Oral Administration of Nicotinamide Mononucleotide with Bioenhancer BioPerine® Increases the Serum Concentration of Nicotinamide Adenine Dinucleotide in Healthy Human Volunteers: A Pilot, Open-Label, Cross-Over Study

Background: Bioenhancers augment the bioavailability of co-administered molecules without showing any significant effect on their own. Piperine, an alkaloid from Piper nigrum, is an established natural bioenhancer. Nicotinamide mononucleotide (NMN), an antiaging supplement, is the precursor of coenzyme nicotinamide adenine dinucleotide (NAD) that plays an important role in intracellular redox reactions. Objective: The study compared the serum concentrations of NAD in normal healthy participants, supplemented with NMN 500 mg and NMN 500 mg + 5 mg BioPerine® (95% piperine). Methods: In a randomized, open-label, crossover study, NMN (500 mg) was compared to NMN + BioPerine® (500 mg + 5 mg) in 6 healthy adults, aged 18 - 45 years. The participants received a single oral dose of NMN or NMN + BioPerine® capsules with 240 mL water, and blood samples were collected over 8hr. After a 4-day washout period, the same procedures were repeated as per the crossover design. Total NAD (NADtotal), including oxidized NAD (the oxidized) and its reduced form NADH, was measured in human serum samples. Results: The maximum concentration (Cmax) of NAD in serum was higher with NMN + BioPerine® (282 pmol/mL) compared to NMN (246 pmol/mL) alone. In the presence of BioPerine®, the NAD concentrations reached 257 pmol/mL during the first 2 hr, whereas a comparable serum concentration (246 pmol/mL) was attained only after 6 hr in NMN alone. The AUC0-8hr was 1738 pmol/mL/hr in NMN compared to 2004 pmol/mL/hr in NMN+ BioPerine®. The time to reach peak concentration (t1/2) was similar (6hr) in both groups. No clinically relevant adverse events (AE) were observed, and safety parameters remained within normal ranges in all the participants with both formulations. Conclusion: These results reveal that BioPerine® can effectively increase the NAD concentrations in the serum following NMN supplementation in healthy volunteers. The present study was registered prospectively with the Clinical Trials Registry-India (CTRI/2023/11/059982).

Nicotinamide adenine dinucleotide treatment confers resistance to neonatal ischemia and hypoxia:effects on neurobehavioral phenotypes

Neonatal hypoxic-ischemic brain injury is the main cause of hypoxic-ischemic encephalopathy and cerebral palsy.Currently,there are few effective clinical treatments for neonatal hypoxic-ischemic brain injury.Here,we investigated the neuroprotective and molecular mechanisms of exogenous nicotinamide adenine dinucleotide,which can protect against hypoxic injury in adulthood,in a mouse model of neonatal hypoxic-ischemic brain injury.In this study,nicotinamide adenine dinucleotide(5 mg/kg)was intraperitoneally administered 30 minutes befo re surgery and every 24 hours thereafter.The results showed that nicotinamide adenine dinucleotide treatment improved body weight,brain structure,adenosine triphosphate levels,oxidative damage,neurobehavioral test outcomes,and seizure threshold in experimental mice.Tandem mass tag proteomics revealed that numerous proteins were altered after nicotinamide adenine dinucleotide treatment in hypoxic-ischemic brain injury mice.Parallel reaction monitoring and western blotting confirmed changes in the expression levels of proteins including serine(or cysteine)peptidase inhibitor,clade A,member 3N,fibronectin 1,5'-nucleotidase,cytosolic IA,microtubule associated protein 2,and complexin 2.Proteomics analyses showed that nicotinamide adenine dinucleotide ameliorated hypoxic-ischemic injury through inflammation-related signaling pathways(e.g.,nuclear factor-kappa B,mitogen-activated protein kinase,and phosphatidylinositol 3 kinase/protein kinase B).These findings suggest that nicotinamide adenine dinucleotide treatment can improve neurobehavioral phenotypes in hypoxic-ischemic brain injury mice through inflammation-related pathways.

Effects of different concentrations of nicotinamide on hematopoietic stem cells cultured in vitro

BACKGROUND In vitro expansion to increase numbers of hematopoietic stem cells(HSCs)in cord blood could improve clinical efficacy of this vital resource.Nicotinamide(NAM)can promote HSC expansion ex vivo,but its effect on hematopoietic stem and progenitor cells(HSPCs,CD34^(+)CD38)and functional subtypes of HSCs-shortterm repopulating HSCs(ST-HSCs,CD34^(+)CD38CD45RACD49f^(+))and long-term repopulating HSCs(LT-HSCs,CD34^(+)CD38CD45RACD49f^(+)CD90^(+))is not yet known.As a sirtuin 1(SIRT1)inhibitor,NAM participates in regulating cell adhesion,polarity,migration,proliferation,and differentiation.However,SIRT1 exhibits dual effects by promoting or inhibiting differentiation in different tissues or cells.We propose that the concentration of NAM may influence proliferation,differentiation,and SIRT1 signaling of HSCs.AIM To evaluate the effects and underlying mechanisms of action of different concentrations of NAM on HSC proliferation and differentiation.METHODS CD34^(+)cells were purified from umbilical cord blood using MacsCD34 beads,and cultured for 10-12 d in a serum-free medium supplemented with cytokines,with different concentrations of NAM added according to experimental requirements.Flow cytometry was used to detect phenotype,cell cycle distribution,and apoptosis of the cultured cells.Real-time polymerase chain reaction was used to detect the transcription levels of target genes encoding stemness-related factors,che mokines,components of hypoxia pathways,and antioxidant enzymes.Dichloro-dihydro-fluorescein diacetate probes were used to evaluate intracellular production of reactive oxygen species(ROS).Determination of the effect of different culture conditions on the balance of cytokine by cytometric bead array.RESULTS Compared with the control group,the proportion and expansion folds of HSPCs(CD34^(+)CD38)incubated with 5 mmol/L or 10 mmol/L NAM were significantly increased(all P<0.05).The ST-HSCs ratio and fold expansion of the 5 mmol/L NAM group were significantly higher than those of the control and 10 mmol/L NAM groups(all P<0.001),whereas the LT-HSCs ratio and fold expansion of the 10 mmol/L NAM group were significantly higher than those of the other two groups(all P<0.05).When the NAM concentration was>10 mmol/L,cell viability significantly decreased.In addition,compared with the 5 mmol/L NAM group,the proportion of apoptotic cells in the 10 mmol/L NAM group increased and the proportion of cells in S and G2 phase decreased.Compared with the 5 mmol/L NAM group,the HSCs incubated with 10 mmol/L NAM exhibited significantly inhibited SIRT1 expression,increased intracellular ROS content,and downregulated expression of genes encoding antioxidant enzymes(superoxide dismutase 1,peroxiredoxin 1).CONCLUSION Low concentrations(5 mmol/L)of NAM can better regulate the balance between proliferation and differentiation,thereby promoting expansion of HSCs.These findings allow adjustment of NAM concentrations according to expansion needs.

Nicotinamide adenine dinucleotide phosphate oxidase in pancreatic diseases:Mechanisms and future perspectives

Pancreatitis and pancreatic cancer(PC)stand as the most worrisome ailments affecting the pancreas.Researchers have dedicated efforts to unraveling the mechanisms underlying these diseases,yet their true nature continues to elude their grasp.Within this realm,oxidative stress is often believed to play a causal and contributory role in the development of pancreatitis and PC.Excessive accumulation of reactive oxygen species(ROS)can cause oxidative stress,and the key enzyme responsible for inducing ROS production in cells is nicotinamide adenine dinucleotide phosphate hydrogen oxides(NOX).NOX contribute to pancreatic fibrosis and inflammation by generating ROS that injure acinar cells,activate pancreatic stellate cells,and mediate macrophage polarization.Excessive ROS production occurs during malignant transformation and pancreatic carcinogenesis,creating an oxidative microenvironment that can cause abnormal apoptosis,epithelial to mesenchymal transition and genomic instability.Therefore,understanding the role of NOX in pancreatic diseases contributes to a more in-depth exploration of the exact pathogenesis of these diseases.In this review,we aim to summarize the potential roles of NOX and its mechanism in pancreatic disorders,aiming to provide novel insights into understanding the mechanisms underlying these diseases.

Research Progress on the Application of β-Nicotinamide Mononucleotides in Cosmetics

Recent studies have found that nicotinamide mononucleotide(nicotinamide mononucleotide,NMN)also has certain anti-aging,photoprotection,anti-oxidation,and soothing to the skin,while affects the level of nicotinamide adenine dinucleotide(nicotinamide adenine dinucleotide,NAD+)to improve aging-related diseases.These skincare features have laid the foundation for its application in cosmetics.NMN has been used in cosmetics and sold on the market in foreign countries,but there have been debates about its safety.It is currently as a new raw material for cosmetics in China,and its safety is also under monitoring.This paper systematically reviewed the basic properties,skin care functions and the safety of its application in cosmetics,aiming to provide theoretical reference for the development and application of NMN.

Nicotinamide adenine dinucleotide phosphate oxidase activation and neuronal death after ischemic stroke

Nicotinamide adenine dinucleotide phosphate oxidase(NOX) is a multisubunit enzyme complex that utilizes nicotinamide adenine dinucleotide phosphate to produce superoxide anions and other reactive oxygen species. Under normal circumstances, reactive oxygen species mediate a number of important cellular functions, including the facilitation of adaptive immunity. In pathogenic circumstances, however,excess reactive oxygen species generated by NOX promotes apoptotic cell death. In ischemic stroke, in particular, it has been shown that both NOX activation and derangements in glucose metabolism result in increased apoptosis. Moreover, recent studies have established that glucose, as a NOX substrate, plays a vital role in the pathogenesis of reperfusion injury. Thus, NOX inhibition has the potential to mitigate the deleterious impact of hyperglycemia on stroke. In this paper, we provide an overview of this research,coupled with a discussion of its implications for the development of NOX inhibition as a strategy for the treatment of ischemic stroke. Both inhibition using apocynin, as well as the prospect of developing more specific inhibitors based on what is now understood of the biology of NOX assembly and activation, will be highlighted in the course of our discussion.

OTOTOXIC MODEL OF OXALIPLATIN AND PROTECTION FROM NICOTINAMIDE ADENINE DINUCLEOTIDE

Oxaliplatin, an anticancer drug commonly used to treat colorectal cancer and other tumors, has a number of serious side effects, most notably neuropathy and ototoxicity. To gain insights into its ototoxic profile, oxaliplatin was applied to rat cochlear organ cultures. Consistent with it neurotoxic propensity, oxaliplatin selectively damaged nerve fibers at a very low dose 1 μM. In contrast, the dose required to damage hair cells and spiral ganglion neurons was 50 fold higher (50 μM). Oxailiplatin-induced cochlear lesions initial-ly increased with dose, but unexpectedly decreased at very high doses. This non-linear dose response could be related to depressed oxaliplatin uptake via active transport mechanisms. Previous studies have demon-strated that axonal degeneration involves biologically active processes which can be greatly attenuated by nicotinamide adenine dinucleotide (NAD+). To determine if NAD+would protect spiral ganglion axons and the hair cells from oxaliplatin damage, cochlear cultures were treated with oxaliplatin alone at doses of 10 μM or 50 μM respectively as controls or combined with 20 mM NAD+. Treatment with 10 μM oxaliplatin for 48 hours resulted in minor damage to auditory nerve fibers, but spared cochlear hair cells. However, when cochlear cultures were treated with 10 μM oxaliplatin plus 20 mM NAD+, most auditory nerve fibers were intact. 50 μM oxaliplatin destroyed most of spiral ganglion neurons and cochlear hair cells with apop-totic characteristics of cell fragmentations. However, 50 μM oxaliplatin plus 20 mM NAD+treatment great-ly reduced neuronal degenerations and hair cell missing. The results suggested that NAD+provides signifi-cant protection against oxaliplatin-induced neurotoxicity and ototoxicity, which may be due to its actions of antioxidant, antiapoptosis, and energy supply.

Effects of nicotinamide and riboflavin on the biodesulfurization activity of dibenzothiophene by Rhodococcus erythropolis USTB-03

Rhodococcus erythropolis USTB-03 is a promising bacterial strain for the biodesulfurization of dibenzothiophene （DBT） via a sulfurspecific pathway in which DBT is converted to 2-hydroxybiphenyl （2HBP） as an end product. The effects of nicotinamide and riboflavin on the sulfur specific activity （SA） of DBT biodesulfurization by R. erythropolis USTB-03 were investigated. Both nicotinamide and riboflavin were found to enhance the expression of SA, which was not previously reported. When R. erythropolis USTB-03 was grown on a medium containing nicotinamide of 10.0 mmol or riboflavin of 50.0 μmol, SA was raised from 68.0 or so to more than 130 mmol 2HBP/（kg dry cells.h）. When R. erythropolis USTB-03 was grown in the presence of both nicotinamide of 5.0 mmol and riboflavin of 25.0 μmol, SA was further increased to 159.0 mmol 2HBP/（kg dry cells.h）. It is suggested that the biological synthesis of reduced form of flavin mononucleotide （FMNH2）, an essential coenzyme for the activities of biodesulfurization enzyme Dsz C and A, might be enhanced by nicotinamide and riboflavin, which was responsible for the increased SA of R. erythropolis USTB-03.

Physiological Role of Humic Acid and Nicotinamide on Improving Plant Growth, Yield, and Mineral Nutrient of Wheat (<i>Triticum durum</i>) Grown under Newly Reclaimed Sandy Soil

Humic acid is not a fertilizer as it does not directly provide nutrients to plants, but is a compliment to fertilizer. Nicotinamide (Vitamin pp) is a stress-associated compound that can induce and regulate secondary metabolic accumulation and/or the manifestation of defense metabolism in plants. A field experiment was conducted at the experimental station of National Research center at El-Nubaria region, Egypt, to study the role of foliar application of humic acid (as soil conditioner 13 cm/l) and/or priming grains in nicotinamide (vitamin pp 5 mg/l) in saving irrigation water, decreasing fertilizer dose of NPK and at the same time increasing durum wheat (Triticum durum) cultivars’ (Beni Sweif-1 and Beni Sweif-3) productivity grown under newly reclaimed sandy soil exposed to drought for three weeks continuously. The results showed that plant treated with humic acid or nicotinamide increased significantly all morphological criteria (plant height, leaves number, fresh and dry weights of shoots), metabolism (photosynthetic pigment, total soluble sugar, total carbohydrates, total amino acids and proline), mineral contents (N, P, K, Ca and Mg) and yield (grain, straw and biology) of both cultivars amended with either recommended or half recommended doses of NPK. Foliar application of humic acid to plant priming in Vitamin pp induced significant increases in all studied parameters (morphology, chemical and yield) of plants amended with recommended or half recommended doses of NPK as compared with control plants. The maximum yields of grain, straw and biology of both cultivars were obtained in response to triple treatment (humic acid + nicotinamide + recommended dose of NPK) or (humic acid + nicotinamide + half recommended dose of NPK) respectively.

Nicotinamide overload may play a role in the development of type 2 diabetes

AIM： To investigate whether nicotinamide overload plays a role in type 2 diabetes. METHODS： Nicotinamide metabolic patterns of 14 diabetic and 14 non-diabetic subjects were compared using HPLC. Cumulative effects of nicotinamide and N^1-methylnicotinamide on glucose metabolism, plasma HzO2 levels and tissue nicotinamide adenine dinucleotide （NAD） contents of adult Sprague-Dawley rats were observed. The role of human sweat glands and rat skin in nicotinamide metabolism was investigated using sauna and burn injury, respectively. RESULTS： Diabetic subjects had significantly higher plasma N^1-methylnicotinamide levels 5 h after a 100-mg nicotinamide load than the non-diabetic subjects （0.89 ± 0.13 μmol/L vs 0.6 ± 0.13 μmol/L, P 〈 0.001）. Cumulative doses of nicotinamide （2 g/kg） significantly increased rat plasma Nl-methylnicotinamide concentrations associated with severe insulin resistance, which was mimicked by Nl-methy-Inicotinamide. Moreover, cumulative exposure to N^1- methylnicotinamide （2 g/kg） markedly reduced rat muscle and liver NAD contents and erythrocyte NAD/ NADH ratio, and increased plasma H2O2 levels. Decrease in NAD/NADH ratio and increase in H2O2 generation were also observed in human erythrocytes after exposure to N^1-methylnicotinamide in vitro. Sweating eliminated excessive nicotinamide （5.3-fold increase in sweat nicotinamide concentration 1 h after a 100-mg nicotinamide load）. Skin damage or aldehyde oxidase inhibition with tamoxifen or olanzapine, both being notorious for impairing glucose tolerance, delayed N^1- methylnicotinamide clearance. CONCLUSION： These findings suggest that nicotinamide overload, which induced an increase in plasma N^1- methylnicotinamide, associated with oxidative stress and insulin resistance, plays a role in type 2 diabetes.

A unique insertion of low complexity amino acid sequence underlies protein-protein interaction in human malaria parasite orotate phosphoribosyltransferase and orotidine 5'-monophosphate decarboxylase

Objective:To investigate the multienzyine complex formation of human malaria parasite Plasmodium falciparum[P.falciparum)orotate phosphoribosyltransferase(OPRT)and orotidine5'-monophosphate decarboxylase(OMPDC),the fifth and sixth enzyme of the de novo pyrimidine biosynthetic palhway.Previously,we have clearly established that the two enzymes in the malaria parasite exist physically as a heterotetrameric(OPRT)_2(OMPDG)_2 complex containing two subunits each of OPRT and OMPDC.and that the complex have catalytic kinetic advantages over the monofunetional enzyme.Methods:Both enzymes were cloned and expressed as recombinant proteins.The protein-protein interaction in the enzyme complex was identified using bifunctionul chemical cross-linker,liquid chromatography-mass spectrometric analysis and homology modeling,Results:The unique insertions of low complexity region at the a 2 and a 5 helices of the parasite OMPDC,characterized by single amino acid repeat sequence which was not found in homologous proteins from other organisms,was located on the OPRT-OMPDC interface.The structural models for the protein-prolein interaction of the helerotetrameric(OPRT)_2(OMPDC)_2multienzyme complex were proposed.Conclusions:Based on the proteomic data and structural modeling,it is surmised that the human malaria parasite low complexity region is responsible for the OPRT-OMPDC interaction.The structural complex of the parasite enzymes,thus,represents an efficient functional kinetic advantage,which in line with co-localization principles of evolutional origin,and allosteric control in protein-protein-interactions.

Thermodynamics of Stability Constant of Binary Complex of Nicotinamide with Mn^(++)

The stability constants of the binary complexes of Mn2+ with nicotinamide (NA) were determined from potentiometric pH titrations data at 15.0 , 25.0 and 35.0oC and I = 0.1,0.2,0.4 mol L-1 (NaClO4). The formation of binary 1:1 , 1:2 NA-Mn complexes at three different temperatures and the influence of three different ionic strength on their stability were reported. The thermodynamic parameters (DGof, DSof, DHof) for the complex formation reaction were estimated from stability constant at different temperatures.

Synthesis and Crystal Structure of an Inclusion Compound Constructed through Self-assembly of Nicotinamide and Dicyanamide Coordination Bridges

A novel inclusion compound of [Mn（dca）2（3-nic）2]n·（3-nic）2n（1,dca = dicyanamide,3-nic = nicotinamide） has been prepared and characterized by single-crystal X-ray diffraction,elemental analysis and IR.This complex crystallizes in the triclinic system,space group P1 with a = 7.5979（8）,b = 7.7128（9）,c = 14.5346（17）,α = 100.094（2）,β = 92.444（2）,γ = 116.736（2）o,V = 741.72（15） 3,Z = 1,Dc = 1.512 g/cm3,Mr = 675.55,μ = 0.507 mm-1,F（000） = 347,S = 1.023,the final R = 0.0400 and wR = 0.1010.The inclusion compound is constructed by a three-dimensional host network that consists of dca,3-nic and Mn2＋ with coordination and hydrogen-bonding interactions.In the network,one-dimensional rectangular channels are formed,and the guest molecules（3-nic） are included in the channels.The guest molecules interlink with the host through strong hydrogen bonds.

Nicotinamide mononucleotide supplementation improves the quality of porcine oocytes under heat stress

Background:Elevated ambient temperature-caused heat stress is a major concern for livestock production due to its negative impact on animal feed intake,growth,reproduction,and health.Particularly,the germ cells are extremely sensitive to the heat stress.However,the effective approach and strategy regarding how to protect mammalian oocytes from heat stress-induced defects have not been determined.Methods:Germinal vesicle(GV)porcine oocytes were cultured at 41.5℃ for 24 h to induce heat stress,and then cultured at 38.5℃ to the specific developmental stage for subsequent analysis.Nicotinamide mononucleotide(NMN)was dissolved in water to 1 mol/L for a stock solution and further diluted with the maturation medium to the final concentrations of 10μmol/L,20μmol/L,50μmol/L or 100μmol/L,respectively,during heat stress.Immunostaining and fluorescence intensity quantification were applied to assess the effects of heat stress and NMN supplementation on the key processes during the oocyte meiotic maturation.Results:Here,we report that NMN supplementation improves the quality of porcine oocytes under heat stress.Specifically,we found that heat stress resulted in oocyte maturation failure by disturbing the dynamics of meiotic organelles,including the cytoskeleton assembly,cortical granule distribution and mitochondrial function.In addition,heat stress induced the production of excessive reactive oxygen species(ROS)and DNA damage,leading to the occurrence of apoptosis in oocytes and subsequent embryonic development arrest.More importantly,we validated that supplementation of NMN during heat stress restored the meiotic defects during porcine oocyte maturation.Conclusions:Taken together,our study documents that NMN supplementation is an effective approach to improve the quality of oocytes under heat stress by promoting both nuclear and cytoplasmic maturation.

Effect of Biophytum sensitivum on streptozotocin and nicotinamideinduced diabetic rats

Objective:To investigate the effect of aqueous solution of Biophytum sensUiuum leaf extract(BSEt)on normal and strcptozotocin(STZ)-nicotinamide-induced diabetic rats.Methods:Diabetes was induced in adult male Wislar rata by the administration of STZ-nicotinamide(40,110 mg/kg b.w.,respectively)intraperitoneally.BSEt(200 mg/kg)was administered to diabetic rats for 28 days.The effect of extract on blood glucose,plasma insulin,total haemoglobin,glycosylated haemoglobin,liver glycogen and carbohydrate melatxilism regulating enzymes of liver was studied in diabetic rats.Results:BSEt significantly reduced the blood glucose and glycosylated haemoglobin levels and significantly increased the total haemoglobin,plasma insulin and liver glycogen levels in diabetic rats.It also increased the hexokinase activity and decreased glucose-6-phosphatase,fructose-1,6-bisphosphatase activities in diabetic rats.Conclusions:The results of our study suggest that BSEt possesses a promising effecl on STZ-nicotinamide-induced diabetes.

Sitagliptin,sitagliptin and metformin,or sitagliptin and amitriptyline attenuate streptozotocin-nicotinamide induced diabetic neuropathy in rats

Diabetic neuropathies are a family of nerve disorders caused by diabetes. Symptoms of the disease include nerve palsy, mononeuropathy, mononeuropathy multiplex, diabetic amyotrophy, painful polyneuropathy, autonomic neu- ropathy, and thoracoabdominal neuropathy. In this study, type 2 diabetes in rats was induced with nicotinamide- streptozotocin. Drug treatment was initiated on the d 15, with the combination regimen of metformin, pioglitazone and glimipiride or metformin and sitagliptin or sitagliptin, amitriptyline and sitagliptin and led to significantly im- proved glycemic control, increased grip strength and paw jumping response on d 21, 28 and 35 （P 〈 0.001）. Signif- icant increases in blood protein levels and decreases in urinary protein levels were observed in the animals treated with the different regimens on d 21, 28 and 35 （P 〈 0.001）. Combined treatment of streptozotocin and nicotinamide caused marked degeneration of nerve cells, while administration of metformin and sitagliptin showed tissue regen- eration and no body weight gain. In conclusion, treatment with sitagliptin and sitagliptin combined with metformin or amitriptyline results in no body weight gain, but causes an increase in grip strength and pain sensitivity, exhibits neural protection, and reverses the alteration of biochemical parameters in rats with streptozotocin-nicotinamide induced type 2 diabetes.

NAD replenishment with nicotinamide mononucleotide protects blood-brain barrier integrity and attenuates delayed tissue plasminogen activator-induced haemorrhagic transformation after cerebral ischaemia

OBJECTIVE Tissue plasminogen activator(tPA) is the only approved pharmaco.logical therapy for acute brain ischaemia;however,a major limitation of tPA is the haemorrhagic trans.formation that follows tPA treatment.Here,we determined whether nicotinamide mononucleotide(NMN),a key intermediate of nicotinamide adenine dinucleotide biosynthesis,affects tPA-induced haemorrhagic transformation.METHODS Middle cerebral artery occlusion(MCAO) was achieved in CD1 mice by introducing a filament to the left MCA for 5 h.When the filament was removed for reperfusion,tPA was infused via the tail vein.A single dose of NMN was injected i.p.(300 mg·kg^(-1)).Mice were killed at 24 h post ischaemia,and their brains were evaluated for brain infarction,oedema,haemoglobin content,apoptosis,neuroinflammation,blood-brain barrier(BBB) permeability,the expression of tight junction proteins(TJPs) and the activity/expression of MMPs.RESULTS In the mice infused with tPA at 5 h post ischaemia,there were significant increases in mortality,brain infarction,brain oedema,brain haemoglobin level,neural apoptosis,Iba-1 staining(microglia activation) and myeloperoxidase staining(neutrophil infiltration).All these tPA-induced alterations were significantly prevented by NMN administration.Mechanistically,the delayed tPA treatment increased BBB permeability by downregulating TJPs,including claudin-1,occludin and zonula occludens-1,and enhancing the activities and protein expression of MMP9 and MMP2.Similarly,NMN administration partly blocked these tPAinduced molecular changes.CONCLUSION Our results demonstrate that NMN ameliorates tPAinduced haemorrhagic transformation in brain ischaemia by maintaining the integrity of the BBB.

Syntheses, Crystal Structures and Antitumor Activities of Two Nicotinamides

Two nicotinamides have been synthesized by coupling aromatic amines with equimolar quantities of substituted nicotinic acids. Their structures have been determined by X-ray single-crystal diffraction. Compound 1, 6-chloro-N-（4-fluorophenyl）nicotinamide, crystallizes in the triclinic space group Pī with a = 4.2188（8） , b = 5.0687（10）, c = 25.956（5）, α = 86.38（3）, β = 87.75（3）, γ = 83.20（3）°, V = 549.75（19） 3 and Z = 2. Compound 2, N-（quinolin-3-yl）nico- tinamide, crystallizes in the monoclinic space group P21 with a = 8.5100（17）, b = 6.4230（13）, c = 11.620（2） , β = 110.39（3）°, V = 595.3（2） 3 and Z = 2. The cytotoxic activities of the two nicotinamides against five human tumor cell lines were tested in vitro. The results indicated that they showed a wide range of antitumor activities.

Structure-function Relationships in Human Hypoxanthine-guanine Phosphoribosyltransferase(HGPRT) by Random Mutagenesis

Hypoxanthine-guanine phosphoribosyltransferase （ HGPRT, EC 2.4.2.8） is a key enzyme of the purine salvage pathway, which allows recycling of purine bases into DNA and RNA. It is widely distributed in nature and has been studied both in prokaryotes and eukaryotes. In humans, a complete lack of HGPRT activity causes the Lesch-Nyhan syndrome, which is characterized by hyperuricaemia and neural disorders,

Study of Tripterygium Associated with Nicotinamide in Treating Late-onset Autoimmune Diabetes Mellitus in Adults

Objective: To explore the effect of Tripterygium polyglycoside (TP) associated with nicotinamide on the islet cell function, immune parameters and lipoperoxide (LPO) in adult patients with late-onset autoimmune diabetes mellitus (LADA). Methods: Thirty-six cases of LADA were randomly divided into three groups: TP group (n=12), treated with TP plus orally taken metformin; combined treatment group (n= 12), treated with TP combined with nicotinamide and metformin, and control group (n=12) treated with metformin alone. They were followed-up for 18 months. Results: (1) Compared with the control group after 9 months of treatment, postprandial plasma glucose and LPO in combined treatment group were decreased (P <0.05), and the postprandial C-peptide was higher (P<0.05). At the 18th month, the value of postprandial C-peptide in the TP and combined treatment group was higher than that in the control group. The slL-2R level of both TP and combined treatment groups were lowered (P<0.01); (2) Islet cell antibody (ICA) positive of 5 cases in the TP group and 6 cases in the combined treatment group got converted to the negative respectively , while only one in the control group at the time (P<0.05); (3) The level of LPO in the combined treatment group was significantly lower than that in the TP group at the 18th month of treatment (P<0. 05). Conclusion: TP combined with nicotinamide played a role in immunity regulation, decreasing the titer of islet cell antibody and slL-2R, which also reduced the production of LPO and had a tendency to improve islet cell function in early LADA patients.