC% pred. in treated patients with SMA type 2 declined annually by 2.3% vs 3.9% in NH (p = 0.08) and in treated patients with type 3 by 2.6% vs 3.4% NH (p = 0.59). Patients treated reached FVC <60% later than untreated (12.1 vs 10 years, p = 0.05). A higher percentage of treated vs untreated patients maintained FVC% pred. equal/above their baseline after 12 (65% vs 36%) and 24 (50% vs 24%) months, respectively. NIV use among treated did not significantly change throughout 1-year follow-up. Discussion: This study included the largest real-world cohort of pediatric patients with milder SMA types. The results suggest a positive role of nusinersen in delaying the respiratory decline in patients treated longer than 1 year when compared with natural history. Larger cohorts and longer observation are planned. Classification of Evidence: This study provided Class III evidence that nusinersen slows progression for patients with SMA types 2 and 3 compared with a natural history cohort.

Risk factors associated with fasting during Ramadan among individuals with diabetes according to IDF-DAR risk score in Atbara city, Sudan: Cross-sectional hospital-based study.

BACKGROUND: Ramadan is a holy month for Muslims. The aim of this study was to assess risk associated with Ramadan fasting among Sudanese individuals with diabetes (high, moderate, and low risk) according to International Diabetes Federation in collaboration with Diabetes and Ramadan International alliance (IDF-DAR) Practical Guidelines 2021 risk score. METHODS: This was a cross-sectional hospital-based study recruited 300 individuals with diabetes (79% have type 2 diabetes) from diabetes centers in Atbara city, the River Nile state, Sudan. RESULTS: The risk score was distributed as low risk (13.7%), Moderate risk (24%), and High risk (62.3%). T-test showed a significant difference in mean risk score by gender, duration and type of diabetes (p values = 0.004, 0.000, & 0.000, respectively). One-way ANOVA revealed a statistically significant difference in the risk score by age groups (p = 0.000). Logistic regression revealed that the odds of being in the 41-60 years age group had lower probability to be categorized in the moderate risk group of fasting rather than low risk by 4.3 times than being in the age more than 60 years. (p = 0.008), the odds of being in the age group 41-60 years lower probability to be categorized in the high risk of fasting rather than low risk by 8 times than being in the age more than 60 years. (p = 0.000). CONCLUSION: The majority of patients in this study have a high risk for Ramadan fasting. IDF-DAR risk score is of great significance in assessing individuals with diabetes for Ramadan fasting.

Antineurofascin IgG2-associated paediatric autoimmune nodopathy.

In this case series of four paediatric patients, we present the first described cases of immunotherapy-responsive autoimmune nodopathy with IgG2 antineurofascin antibodies. In three cases, the antineurofascin antibodies were predominantly of the IgG2 subclass, a novel finding in comparison to previously described adult cases where IgG4 and/or IgG1/3 have typically been described. One patient had low signal for IgG2 with predominant IgG1 and IgG4 antibodies, a pattern commonly seen in adult patients. Two patients had antibodies targeting all three neurofascin isoforms (155, 186, and 140), whereas antibodies in the sera from the third targeted only the nodal isoforms 186 and 140, and the fourth patient only neurofascin 155. The three patients with IgG2 predominant antibodies appear to be responsive to intravenous immunoglobulin (IVIG) to varying degrees thus far, whereas the patient with IgG1/4 antibodies had poor response to IVIG but good response to steroids. Although the full clinical significance of IgG2 predominant antineurofascin antibodies in the context of childhood polyneuropathy remains unclear, emerging evidence of serological-phenotypic correlation may inform prognostication and therapeutic decision-making, warranting further study into this area. WHAT THIS PAPER ADDS: Paediatric immunotherapy-responsive nodopathies were associated with antineurofascin antibodies predominantly of the IgG2 subclass in 3 out of 4 patients. Identification of antibodies and understanding their phenotypic relevance could predict response to treatment and guide therapeutic decision-making in children.

Pharmacological characterization of three chicken melanocortin-3 receptor mutants.

The melanocortin-3 receptor (MC3R) is a G protein-coupled receptor and potentially important in production traits. Three naturally occurring mutations (M54L, G104S, and L151R) in chicken MC3R (cMC3R) were reported previously to be associated with production traits. Here, we inserted the full-length cMC3R coding sequence into pcDNA3.1(+) and generated the 3 mutations by site-directed mutagenesis. The total and cell surface expression of the receptors was measured by flow cytometry. We analyzed the pharmacological characteristics, including binding and cyclic adenosine monophosphate (cAMP) and mitogen-activated protein kinase (MAPK) signaling, using 6 ligands ([Nle4, D-Phe7]-α-melanocyte stimulating hormone (MSH), α-, ß-, γ-, and D-Trp8-γ-MSHs, and agouti-related peptide). All mutants had similar total and cell surface expression as the wild-type (WT) cMC3R. M54L had similar pharmacological properties as the WT cMC3R. G104S did not exhibit any specific binding but had minimal response to α-, ß-, γ-, and D-Trp8-γ-MSH, although it generated 24% WT response when stimulated by NDP-MSH. Although L151R had normal binding, the responses to agonists were reduced to approximately 25% of that of the WT. In MAPK signaling, all 3 mutants showed significantly increased agonist-stimulated phosphorylation of extracellular signal-regulated protein kinases 1/2, indicating the existence of biased signaling at G104S and L151R. In summary, our studies demonstrated that although all 3 mutations are significantly associated with production traits, only G104S and L151R had severe defects in receptor pharmacology. How M54L might cause production trait differences remains to be investigated.

Label-free proteomics study on Shewanella putrefaciens regulated by ε-poly-lysine treatment.

AIMS: The purpose of this study was to investigate the regulatory mechanism of ε-PL on Shewanella putrefaciens. METHODS AND RESULTS: Proteomics analysis of inhibitory effect of ε-PL against S. putrefaciens was performed by label-free quantitative assay based on high-resolution mass spectrometry (MS). Quantification of 2206 proteins was obtained with high confidence, and a total of 36 differentially expressed proteins (DEPs), with 10 and 26 proteins showing upregulation and downregulation, respectively, were identified. Upon Go functional enrichment, 11, 5 and 8 specific Go terms in biological processes, molecular functions and cellular components were identified, respectively. Six KEGG pathways, including 'ribosome', were significantly enriched. Among the ribosome pathway, there were seven DEPs and all of them were distributed on large and small subunits of ribosome. CONCLUSIONS: The significant downregulation of proteins, large subunits of ribosomal proteins RP-L18, L30 and L27, small subunits ribosomal proteins S16 and S20, and RNA polymerase ß' subunit protein rpoC were the critical action sites of ε-PL to inhibit S. putrefaciens growth. SIGNIFICANCE AND IMPACT OF THE STUDY: Shewanella putrefaciens is one of the representative fish-spoilage bacteria regardless of fish type, and poses significant problems for the fish brewery. A better understanding of the antibacterial mechanism of ε-PL on S. putrefaciens could make important contributions to development of biological control strategies of these economically important pathogens.

Retention Time and Ion Abundance Ratio of 8 Common Drugs （Poisons） by LC-MS/MS Qualitative Analysis.

ABSTRACT: Objective To investigate the maximum allowable deviations of retention time and ion abundance ratio of the 8 common drugs （poisons） from 3 categories, poisons （methamphetamine, morphine, ketamine）, benzodiazepines （estazolam, midazolam, diazepam, clonazepam） and barbiturates （phenobarbital） in blood, by liquid chromatography-tandem mass spectrometry （LC-MS/MS） in forensic toxicology analysis. Methods The deviations of retention time and ion abundance ratio at 7 low mass concentrations, limit of detection （LOD）, 2LOD, limit of quantitation （LOQ）, 1.5LOQ, 2LOQ, 4LOQ and 6LOQ, were tested by LC-MS/MS after liquid-liquid extraction under the conditions of two chromatographic columns and three chromatographs. Results The deviation of absolute retention time of 98.11% of 8 drugs （poisons） in the blood samples was within the range of ±0.05 min, and that of the relative retention time of 96.21% was within the range of ±0.4%. The maximum deviation of the ion abundance ratio was highly correlated with the mass concentration. When the mass concentration of drugs （poisons） was LOQ or above, more than 95% of the absolute deviation and relative deviation of the ion abundance ratio were in the range of ±25% and ±40%, respectively; when the mass concentration was below LOQ, the range could be expanded to ±35% and ±50%, respectively. Conclusion It is recommended for the determination range of the absolute retention time deviation of 8 common drugs （poisons） to be ±0.1 min and that of the relative retention time deviation to be ±1.0%. The determination range of absolute deviation of the ion abundance ratio should be ±25% when the mass concentration is LOQ or above, and the relative deviation should be ±40%. When the mass concentration is below LOQ, the deviation determination range can be expanded to ±35% and ±50%, respectively.

Pre-existing chronic medical illnesses and follow up status among active pulmonary tuberculosis cases in a district population.

INTRODUCTION: The knowledge of pre-existing medical illnesses and their follow up status among active pulmonary tuberculosis (PTB) subjects can help in tuberculosis (TB) control programme. The aims of our study were to examine: the prevalence of pre-existing chronic medical illnesses, the follow up status of known pre-existing co-morbid and to distinguish between diagnosed and undiagnosed preexisting tuberculosis related chronic medical illnesses among our active PTB subjects. METHODS: We conducted a retrospective review of demographic and clinical data of active PTB subjects that were diagnosed between January 2015 and June 2017 in the district of Manjung, Perak, Malaysia. Among the 302 TB clinical notes reviewed, 253 patients were included. Subjects below the age of 18 years and whose follow up centres for their medical illnesses that were located outside of Manjung were excluded. Demographic and clinical data were collected using pre-tested data collection form by trained investigators. The data was analysed using SPSS Version 20.0. RESULTS: We identified diabetes mellitus as the most prevalent pre-existing co-morbid (77 cases) and almost 90% (68 cases) of these diabetic subjects were diagnosed prior to active PTB diagnosis. This was followed by Human Immunodeficiency Virus and Hepatitis C infection which accounted for 12.0% (30 cases) of the study populations. Among 132 subjects who had pre-existing chronic medical illnesses, only 74 subjects (29%) were under regular follow up at healthcare facilities in Manjung prior to PTB diagnosis. CONCLUSION: Overall, our research provides evidence on the existence of wide variation of clinical background among active PTB subjects.

Temporal Effects of Sleeve Gastrectomy on Glucose-Insulin Homeostasis and Incretin Hormone Response at 1 and 6 Months.

BACKGROUND: Bariatric surgery is an effective treatment for morbid obesity and glycaemic dysfunction. OBJECTIVES: The aim of the work was to examine both the static and dynamic changes of glucose-insulin homeostasis and incretin hormone response following sleeve gastrectomy (SG) in a sample of 55 participants preoperatively and 1 month and 6 months postoperatively. The focus was on a sample of patients with impaired glucose tolerance and type 2 diabetes (T2D). SETTING: Morriston Hospital, UK. METHODS: Prospective study comprising of 55 participants with impaired glucose homeostasis and T2D undergoing SG (mean body mass index [BMI] 50.4 kg/m2, mean glycated haemoglobin [A1C] 7.4%). Serial measurements of glucose, insulin, C-peptide, glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic hormone (GIP) were performed during oral glucose tolerance testing preoperatively and 1 and 6 months postoperatively. Areas under the curve (AUC) were examined at 30, 60, and 120 min. RESULTS: We observed significant improvements in measures of obesity, as well as static and dynamic measures of glucose, insulin, C-peptide and HOMA. Furthermore, significant increases in GLP-1 response as early as 6 months postoperatively were also seen. CONCLUSIONS: To our knowledge, no study has examined the detailed dynamic changes in glucose and insulin homeostasis in this number of participants undergoing SG in relation to incretin hormones GIP and GLP-1. This current study supports the role of SG for the treatment of obesity-related glucose dysregulation.

Ion Abundance Ratios of Qualitative Analysis by GC-MS of 4 Common Drugs （Poisons.

ABSTRACT: Objective To investigate the maximum allowable deviation of ion abundance ratios of characteristic fragment ions in common drugs （poisons） in blood by gas chromatography-mass spectrometry （GC-MS） method. Methods Four common drugs （poisons） （dichlorvos, phorate, diazepam and estazolam） were detected by GC-MS full scan mode after liquid-liquid extraction in two laboratories and under three chromatographic conditions. The deviations of ion abundance ratios of the four common drugs （poisons） in marked blood samples with concentrations of 0.5, 1.0, 2.0, 5.0 and 10.0 µg/mL were analyzed. At the same time, the false negative rates of ion abundance ratios were analyzed when the mass concentration was limit of detection （LOD）, 2LOD, limit of quantitation （LOQ） and 2LOQ, and the false positive rates of ion abundance ratios were analyzed with blank blood samples. Results Under the two laboratories, four common drugs （poisons） and three kinds of chromatography conditions, the differences in deviations of the ion abundance ratios of marked blood samples were not statistically significant （P>0.05）. More than 95% of the absolute deviations of the ion abundance ratios of the marked blood samples were within the range of ±10%, and more than 95% of the relative deviations were within the range of ±25%. In cases of low concentration （concentration less than 2LOQ） or low signal to noise ratio （3-15）, the false negative rate was less than 5% and the false positive rate was 0% when the relative deviation was greater than 50%. Conclusion The absolute deviations of ion abundance ratios of four common drugs （poisons） in marked blood samples are advised to have a determination range within ±10%, and the determination range of relative deviations within ±25%.

Retention Time of Four Common Poisons （Drugs） by GC-MS Qualitative Analysis.

OBJECTIVES: To investigate the maximum allowable deviation of retention time （RT） or relative retention time （RRT） between the common poisons （drugs） and standard solvent by gas chromatography-mass spectrometry （GC-MS）. METHODS: After pretreatment with liquid-liquid extraction, four common poisons （drugs）-dichlorvos, phorate, diazepam and estazolam-were detected by full scan mode GC-MS. RT and RRT were analyzed according to combined uncertainty and expanded uncertainty. RESULTS: The expanded uncertainty of RT and RRT were 6.0×10-4-14.1×10-3 and 2.5×10-6-5.9×10-5 （k=3）, respectively. The RT of poisons （drugs） was relatively stable in blood samples with different mass concentrations. Among dichlorvos, phorate, diazepam and estazolam, the absolute deviation and relative deviation of RT were ≤0.03 min and ≤0.4%, respectively, and those of RRT were ≤0.003 min and ≤0.3%, respectively. CONCLUSIONS: The maximum allowable deviations of RT and RRT for common poisons （drugs） in blood samples are recommended to be ±0.05 min and ±0.5%.

Targeting Wall Teichoic Acid in Situ with Branched Polyethylenimine Potentiates ß-Lactam Efficacy against MRSA.

Methicillin-resistant Staphylococcus aureus (MRSA) is a medical concern. Here, we show that branched polyethylenimine (BPEI), a nontoxic, cationic polymer, restores MRSA's susceptibility to ß-lactam antibiotics. Checkerboard assays with MRSA demonstrated synergy between BPEI and ß-lactam antibiotics. A time-killing curve showed BPEI to be bactericidal in combination with oxacillin. BPEI did not potentiate efficacy with vancomycin, chloramphenicol, or linezolid. When exposed to BPEI, MRSA increased in size and had difficulty forming septa. BPEI electrostatically binds to wall teichoic acid (WTA), a cell wall anionic polymer of Gram-positive bacteria that is important for localization of certain cell wall proteins. Lack of potentiation in a WTA knockout mutant supports the WTA-based mechanism. These data suggest that BPEI may prevent proper localization of cell wall machinery by binding to WTA; leading to cell death when administered in combination with ß-lactam antibiotics. Negligible in vitro toxicity suggests the combination could be a viable treatment option.

Subcortical Shape Changes, Hippocampal Atrophy and Cortical Thinning in Future Alzheimer's Disease Patients.

Efficacy of future treatments depends on biomarkers identifying patients with mild cognitive impairment at highest risk for transitioning to Alzheimer's disease. Here, we applied recently developed analysis techniques to investigate cross-sectional differences in subcortical shape and volume alterations in patients with stable mild cognitive impairment (MCI) (n = 23, age range 59-82, 47.8% female), future converters at baseline (n = 10, age range 66-84, 90% female) and at time of conversion (age range 68-87) compared to group-wise age and gender matched healthy control subjects (n = 23, age range 61-81, 47.8% female; n = 10, age range 66-82, 80% female; n = 10, age range 68-82, 70% female). Additionally, we studied cortical thinning and global and local measures of hippocampal atrophy as known key imaging markers for Alzheimer's disease. Apart from bilateral striatal volume reductions, no morphometric alterations were found in cognitively stable patients. In contrast, we identified shape alterations in striatal and thalamic regions in future converters at baseline and at time of conversion. These shape alterations were paralleled by Alzheimer's disease like patterns of left hemispheric morphometric changes (cortical thinning in medial temporal regions, hippocampal total and subfield atrophy) in future converters at baseline with progression to similar right hemispheric alterations at time of conversion. Additionally, receiver operating characteristic curve analysis indicated that subcortical shape alterations may outperform hippocampal volume in identifying future converters at baseline. These results further confirm the key role of early cortical thinning and hippocampal atrophy in the early detection of Alzheimer's disease. But first and foremost, and by distinguishing future converters but not patients with stable cognitive abilities from cognitively normal subjects, our results support the value of early subcortical shape alterations and reduced hippocampal subfield volumes as potential markers for the early detection of Alzheimer's disease.

Post-Anoxic Reticular Reflex Myoclonus in a Child and Proposed Classification of Post-Anoxic Myoclonus.

OBJECTIVE: We describe a child with post-anoxic myoclonus of the reticular reflex type and discuss the classification of post-anoxic myoclonus. PATIENT DESCRIPTION: A nine-year-old boy with severe hypoxic-ischemic encephalopathy due to submersion developed early epileptic spasms followed by stimulus sensitive multifocal generalized myoclonus and later dystonia. Video electromyography (EMG) polygraphy performed before treatment demonstrated that the discharges associated with the myoclonus lasted less than 50 milliseconds. Cortical myoclonus was excluded by jerk-locked averaging using arm muscles, which showed no cortical correlates. The recruitment order on EMG polygraphy was consistent with a brainstem generator for the myoclonus, characteristic of reticular reflex myoclonus. Both myoclonus and dystonia responded to clonazepam. He remains in a persistent vegetative state. CONCLUSIONS: Reticular reflex myoclonus can be demonstrated by detailed neurophysiological assessment in children as in adults, and it has a similar poor prognosis in children. Post-anoxic myoclonus can have several mechanisms and should not be considered synonymous with Lance-Adams myoclonus.

A multicenter study on anaphylaxis caused by peanut, tree nuts, and seeds in children and adolescents.

Peanut (PN) and tree nuts (TNs) are common causes of anaphylaxis in Western countries, but no information is available in Korea. To feature clinical characteristics of anaphylaxis caused by PN, TNs, and seeds, a retrospective medical record review was performed in 14 university hospitals in Korea (2009-2013). One hundred and twenty-six cases were identified, with the mean age of 4.9 years. PN, walnut (WN), and pine nut accounted for 32.5%, 41.3%, and 7.1%, respectively. The median values of specific IgE (sIgE) to PN, WN, and pine nut were 10.50, 8.74, and 4.61 kUA /l, respectively. Among 50 cases managed in the emergency department, 52.0% were treated with epinephrine, 66.0% with steroid, 94.0% with antihistamines, 36.0% with oxygen, and 48.0% with bronchodilator. In conclusion, WN, PN, and pine nut were the three most common triggers of anaphylaxis in Korean children, and anaphylaxis could occur at remarkably low levels of sIgE.

Efficacy of ampicillin against methicillin-resistant Staphylococcus aureus restored through synergy with branched poly(ethylenimine).

ß-Lactam antibiotics kill Staphylococcus aureus bacteria by inhibiting the function of cell wall penicillin-binding proteins (PBPs) 1 and 3. However, ß-lactams are ineffective against PBP2a, used by methicillin-resistant S. aureus (MRSA) to perform essential cell wall crosslinking functions. PBP2a requires teichoic acid to properly locate and orient the enzyme, and thus MRSA is susceptible to antibiotics that prevent teichoic acid synthesis in the bacterial cytoplasm. As an alternative, we have used branched poly(ethylenimine), BPEI, to target teichoic acid in the bacterial cell wall. The result is restoration of MRSA susceptibility to the ß-lactam antibiotic ampicillin with a MIC of 1 µg ml-1, superior to that of vancomycin (MIC=3.7 µg ml-1). A checkerboard assay shows synergy of BPEI and ampicillin. NMR data show that BPEI alters the teichoic acid chemical environment. Laser scanning confocal microscopy images show BPEI residing on the bacterial cell wall, where teichoic acids and PBPs are located.

A superhydrophilic nitinol shape memory alloy with enhanced anti-biofouling and anti-corrosion properties.

This work reports on a nitinol (NiTi) surface modification scheme based on a chemical oxidation method, and characterizes its effects on wetting, biofouling and corrosion. The scheme developed is also compared with selected previous oxidation methods. The proposed method turns NiTi into superhydrophilic in ~5 min, and the static contact angle and contact angle hysteresis were measured to be ~7° and ~12°, respectively. In the PRP (platelet rich plasma) test, platelet adhesion was reduced by ~89% and ~77% respectively, compared with the original NiTi and the NiTi treated with the previous chemical oxidation scheme. The method developed provides a high (~1.1 V) breakdown voltage, which surpasses the ASTM standard for intervascular medical devices. It also provides higher superhydrophilicity, hemo-compatibility and anti-corrosion resistance than previous oxidation schemes, with a significantly reduced process time (~5 min), and will help the development of high performance NiTi devices.

A new method for sudden mechanical perturbation with axial load, to assess postural control in sitting and standing.

Sudden application of load along a sagittal or coronal axis has been used to study trunk stiffness, but not axial (vertical) load. This study introduces a new method for sudden-release axial load perturbation. Prima facie validity was supported by comparison with standard mechanical systems. We report the response of the human body to axial perturbation in sitting and standing and within-day repeatability of measures. Load of 20% of body weight was released from light contact onto the shoulders of 22 healthy participants (10 males). Force input was measured via force transducers at shoulders, output via a force plate below the participant, and kinematics via 3-D motion capture. System identification was used to fit data from the time of load release to time of peak load-displacement, fitting with a 2nd-order mass-spring-damper system with a delay term. At peak load-displacement, the mean (SD) effective stiffness measured with this device for participants in sitting was 12.0(3.4)N/mm, and in standing was 13.3(4.2)N/mm. Peak force output exceeded input by 44.8 (10.0)% in sitting and by 30.4(7.9)% in standing. Intra-class correlation coefficients for within-day repeatability of axial stiffness were 0.58 (CI: -0.03 to 0.83) in sitting and 0.82(0.57-0.93) in standing. Despite greater degrees of freedom in standing than sitting, standing involved lesser time, downward displacement, peak output force and was more repeatable in defending upright postural control against the same axial loads. This method provides a foundation for future studies of neuromuscular control with axial perturbation.