Biomedical subjects

T Uchida

Publications and source records attributed to T Uchida.

At least 325 records · Page 18Linked to original sources

PubMed1997

Meandering and unstable reentrant wave fronts induced by acetylcholine in isolated canine right atrium.

The mechanism(s) by which acetylcholine (ACh) increases atrial vulnerability to reentry and maintains its activity for longer durations remains poorly defined. In the present study we used high-resolution activation maps to test the hypothesis that ACh promotes meandering of atrial reentrant wave fronts, resulting in breakup and the generation of new wave fronts that sustain the activity. Reentry was induced in 11 isolated canine right atrial tissues (3.8 x 3.2 cm) by a premature point stimulus (S2) before and after superfusion with ACh (15 x 10(-6) M). Endocardial isochronal activation maps were constructed with the use of 509 bipolar electrodes (1.6-mm spatial resolution), and the dynamics of the activation wave fronts were visualized with animation. A vulnerable period was found during which an S2 current strength > 4.4 +/- 2.5 mA [lower limit of vulnerability (LLV)] and < 26 +/- 13 mA [upper limit of vulnerability (ULV)] induced a single stationary reentrant wave front that lasted 3 +/- 2.5 s with a period of 159 +/- 17 ms (16 episodes). AC shortened the refractory period from 100 +/- 12 to 59 +/- 9 ms (P < 0.001) and increased vulnerability to reentry induction by simultaneous decrease in the LLV (0.7 +/- 0.2 mA, P < 0.001) and an increase in the ULV (82 +/- 24 mA, P < 0.01). ACh accelerated the rate (period of 110 +/- 16 ms, P < 0.001) and converted the stationary reentrant wave front to a nonstationary (meandering) reentrant wave front showing polymorphic electrograms, i.e., "fibrillation-like" activity (22 episodes). Rapid meandering of the reentry tip led to wave front breakup (18 episodes) and the generation of new wave fronts (19 episodes). These wave front dynamics also led to sustained (76 +/- 224 s, P < 0.001) fibrillation-like electrograms. We conclude that ACh increases the ULV and promotes meandering of a single reentrant wave front, leading to breakup and the generation of new wave fronts. Single meandering and complex wave front dynamics cause fibrillation-like activity and sustain the activity for longer duration.

Acetylcholine↗

PubMed1997

Protection against verocytotoxin in mice induced by liposome-coupled verocytotoxin.

Purified verocytotoxins (VTs), VT1 and VT2, were coupled to liposomes via glutaraldehyde. During the coupling procedure, both VT1 and VT2 were detoxified. Intraperitoneal injection in BALB/c mice with either VT1-liposome or VT2-liposome induced a substantial amount of anti-VT1 or anti-VT2 IgG antibody production, respectively. Mice immunized with VT2-liposome were protected against intravenous challenge with a lethal dose of VT2 and the degree of protection correlated well with the amount of IgG induced against VT2. Although VT1-liposome failed to induce protection against VT1, the decrease of the body weight observed after the toxin challenge correlated inversely with the amount of anti-VT1 IgG induced, suggesting that VT1 neutralizing antibody was present in VT1-liposome-immune mice. In addition, VT-liposome conjugate induced no detectable anti-VT IgE antibody production. These results demonstrate the potential ability of VT-liposome conjugates for the production of VT vaccine which induces protection against VTs.

Animals↗

PubMed1997

Attachment of meandering reentrant wave fronts to anatomic obstacles in the atrium. Role of the obstacle size.

Acetylcholine chloride (ACh) induces nonstationary meandering reentrant wave fronts in the atrium. We hypothesized that an anatomic obstacle of a suitable size prevents meandering by causing attachment of the reentrant wave front tip to the obstacle. Eight isolated canine right atrial tissues (area, 3.8 x 3.2 cm) were mounted in a tissue bath and superfused with Tyrode's solution containing 10 to 15 mumol/L ACh. Holes with 2- to 10-mm diameters were sequentially created in the center of the tissue with biopsy punches. Reentry was induced by a premature stimulus after eight regular stimuli at 400-ms cycle length. The endocardial activation maps and the motion of the induced reentry were visualized dynamically before and after each test lesion using 509 bipolar electrodes. In the absence of a lesion (n = 8), the induced single reentrant wave front, in the form of a spiral wave, meandered irregularly from one site to another before terminating at the tissue border. Holes with 2- to 4-mm diameters (n = 6) had no effect on meandering. However, when the hole diameters were increased to 6 mm (n = 8), 8 mm (n = 8), and 10 mm (n = 6), the tip of the spiral wave attached to the holes, and reentry became stationary. Transition from meandering to an attached state converted the irregular and polymorphic electrogram to a periodic and monomorphic activity with longer cycle lengths (101 +/- 11 versus 131 +/- 9 ms for no hole versus 10-mm hole, respectively; P < .001). Regression analysis showed a significant positive linear correlation between the cycle length of the reentry and the hole diameter (r = .89, P < .01) and between the cycle length of the reentry and the excitable gap (r = .89, P < .05). We conclude that a critically sized anatomic obstacle converts a nonstationary meandering reentrant wave front to a stationary one. This transition converts an irregular "fibrillation-like" activity into regular monomorphic activity.

Acetylcholine↗

PubMed1997

A case of favorable dilation of protected left main coronary artery lesion achieved through performance of adjunctive DCA on residual stenosis following use of rotablator.

It is believed that directional coronary atherectomy (DCA) is more suitable than percutaneous transluminal coronary angioplasty for lesions such as severe eccentric lesions, ostial lesions, and branch lesions. However, it remains a fact that lesions that are also highly calcified are often suboptimal, since there may be difficulties such as in insertion of the housing and in sufficient cutting and removal. On the other hand, Rotablator is effective on calcified lesions, but afterward, dilation by balloon angioplasty for residual stenosis becomes necessary in many cases. This is a report of the authors' experience on an interesting case in which favorable dilation of a lesion in the protected left main coronary artery (LMCA) was achieved by using Rotablator after confirmation of a high degree of calcification by means of intravascular ultrasound (IVUS) echocardiograhy, followed by the performance of DCA on the residual stenosis.

Angioplasty, Balloon↗

PubMed1997

Sheathlin: cloning, cDNA/polypeptide sequences, and immunolocalization of porcine enamel sheath proteins.

Sheath proteins designate low-molecular-weight non-amelogenin enamel polypeptides and their parent protein, which concentrate in the sheath space separating rod and inter-rod enamel (Uchida et al., 1995). Two porcine sheath proteins, with apparent molecular weights of 13 and 15 kDa, are characterized by protein sequencing. The primary structures of these polypeptides match a portion of the derived amino acid sequences of clones isolated from a porcine enamel organ epithelia-specific cDNA library. Sheath protein RNA messages differ by the inclusion or deletion of a 45-nucleotide segment and by the use of three alternative polyadenylation/cleavage sites. The secreted proteins are 395 and 380 residues in length, with molecular masses of 42,358 and 40,279 Daltons and calculated isoelectric points of 6.3 and 6.7, respectively. Polyclonal antibodies were raised against a synthetic peptide having the sheathlin-specific sequence EHETQQYEYSGGC. Immunohistochemistry with this antibody demonstrates that the protein encoded by the sheathlin cDNA is preferentially localized in the sheath space. We propose that the porcine sheath proteins and their proteolytic cleavage products be designated "sheathlin".

Amino Acid Sequence↗

PubMed1997

Cloning and characterization of porcine enamelin mRNAs.

Dental enamel forms by matrix-mediated biomineralization. The components of the developing enamel matrix are generally specific for that matrix. The primary structures of three enamel proteins-amelogenin, tuftelin, and sheathlin (ameloblastin/amelin)-have been derived from cDNA sequences. Here we report the cloning and characterization of mRNA encoding a fourth enamel protein: enamelin. The longest porcine enamelin cDNA clone has 3907 nucleotides, exclusive of the poly(A) tail. The primary structure of the secreted protein is 1104 amino acids in length. Without post-translational modifications, the secreted protein has an isotope-averaged molecular mass of 124.3 kDa and an isoelectric point of 6.5. Polymerase chain-reaction phenotyping of enamelin cDNA suggests that porcine enamelin transcripts are not alternatively spliced and use a single polyadenylation/cleavage site. Immunohistochemical and Western blot analyses with an affinity-purified antipeptide antibody specific for the enamelin carboxyl terminus demonstrate that enamelin is synthesized and secreted by secretory-phase ameloblasts. The parent protein is a 186-kDa glycoprotein that concentrates along the secretory face of the ameloblast Tomes' process. Intact enamelin and proteolytic cleavage products containing its carboxyl terminus are limited to the most superficial layer of the developing enamel matrix, while other enamelin cleavage products are observed in deeper enamel.

Amino Acid Sequence↗

PubMed1997

Synthesis, secretion, degradation, and fate of ameloblastin during the matrix formation stage of the rat incisor as shown by immunocytochemistry and immunochemistry using region-specific antibodies.

Rat ameloblastin is a recently cloned tooth-specific enamel matrix protein containing 422 amino acid residues. We investigated the expression of this protein during the matrix formation stage of the rat incisor immunohistochemically and immunochemically, using anti-synthetic peptide antibodies that recognize residues 27-47 (Nt), 98-107 (M-1), 224-232 (M-2), 386-399 (M-3), and 406-419 (Ct) of ameloblastin. Immunohistochemical preparations using antibodies Nt and M-1 stained the Golgi apparatus and secretory granules of the secretory ameloblast and the entire thickness of the enamel matrix. Only M-1 intensely stained the peripheral region of the enamel rods. Immunostained protein bands were observed near 65, 55, and below 22 kD. Immunohistochemical preparations using antibodies M-2 and Ct stained the Golgi apparatus and secretory granules of the ameloblast and the immature enamel adjacent to the secretion sites, but not deeper enamel layers. Immunostaining using M-2 and Ct revealed protein bands near 65 and 40-56 kD, and 65, 55, 48, 36, and 25 kD, respectively. M-3 stained the cis side of the Golgi apparatus but not the enamel matrix. This antibody recognized a protein band near 55 kD, but none larger. After brefeldin A treatment, immunoreaction of the 55-kD protein band intensified, and dilated cisternae of rER of the secretory ameloblast contained immunoreactive material irrespective of the antibodies used. These data indicate that ameloblastin is synthesized as a 55-kD core protein and then is post-translationally modified with O-linked oligosaccharides to become the 65-kD secretory form. Initial cleavages of the 65-kD protein generate N-terminal polypeptides, some of which concentrate in the prism sheath, and C-terminal polypeptides, which are rapidly degraded and lost from the enamel matrix soon after secretion.

Ameloblasts↗

PubMed1997

Enhanced myopathy following administration of hypolipidemic agents under urethane anesthesia.

The enhanced effect of urethane anesthesia on the serum creatine kinase (CPK) level following administration of hypolipidemic agents was examined to develop a convenient experimental screening method for drug-induced myopathy. After oral administration of a hypolipidemic agent to rats, 25% urethane solution was infused intravenously at a very low rate using a microinfusion pump. Blood samples were collected 7 h after drug administration and the risk of myopathy was evaluated based on the CPK level. When bezafibrate (BF), simvastatin (SV), or pravastatin (PV) (50-500 mg/kg) was orally administered under urethane infusion, enhanced elevation of the serum CPK level was observed dose dependently for BF and SV, but not for PV. The elevation of serum CPK was much higher with BF than with SV or PV. In addition, when SV or PV (50-500 mg/kg) was coadministered with 50 mg/kg of BF, there was a striking increase in the serum CPK level as compared with the drug alone. Without urethane infusion, no significant elevation in serum CPK level was observed even at a high dose of these hypolipidemic agents. These phenomena suggest that the urethane anesthesia enhanced the elevation of the serum CPK level following administration of hypolipidemic agents. We propose that this method is a simple and speedy screening test for drug-induced myopathy.

Anesthesia↗

PubMed1997

Preparation and characterization of polylactic acid microspheres containing bovine insulin by a w/o/w emulsion solvent evaporation method.

The objective of this study was to produce polylactic acid (PLA) microspheres containing bovine insulin as a sparingly water soluble model drug using a water-in-oil-in-water (w/o/w) emulsion solvent evaporation method. The preparative conditions were optimized. Employment of smaller internal aqueous phase volume (50 microliters or 100 microliters) in the manufacturing process, resulted in the high loading efficiency (over 95% of theoretical insulin loading efficiency). The addition of 10% (w/v) NaCl to the external aqueous phase (0.5% polyvinyl alcohol solution) reduced loading efficiency compared to the case where no NaCl was added to the external phase. The mean volume diameter for prepared PLA microspheres was in the region of 15-25 microns in all cases. PLA microspheres containing 5% and 10% insulin theoretically exhibited burst release in the initial stage. After a three week dissolution test, the surface of the microspheres became more porous due to the degradable characteristics of PLA polymer itself. Nevertheless, about 80% of the insulin still remained undegraded in PLA microspheres. Finally, insulin-loaded PLA microspheres (corresponding to 4 I.U. insulin) were administered to normal rats subcutaneously, and the pharmacological effect (a decrease in serum glucose level) was demonstrated.

Animals↗

PubMed1997

[Antihypertensive effects of a novel calcium antagonist, AE0047, in various hypertensive models].

The antihypertensive effects of oral or intravenous administration of AE0047, a novel 1,4-dihydropyridine-type calcium antagonist, were investigated in spontaneously hypertensive rats (SHR/crj), one kidney-one clip renal hypertensive rats (RHR), deoxycorticosterone acetate (DOCA)-salt hypertensive rats (DHR) and two kidney-one clip renal hypertensive dogs (RHD). AE0047 (1, 3, 10 mg/kg, p.o.) caused a dose-related reduction of systolic blood pressure (SBP) with low reflex tachycardia in SHR/crj and RHR. The effect reached its maximum at 2-4 hr after administration and was sustained for a long time. In DHR, AE0047 (0.3, 1, 3 mg/kg, p.o.) similarly showed the antihypertensive effects at 2-7 hr with no significant changes in heart rates (HR). The doses (ED30) of AE0047 required to decrease SBP by 30% were 2.6, 3.4 and 0.68 mg/kg in SHR/crj, RHR and DHR, respectively. In RHD, and AE0047 capsule (GJ-0956: 4, 8, 16, 32 mg/body, p.o.) produced dose-dependent and long lasting effects with a transient and slight increase in HR. Furthermore, the intravenous administration of AE0047 (10, 30, 100 micrograms/kg) produced the antihypertensive action slowly, reached a plateau 10 min later and then maintained for many hours. In contrast, nitrendipine (3-100 mg/kg, p.o., 3-30 micrograms/kg, i.v.) and nicardipine (1-30 mg/kg, p.o., 3-30 micrograms/kg, i.v.) exhibited a similar potency to AE0047, but these maximal effects were produced at 1-2 hr and 0.5-1 min in the case of oral and intravenous administration, respectively, with a rapid recovery in the above hypertensive rats. These results indicate that AE0047 exhibits an antihypertensive effect with a slow onset and long-lasting profile.

Administration, Oral↗

PubMed1997

Pharmacological properties of YM17E, an acyl-CoA:cholesterol acyltransferase inhibitor, and diarrheal effect in beagle dogs.

YM17E (1,3-bis[[1-cycloheptyl-3-(p-dimethylaminophenyl)ureido]methyl]ben zene dihydrochloride) was found to be a potent inhibitor of acyl-CoA:cholesterol acyltransferase (ACAT) in rabbit liver and intestine microsomes. Dixon plot analysis revealed that YM17E inhibited microsomal ACAT in a non-competitive manner. YM17E induced a marked decrease in serum cholesterol, especially in non-high-density lipoprotein (HDL) fractions, in cholesterol-fed rats and rats fed normal chow. Measurement of bile secretion after oral administration of YM17E in cholesterol-fed rats showed that the drug markedly accelerated the secretion of bile acids and neutral sterols. Furthermore, absorption of [3H]cholesterol from the gut of cholesterol-fed rats was significantly inhibited by YM17E. From these results, the hypocholesterolemic activity of YM17E in these animals resulted from both a decrease in cholesterol absorption from the gut and the stimulation of excretion of cholesterol from the liver into bile. However, YM17E caused secretory diarrhea in beagle dogs at near lipid lowering doses. When YM17E was administered at the same total dosage but divided into 5 daily administrations, the incidence of diarrhea was significantly reduced while its cholesterol lowering effect became stronger. These results suggest that the inhibition of intestinal and/or liver ACAT increases the risk of diarrhea development which, however, can be avoided by controlled drug administration in beagle dogs.

Animals↗

PubMed1997

High attenuation lymphadenopathy caused by amyloidosis.

We describe a patient with generalized lymphadenopathy due to amyloidosis which showed on computed tomography as masses with high attenuation. Haemorrhage within lymph nodes caused by vascular amyloid deposition was confirmed at autopsy.

Amyloidosis↗

PubMed1997

Monoclonal antibodies inhibiting invasion of cultured cells by Eimeria tenella sporozoites.

Monoclonal antibodies (MoAbs) secreted by 12 hybridomas and reactive with the surface antigens of E. tenella sporozoites were produced. These MoAbs designated as KC-1 to KC-12 were characterized as IgG3 kappa. In Western blot analysis, these MoAbs reacted with only one polypeptide band of sporozoite antigens having a molecular mass of 25 kDa. All the MoAbs were reactive with sporozoites, trophozoites, immature and mature first generation schizonts, first generation merozoites, and the interior structure of sporulating oocysts of E. tenella, however, not with any of the methanol-fixed sporozoites of E. acervulina, E. mitis, E. maxima, E. brunetti, E. necatrix and E. praecox. Invasion of primary culture of chicken kidney cells by E. tenella sporozoites was significantly inhibited by several of the MoAbs. These results suggest that these MoAbs recognizing the E. tenella-specific surface molecule are involved in the invasion of sporozoites into host cells.

Animals↗

PubMed1997

Treatment of sleep apnea with a new separated type of dental appliance (mandibular advancing positioner).

We investigated a new separated type of dental appliance (mandibular advancing positioner: MAP) that is mobile and allows free adjustment of mandibular advancement. In 8 adult male patients with sleep apnea syndrome (SAS), the mean apnea index (AI) decreased from 31.1 to 4.2, and the mean apnea hypopnea index (AHI) decreased from 44.2 to 11.7. The distance of mandibular advancement using the Flankfort horizontal plane as a standard ranged in the 8 SAS patients from 1.8 to 5.0 mm by lateral cephalograms. A high positive correlation was observed between the distance of mandibular advancement and the rate of improvement in AI (R2 = 0.878), or the rate of AHI (R2 = 0.861), showing a higher improvement rate with more marked mandibular advancement.

Adult↗

PubMed1997

CDKN2 (MTS1/p16INK4A) gene alterations in hematological malignancies.

Cyclin dependent kinases (CDKs) make complexes with cyclins, and regulate cell cycle progression by their serine/threonine kinase activities. CDK inhibitors (CDKIs) arrest the inappropriate progression of the cell cycle by combining with CDKs. Because the functional loss of CDKIs may permit unlimited cell growth, their disruptions are thought to be associated with tumorigenesis. Recently, one CDKI, p16, was found, and its gene, CDKN2 (MTS1/p16INK4A), was identified on chromosome 9p21. Intensive investigations of the CDKN2 gene in various tumors have shown that alterations frequently occur in this gene, thus suggesting that the CDKN2 gene is a tumor suppressor gene. In hematological malignancies, CDKN2 gene alterations may be limited to lymphoid malignancies, especially T-cell type acute lymphocytic leukemias, in which frequent chromosomal abnormalities in the 9p21 region have been reported. The CDKN2 gene is also inactivated in some patients with non-Hodgkin's lymphomas, adult T-cell leukemias, and lymphoid blastic crisis of chronic myelogenous leukemias. The main mechanism of CDKN2 gene inactivation is thought to be homozygous deletion, but point mutations may also inactivate it in some cases. The CDKN2 gene appears to be the major tumor suppressor gene on chromosome 9p21, and it is thought to be involved in the tumorigenesis of various lymphoid malignancies.

Carrier Proteins↗

PubMed1997

[Androgen receptor gene mutations in prostate cancer].

PURPOSE: Androgens are required for the development of normal prostate and prostate cancer, through their action via the androgen receptor (AR). Although prostate cancer is potentially curable in the early stages by radical prostatectomy, androgen ablation is standard treatment for metastatic prostate cancer. Metastatic prostate cancer is incurable despite temporary remission commonly achieved by androgen ablation therapy. To investigate the mechanism for the development of human prostate cancer, examination was made of AR gene mutations. MATERIALS AND METHODS: Thirty-two samples including 29 primary prostate cancers and 3 metastatic lymph nodes were examined from exons B to H of the AR gene by polymerase chain reaction of single-strand conformation polymorphism (PCR-SSCP) and direct-sequencing analysis. Three metastatic lymph nodes were removed from non-hormone treated stage D1 patients by radical prostatectomy. Six of 11 stage D2 patients were hormone-independent stage following androgen ablation therapy. RESULTS: One of 29 (3.4%) primary prostate cancers and 1 of 6 (16.7%) hormone-independent stage D2 patients showed the AR mutation. This AR mutation is a G to A transition at nucleotide 2677 that leads to substitution of glutamine (CAG) for the wild type arginine (CGG) at codon 629. The serum prostate specific antigen level of the patients increased to 480 ng/ml. Drugs for hormone therapy and duration of treatment had the same effects on the remaining 5 hormone-independent patients. No mutation was found in the other 28 primary prostate cancer or 3 metastatic lymph node samples. CONCLUSIONS: The AR mutation may possibly be involved in the development of prostate cancer from the androgen-dependent to -independent stage during androgen ablation therapy.

Aged↗

PubMed1997

[A case of fulminant hepatitis caused by antiandrogen, flutamide in a patient with prostate cancer].

Luteinizing hormone-releasing hormone analogue, 3.6 mg goserelin acetate and flutamide, 375 mg per day were used to treat metastatic prostate cancer in a 66-year-old male. Marked increase in serum aminotransferases and total bilirubin levels was noted after 7 weeks of treatment. The enzyme levels returned to normal at 2 months following discontinuation of the treatment. Though incidences of severe hepatotoxicity are rare (0.003-0.18%), cases of flutamide-induced death have been reported. Careful monitoring of patients is mandatory while administering flutamide.

Aged↗

PubMed1997

Physiology and pathology of tau protein kinases in relation to Alzheimer's disease.

Alzheimer's disease (AD) is characterized by neuronal cell death and two kinds of deposits, neurofibrillary tangles (NFT) and senile plaques. The main component of NFT is paired helical filaments (PHF), which mainly consist of hyperphosphorylated tau protein. Tau protein kinases I and II were found as candidate enzymes responsible for hyperphosphorylation of tau to induce the formation of PHF. Since prior phosphorylation of tau by TPKII strongly enhanced the action of TPKI, it was thought that TPKII was involved in the formation of PHF-tau in concert with TPKI. After cloning, TPKI was found to be identical with glycogen synthase kinase 3 beta (GSK3 beta), while TPKII consists of a novel 23 kDa protein activator and a catalytic subunit that is identical with cyclin-dependent kinase 5 (CDK5). The phosphorylation sites on tau by TPKI and TPKII could account for the most, but not all, of the major phosphorylation sites of fetal tau and PHF-tau. An antibody for a site specifically phosphorylated by TPKI (Ser413) could identify all three neurofibrillary lesions in the AD brain, and double staining for either TPKI or TPKII and NFT in the brain of Down's syndrome patients clearly demonstrated that TPKI and TPKII are both associated with NFT in vivo, suggesting that the level of TPKI or TPKII is elevated in AD brain by some mechanism. On the other hand, the levels of both TPKs change developmentally, being high in the neonatal period when the phosphorylation of fetal tau proceeds actively, suggesting that the TPKI/TPKII cooperative system has an important physiological role in the formation of neural networks. In AD brain, aberrant accumulation of amyloid-beta protein (A beta) occurs ahead of the accumulation of PHF in NFT. When a primary culture of embryonic rat hippocampus was treated with 20 microM A beta, induction of TPKI, extensive phosphorylation of tau and then programmed cell death were observed, indicating that TPKI induced by A beta phosphorylates tau, followed by disruption of axonal transportation and finally cell death. By using a yeast two hybrid system, TPKI was found to interact with pyruvate dehydrogenase (PDH), which is a key enzyme in the glycolytic pathway. PDH was phosphorylated in vitro by TPKI to reduce the activity converting pyruvate into acetyl-CoA, which is required for acetylcholine synthesis. In a primary culture of rat hippocampal cells treated with A beta, PDH was inactivated in inverse relation to the activation of TPKI, resulting in accumulation of pyruvate or lactate, energy failure induced by the disturbance of glucose metabolism, and a shortage of acetylcholine owing to deficiency of acetyl-CoA, all of which are characteristic of AD brain. In cholinergic neurons such as those of the septum, non-aggregated A beta, specifically A beta (1-42), not A beta (1-40), caused a shortage of acetylcholine by activation of TPKI and inactivation of PDH without cell death.

Alzheimer Disease↗