Impact of homocysteine on acute ischemic stroke severity: possible role of aminothiols redox status.

BACKGROUND: Acute ischemic stroke (AIS) is one of the most common cerebrovascular diseases which accompanied by a disruption of aminothiols homeostasis. To explore the relationship of aminothiols with neurologic impairment severity, we investigated four aminothiols, homocysteine (Hcy), cysteine (Cys), cysteinylglycine (CG) and glutathione (GSH) in plasma and its influence on ischemic stroke severity in AIS patients. METHODS: A total of 150 clinical samples from AIS patients were selected for our study. The concentrations of free reduced Hcy (Hcy), own oxidized Hcy (HHcy), free reduced Cys (Cys), own oxidized Cys (cysteine, Cyss), free reduced CG (CG) and free reduced GSH (GSH) were measured by our previously developed hollow fiber centrifugal ultrafiltration (HFCF-UF) method coupled with high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). The concentration ratio of Hcy to HHcy (Hcy/HHcy), Cys to Cyss (Cys/Cyss) were also calculated. The neurologic impairment severity of AIS was evaluated using National Institutes of Health Stroke Scale (NIHSS). The Spearman correlation coefficient and logistic regression analysis was used to estimate and perform the correlation between Hcy, HHcy, Cys, Cyss, CG, GSH, Hcy/HHcy, Cys/Cyss and total Hcy with NIHSS score. RESULTS: The reduced Hcy and Hcy/HHcy was both negatively correlated with NIHSS score in AIS patients with P = 0.008, r=-0.215 and P = 0.002, r=-0.249, respectively. There was no significant correlation of Cys, CG, GSH, HHcy, Cyss, Cys/Cyss and total Hcy with NIHSS score in AIS patients with P value > 0.05. CONCLUSIONS: The reduced Hcy and Hcy/HHcy, not total Hcy concentration should be used to evaluate neurologic impairment severity of AIS patient.

Individuals Diagnosed With Type 2 Diabetes Mellitus and the Status of Vitamin B12 Deficiency: A Review.

Type 2 diabetes mellitus (T2DM) is a complex metabolic disorder with a multifactorial etiology and a significant global burden. In recent years, emerging evidence has suggested a potential link between T2DM and vitamin B12 deficiency, raising concerns about its impact on disease progression, management, and associated complications. This comprehensive review critically examines the current understanding of the prevalence, risk factors, clinical implications, and management strategies related to vitamin B12 deficiency in individuals diagnosed with T2DM. The review begins by providing an overview of the epidemiology of T2DM and its associated complications, underscoring the need for comprehensive management approaches. Subsequently, it delves into the physiology of vitamin B12, including its sources, absorption mechanisms, and biological functions, laying the groundwork for understanding the potential implications of deficiency in T2DM. A thorough analysis of the literature is conducted to elucidate the prevalence and risk factors of vitamin B12 deficiency in individuals with T2DM, considering factors such as age, duration of diabetes, medication use (e.g., metformin), dietary patterns, and comorbidities. Special attention is given to the role of metformin, the first-line therapy for T2DM, in precipitating or exacerbating vitamin B12 deficiency through mechanisms involving alterations in the gut microbiota and intestinal absorption. The review further explores the clinical manifestations and diagnostic challenges associated with vitamin B12 deficiency in the context of T2DM, emphasizing the importance of recognizing subtle symptoms and implementing appropriate screening protocols. It discusses the potential implications of vitamin B12 deficiency on glycemic control, diabetic neuropathy, cognitive function, cardiovascular health, and overall quality of life in individuals with T2DM. In addressing the management of vitamin B12 deficiency in T2DM, the review examines various therapeutic strategies, including oral and parenteral supplementation, dietary modifications, and lifestyle interventions. It critically evaluates the evidence supporting routine screening for vitamin B12 deficiency in individuals with T2DM and discusses controversies surrounding optimal supplementation protocols, dosing regimens, and monitoring strategies. Furthermore, the review highlights gaps in current knowledge and identifies areas for future research, such as the long-term effects of vitamin B12 supplementation on clinical outcomes in T2DM, the impact of genetic factors on vitamin B12 metabolism, and the potential role of personalized interventions. Overall, this review consolidates existing evidence and provides insights into the complex relationship between T2DM and vitamin B12 deficiency, aiming to inform clinical practice, enhance patient care, and guide future research endeavors in this important area of metabolic medicine.

Yoga: A Natural Solution to Decrease Disease Burden in Children of MTHFR Deficient Parents.

Introduction: MTHFR being a key regulatory enzyme of 1-carbon metabolism pathway serves critical function of generation of SAM, replenishment of glutathione and nucleotide synthesis and finally methylation of the bio molecules. MTHFR gene mutation is a rare au-tosomal recessive inborn error of metabolism and presents with severe hyperhomocysteinemia. MTHFR polymorphisms on the other hand are commonly encountered of which two 677 C>T and 1298 A>C have been most widely studied and reported to increase the vulnerability to neural tube defects, congenital heart disease, various neuropsychiatric disorders like autism spectrum diseases and attention deficit hyperactiv-ity disease, cleft lip/ palate, acute leukaemia, cardiovascular diseases, occlusive vascular disease in children. Methods: We conducted this prospective clinical trial to examine whether yoga practice can up regulate MTHFR gene expression. Considering the prevalence of MTHFR polymorphism, varied spectrum of its implications in disease causation including male infertility, we conducted the trial involving 30 infertile men who underwent 3 weeks of supervised YBLI. Pre and post intervention assessment of the blood and semen sample was done to see the effects. Results: We have found more than fivefold up-regulation in the expression of MTHFR gene with significant reduction of seminal free radical levels after 3 weeks of yoga practice. Interestingly we noticed significantly higher MTHFR polymorphic variants in infertile male patients compared to healthy fertile controls. Conclusion: MTHFR polymorphisms are also independently associated with many paediatric diseases. Diagnosing MTHFR deficiency in children is a challenging job and requires high index of suspicion and continuous vigilance. Yoga based lifestyle may be adopted both by parents planning conception and also by adolescent children who are sufferers of this condition to halt the consequences of mild to moderate MTHFR deficiency.

Selective Homocysteine Assay with Cucurbit[7]uril by pH Regulation.

We report the effect of pH on the supramolecular complexation of two biothiols, viz., homocysteine (Hcy) and cysteine (Cys), with cucurbit[7]uril (CB[7]). Under basic pH conditions, Cys did not complex with CB[7], whereas Hcy efficiently complexed with CB[7], as confirmed by 1H NMR spectroscopy and Ellman's reagent (5,5'-dithio-bis(2-nitrobenzoic acid), DTNB) assay. 1H NMR and Raman spectroscopic studies revealed that, in the absence of CB[7], Hcy auto-oxidized slowly (~36 h) to homocystine (HSSH) under basic pH conditions. However, the rate of Hcy oxidation increased by up to 150 fold in the presence of CB[7], as suggested by the DTNB assay. Thus, supramolecular complexation under basic pH conditions led to the formation of a HSSH-CB[7] complex, and not Hcy-CB[7]. The results indicate that Hcy is rapidly oxidized to HSSH under the catalysis of CB[7], which acts as a reaction chamber, in basic pH conditions. Our studies suggest that Hcy concentration, a risk factor for cardiovascular disease, can be selectively and more easily quantified by supramolecular complexation with CB [7].

Sensitive determination of tobramycin using homocystine capped gold nanoclusters as probe by second-order scattering.

A novel photoluminescent Hcy-AuNCs has been developed through one-pot reduction method, to establish a tobramycin sensing by second-order scattering (SOS). Hcy-AuNCs could spontaneously assemble to small-scaled aggregation, resulting in remarkable intensity enhancement of scattered luminescence signals. The luminescence of Hcy-AuNCs could be clearly observed under ultraviolet lamp, when excited at 365 nm, a significant luminescent intensity at 741 nm was monitored in SOS spectra. The introduction of AuNPs would cause large-scaled aggregation of Hcy-AuNCs that was rapidly settled in the solution, resulting in the decrease of SOS intensity. Besides, the non-radiative energy transfer between AuNPs and Hcy-AuNCs would also reduce the luminescent intensity. However, the addition of tobramycin would cause the aggregation of AuNPs due to the electrostatic and covalent bonding between AuNPs and tobramycin, thus eliminating the interference of AuNPs. The luminescence of Hcy-AuNCs reappeared, exhibiting an optical response toward tobramycin. The good linearity was obtained in a wide range from 4 nM to 300 nM with a low detection limit of 0.27 nM. The selectivity was acceptable toward different types of antibiotics. Finally, the proposed method was successfully applied to the widely used tobramycin eye drops.

Assessing Diagnostic Accuracy of Serum Holotranscobalamin (Active-B12) in Comparison with Other Markers of Vitamin B12 Deficiency.

About 15-40% India is Vitamin B12 deficient (commonly diagnosed by total Vitamin B12) but, as only holoTC (active form) is taken up by body cells, thus measuring holoTC is more reflective of Vitamin B12 status than the former. We aimed to assess diagnostic accuracy of serum holoTC in comparison with total Vitamin B12 and total Homocysteine (HCY) as indicator of serum Vitamin B12 status. 217 human subjects (99 males and 118 females) ranging from 17 to 83 years were divided into Vitamin B12 deficient (n = 70), borderline (n = 100) and sufficient groups (n = 47) who were further assessed for markers of Vitamin B12 deficiency-holoTC, HCY, Mean Corpuscular Volume (MCV), Folate, heamoglobin and creatinine. Samples were analysed using Siemens Advia Centaur Xpi. Total Vitamin B12 deficient group had - 84.3% holoTC deficient; 15.7% holoTC sufficient; 72.9% with elevated HCY; 27.1% with normal HCY; 11.4% with megaloblastic anaemia. Borderline group had - 34% holoTC deficient; 28% elevated HCY. A strong positive correlation was found between Total Vitamin B12 and holoTC (r = 0.754, p = <0.001) but strong negative correlation existed between holoTC and HCY (r = - 0.471, p = <0.001). Concordance between Total Vit B12 and HCY (Kappa index = 0.51, p < 0.001); between holoTC and HCY (Kappa index = 0.52, p = <0.001) were statically significant but the latter had a better sensitivity and specificity. Also, statically significant association exists between Total Vitamin B12 and holoTC with HCY (p = <0.001). Therefore, it is ascertained that Active Vitamin B12 assay is a better test and can be considered as an early marker of vitamin B12 deficiency.

The Staphylococcus aureus Cystine Transporters TcyABC and TcyP Facilitate Nutrient Sulfur Acquisition during Infection.

Staphylococcus aureus is a significant human pathogen due to its capacity to cause a multitude of diseases. As such, S. aureus efficiently pillages vital nutrients from the host; however, the molecular mechanisms that support sulfur acquisition during infection have not been established. One of the most abundant extracellular sulfur-containing metabolites within the host is cysteine, which acts as the major redox buffer in the blood by transitioning between reduced and oxidized (cystine) forms. We therefore hypothesized that S. aureus acquires host-derived cysteine and cystine as sources of nutrient sulfur during systemic infection. To test this hypothesis, we used the toxic cystine analogue selenocystine to initially characterize S. aureus homologues of the Bacillus subtilis cystine transporters TcyABC and TcyP. We found that genetic inactivation of both TcyA and TcyP induced selenocystine resistance. The double mutant also failed to proliferate in medium supplemented with cystine, cysteine, or N-acetyl cysteine as the sole sulfur source. However, only TcyABC was necessary for proliferation in defined medium containing homocystine as the sulfur source. Using a murine model of systemic infection, we observed tcyP-dependent competitive defects in the liver and heart, indicating that this sulfur acquisition strategy supports proliferation of S. aureus in these organs. Phylogenetic analyses identified TcyP homologues in many pathogenic species, implying that this sulfur procurement strategy is conserved. In total, this study is the first to experimentally validate sulfur acquisition systems in S. aureus and establish their importance during pathogenesis.

Effect of Yiqi-Yangyin-Quyu decoction combined with acupuncture on serum homocystine level and blood coagulation function of patients with cerebral infarction in convalescent period / 国际中医中药杂志

Objective To observe the effect of Yiqi-Yangyin-Quyu decoction combined with acupuncture on serum homocystine level and blood coagulation function of patients with cerebral infarction in convalescent period. Methods A total of 80 patients with cerebral infarction convalescence were divided into the observation group and the control group according to the random number table method, with 40 cases in each group. The control group was given routine treatment of western medicine. Based on this, the observation group was treated with Yiqi-Yangyin-Quyu decoction combined with acupuncture. After 4 weeks of treatment, the Barthel index (BI), neurological impairment National Institutes of Health Stroke Scale (NIHSS), C-reactive protein (CRP), and the level of homocystine (HCY) and blood coagulation function index of plasma prothrombin time (PT), thrombin time (TT), activated partial thromboplastin time (APTT), fibrinogen (FIB)] of the two groups were compared and analyzed. Results The total effective rate of the observation group was 90.0％ (36/40), while the total effective rate of the control group was 70.0％ (28/40). The total effective rate of the 2 groups was statistically significant (Z=-3.184, P=0.004). After treatment, the NIHSS score of the observation group were lower than those of the control group,and the BI score of the observation group were higher than those of the control group(t value were 4.528, 4.337, P<0.05). After treatment, the content of CRP (9.25 ± 1.21 mg/L vs. 15.24 ± 1.74 mg/L, t=4.905) and HCY (12.59 ± 2.05 μmol/L vs. 16.52 ± 2.05 μmol/L, t=4.821) of the observation group were significantly lower than those of the control group (P<0.05). After treatment, the level of PT (15.20 ± 1.17 s vs. 12.95 ± 1.02 s, t=4.891), APTT (36.72 ± 1.98 s vs. 33.02 ± 1.68 s, t=4.553), and TT (14.67 ± 0.95 s vs. 12.21 ± 1.10 s, t=4.210) of the observation group were significantly higher than those of the control group (P<0.05), while the level of FIB (2.55 ± 0.25 g/L vs. 3.03 ± 0.51 g/L, t=4.027) of the observation group was significantly lower than this of the control group (P<0.05). Conclusions The application of Yiqi-Yangyin-Quyu decoction combined with acupuncture can improve the blood coagulation function, reduce the injury of brain tissue, promote the recovery of cerebral nerve function, and improve the quality of life of the patients.

Quantitative Analysis of Total Plasma Homocysteine by LC-MS/MS.

Homocysteine is a nonessential, sulfur-containing amino acid involved in one-carbon (folate) metabolism. A number of inherited and acquired conditions cause increased accumulation of this metabolite in blood (homocysteinemia) and other biofluids. Homocysteinemia is a risk factor for cardiovascular disease, including recurrent thrombosis. Accurate measurement of total plasma homocysteine is an important element in the diagnostic evaluation of these disorders. While a number of different methods have been developed for this purpose, the focus of this unit will be on a specific technique utilizing liquid chromatography-tandem mass spectrometry, which provides several advantages in terms of speed, sensitivity, and specificity.

Association between homocystine and C677T polymorphism with gestational diabetes mellitus / 国际检验医学杂志

Objective To investigate the relationship between homocystine (Hcy) ,methylenetetra hydrofolate reductase(M T H-FR) C677T and gestational diabetes mellitus (GDM ) .Methods A total of 91 GDM cases(GDM group) and 123 cases with normal pregnancy(control group) were detected for the C677T polymorphism of M T HFR and serum levels of Hcy and glucose .Results Serum Hcy level in GDM group was remarkable higher than that of control group (P< 0 .05) .Hcy level was positively correlated with fasting plasma glucose ( P < 0 .05) .The genotype frequencies (CC ,CT ,TT ) of M T HFR C677T in GDM group and control group were with significantly difference(P< 0 .05) .Hcy was significantly higher in women with C677T TT genotype than those with CC genotype(P< 0 .05) .Conclusion Hcy could be related to GDM .The mutation of M T HFR might affect serum Hcy level , and be involved in the occurrence and development of GDM .

Determination of cysteine, homocysteine, cystine, and homocystine in biological fluids by HPLC using fluorosurfactant-capped gold nanoparticles as postcolumn colorimetric reagents.

We have demonstrated for the first time the suitability of fluorosurfactant-capped spherical gold nanoparticles as HPLC postcolumn colorimetric reagents for the direct assay of cysteine, homocysteine, cystine, and homocystine. The success of this work was based on the use of an on-line tris(2-carboxyethyl)phosphine reduction column for cystine and homocystine. Several parameters affecting the separation efficiency and the postcolumn colorimetric detection were thoroughly investigated. Under the optimized conditions, cysteine, homocysteine, cystine, and homocystine in human urine and plasma samples were determined. Detection limits for cysteine, homocysteine, cystine, and homocystine ranged from 0.16-0.49 µM. The accuracy in terms of recoveries ranged between 94.0-102.1%. This proposed method was rapid, inexpensive, and simple.

Cu(II)-disulfide complexes display simultaneous superoxide dismutase- and catalase-like activities.

Superoxide is a potentially toxic by-product of cellular metabolism. We have addressed here the in vitro ability of complexes formed between copper(II) ions and various biologically-occurring disulfides (RSSR: oxidized glutathione, cystine, homocystine and α-lipoic acid) to react with superoxide. The studied complexes were found to react with superoxide (generated by a xanthine/xanthine oxidase system) at rate constants (kCu(II)-RSSR) close to 10(6)M(-1)s(-1), which are three orders of magnitude lower than that reported for superoxide dismutase (SOD) but comparable to that of several other copper-containing complexes reported as SOD mimetics. The interaction between the tested Cu(II)-RSSR and superoxide, led to the generation and recovery of concentrations of hydrogen peroxide and oxygen that were, respectively, below and above those theoretically-expected from a sole SOD mimetic action. Interestingly, oxygen was generated when the Cu(II)-RSSR complexes were directly incubated with hydrogen peroxide. Taken together, these results reveal that the Cu(II)-RSSR complexes not only have the capacity to dismutate superoxide but also to simultaneously act like catalase mimetic molecules. When added to superoxide-overproducing mitochondria (condition attained by its exposure to diclofenac), three of the tested complexes were able (2-4µM), not only to totally restore, but also to lower below the basal level the mitochondrial production of superoxide. The present study is first in reporting on the potential of Cu(II)-disulfide complexes to act as SOD and catalase like molecules, suggesting a potential for these types of molecules to act as such under physiological and/or oxidative-stress conditions.

Toward decentralized analysis of mercury (II) in real samples. A critical review on nanotechnology-based methodologies.

In recent years, it has increased the number of works focused on the development of novel nanoparticle-based sensors for mercury detection, mainly motivated by the need of low cost portable devices capable of giving fast and reliable analytical response, thus contributing to the analytical decentralization. Methodologies employing colorimetric, fluorometric, magnetic, and electrochemical output signals allowed reaching detection limits within the pM and nM ranges. Most of these developments proved their suitability in detecting and quantifying mercury (II) ions in synthetic solutions or spiked water samples. However, the state of art in these technologies is still behind the standard methods of mercury quantification, such as cold vapor atomic absorption spectrometry and inductively coupled plasma techniques, in terms of reliability and sensitivity. This is mainly because the response of nanoparticle-based sensors is highly affected by the sample matrix. The developed analytical nanosystems may fail in real samples because of the negative incidence of the ionic strength and the presence of exchangeable ligands. The aim of this review is to critically consider the recently published innovations in this area, and highlight the needs to include more realistic assays in future research in order to make these advances suitable for on-site analysis.

Clinical Study on Relationship between Polymorphism in Methylenetetrahydrofolate Reductase and Hcy and Acute Cerebral Infarction / 中国康复理论与实践

@#Objective To investigate the relationship between polymorphism in methylenetetra-hydrofolate reductase (MTHFR ) and acute cerebral infarction (CI), observe the variation regular of fasting plasma homocysteine (Hcy) level.Methods Using Homocysteine Microplate STE Assay to examine the fasting plasma homocysteine level of 28 CI patients during their initial stage (flaring up between 1 to 3 days) and later stage (flaring up 10 to 15 days) of acute period and 27 healthy controls. The presence of the MTHFR genetic type was determined by polymerase chain reaction (PCR) assay and subsequent restriction enzyme digestion.Results There was no significant difference among the three MTHFR genotypes in distributed frequency of the CI group, normal controls and the 677 allelic gene (P>0.05). The discrepancy of Hcy level in various kinds of genotypes: heterozygote mutation and homozygoto mutation were much higher than wild type (P<0.01). Homozygoto mutation was higher than heterozygote mutation, but there was no significant difference between them (P>0.05). The high homocysteine of group CI during the acute early stage were found out more frequent than normal control (P<0.05). There was no significant difference of fasting plasma Hcy level between the initial stage and later stage of CI group which were in acute period (P>0.05), both of the Results were higher than normal control (P<0.01). There was no significant difference among the Hcy level of various genetypes in CI group during the initial stage and later stage of acute period (P<0.05).Conclusion MTHFR gene C677T mutation is one of the cause of high homocystinemia, while it dose not lead to CI directly. High Hcy level is the independent risk factor of CI, but has no concern to the course of acute CI.

Relationship between serum homocystine and diabetic nephropathy and uremia in the patients with type 2 diabets mellitus / 中国基层医药

Objective To explore the relationship between homocystine(HCY) and diabetic nephropathy and uremia in the patients with type 2 diabetes mellitus.Methods 20 cases with type 2-DM,20 cases with incipient diabetic nephropathy and 17 cases with uremia were collected.The plasma levels of HCY were measured by FPIA,the fasting blood sugar were measured by standard automated clinical chemistry laboratory methods.The plasma levels of HCY were compared with each other with t test.Results The plasma levels of HCY between the type 2-DM and diabetic nephropathy were significantly different,(26.16?12.31)?mol/L vs (8.79?3.51)?mol/L,P

Relationship of plasma homocysteine level and pathological change of large artery in stroke patients / 基础医学与临床

Objective To determine the association between homocysteine and severity of large cerebral artery atherosclerosis.Methods Total 152 ischemic stroke patients were evaluated using transcranial Doppler(TCD),magnetic resonance angiography(MRA) or digital substraction angiography(DSA).Severity of cerebral artery atherosclerosis was scaled by number of stenosed or occluded artery.Results There were 83 patients in low level homocysteine group with mean Hcy level(15.31?3.08)?mol/L,while 69 patients in high level homocysteine group with mean Hcy(34.47?21.38)?mol/L.Number of abnormal intra-and extra-artery was 1.86?1.51 and 1.52?1.46 respectively.No significant difference was demonstrated in the number of abnormal artery between two groups.No correlation was noted between Hcy and number of abnormal artery.Conclusion There is no relationship between high homocysteine and severity of large cerebral artery atherosclerosis in ischemic stroke patients.Homocysteine might be the marker of the disease.

Influence of plasma homocystine level on the prognosis of acute cerebral infarction and the effect of interventional therapy / 临床神经病学杂志

Objective To explore the influence of plasma homocystine(Hcy) level on the prognosis of acute cerebral infarction(ACI) and the effect of interventional therapy.Methods The plasma Hcy levels of 152 patients with ACI were measured by fluorometric method at

A case of homocystinuria

No abstract available.