Prevalence of DDC genotypes in patients with aromatic L-amino acid decarboxylase (AADC) deficiency and in silico prediction of structural protein changes

Aromatic L-amino acid decarboxylase (AADC) deficiency is a rare autosomal recessive genetic disorder affecting the biosynthesis of dopamine, a precursor of both norepinephrine and epinephrine, and serotonin. Diagnosis is based on the analysis of CSF or plasma metabolites, AADC activity in plasma and genetic testing for variants in the DDC gene. The exact prevalence of AADC deficiency, the number of patients, and the variant and genotype prevalence are not known. Here, we present the DDC variant (n = 143) and genotype (n = 151) prevalence of 348 patients with AADC deficiency, 121 of whom were previously not reported. In addition, we report 26 new DDC variants, classify them according to the ACMG/AMP/ACGS recommendations for pathogenicity and score them based on the predicted structural effect. The splice variant c.714+4A>T, with a founder effect in Taiwan and China, was the most common variant (allele frequency = 32.4%), and c.[714+4A>T];[714+4A>T] was the most common genotype (genotype frequency = 21.3%). Approximately 90% of genotypes had variants classified as pathogenic or likely pathogenic, while 7% had one VUS allele and 3% had two VUS alleles. Only one benign variant was reported. Homozygous and compound heterozygous genotypes were interpreted in terms of AADC protein and categorized as: i) devoid of full-length AADC, ii) bearing one type of AADC homodimeric variant or iii) producing an AADC protein population composed of two homodimeric and one heterodimeric variant. Based on structural features, a score was attributed for all homodimers, and a tentative prediction was advanced for the heterodimer. Almost all AADC protein variants were pathogenic or likely pathogenic

Prevalence of DDC genotypes in patients with aromatic L-amino acid decarboxylase (AADC) deficiency and in silico prediction of structural protein changes

Aromatic L-amino acid decarboxylase (AADC) deficiency is a rare autosomal recessive genetic disorder affecting the biosynthesis of dopamine, a precursor of both norepinephrine and epinephrine, and serotonin. Diagnosis is based on the analysis of CSF or plasma metabolites, AADC activity in plasma and genetic testing for variants in the DDC gene. The exact prevalence of AADC deficiency, the number of patients, and the variant and genotype prevalence are not known. Here, we present the DDC variant (n = 143) and genotype (n = 151) prevalence of 348 patients with AADC deficiency, 121 of whom were previously not reported. In addition, we report 26 new DDC variants, classify them according to the ACMG/AMP/ACGS recommendations for pathogenicity and score them based on the predicted structural effect. The splice variant c.714+4A>T, with a founder effect in Taiwan and China, was the most common variant (allele frequency = 32.4%), and c.[714+4A>T];[714+4A>T] was the most common genotype (genotype frequency = 21.3%). Approximately 90% of genotypes had variants classified as pathogenic or likely pathogenic, while 7% had one VUS allele and 3% had two VUS alleles. Only one benign variant was reported. Homozygous and compound heterozygous genotypes were interpreted in terms of AADC protein and categorized as: i) devoid of full-length AADC, ii) bearing one type of AADC homodimeric variant or iii) producing an AADC protein population composed of two homodimeric and one heterodimeric variant. Based on structural features, a score was attributed for all homodimers, and a tentative prediction was advanced for the heterodimer. Almost all AADC protein variants were pathogenic or likely pathogenic

Impact of primary kidney disease on the effects of empagliflozin in patients with chronic kidney disease: secondary analyses of the EMPA-KIDNEY trial

Background: The EMPA KIDNEY trial showed that empagliflozin reduced the risk of the primary composite outcome of kidney disease progression or cardiovascular death in patients with chronic kidney disease mainly through slowing progression. We aimed to assess how effects of empagliflozin might differ by primary kidney disease across its broad population. Methods: EMPA-KIDNEY, a randomised, controlled, phase 3 trial, was conducted at 241 centres in eight countries (Canada, China, Germany, Italy, Japan, Malaysia, the UK, and the USA). Patients were eligible if their estimated glomerular filtration rate (eGFR) was 20 to less than 45 mL/min per 1·73 m2, or 45 to less than 90 mL/min per 1·73 m2 with a urinary albumin-to-creatinine ratio (uACR) of 200 mg/g or higher at screening. They were randomly assigned (1:1) to 10 mg oral empagliflozin once daily or matching placebo. Effects on kidney disease progression (defined as a sustained ≥40% eGFR decline from randomisation, end-stage kidney disease, a sustained eGFR below 10 mL/min per 1·73 m2, or death from kidney failure) were assessed using prespecified Cox models, and eGFR slope analyses used shared parameter models. Subgroup comparisons were performed by including relevant interaction terms in models. EMPA-KIDNEY is registered with ClinicalTrials.gov, NCT03594110. Findings: Between May 15, 2019, and April 16, 2021, 6609 participants were randomly assigned and followed up for a median of 2·0 years (IQR 1·5–2·4). Prespecified subgroupings by primary kidney disease included 2057 (31·1%) participants with diabetic kidney disease, 1669 (25·3%) with glomerular disease, 1445 (21·9%) with hypertensive or renovascular disease, and 1438 (21·8%) with other or unknown causes. Kidney disease progression occurred in 384 (11·6%) of 3304 patients in the empagliflozin group and 504 (15·2%) of 3305 patients in the placebo group (hazard ratio 0·71 [95% CI 0·62–0·81]), with no evidence that the relative effect size varied significantly by primary kidney disease (pheterogeneity=0·62). The between-group difference in chronic eGFR slopes (ie, from 2 months to final follow-up) was 1·37 mL/min per 1·73 m2 per year (95% CI 1·16–1·59), representing a 50% (42–58) reduction in the rate of chronic eGFR decline. This relative effect of empagliflozin on chronic eGFR slope was similar in analyses by different primary kidney diseases, including in explorations by type of glomerular disease and diabetes (p values for heterogeneity all >0·1). Interpretation: In a broad range of patients with chronic kidney disease at risk of progression, including a wide range of non-diabetic causes of chronic kidney disease, empagliflozin reduced risk of kidney disease progression. Relative effect sizes were broadly similar irrespective of the cause of primary kidney disease, suggesting that SGLT2 inhibitors should be part of a standard of care to minimise risk of kidney failure in chronic kidney disease. Funding: Boehringer Ingelheim, Eli Lilly, and UK Medical Research Council

Evaluation of appendicitis risk prediction models in adults with suspected appendicitis

Background Appendicitis is the most common general surgical emergency worldwide, but its diagnosis remains challenging. The aim of this study was to determine whether existing risk prediction models can reliably identify patients presenting to hospital in the UK with acute right iliac fossa (RIF) pain who are at low risk of appendicitis. Methods A systematic search was completed to identify all existing appendicitis risk prediction models. Models were validated using UK data from an international prospective cohort study that captured consecutive patients aged 16–45 years presenting to hospital with acute RIF in March to June 2017. The main outcome was best achievable model specificity (proportion of patients who did not have appendicitis correctly classified as low risk) whilst maintaining a failure rate below 5 per cent (proportion of patients identified as low risk who actually had appendicitis). Results Some 5345 patients across 154 UK hospitals were identified, of which two‐thirds (3613 of 5345, 67·6 per cent) were women. Women were more than twice as likely to undergo surgery with removal of a histologically normal appendix (272 of 964, 28·2 per cent) than men (120 of 993, 12·1 per cent) (relative risk 2·33, 95 per cent c.i. 1·92 to 2·84; P < 0·001). Of 15 validated risk prediction models, the Adult Appendicitis Score performed best (cut‐off score 8 or less, specificity 63·1 per cent, failure rate 3·7 per cent). The Appendicitis Inflammatory Response Score performed best for men (cut‐off score 2 or less, specificity 24·7 per cent, failure rate 2·4 per cent). Conclusion Women in the UK had a disproportionate risk of admission without surgical intervention and had high rates of normal appendicectomy. Risk prediction models to support shared decision‐making by identifying adults in the UK at low risk of appendicitis were identified

Spirulina platensis mitigates the neuroinflammatory effect of LPS-Induced BV2 Microglia

Neurodegenerative diseases are diseases attributable to neuronal loss through progressive degeneration, and one of the pathophysiologies is neuroinflammation. As one of the neuroinflammation regulatory cell, uncontrolled microglia activation promotes excessive secretion of proinflammatory mediators that will lead to neuronal cell death in neurodegenerative diseases. Current therapeutic options can only alleviate symptoms and were accompanied by multiple adverse effects. Furthermore, the socio-economic burden caused by neurodegenerative disease also increases due to an increase in life expectancy. Therefore, research efforts have been focusing on the discovery of potential therapeutic agents from natural resources for minimal adverse effects. Antioxidative and anti-inflammatory activities of S. platensis give rise to protection on dopaminergic neurons. In addition, S. platensis alleviates the morphological impairment on spinal cord and aids in the post-injury recovery. Therefore, we hypothesise that S. platensis UMACC 159 extracts can promote anti-neuroinflammatory activity by attenuating the lipopolysaccharide (LPS)-stimulated neuroinflammation in BV2 microglia. Objectives of this study include: (1) to investigate the presence of phytochemical constituents and antioxidant capacity of S. platensis UMACC 159 extracts; (2) to study the anti-neuroinflammatory effect of S. platensis UMACC 159 extracts in LPS-induced BV2 microglia; and (3) to determine the potential bioactive compound(s) profile of the most potent S. platensis UMACC 159 extract using Liquid Chromatography-Mass Spectrometry (LC-MS). S. platensis UMACC 159 culture strain was obtained from the Algae Research Laboratory in the University of Malaya. Four S. platensis UMACC 159 extracts (hexane, ethyl acetate, ethanol, and water) were screened for total phenolic content (TPC) and total flavonoid content (TFC) using Folin Ciocalteu reagent and aluminium chloride respectively. Antioxidant capacity of all extracts were determined using 2,2’-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 2,2-Diphenyl-1-pircrylhydrazyl (DPPH) and reducing power. Cell-based assays using BV2 microglia were used to screen for (1) the effect on cell viability via 3-(4,5-dimethylthiazol-2-yl)-2,5-iphenyltetrazolium bromide (MTT) assay and (2) the NO inhibitory activity via Griess assay. Ethanol extract with the highest antioxidative and NO inhibitory activities was selected for subsequent study on (1) the production of PGE2, IL-6 and TNF-α by BV2 microglia using ELISA, and (2) the protein expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) by BV2 microglia using western blot. The list of compound(s) present in ethanol extract was tentatively identified via LC-MS and the potential bioactive compounds related to neurodegenerative disease were identified based on literature search. Screening of the four S. platensis UMACC 159 extracts demonstrated that ethanol extract had the highest flavonoid content, antioxidative and NO inhibitory activities. Ethanol extract inhibits the NO production via downregulation of iNOS and reduces TNF-α and IL-6 secretion. LC-MS analysis revealed that ethanol extract contained 35 compounds, with 21 compounds being identified and 14 compounds remaining unknown. Based on literature search, emmotin A, palmitic amide, and 1-monopalmitin were proposed as the bioactive compounds in ethanol extract. In conclusion, S. platensis UMACC 159 ethanol extract possess neuroprotective property through antioxidative and anti-neuroinflammatory mechanisms, with emmotin A, palmitic amide, and 1-monopalmitin being the potential bioactive compounds. The current study would certainly contribute to the identification of bioactive compound(s) for the development on functional food targeting neurodegenerative disease. However, further isolation and bioassay-guided identification on the bioactive compound(s) are essential to conclude the identity of the bioactive compound(s) that contribute to the neuroprotective activity of S. platensis UMACC 159 ethanol extract. In addition, molecular study on the signalling pathway(s) involved in the neuroprotective activity of ethanol extract is also important in the effort of managing neurodegenerative disease with complex pathologies, as the signalling pathway(s) may be involved in more than one of the pathological processes in neurodegenerative disease

Development of An IoT-Enabled Photovoltaic-Battery Renewable Energy System

Solar energy is considered as a prominent source of renewable energy, mainly due to the vast abundance of sunlight and rapid advancements of photovoltaic (PV) technology. The performance, reliability and lifespan of PV systems are severely affected by numerous environmental factors and fault occurrences, which include: (1) extreme swing in the operating temperature; (2) low solar irradiation levels which appear undetected in PV systems, resulting in energy losses and degradation of PV panels; and (3) non-homogenous shading and accumulation of dirt on PV panels, causing thermal imbalance and hotspots on the panels. Therefore, it is important to monitor the operating temperature and homogeneous detection of sunlight on the PV modules to guarantee efficient energy production. In this paper, we present the development and demonstration of a sensor-assisted Internet of Things (IoT)-based photovoltaic-battery renewable energy system. The adoption of the IoT solution for monitoring the real-time variations in environmental factors and system performance is discussed here. For the PV-battery hardware module, solar panels along with rechargeable batteries are constructed to supply the system. Inverters and controllers are used to synchronize the voltage level and transformation of AC power from DC power. In the design of the IoT system, the Arduino Mega microcontroller and ESP32 TTGO board are used along with sensors for recording the temperature, presence of dust/dirt, and voltage and current levels. The working prototype enables real-time data to be captured and sent to the cloud database for data collection, performance analysis, and diagnosis/detection of faults in the proposed system

Spirulina platensis Suppressed iNOS and Proinflammatory Cytokines in Lipopolysaccharide-Induced BV2 Microglia

The disease burden of neurodegenerative diseases is on the rise due to the aging population, and neuroinflammation is one of the underlying causes. Spirulina platensis is a well-known superfood with numerous reported bioactivities. However, the effect of S. platensis Universiti Malaya Algae Culture Collection 159 (UMACC 159) (a strain isolated from Israel) on proinflammatory mediators and cytokines remains unknown. In this study, we aimed to determine the anti-neuroinflammatory activity of S. platensis extracts and identify the potential bioactive compounds. S. platensis extracts (hexane, ethyl acetate, ethanol, and aqueous) were screened for phytochemical content and antioxidant activity. Ethanol extract was studied for its effect on proinflammatory mediators and cytokines in lipopolysaccharide (LPS)-induced BV2 microglia. The potential bioactive compounds were identified using liquid chromatography-mass spectrometric (LC-MS) analysis. Ethanol extract had the highest flavonoid content and antioxidant and nitric oxide (NO) inhibitory activity. Ethanol extract completely inhibited the production of NO via the downregulation of inducible NO synthase (iNOS) and significantly reduced the production of tumor necrosis factor (TNF)-&alpha; and interleukin (IL)-6. Emmotin A, palmitic amide, and 1-monopalmitin, which might play an important role in cell signaling, have been identified. In conclusion, S. platensis ethanol extract inhibited neuroinflammation through the downregulation of NO, TNF-&alpha; and IL-6. This preliminary study provided insight into compound(s) isolation, which could contribute to the development of precision nutrition for disease management

Carrageenophyte Kappaphycus malesianus Inhibits Microglia-Mediated Neuroinflammation via Suppression of AKT/NF-&kappa;B and ERK Signaling Pathways

Neuroinflammation is an inflammatory response in any part of the central nervous system triggered by the activation of microglia and astrocytes to produce proinflammatory cytokines in the brain. However, overproduction of proinflammatory cytokines further contributes to the development of neurodegenerative disorders. Red seaweed, Kappaphycus malesianus, is a predominant carrageenophyte commercially cultivated in Semporna, Sabah, Malaysia. It is an important source of raw material for kappa-carrageenan productions in the food, pharmaceutical and cosmetics industries. However, no studies have been conducted focusing on the antineuroinflammatory effects of K. malesianus. The aim of the present study was to investigate the effect of the antineuroinflammatory activity of K. malesianus extracts (ethyl acetate, ethanol and methanol) on lipopolysaccharide-stimulated BV2 microglia and the underlying mechanisms involved in the regulation of neuroinflammatory pathways. Extract with the most promising antineuroinflammatory activity was analyzed using liquid chromatography-mass spectrometry (LC-MS). Our results show that methanol extract has a convincing antineuroinflammatory effect by suppressing both AKT/NF-&kappa;B and ERK signaling pathways to inhibit the expression of all proinflammatory cytokines without causing a cytotoxicity effect. LC-MS analysis of methanol extract revealed two compounds: prosopinine and eplerenone. Our findings indicated that metabolites of K. malesianus are potent antineuroinflammatory agents with respect to prevention of neurological disorders