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Objective: To compare the reduction in mean pain score with local Methylcobalamin injection versus local Bupivacaine injection in patients with acute herpetic neuralgia. Study Design: Quasi experimental study. Place and Duration of Study: Dermatology Outpatient Department, Pak Emirates Military Hospital, Rawalpindi, from Jun to Dec 2019. Methodology: Total 100 patients, having pain score more than 3, fulfilling the selection criteria were divided into two groups. Group A was treated with daily subcutaneous injection Bupivacaine, whereas Group B was treated with daily subcutaneous injection Methycobalamin at the site of neuralgia. Patients were followed up for 4 weeks. The pain score was noted. All the data was entered and analyzed on SPSS version 21. Results: In this study mean age of patients in group A was 43.82 ± 15.76 years and in group B was 44.76 ± 16.92 years. The mean visual analogue pain score at 4th week in the group A patients was 1.14 ± 0.32 and in group B was 1.90 ± 0.97. Statistically significant difference was found in group A (local Bupivacaine) with visual analogue score (VAS) at 4th week (pvalue=0.002). Conclusion: The local Bupivacaine injection showed significant reduction in mean pain score than local Methylcobalamin injection in patients with acute herpetic neuralgia.

A new rapid, accurate, precise and economical reverse phase high performance liquid chromatographic method has been developed and validated for simultaneous estimation of Methylcobalamin, Epalrestat and Pregabalin in bulk and pharmaceutical dosage form. The separation was accomplished utilizing Agilent C18, 150 x 4.6mm, 5 column at a detection wavelength of 210nm utilizing the mobile phase water and acetonitrile 60: 40 v/v at the flow rate of 0.8ml/min and injection volume of 10µl. The total run time was 6.0 min. Validation discovered the method was specific, rapid, accurate, precise, reliable and reproducible. The calibration curve was linear over the concentration range of 0.37 – 2.25μg/ml of Methylcobalamin, 37.5-225μg/ml of Epalrestat and 37.5-225μg/ml of Pregabalin respectively with correlation coefficient of 0.999. The accuracy was determined by recovery studies and was found to be 99.5-100%. The precision of the results stated that the %RSD was

Актуальність. Діабетична нейропатія — найбільш часте ускладнення цукрового діабету (ЦД), що зустрічається у 30–90 % пацієнтів. Актуальність дослідження обумовлена поширеністю діабетичної нейропатії, а також оцінкою ефективності використання метильованої форми вітаміну В12 в лікуванні пацієнтів із ЦД. Метою даного дослідження було визначення ефективності метилкобаламіну (препарат Діакобал, «Кусум Фарм») у комплексному лікуванні пацієнтів із ЦД, ускладненим діабетичною периферичною нейропатією (ДПН). Матеріали та методи. Дослідження проведене простим відкритим методом. Для скринінгу та визначення вираженості симптомів ДПН використовували Мічиганський опитувальник для скринінгу нейропатії (The Michigan Neuropathy Screening Instrument, MNSI), шкалу неврологічних симптомів (Neurological Symptoms Score, NSS), Європейський опитувальник оцінки якості життя (European Quality of Life Questionnaire, EQ-5D). Неврологічне обстеження пацієнтів включало оцінку різних видів чутливості (больової, тактильної, вібраційної, холодової, теплової, пропріоцептивної, відчуття тиску). Усім пацієнтам визначали рівень у крові вітаміну В12 (ціанокобаламіну). Пацієнтам із ДПН, незалежно від показників вмісту вітаміну В12, до схеми традиційного лікування ДПН додавали Діакобал. Результати. Показано, що ДПН є поширеним ускладненням ЦД: за суб’єктивними характеристиками (шкали MNSI та NSS) її симптоми виявлено в 84,3 % випадків серед пацієнтів із ЦД. Суб’єктивних шкал, таких як EQ-5D, MNSI, NSS, недостатньо для оцінки наявності та ступеня полінейропатії у хворих на ЦД. Дані цих шкал не корелюють із результатами фізикального обстеження, але можуть застосовуватися як допоміжний метод для аналізу суб’єктивних симптомів пацієнта. Неврологічне обстеження рекомендоване пацієнтам із ЦД незалежно від того, чи пред’являють вони активні скарги. ДПН виявлена в 64,8 % випадків серед пацієнтів із ЦД за об’єктивними критеріями діагностики та визначенням різних видів чутливості (больової, тактильної, вібраційної, холодової, теплової, пропріоцептивної, відчуття тиску). Визначення рівня ціанокобаламіну має використовуватися в комплексі з визначенням гомоцистеїну та фолієвої кислоти. Висновки. Застосування метилкобаламіну (препарат Діакобал у добовій дозі 1500 мкг протягом 3 місяців) у комплекс­ному лікуванні діабетичної нейропатії в пацієнтів із ЦД є ефективним і безпечним, сприяє усуненню проявів діабетичної нейропатії та поліпшенню якості життя пацієнтів.

Актуальность. К нежелательным побочным эффектам при использовании метформина относится нарушение обмена витамина В12, ассоциирующееся со снижением уровня В12 в сыворотке крови. Цель: установить эффективность применения метилкобаламина (препарат Диакобал производства ООО «Кусум Фарм», Украина) в коррекции дефицита витамина В12 у больных сахарным диабетом (СД) 2­го типа с метформин­ассоциированным дефицитом витамина В12, а также установить его влияние на проявления диабетической нейропатии. Материалы и методы. Под наблюдением находилось 30 больных СД 2­го типа с метформин­ассоциированным дефицитом витамина В12 (уровень витамина В12 ниже 190 пг/мл). Продолжительность приема метформина составляла в среднем 6,1 года, средняя доза — 2100 мг/сут. Метилкобаламин назначали в дозе 500 мг три раза в сутки в течение трех месяцев. Определение содержания витамина В12 и оценку проявлений нейропатии проводили до и после курса лечения. Результаты. После завершения курса лечения установлено, что среднее содержание витамина В12 в сыворотке крови достигло референсных значений (312,4 ± 39,8 пг/мл). Выявлено положительное влияние терапии на выраженность клинических проявлений периферической сенсорно­моторной нейропатии у обследованных больных: у 17 (56,7 %) пациентов значительно уменьшилась выраженность онемения стоп, у шести (20 %) больных было отмечено полное исчезновение онемения. Интенсивность болевого синдрома уменьшилась у 19 (63,3 %) пациентов, болевой синдром удалось полностью снять у троих (10 %) больных. Выводы. Подтверждена взаимосвязь приема метформина и гиповитаминоза В12. Диакобал производства ООО «Кусум Фарм» (Украина) является эффективным и безопасным препаратом метилкобаламина в комплексной терапии сахарного диабета для профилактики и лечения метформин­индуцированного дефицита витамина В12 и диабетической нейропатии.

Objective: To observe the clinical effect of hyperbaric oxygen in the treatment of sudden deafness. Methods: Ninety-six patients with sudden deafness diagnosed by the otolaryngology department were divided into 2 groups which comprised of 48 patients in the conventional treatment group and the other 48 patients in the hyperbaric oxygen treatment group. Both groups were treated with methylcobalamin, vitamin B1, and ginkgo biloba extract. The patients in the hyperbaric oxygen treatment group were given hyperbaric oxygen therapy of 2.0 ATA once a day. Each course of treatment lasted 10 days and after two courses of treatment, the clinical efficacies of the two groups were compared. Results: After two courses of treatment, the effect of treatment in the hyperbaric oxygen treatment group was significantly better than that of the conventional treatment group. Conclusion: Hyperbaric oxygen therapy can effectively improve the hearing level of patients with sudden deafness and the overall effective rate of treatment.

Introduction: Age related hearing loss is defined as a bilateral, progressive, symmetrical age-related sensorineural hearing loss, which is most pronounced at the higher frequencies. It is considered that oxidative stress has a central role in the pathology of age-related hearing loss so, this study was conducted to test the efficacy and safety for the combination of antioxidants for the treatment of age-related hearing loss. Methodology: This was a multi-centric study conducted at 11 clinical trial sites. Out of total 176 enrolled, 160 trial subjects completed the study. For efficacy assessment, 2 parameters were used including hearing related quality of life score and score obtained from hearing impairment questionnaire on day 0 (visit 1), 45 (visit 2) and 90 (visit 3). Results: At visit 1, the mean hearing related quality of life score was 4.437 increased to 5.725 at visit 2 and further increased to 6.906 at visit 3. At visit 2 and 3, the increase in hearing related quality of life score was 29.028% and 55.645% respectively as compared to baseline. The score obtained from hearing impairment questionnaire was 25.581 at visit 1 reduced to 16.200 and further reduced to 9.943 at visit 3. Conclusion: The combination of Alpha lipoic acid, Gingko biloba, Vitamin C, Zinc, Magnesium, Vitamin B6, Methyl cobalamin, Vitamin E and Chromium Picolinate can be used for the medical management of age related hearing loss.

Introduction: Sudden sensorineural hearing loss (SSNHL) is defined as sensorineural hearing loss of 30 dB or more over 3 contiguous audiometric frequencies occurring over 72hrs. Conventionally the SSNHL is treated by corticosteroids but it is considered that, addition of antioxidants to the treatment can increase the percentage of patients with the complete recovery. So, this study was conducted to test the efficacy and safety for the combination of Alpha lipoic acid, Gingko biloba, Vitamin C, Zinc, Magnesium, Vitamin B6, Methylcobalamin, Vitamin E and Chromium Picolinate for the treatment of SSNHL and high Intensity noise and vibration induced hearing loss as a concomitant therapy in addition to the standard medical management. Methodology: Out of total 225 enrolled, 208 patients completed the study. Efficacy and safety evaluation was done on day 7 and 14 considering the baseline visit at day 0. Efficacy assessment was done by hearing Impairment questionnaire and hearing related quality of Life. Results: Mean score obtained from hearing impairment questionnaire at day 0 was 19.59 which was reduced to 12.56 at day 7 and was further reduced to 7.70 at day 14. On Visit 1 the mean hearing related quality of life score was 5.24 which was increased to 6.4 at day 7 and was found to be further increased to 7.41 at day 14. Conclusion: The investigational product was efficacious and safe for the treatment of SSNHL and high Intensity noise and vibration induced hearing loss when given as a concomitant therapy to the standard medical management.

Introduction: Tinnitus is the false perception of sound. Normally it is considered to be developed due to oxidative stress to the inner ear. This study was conducted to test the efficacy and safety for the combination of Alpha lipoic acid, Gingko biloba, Vitamin C, Zinc, Magnesium, Vitamin B6, Methylcobalamin, Vitamin E and Chromium Picolinate in the patients of tinnitus. Method: The study was conducted on 165 patients out of which 142 completed the study. Efficacy was evaluated by tinnitus symptom score (TSS). Patients were asked to rate the TSS ranging from 0 to 10 where 0 means no symptom and 10 means maximum tolerable symptoms. Safety assessment was made by analysing the adverse events reported by the patient. Efficacy and safety evaluation was done on day 0, 10 and 22. Results: Patients had TSS 6.26 on baseline visit which was reduced to 4.50 at day 10 and was further reduced to 2.47 at day 22. Also, in the clinical trial duration of 22 days, only 5 episodes of the adverse drug reactions were reported by the patient and all of them were of non-serious in nature and mild in intensity. Conclusion: The fixed dose combination of Alpha lipoic acid 200 mg, Gingko biloba 120 mg, Vitamin C 30 mg, Zinc 12 mg, Magnesium 10 mg, Vitamin B6 3 mg, Methylcobalamin 1500 mcg, Vitamin E 10 IU and Chromium Picolinate 1.66 mg equivalent to elemental chromium 200 mcg was found to be efficacious and safe for the treatment of Tinnitus.

The objective of this study wasto assess the effect of methylcobalaminonmechanical allodynia and the voltage-gated sodium channels (VGSCs) expression of injured nerves in spinal nerve ligation-induced neuropathic pain model in animals.Three different doses of methylcobalamin were administrated intramuscularly into neuropathic pain rat model, twice a week for 14 weeks. The effect of methylcobalamin on neuropathic pain was assessed using mechanical allodynia (using the von Frey filaments) while its effect on VGSC expression was assessed using immunohistochemistry. ANOVA and independent t-test were employed to compared the effect of methylcobalamin on mechanical allodynia between groups.The size of von Frey filament that induced the first onset of mechanical allodynia was smaller in control group compared to 50µg methylcobalamin group (p=0.013) and methylcobalamin 100µg group (p=0.019). There is a dose–response relationship between methylcobalamin dose and the average duration of mechanical allodynia (43.8, 38.2, 30.6 and 29.6 days for control, 50µg, 100µg, and 150µg methylcobalamin group, respectively) with a significant different observed between control and 150µg methylcobalamin group only (p=0.027). Nerve tissues from all animals within control group expressed VGSC while all nerve tissues from both 100µg, and 150µg methylcobalamin, had no VGCS expression. In conclusion, methylcobalamin is potentially shorten the duration of mechanical allodynia and increase pain threshold in neuropathic pain animal model. These effects might associate with reduction of VGSC expression on the injured neurons.

Objective: To assess relative comparison of stability and degradation of Methylcobalamin tablets of different brands at various storage circumstances. Methods: The comparative in vitro study of Methycobal (innovator brand) with its other 5 different brands Cobalamin, Neuromet, Incobal, Qbal and Mecobal was organized for evaluation of physicochemical features of hardness, thickness, friability, weight variation, disintegration time and accelerated stability at 3 temperatures, 25 °C, 30 °C±65 % and 40 °C±75 % respectively for 3 mo. Later all brands were passed through HPLC for checking the extent of degradation of drug products. Results: All tablet brands were within the weight variation specified limits except Mecobal with a relative standard deviation of 6.83%. The weight variation values of Methycobal, Cobalamin, Neuromet, Incobal, Qbal and Mecobal were 0.29%, 0.11%, 0.09%, 0.13%, 0.09% and 0.14% after friability test respectively as per standard limits. The average thickness of Cobalamin, Incobal and Mecobal were not within specified limits. The average hardness of all trades was within limits except Cobalamin and Mecobal exceeding 6kp. The disintegration time of all companies was as per specifications. Conclusion: Qbal was found economical and cost-effective. However, study facts unveiled no noteworthy variety in the Q. C assessments of Methylcobalamin brands.

Methylcobalamin-dependent radical S-adenosylmethionine (SAM) enzymes methylate non-nucleophilic atoms in a range of substrates. The mechanism of the methyl transfer from cobalt to the receiving atom is still mostly unresolved. Here we determine the stereochemical course of this process at the methyl group during the biosynthesis of the clinically used antibiotic fosfomycin. In vitro reaction of the methyltransferase Fom3 using SAM labeled with 1H, 2H, and 3H in a stereochemically defined manner, followed by chemoenzymatic conversion of the Fom3 product to acetate and subsequent stereochemical analysis, shows that the overall reaction occurs with retention of configuration. This outcome is consistent with a double-inversion process, first in the SN2 reaction of cob(I)alamin with SAM to form methylcobalamin and again in a radical transfer of the methyl group from methylcobalamin to the substrate. The methods developed during this study allow high-yield in situ generation of labeled SAM and recombinant expression and purification of the malate synthase needed for chiral methyl analysis. These methods facilitate the broader use of in vitro chiral methyl analysis techniques to investigate the mechanisms of other novel enzymes.

Статтю присвячено питанням лікування полінейропатії діабетичного генезу. Приділено увагу препаратам вітамінів групи В, а саме метилкобаламіну, при даній патології. Продемонстровано дослідження ефективності метилкобаламіну (препарат Нейрокобал®) у 18 пацієнтів із діабетичною полінейропатією. Показано, що застосування даного препарату дозволило зменшити больовий синдром і підвищити поріг вібраційної чутливості в пацієнтів.

Vitamin B12 (cobalamin) is an essential cofactor for two metabolic pathways. It is obtained principally from food of animal origin. Cobalamin becomes bioavailable through a series of steps pertaining to its release from dietary protein, intrinsic factor-mediated absorption, haptocorrin or transcobalamin-mediated transport, cellular uptake, and two enzymatic conversions (via methionine synthase and methylmalonyl-CoA-mutase) into cofactor forms: methylcobalamin and adenosylcobalamin. Vitamin B12 deficiency can masquerade as a multitude of illnesses, presenting different perspectives from the point of view of the hematologist, neurologist, gastroenterologist, general physician, or dietician. Increased physician vigilance and heightened patient awareness often account for its early presentation, and testing sometimes occurs during a phase of vitamin B12 insufficiency before the main onset of the disease. The chosen test often depends on its availability rather than on the diagnostic performance and sensitivity to irrelevant factors interfering with vitamin B12 markers. Although serum B12 is still the most commonly used and widely available test, diagnostics by holotranscobalamin, serum methylmalonic acid, and plasma homocysteine measurements have grown in the last several years in routine practice. The lack of a robust absorption test, coupled with compromised sensitivity and specificity of other tests (intrinsic factor and gastric parietal cell antibodies), hinders determination of the cause for depleted B12 status. This can lead to incorrect supplementation regimes and uncertainty regarding later treatment. This review discusses currently available knowledge on vitamin B12, informs the reader about the pitfalls of tests for assessing its deficiency, reviews B12 status in various populations at different disease stages, and provides recommendations for interpretation, treatment, and associated risks. Future directions for diagnostics of B12 status and health interventions are also discussed.

The oral availability of many drugs is problematic due to the pH of the stomach, enzymes, and first-pass effects through the liver. However, especially geriatric, pediatric, bedridden, or mentally handicapped patients and those with dysphagia have difficulty swallowing or chewing solid dosage forms. Oral Thin Films (OTFs) are one of the new drug delivery systems that can solve these problems. Pregabalin (PG) and Methylcobalamin (MC), which are frequently preferred for pain originating in the central nervous system, were brought together for the first time using OTF technology in this study. In this study, a quantification method for PG and MC was developed and validated simultaneously. Optimum formulations were selected with organoleptic and morphological controls, moisture absorption capacity, swelling capacity, percent elongation, foldability, pH, weight variability, thickness, disintegration time, and transparency tests on OTFs prepared by the solvent pouring method. Content uniformity, dissolution rate, determination of release kinetics, SEM, XRD, FT-IR, DSC, long-term stability, and cytotoxicity studies on the tongue epithelial cell line (SCC-9) were performed on selected OTFs. As a result, OTFs containing PG-MC, which are non-toxic, highly flexible, transparent, compatible with intraoral pH, with fast disintegration time (

Background: The cerebellum principally the motor organ is involved in the regulation of muscular tone and skilled motor movements. The cerebellar histology consists of three layers and the middle is the Purkinje cell layer which consists of pyramidal shaped purkinje cells. Clinical research shows scanty literature on the beneficial effects of Methylcobalamin on Purkinje cells layer. Aim: Our aim was to bring to light the need for prescribing Methylcobalamin in the masses and patient suffering from motor incoordination. Method: 15 animals were given Methylcobalamin and the changes in the thickness of Purkinje cell layer ware recorded at twelve weeks Result: The morphometric analysis showed restored thickness of Purkinje cell layer Conclusion: The recorded data of the regenerated purinje cell layer thickness proved that the use of Methylcobalamin is mandatory as protective drug in damaged neuronal tissue. Key words: Proliferative, Cytostatic, Purkine cell layer

In order to propose a course of action to be taken in the face of any hyperhomocysteinemia, we have reported for the first time in a French journal the recommendations made within the framework of the European E-HOD project for the diagnosis and treatment of remethylation disorders. The remethylation route ensures homocysteine-methionine conversion. It is linked to the folate cycle and the intracellular metabolism of cobalamins. Remethylation disorders can be classified into three groups: 1) isolated disorders (cblD-HC, cblE, cblG) corresponding to an isolated deficit in the production of methylcobalamin, cofactor of methionine synthase; 2) combined disorders (cblC, cblD-MMA/HC, cblF, cblJ) corresponding to an alteration of the transport and intracellular metabolism of cobalamins, which causes a defect in the synthesis of the two functional forms of cobalamin: methylcobalamin and adenosylcobalamin, a cofactor for methyl malonylCoA mutase; 3) MTHFR deficit, an abnormality of the folate cycle. The biological anomalies observed are hyperhomocysteinemia and hypomethioninaemia associated in the case of disorders combined with increased urinary excretion of methylmalonic acid. The clinical presentation is however heterogeneous according to the remethylation disorder but also for the same pathology according to the age. Given the large number of pathologies grouped together in remethylation disorders, this point is illustrated by only two clinical cases concerning the same deficit (deficit in MTHFR) but with different discovery circumstances: a neonatal form and a late form.

Background Vitamin B-12 deficiency is widespread in many parts of the world, affecting all age groups and increasing with age. It is primarily due to a low intake of animal source foods or malabsorption. The measurement of bioavailability of vitamin B-12 is etiologically important in deficiency but is limited due to the use of radioactive isotopes like [57Co]- or [14C]-cyanocobalamin. Objectives The aim of this study was to measure the bioavailability of [13C]-cyanocobalamin in humans and to assess the effect of parenteral replenishment of vitamin B-12 on the bioavailability. Methods We synthesized a stable isotope-labeled vitamin B-12, [13C]-cyanocobalamin, using Salmonella enterica by providing [13C2]-ethanolamine as a sole carbon source. After purification and mass spectrometry-based characterization, its oral bioavailability was measured in the fasted state with high and low oral doses, before and after parenteral replenishment of vitamin B-12 stores, from the kinetics of its plasma appearance in a 2-compartment model. Results [13C]-cyanocobalamin was completely decyanated to [13C]-methylcobalamin describing metabolic utilization, and its plasma appearance showed early and late absorption phases. At a low dose of 2.3 µg, the mean bioavailability was 46.2 ± 12.8 (%, mean ± SD, n = 11). At a higher dose of 18.3 µg, the mean bioavailability was 7.6 ± 1.7 (%, mean ± SD, n = 4). Parenteral replenishment of the vitamin B-12 store in deficient individuals prior to the measurement resulted in a 1.9-fold increase in bioavailability. Conclusions Vitamin B-12 bioavailability is dose dependent and at a low dose that approximates the normal daily requirement (46%). The stable isotope method described here could be used to define the etiology of deficiency and to inform the dietary requirement in different physiologic states as well as the dose required for supplementation and food fortification. This trial was registered at the Clinical Trials Registry of India as CTRI/2018/04/012957.

SARS‐CoV‐2 is the causative agent for the ongoing COVID19 pandemic, and this virus belongs to the Coronaviridae family. Like other members of this family, the virus possesses a positive‐sense single‐stranded RNA genome. The genome encodes for the nsp12 protein, which houses the RNA‐dependent‐RNA polymerase (RdRP) activity responsible for the replication of the viral genome. A homology model of nsp12 was prepared using the structure of the SARS nsp12 (6NUR) as a model. The model was used to carry out in silico screening to identify molecules among natural products, or Food and Drug Administration‐approved drugs that can potentially inhibit the activity of nsp12. This exercise showed that vitamin B12 (methylcobalamin) may bind to the active site of the nsp12 protein. A model of the nsp12 in complex with substrate RNA and incoming NTP showed that vitamin B12 binding site overlaps with that of the incoming nucleotide. A comparison of the calculated energies of binding for RNA plus NTP and methylcobalamin suggested that the vitamin may bind to the active site of nsp12 with significant affinity. It is, therefore, possible that methylcobalamin binding may prevent association with RNA and NTP and thus inhibit the RdRP activity of nsp12. Overall, our computational studies suggest that methylcobalamin form of vitamin B12 may serve as an effective inhibitor of the nsp12 protein.

Methylcobalamin, which is used for the clinical treatment of patients with neuropathy, can have an impact on the sensorineural components associated with the cochlea, and it is possible that the auditory threshold in a certain population of patients with deafness may be recovered. Nonetheless, it remains uncertain whether the action site of methylcobalamin is localized inside or outside the cochlea and which cellular or tissue element is targeted by the drug. In the present work, we developed a method to realize in vivo real-time simultaneous examination of the drug kinetics in two separate locations using boron-doped diamond microelectrodes. First, the analytical performance of methylcobalamin was studied and the measurement protocol was optimized in vitro. Then, the optimized protocol was applied to carry out real-time measurements inside the cochlea and the leg muscle in live guinea pigs while systemically administering methylcobalamin. The results showed that the methylcobalamin concentration in the cochlea was below the limit of detection for the microelectrodes or the drug did not reach the cochlea, whereas the compound clearly reached the leg muscle.

In 44 years of practicing stroke prevention, I have learned many lessons; in this article, I hope to impart some of them. Three areas of my research are discussed. Controlling resistant hypertension is markedly improved by physiologically individualized therapy based on renin/aldosterone phenotyping; this is particularly important in black patients. Measurement of carotid plaque burden strongly predicts cardiovascular risk and is useful for genetic research and for a process called treating arteries instead of risk factors. Doing so in high-risk patients with asymptomatic carotid stenosis was associated with a >80% reduction in the 2-year risk of stroke and myocardial infarction. It also permitted the identification of extremes of atherosclerosis that are useful for studying both the genetics and the biology of atherosclerosis. Patients with very high plaque burden despite low levels of risk factors have an unexplained phenotype; those with little or no plaque despite high levels of risk factors are protected. Patients with unexplained atherosclerosis have higher plasma levels of toxic metabolites produced by the intestinal microbiome largely from egg yolk, red meat, and protein, and those metabolites are renally excreted. This has important dietary implications for stroke prevention. Lowering of plasma total homocysteine with B vitamins significantly reduces the risk of stroke. That was not apparent in early studies because harm from cyanocobalamin among participants with renal failure obscured the benefit among those with good renal function. We should be using B vitamins to prevent stroke but should use methylcobalamin or oxocobalamin instead of cyanocobalamin.

A Simple, accurate precise and validated FTIR method was developed for the determination ofpregabalin (PRG) and methylcobalamin (MCA) in bulk as well as in tablet dosage form. The drug was analyzed by FTIR spectrophotometry with DRIFT sampling technique. wavenumber ranges 1660-1600 cm-1 (-COOH) and3400-3250 cm−1(-CONH2) were selected for pregabalin and methylcobalamin, respectively. The method was found to be linear over the range of PRG 1–6% w/w and MCA 0.2–1.2% w/w with good regression coefficient (r2) of PRG 0.9960 and MCA 0.9976. The percent recovery of pregabalin and methylcobalamin in marketed tablet dosage form was in the range of 98.25 – 99.00% and 98.62 – 99.60%, respectively. The LOD & LOQ were found 1.4003 & 4.2434 of PRG and 0.2027 & 0.6142 of MCA, respectively.The developed FTIR method was validated as per ICH guidelines and it can be used for the estimation of both drugs in combined dosage form.

A high-performance liquid chromatographic method was developed for the separation and determination of 13 vitamin B compounds, including thiamin, riboflavin, riboflavin 5′-monophosphate, pyridoxine, hydroxocobalamin, cyanocobalamin, adenosylcobalamin, methylcobalamin, niacin, niacinamide, pantothenic acid, folic acid, and biotin. The method consisted of an ODS column, a gradient elution using pH 3.0 phosphate buffer–acetonitrile, ion-pairing reagent as mobile phase at 1.0 mL min−1, and UV detection at 210 nm. As a result, the 13 vitamin B compounds were separated in 60 min, and the specificity, linearity, range, limit of detection, limit of quantitation, intra- and inter-day precision, and recovery were satisfactory. The limit of detections ranged from 0.03 to 0.41 µg mL−1. Intra- and inter-day precision of peak area, expressed as RSD, were 0.542–5.144% and 0.216–6.367%, respectively. The method was used to evaluate vitamin B compounds in energy drinks and multivitamin pills. In case of energy drink analysis, the method identified and evaluated not only vitamin B compounds but also major ingredients, such as caffeine, vanillin, and benzoic acid. The vitamin B compounds in energy drinks and multivitamin pills were also quantitated and compared with values listed on each product label. Each quantitative value was close to the label values, suggesting high quantitative ability of the developed method.

The industrial solvent trichloroethylene (TCE) and its two major metabolites trichloroethanol (TCE-OH) and trichloroacetic acid (TCA) cause formic aciduria in male F344 rats. Prior treatment of male F344 rats with 1-aminobenzotriazole a cytochrome P450 inhibitor, followed by TCE (16mk/kg, po), completely prevented formic aciduria, but had no effect on formic acid excretion produced by TCA (8 or 16 mg/kg, po), suggesting TCA may be the proximate metabolite producing this response. Dow and Green reported an increase in the concentration of 5-methyltetrahydrofolate (5-MTHF) in the plasma of rats treated with TCE-OH, suggesting a block in the cycling of 5-MTHF to tetrahydrofolate (THF). This pathway is under the control of the vitamin B12-dependent methionine salvage pathway. We therefore treated rats with three daily doses of methylcobalamin (CH3Cbl) or hydroxocobalamin (OHCbl), a cofactor for methionine synthase, or L-methionine, followed by TCE (16 mg/kg) to determine if they could alleviate the formic aciduria. These pretreatments only partially reduced the excretion of formic acid in the urine. Although prior treatment with S-adenosyl-L-methionine had no effect on formic acid excretion. Consistent with these findings, the activity of methionine synthase in the liver of TCE-treated rats was not inhibited. Transcriptomic analysis of the liver-identified nine differential expressed genes, of note, was downregulation of Lmbrd1 involved in the conversion of vitamin B12 into CH3Cbl, a cofactor for methionine synthase. Our findings indicate that the formic aciduria produced by TCE-OH and TCA may be the result of a block in the recycling of 5-MTHF to THF, the effect on the methionine salvage pathway being a secondary response following acute exposure.

Background: Nanocomposites are of great interest due to their competency to show multifunctional properties. They have been recently given much attention due to their credibility to offer the synergistic feature of organic material with those of inorganic constituents. Different types of nanocomposites have been prepared to date and are being used for different applications. The delivery of drugs in the human body at a particular site was one of the major problems in the medicinal field. The nanocomposite formulations can be used to provide controlled release and they can be combined with ligands for targeted drug delivery. Applications of the nanocomposites as ion exchangers are also increasing at a faster rate. Due to this, they help in the softening of the water. They can also be easily recharged by washing them with a solution containing a high concentration of sodium ions. In the present paper, we have worked on the synthesis and applications of the polysorbate/ironmolybdophosphate (PS/FMP) nanocomposite. Methods: Polysorbate/ironmolybdophosphate (PS/FMPS) was synthesized by co-precipitation method in the presence of polysorbate. The material was well characterized using X-ray diffraction (XRD) analysis, Fourier transform infrared spectroscopy, (FTIR) scanning transmission microscopy (SEM), and transmission electron microscopy (TEM). Physicochemical properties of material were studied in detail. Drug delivery behavior of polysorbate/ironmolybdophosphate was investigated by using methylcobalamin as a test drug. Results: The polysorbate/ironmolybdophosphate nanocomposite show enhanced Na+ ion exchange capacity of 2.1 meq/g. It has been revealed that PS/FMP nanocomposite was thermally stable as it retained the ion exchange capacity of 40.4 % at 400°C. An optimum concentration of sodium nitrate (eluent) was found to be 1.0 M for the complete removal of H+ ions from the PS/FMP column. The optimum volume of sodium nitrate (eluent) was found to be 230 mL. The FTIR spectra showed the changes in intensities of characteristic peaks in PS/FMP and in drug loaded on PS/FMP nanocomposite. The characteristic peak at 1043-1061 cm-1 was observed for ionic phosphate stretching, 560-567 cm-1 for iron group and 959 cm-1 due to molybdate present in the material. The additional peak at 3390 cm-1 and 1711 cm-1 were due to -OH and C=O stretching due to the presence of these groups in the structure of polysorbate. The peak present at 430 cm-1 might be due to the presence of Co-O stretching of methylcobalamin. The XRD results confirmed the semicrystalline structure of FMP and PS/FMP. Scanning electron micrographs results revealed the beaded surface of FMP changes to fibrous surface in case of PS/FMP nanocomposite. The TEM images indicate the appearance of smooth surfactant layer on the surface of FMP. The size of the nanocomposite is between 10- 70 nm. The drug loading efficiency and encapsulation efficiency were found to be 35.2%. and 60.4%, respectively. The cumulative drug release of methylcobalamin was studied for the PS/FMP nanocomposite. The order of drug release was found to be pH 9.4 (54.6%) > pH 7.4 (46.4%) > saline (pH 5.7) (36.2%) > pH 2.2 (33.9%). The release at pH 9.4 was higher. As the pH of medium changes from acidic to basic i.e. 2.2 - 9.4, there is an appreciable increase in drug release from the PS/FMP nanocomposite due to the presence of more OH- ions resulting in neutralization of cationic nanocomposite and thus increasing the rate of drug release by ion exchange process and matrix deterioration. : The novel nanocomposite PS/FMP has been synthesized by a simple co-precipitation method. The increase in Na+ ion exchange capacity for nanocomposite is due to the binding of organic part (Polysorbate) with inorganic ironmolybdophosphate. The physiochemical properties of PS/FMP were found to be superior. Fourier transform infrared spectra of PS/FMP and drug loaded PS/FMP confirmed the formation of materials. The SEM results indicated the surface of synthesized FMP is bead-like appearance whereas the beaded surface of FMP changes to fibrous surface on the addition of polysorbate thus indicated the fabrication of nanocomposite. The cumulative drug release of methylcobalamin was studied and the order of drug release was found to be pH 9.4 > pH 7.4 > saline (pH 5.7) > pH 2.2. Thus PS/FMP is a promising multifunctional nanocomposite.

Photolysis of methylcobalamin (MeCbl) in the presence of molecular oxygen (O2) has been investigated using density functional theory (DFT) and time-dependent DFT (TD-DFT). The key step involves the formation of the Cbl–O–O–CH3 intermediate as a result of triplet O2 insertion in the Co–C bond in the presence of light. Analysis of low-lying excited states shows that the presence of light is only needed to activate the Co–C bond via the formation of the ligand field (LF) state. The insertion of O2, as well as the change in the spin state, takes place in the ground state. The analysis of the structural and electronic properties of the Cbl–O–O–CH3 intermediate is presented and possible decomposition also discussed.

Vitamin B12 is a vital water-soluble micronutrient required for maintaining the human health. However, one of the major encountered problems related to human health is a deficiency of vitamin B12 due to a strict vegetarian diet, malabsorption, pernicious anemia, surgical resection, etc. The deficiency of vitamin B12 can be overcome by taking vitamin supplements like methylcobalamin. Oral delivery of vitamin supplements is the most widely preferred route for administration as well as it gives good patient compliance. Particle size reduction of active ingredient enhances the dissolution rate which is very important in oral delivery. In the present study, methylcobalamin was re-crystallized using antisolvent crystallization technique and effect of various parameters like time (5–15 min), volume of solvent (0.5–1.5 mL), rate of addition (0.056–0.16 mL/min) and temperature (15–31 °C) on particle size was investigated. The aciculate shape of methylcobalamin was observed when re-crystallized using water as a solvent and acetone as an antisolvent. Increase in time from 5 min to 15 min increased the particle size (4.85 μm to 8.18 μm) while the increase in the volume of solvent from 0.5 mL to 1.5 mL has decreased the particle size (8.18 μm to 5.32 μm) and less effect was observed on particle size with a decrease in temperature (31 °C to 15 °C). The maximum impact on particle size (8.18 μm to 3.21 μm) was observed with the rate of addition from 0.16 mL/min to 0.056 mL/min. For water-acetone system, the particle size of methylcobalamin was found to decrease from 40.5 µm to 2.37 µm at 31 °C, 10 mL antisolvent, 0.5 mL solvent, 0.067 mL/min and 15 min. Hence, ∼18 times reduction in particle size was observed using antisolvent crystallization.

The first multiple stable isotope dilution assay method was developed for the simultaneous determination of four cobalamins, namely, hydroxocobalamin (OHCbl), adenosylcobalamin (AdoCbl), methylcobalamin (MeCbl), and cyanocobalamin (CNCbl), in their native forms. The sample preparation was optimized with enzyme treatment and immunoaffinity purification. The analysis was performed by LC-MS/MS using respective 15N-labeled cobalamins as internal standards. Method validation resulted in limits of detection ranging from 0.19 to 0.58 ng/g and limits of quantification ranging from 0.68 to 1.73 ng/g. Recoveries at three levels were between 82 and 121%. Intra-day and inter-day precisions were below 6% and 11% RSD, respectively. The analysis of a reference material resulted in a variance of

Purpose Oxidative stress is considered a major determinant in the pathogenesis of vitiligo. Methylcobalamin (MeCbl) is an activated form of vitamin B12 that regulates inflammatory factors, counters oxidative stress, and reduces apoptosis in many disease models. However, the specific mechanism of MeCbl repigmentation against vitiligo is unknown. In this study, we explored the effect of MeCbl on melanocytes following hydrogen peroxide (H2O2)-induced oxidative stress. Methods We established an oxidative stress model using the immortalized human normal melanocyte cell line PIG1. We used a Cell Counting Kit-8 (CCK-8) to detect drug cytotoxicity, and we measured the melanin content of cells using the NaOH method. Intracellular oxidative damage was assessed by flow cytometry and antioxidant enzyme detection kits. In addition, we assessed the presence of apoptosis by flow cytometry and Western blots. We explored the underlying mechanisms of MeCbl during oxidative stress in melanocytes by analyzing the results of experiments based on real-time quantitative polymerase chain reaction (RT-qPCR), Western blotting, and laser scanning confocal immunofluorescence microscopy. Finally, we repeated the experiments after applying an inhibitor to block the Nrf2 pathway. Results We found that MeCbl treatment enhanced cell viability, increased melanin content, reduced intracellular reactive oxygen species (ROS) accumulation, increased the activities of antioxidant enzyme superoxide dismutase (SOD) and catalase (CAT), reduced melanocyte apoptosis, and up-regulated the expression of the Nrf2/HO-1 pathway. Moreover, the protective effects of MeCbl were significantly weakened after inhibiting the Nrf2/HO-1 pathway. Conclusion Our results indicate that MeCbl attenuated the H2O2-induced oxidative stress in melanocytes by activating the Nrf2/HO-1 pathway, this suggests that MeCbl may be an effective treatment against vitiligo.

Abstract. Objective: To evaluate the efficiency of the sublingual route for the treatment of vitamin B12 deficiency in infants. Background: Vitamin B12 deficiency is common in children. In breastfed infants, the main reason is maternal B12 deficiency. Parenteral administration is commonly prescribed. However, patient compliance is not satisfactory due to repeated painful parenteral applications. It is also known that the oral route is efficient in high doses. In recent years, the sublingual route has been tried. This route stands out due to its easy applicability and low cost. However, there are few efficacy studies in infants for the sublingual route. Materials and methods: The study included 49 infants aged 6–12 months. All infants with marginal or deficient B12 levels (

Plant-based nutrition has become extremely popular in the contemporary era. Its positive effects are sustained by many studies, but one of its risks is that it is almost completely devoid of vitamin B12. In the present study, we analyzed the effects of two types of vitamin B12 supplements, cyancobalamin and methylcobalamin on the level of active serum vitamin (holotranscobalamin) in a group of Romanian individuals (n=42) following a (vegan) plant-based diet. The results revealed that cyancobalamin gives better results in maintaining B12, as quantified by the holotranscobalamin value (median=150 pcg/l) when compared with methylcobalamin (median=78.5 pcg/l). The frequency of administration, regardless of the quantity in one dose, is another important factor in maintaining the holotranscobalamin level within suitable limits. More frequent intakes give more optimal results. Vegans trying to supplement with alternative products (algae, kombucha, other fermented products), had the lowest levels of holotranscobalamin, always bellow the recommended level of 35 pcg/l (median=29 pcg/l). Vegans must be educated on B12 supplementation, about the pharmaceutical forms on the market and their performances and on choosing the optimal plan in order to avoid the onset of B12 deficiency.

Background Vincristine leads to development of debilitating neuropathy in 40-45% patients with resultant compromised efficacy of chemotherapy, suboptimal treatment and worse prognostic outcome. Vitamin B6 and vitamin B12 improves non-oncological neuropathies. Therefore, this study investigated vitamin B6 and vitamin B12 to prevent vincristine- induced peripheral neuropathy (VIPN) by reducing incidence, absolute risk, relative risk, severity as well as delaying the onset. Methods Patients with ALL undergoing induction phase were randomly assigned into intervention or placebo arm in a double- blind manner. Vitamin B6 (25 mg Pyridoxine) two tablets were given three times daily for 5 weeks. Vitamin B12 (500 μg/ml Methylcobalamin) was administered intravenously on day 1, 3 and 5 of every week for 5 weeks during induction period. Placebo arm received oral and intravenous placebo for same duration. Patients were evaluated on the outset of every week by FACT/GOG-NTX questionnaire. Severity was assessed per NCI-CTCAE grading scale. Results 102 patients were enrolled. Among them 81 completed the study, where 42 received vitamin B6 and B12 and 39 received placebo. There was significant difference in incidence of neuropathy between arms (26.19% intervention arm, 56.41% placebo; P-0.01). Relative risk of neuropathy was significantly (RR-0.46) lower in intervention arm. Besides, absolute risk reduction (ARR) was 30% and relative risk reduction (RRR) was 54%. NNT was 3.33. Significant trend was observed in difference of severity of VIPN between groups (P-0.03). No significant difference observed in between arms for time to onset of neuropathy. Conclusion Vitamin B6 and vitamin B12 significantly reduced the incidence, relative risk and severity of VIPN. NNT was encouraging too. Henceforth, the status of vitamin B6 and vitamin B12 as neuroprotective agent against VIPN can be recommended as a promising one. Clinicaltrials.gov ID: NCT02923388

Background: Diabetic peripheral neuropathy (DPN) characterized by nerve damage is a common and disabling chronic microvascular complication in patients with type 2 diabetic mellitus (T2DM), affecting at least half of patients diagnosed with T2DM. Unfortunately, the current treatment for DPN is not ideal. Traditional Chinese medicine (TCM), with a unique theoretical system, has made outstanding contributions in the treatment of T2DM and related complications. Mudan granule, a Chinese patent medicine, has been previously validated that could ameliorate the symptoms, promote nerve tissue repair, increase nerve conduction velocity (NCV) in patients with DPN. However, the previous studies are of variable quality, which limits the clinical application of Mudan granule. Therefore, we designed a double-blind, randomized, placebo-controlled, parallel-arm, multi-center trial to estimate the safety and efficacy of Mudan granule in conjunction with methylcobalamin in individuals suffering from type 2 diabetic peripheral neuropathy.Methods: This work is conducted as a 14-center, double-blind, randomized, placebo-controlled, parallel-arm trial. In all, 402 subjects (aged 30–70 years) will be recruited and randomized in a 1:1 ratio to an intervention group (n = 201; Mudan granule + methylcobalamin) and a control group (n = 201; placebo + methylcobalamin). Treatment is administered in 24 weeks cycles without any treatment interruption between cycles. Michigan Diabetic Neuropathy Score (MDNS) as the primary outcome will be evaluated at baseline, 12 weeks during the intervention period, and after 24 weeks of the intervention. Adverse events and safety assessments will be also documented. The analysis of all data will be carried out based on a predefined statistical analysis plan.Discussion: The outcomes from this study will offer important evidence regarding the safety and efficacy that Mudan granule can be used as an alternative and complementary therapeutic intervention in patients with type 2 diabetic peripheral neuropathy.Clinical trial registration: Registered at https://clinicaltrials.gov/. Trial registration number: NCT04711980. Registered January 2021.

Background. Diabetic neuropathy is the most common complication of diabetes mellitus (DM), found in 30–90 % of patients. The relevance of the study is due to the prevalence of diabetic neuropathy, as well as assessing the effectiveness of the use of methylated form of vitamin B12 in the treatment of patients with DM. The aim of this study was to determine the effectiveness of methylcobalamin (Diacobal, Kusum Pharm) in the complex treatment of patients with DM complicated by diabetic peripheral neuropathy (DPN). Materials and methods. The study was conducted by a simple open method. The Michigan Neuropathy Screening Instrument (MNSI), the Neurological Symptoms Score (NSS), the European Quality of Life Questionnaire (EQ-5D) were used to screen and determine the severity of symptoms of DPN. The neurological examination of patients included an assessment of various types of sensitivity (pain, tactile, vibrational, cold, heat, proprioceptive, pressure sensation). Blood levels of vitamin B12 (cyanocobalamin) were determined in all patients. Diacobal was added to the regimen of traditional treatment for DPN regardless of the levels of vitamin B12. Results. It was shown that DPN is a common complication of DM: according to subjective characteristics (MNSI and NSS scales), its symptoms were detected in 84.3 % of cases among DM patients. Subjective scales, such as EQ-5D, MNSI, NSS, are not enough to assess the presence and degree of polyneuropathy in DM patients. The data of these scales do not correlate with the results of a physical examination, but can be used as an auxiliary method for the analysis of subjective symptoms of a patient. A neurological examination is recommended for DM patients, regardless of whether they present active complaints. DPN was detected in 64.8 % of cases among diabetic patients according to objective criteria for the diagnosis and determination of various types of sensitivity (pain, tactile, vibrational, cold, thermal, proprioceptive, pressure sensation). Determining the level of cyanocobalamin should be used in conjunction with the determination of homocysteine and folic acid. Conclusions. The use of methylcobalamin (Diacobal in a daily dose of 1,500 μg for 3 months) in the complex treatment of diabetic neuropathy in diabetic patients is effective and safe, helps to eliminate the manifestations of diabetic neuropathy and improve the quality of life of patients.

Background This study aimed to report the case of a patient who presented with depression, cognitive impairment, ataxic gait, and urinary incontinence associated with vitamin B12 deficiency. Case Description Serum vitamin B12 level was low in this patient, and anti-intrinsic factor antibody was positive. Neuroimaging revealed abnormal hyperintense signals in the cerebellum and dorsal and lateral columns of the spinal cord, and obstructive hydrocephalus. A biopsy of the stomach revealed chronic gastritis, intestinal metaplasia, and atrophy. After 3 months of initiating methylcobalamin therapy, significant improvement was noticed clinically, and brain magnetic resonance imaging was near to normal. Conclusions This study was novel in reporting subacute combined degeneration of the spinal cord and hydrocephalus associated with vitamin B12 deficiency in adults.

A simple, selective, linear, precise, and accurate RP-HPLC method was developed and validated for the simultaneous estimation of Gabapentin & Methylcobalamin from bulk and formulation. Chromatographic separation was achieved Isocratically on an Inertsil C18 column (150x4.6, 5µ particle size) using a mobile phase Buffer: Acetonitrile in the ratio of 60:40 v/v. The flow rate was 1.0 ml/min, effluents were detected at 264 nm and 10µl of sample was injected. Retention time of Gabapentin & Methylcobalamin was found to be 2.7 and 4.13 min respectively. Linearity of the method was in the concentration range of 25-150 µg for Gabapentin & 0.125-0.750 µg for Methylcobalamin. Percent recoveries obtained for both the drugs were 100.00%. The percentage RSD for precision of the method was found to be less than 2%. The method was validated according to the ICH guidelines. The method developed was successfully applied for the analysis of simultaneous estimation of Gabapentin & Methylcobalamin tablets and was fairly good in comparison with other methods. Keywords: Gabapentin, Methylcobalamin, HPLC.

Chub mackerel (Scomber colias), an underutilised fish species, has been identified as an excellent source of selenium and vitamin B12, which may prevent and delay cognitive decline and dementia. Given these nutrients’ contents seasonal variability, a complete monthly sampling, involving a specifically adjusted methodology for the determination of vitamin B12 (LC-MS/MS) and its main vitamers and a sensitive analytical technique (ICPMS) for Se, was carried out. Regarding total vitamin B12 in the chub mackerel muscle, its content varied from 10.0 ± 1.0 μg/100 g in February to 18.9 ± 1.9 μg/100 g in August. It was also shown that methylcobalamin content ranged from 1.12 ± 0.43 (March) to 6.77 ± 0.80 μg/100 g (August), a sixfold increase. Hydroxocobalamin content varied between 3.49 ± 0.12 (February) and 9.62 ± 0.41 μg/100 g (August). The content of Se also had seasonal variability, ranging from 451 ± 14 μg/kg (December) to 606 ± 11 μg/kg (July). In the worst-case scenario, a weekly 160 g meal of this fish would almost guarantee B12 requirements. Three weekly 160 g meals of S. colias would cover, at least, 56% of the Se requirements. Therefore, as first explored here, it is highly relevant a complete, year-round assessment of the chub mackerel’s nutritional value, in order to elaborate finely tuned dietary recommendations. Highlights: Methylcobalamin rose with temperature from 1.12 ± 0.43 μg/100 g to 6.77 ± 0.80 μg/100 g; Hydroxocobalamin rose with temperature from 3.49 ± 0.12 μg/100 g to 9.62 ± 0.41 μg/100 g; A weekly meal of at least 160 g chub mackerel would cover vitamin B12 requirement; 3 weekly meals of at least 160 g chub mackerel may cover 56% of the Se requirement. This work was supported by national funds through FCT - Fundação para a Ciência e a Tecnologia, I.P., within the scope of projects New- Food4Thought PTDC/SAU-NUT/30455/2017 (Exploitation of the bioactive nutrients’ potential for the prevention of Alzheimer’s Disease and cognitive ageing through in vitro assays, in vivo verification, and in silico quantification of the health benefits) and CIISA (Project UIDB/ 00276/2020). Researchers were supported by the following Post- Doctoral Grants: Ref.: SFRH/BPD/102689/2014 (“Fundação para a Ciência e a Tecnologia”, FCT) for the author Carlos Cardoso and DIVERSIAQUA (MAR2020, Ref.: 16-02-01-FEAM-66) for the author Cláudia Afonso. The author Ana Gomes-Bispo also acknowledges FCT and IPMA for the DL57/2016 Junior Researcher Contract. info:eu-repo/semantics/publishedVersion

Patients with inflammatory bowel disease (IBD) are usually advised to supplement various types of vitamin B12, because vitamin B12 is generally absorbed in the colon. Thus, in the current study, the influence of cyanocobalamin (CNCBL) or methylcobalamin (MECBL) ingestion on IBD symptoms will be investigated. Then, whether and how the application of various cobalamins would modify the taxonomic and functional composition of the gut microbiome in mice will be examined carefully. Dextran-sulfate-sodium-induced IBD mice were treated with MECBL or CNCBL; disease activity index (DAI) scores and intestinal inflammatory conditions of mice were evaluated. Fecal samples were collected; microbiota composition was determined with a 16s rRNA analysis; functional profiles were predicted by phylogenetic investigation of communities by reconstruction of unobserved states (PICRUSt); and short-chain fatty acids were measured. The consequence of higher relative abundances of Enterobacteriaceae and isomeric short-chain fatty acids by cobalamin treatment revealed that a high concentration of CNCBL but not MECBL supplementation obviously aggravated IBD. A microbial ecosystem rich in Escherichia/ Shigella and low in Lactobacillus, Blautia, and Clostridium XVIII was observed in IBD mice after a high concentration of CNCBL supplementation. In cobalamin-dependent enzymes, CNCBL was more efficient in the adenosylcobalamin system than MECBL and vice versa in the MECBL system. The distinct effects of various cobalamins were associated with the distribution and efficiency of vitamin-B12-dependent riboswitches. CNCBL had a strong inhibitory effect on all riboswitches, especially on btuB and pocR riboswitches from Enterobacteriaceae. CNCBL aggravated IBD via enhancing the proportion of Enterobacteriaceae organisms through riboswitch and enzyme systems. The present study provides a critical reference for offering a suitable amount and type of cobalamin during a symbiotic condition.

Objective To observe the differences in curative effects between prophylactic and therapeutic administrations of Gliclazide (GLZ) or Methylcobalamin (MCA) on diabetic peripheral neuropathy in rats. Methods GLZ (25 mg/kg/day) or MCA (175 μg/kg/day) was orally administrated prophylactically to streptozotocin-induced diabetic rats for 8 weeks before diabetic peripheral neuropathy developed or administrated therapeutically after diabetic peripheral neuropathy developed, respectively. The motor nerve conduction velocities (MNCV), aldose reductase (AR) activities, the polyol contents and antioxidative enzyme activities in the sciatic never tissues were determined. The morphology of sciatic never tissues was observed. Results In comparison to vehicle, most of the changes in the sciatic nerves of the diabetic rats (e. g., delayed MNCV, altered/damaged nerve structure, enhanced AR activity, increased polyol contents, altered Cu, Zn-superoxide dismutase, glutathione-peroxidase activities, and elevated malondialdehyde level) were significantly ameliorated by prophylactic administration with either GLZ or MCA. In contrast, only few of above-mentioned parameters were alleviated in DPN rats by therapeutic administration with GLZ or MCA as compared to vehicle. The curative effects of GLZ or MCA prophylactic administration on MNCV, AR activity, polyol contents and antioxidative enzyme activities were markedly stronger than therapeutic administration. Conclusion Prophylactic administration of GLZ or MCA was superior to the therapeutic administration in alleviation of diabetic neuropathy in STZ-rats, suggesting that pharmacotherapy should be initiated at a much earlier stage before diabetic neuropathy developed, but not at a later stage after never damage reached.

Objective To investigate the clinical effect of mouse nerve growth factor (mNGF) and methylcobalamin (MeCbl) for the treatment of lumbar disk herniation (LDH) with foot drop. Methods A total of 46 patients suffering from LDH with foot drop who underwent transforaminal lumbar interbody fusion (TLIF) surgery in our department from January 2015 to December 2017 were retrospectively analyzed. We divided these patients into two groups according to the different postoperative treatment which independently selected by patients after signing informed consent form: one group of 25 patients was treated with MeCbl alone (Group MeCbl), the other group of 21 patients was treated with a combination of mNGF and MeCbl (Group MeCbl+mNGF). Patient demographics, the visual analogue scale (VAS) scores, sensory and muscular strength improvement statistics at 1 week, 4 weeks, 12 weeks, and 12 months postoperatively were recorded. Motor/sensory deficits, sciatica and overall neurological outcome after treatment of MeCbl alone and combination of mNGF and MeCbl were retrospectively analyzed. Results The follow‐up ranged between 12 and 42 months (mean 20.8 months). There were no significant differences between these two groups of patients with respect to sex ratio, age, smoking, diabetes, disease course, section of protruding disc(s), muscular strength of foot dorsiflexion or preoperative visual analogue scale (VAS) score (P > 0.05). The VAS scores of Group MeCbl+mNGF were significantly lower than Group MeCbl at 1 week, 4 weeks, 12 weeks, and 12 months postoperatively (4.32 ± 0.75 vs 5.25 ± 0.79,2.65 ± 0.48 vs 3.42 ± 0.52, 1.72 ± 0.36 vs 2.45 ± 0.39, 1.12 ± 0.22 vs 1.52 ± 0.24, P < 0.05). The effective rates of sensory improvement were significantly higher in Group MeCbl+mNGF compared with Group MeCbl at 12‐week/12‐month follow‐up time point (90.48% vs 52.00%,95.24% vs 68.00%, P, Application of mNGF in combination with MeCbl had clinical effects on promoting the recovery of neurological function in patients suffering from LDH with foot drop.

A 3D PNS disease–on–a–chip enables the monitoring of pathophysiological processes, including demyelination and remyelination., Demyelinating diseases involve loss of myelin sheaths and eventually lead to neurological problems. Unfortunately, the precise mechanisms remain unknown, and there are no effective therapies. To overcome these limitations, a reliable and physiologically relevant in vitro model is required. Here, we present a three-dimensional peripheral nervous system (PNS) microfluidic platform that recapitulates the full spectrum of myelination, demyelination, and remyelination using primary Schwann cells (SCs) and motor neurons (MNs). The platform enables reproducible hydrogel patterning and long-term stable coculture of MNs and SCs over 40 days in vitro based on three distinct design factors. Furthermore, the on-demand detachable substrate allows in-depth biological analysis. We demonstrated the possibility of mimicking segmental demyelination by lysophosphatidylcholine, and recovery of myelin structure by application of two drugs: benzatropine or methylcobalamin. This 3D PNS disease–on–a–chip may serve as a potential platform for understanding the pathophysiology of demyelination and screening drugs for remyelination.

Background & Aims: Methylcobalamin nasal spray (NasoB12® ) has been developed to overcome the drawbacks of currently available therapeutic options like the poorly absorbed oral route and the painful intramuscular route. The present study was designed to evaluate the safety and efficacy of NasoB12® in patients with Vitamin B12 deficiency, with serum vitamin B12 (svB12)

Background/Objective: Methylcobalamin Nasal Spray (NasoB12®, Troikaa, India) has been recently developed to overcome the drawbacks of currently available therapeutic routes. This study was designed to compare pharmacokinetics, safety and tolerability of methylcobalamin nasal spray with intramuscular injection.

Background: Methylcobalamin is essential vitamin required for DNA synthesis during cell division therefore maintain the architecture of nervous tissue distorted by soft metals such as Lithium Carbonate. Accurate documentation of the thickness cerebellar cortical thickness was required in subjects who were injected with methylcobalamin distorted by Lithium Carbonate. Aim: To provide data of cerebellar gray matter thickness distorted by Lithium Carbonate by the anti-oxidant effect of methylcobalamin. Methods: Fifteen albino rats were maintained on food and diet in Animal House of the Basic Medical Sciences Institute, JPMC Karachi for a period of 6 weeks. Results: The results obtained of the thickness of cerebellar gray matter distorted by Lithium Carbonate was restored by methylcobalamin in our study. Conclusion: To observe the neuroprotective effect of B12 on distorted cerebellar cortex treated by Lithium Carbonate. Keywords: Methylcobalamin, Lithium Carbonate, Gray Matter, Cerebellum

Thiolatocobalamins are a class of cobalamins comprised of naturally occurring and synthetic ligands. Glutathionylcobalamin (GSCbl) occurs naturally in mammalian cells, and also as an intermediate in the glutathione-dependent dealkylation of methylcobalamin (MeCbl) to form cob(I)alamin by pure recombinant CblC from C. elegans. Glutathione-driven deglutathionylation of GSCbl was demonstrated both in mammalian as well as in C. elegans CblC. Dethiolation is orders of magnitude faster than dealkylation of Co–C bonded cobalamins, which motivated us to investigate two synthetic thiolatocobalamins as substrates to repair the enzymatic activity of pathogenic CblC variants in humans. We report the synthesis and kinetic characterization of cysteaminylcobalamin (CyaCbl) and 2-mercaptopropionylglycinocobalamin (MpgCbl). Both CyaCbl and MpgCbl were obtained in high purity (90–95%) and yield (78–85%). UV–visible spectral properties agreed with those reported for other thiolatocobalamins with absorbance maxima observed at 372 nm and 532 nm. Both CyaCbl and MpgCbl bound to wild type human recombinant CblC inducing spectral blue-shifts characteristic of the respective base-on to base-off transitions. Addition of excess glutathione (GSH) resulted in rapid elimination of the β-ligand to give aquacobalamin (H2OCbl) as the reaction product under aerobic conditions. Further, CyaCbl and MpgCbl underwent spontaneous dethiolation thereby repairing the loss of activity of pathogenic variants of human CblC, namely R161G and R161Q. We posit that thiolatocobalamins could be exploited therapeutically for the treatment of inborn errors of metabolism that impair processing of dietary and supplemental cobalamin forms. While these disorders are targets for newborn screening in some countries, there is currently no effective treatment available to patients.

Patients with renal failure have extremely high cardiovascular risk; in dialysis patients the risk of stroke is increased approximately 10-fold over that in the general population. Reasons include not only a high prevalence of traditional risk factors such as diabetes, hypertension and dyslipidemia, but also the accumulation of toxic substances that are eliminated by the kidneys, so have very high levels in patients with renal failure. These include plasma total homocysteine, asymmetric dimethylarginine, thiocyanate, and toxic products of the intestinal microbiome (Gut-Derived Uremic Toxins; GDUT), which include trimethylamine N- oxide (TMAO), produced from phosphatidylcholine (largely from egg yolk) and carnitine (largely from red meat). Other GDUT are produced from amino acids, largely from meat consumption. Deficiency of vitamin B12 is very common, raises plasma tHcy, and is easily treated. However, cyanocobalamin is toxic in patients with renal failure. To reduce the risk of stroke in renal failure it is important to limit the intake of meat, avoid egg yolk, and use methylcobalamin instead of cyanocobalamin, in addition to folic acid.

PURPOSE To investigate conjunctival microvascular responses in patients with mild diabetic retinopathy (MDR) and methylenetetrahydrofolate reductase (MTHFR) polymorphisms (D + PM) after administration of Ocufolin™, a medical food containing 900 μg l-methylfolate (levomefolate calcium or [6S]-5-methyltetrahydrofolic acid, calcium salt), methylcobalamin, and other ingredients. METHODS Eight D + PM patients received Ocufolin™ for six months (6 M). Bulbar conjunctival microvasculature and microcirculation metrics, including vessel diameter (D), axial blood flow velocity (Va), cross-sectional blood flow velocity (Vs), flow rate (Q), and vessel density (VD, Dbox), were measured at baseline, 4 M, and 6 M. RESULTS The mean age was 54 ± 7 years. No significant demographic differences were found. Conjunctival microcirculation, measured as Va, Vs, and Q was significantly increased at 4 M and 6 M, compared to baseline. Va was 0.44 ± 0.10 mm/s, 0.58 ± 0.13 mm/s, 0.59 ± 0.13 mm/s in baseline, 4 M, and 6 M, respectively (P