聚腺苷二磷酸核糖聚合酶抑制剂

综述近年来聚腺苷二磷酸核糖聚合酶抑制剂的研究进展，重点评述新发现的该酶抑制剂的结构类型、生物活性、构效关系及其潜在的临床应用价值。聚腺苷二磷酸核糖聚合酶是人体重要的酶之一，参与多种生理病理过程。     

聚腺苷二磷酸核糖聚合酶抑制剂药物不良反应信号挖掘与分析

目的：挖掘、分析聚腺苷二磷酸核糖聚合酶（PARP）抑制剂的药物不良反应（ADR）信号,为提高临床用药安全提供参考。方法：应用英国药品和保健品管理局（MHRA）的综合标准法挖掘美国FDA不良事件报告系统（FAERS）,调取2014年1月1日 ~ 2021年12月31日的不良反应事件数据,使用SQL SERVER（2008 R2）数据库整理信息,以4种PARP抑制剂的名称作为检索关键词。结果：截至2021年12月31日,总共收集9 849 888例不良反应事件信息,其中与尼拉帕利相关的不良反应事件9849例,奥拉帕利6655例,他拉唑帕利420例,卢卡帕利3327例;经挖掘后分别得到尼拉帕利的ADR信号597个,奥拉帕利168个,他拉唑帕利35个,卢卡帕利132个。所有信号涉及25个系统,以各类检查指标、胃肠系统疾病症状、血液及淋巴系统疾病症状和良性、恶性及性质不明的肿瘤这4个系统的信号最多。汇总4种PAPR抑制剂共有的前7种信号分别为糖类抗原125升高、血小板计数下降、卵巢癌复发、恶心、肿瘤标志物升高、恶性肿瘤进展和贫血症,其中尼拉帕利和奥拉帕利相关血小板计数下降和恶心信号的时间强度曲线均不稳定。结论：临床使用PARP抑制剂应重点关注血液学检验指标异常和胃肠系统不良反应,尤其对于低体重或基线血小板计数下降的患者,应加强尼拉帕利的药学监护。     

Conformational switches in the VPAC(1) receptor

The vasoactive intestinal peptide receptor 1 (VPAC(1) ) belongs to family B of GPCRs and is activated upon binding of vasoactive intestinal peptide (VIP) and pituitary AC-activating polypeptide neuropeptides. Widely distributed throughout body, VPAC(1) plays important regulatory roles in human physiology and physiopathology. Like most members of the GPCR-B family, VPAC(1) receptor is predicted to follow the actual paradigm of a common 'two-domain' model of natural ligand action. However the precise structural basis for ligand binding, receptor activation and signal transduction are still incompletely understood due in part to the absence of X-ray crystal structure of the whole receptor and to significant structural differences with the most extensively studied family of receptor, the GPCR-A/rhodopsin family. Here, we try to summarize the current knowledge of the molecular mechanisms involved in VPAC(1) receptor activation and signal transduction. This includes search for amino acids involved in the two-step process of VIP binding, in the stabilization of VPAC(1) inactive and active conformations, and in binding and activation of G proteins.     

Real-time characterization of cannabinoid receptor 1 (CB1) allosteric modulators reveals novel mechanism of action

BACKGROUND AND PURPOSE

The cannabinoid receptor type 1 (CB₁) has an allosteric binding site. The drugs {"type":"entrez-protein","attrs":{"text":"ORG27569","term_id":"1179174593"}}ORG27569 {5-chloro-3-ethyl-N-[2-[4-(1-piperidinyl)phenyl]ethyl]-1H-indole-2-carboxamide} and PSNCBAM-1 {1-(4-chlorophenyl)-3-[3-(6-pyrrolidin-1-ylpyridin-2-yl)phenyl]urea} have been extensively characterized with regard to their effects on signalling of the orthosteric ligand CP55,940 {(−)-cis-3-[2-hydroxy-4-(1,1-dimethylheptyl)phenyl]-trans-4-(3-hydroxypropyl)cyclohexanol}, and studies have suggested that these allosteric modulators increase binding affinity but act as non-competitive antagonists in functional assays. To gain a deeper understanding of allosteric modulation of CB₁, we examined real-time signalling and trafficking responses of the receptor in the presence of allosteric modulators.

EXPERIMENTAL APPROACH

Studies of CB₁ signalling were carried out in HEK 293 and AtT20 cells expressing haemagglutinin-tagged human and rat CB₁. We measured real-time accumulation of cAMP, activation and desensitization of potassium channel-mediated cellular hyperpolarization and CB₁ internalization.

KEY RESULTS

{"type":"entrez-protein","attrs":{"text":"ORG27569","term_id":"1179174593"}}ORG27569 and PSNCBAM-1 produce a complex, concentration and time-dependent modulation of agonist-mediated regulation of cAMP levels, as well as an increased rate of desensitization of CB₁-mediated cellular hyperpolarization and a decrease in agonist-induced receptor internalization.

CONCLUSIONS AND IMPLICATIONS

Contrary to previous studies characterizing allosteric modulators at CB_1, this study suggests that the mechanism of action is not non-competitive antagonism of signalling, but rather that enhanced binding results in an increased rate of receptor desensitization and reduced internalization, which results in time-dependent modulation of cAMP signalling. The observed effect of the allosteric modulators is therefore dependent on the time frame over which the signalling response occurs. This finding may have important consequences for the potential therapeutic application of these compounds.     

Poly(ADP-ribose) polymerase (PARP) inhibition counteracts multiple manifestations of kidney disease in long-term streptozotocin-diabetic rat model

Evidence for the important role for poly(ADP-ribose) polymerase (PARP) in the pathogenesis of diabetic nephropathy is emerging. We previously reported that PARP inhibitors counteract early Type 1 diabetic nephropathy. This study evaluated the role for PARP in kidney disease in long-term Type 1 diabetes. Control and streptozotocin-diabetic rats were maintained with or without treatment with the PARP inhibitor 10-(4-methyl-piperazin-1-ylmethyl)-2H-7-oxa-1,2-diaza-benzo[de] anthracen-3-one (GPI-15,427, Eisai Inc.), 30 mg kg⁻¹ d⁻¹, for 26 weeks after first 2 weeks without treatment. PARP activity in the renal cortex was assessed by Western blot analysis of poly(ADP-ribosyl)ated proteins. Urinary albumin, isoprostane, and 8-hydroxy-2′-deoxyguanosine excretion, and renal concentrations of transforming growth factor-β₁, vascular endothelial growth factor, soluble intercellular adhesion molecule-1, fibronectin, and nitrotyrosine were evaluated by ELISA, and urinary creatinine and renal lipid peroxidation products by colorimetric assays. PARP inhibition counteracted diabetes-associated increase in renal cortex poly(ADP-ribosyl)ated protein level. Urinary albumin, isoprostane, and 8-hydroxy-2′-deoxyguanosine excretions and urinary albumin/creatinine ratio were increased in diabetic rats, and all these changes were at least partially prevented by GPI-15,427 treatment. PARP inhibition counteracted diabetes-induced renal transforming growth factor-β₁, vascular endothelial growth factor, and fibronectin, but not soluble intercellular adhesion molecule-1 and nitrotyrosine, accumulations. Lipid peroxidation product concentrations were indistinguishable among control and diabetic rats maintained with or without GPI-15,427 treatment. In conclusion, PARP activation plays an important role in kidney disease in long-term diabetes. These findings provide rationale for development and further studies of PARP inhibitors and PARP inhibitor-containing combination therapies, for prevention and treatment of diabetic nephropathy.     

PARP抑制剂对糖尿病大鼠早期神经病变的影响

目的：探讨腺苷二磷酸-核酸聚合酶（PARP）抑制剂3-氨基苯甲酰胺（3-aminobenzamide，3-AB）对实验性糖尿病大鼠神经病变的影响及可能机制。方法：实验动物分为糖尿病对照组（DC组）及糖尿病3-AB治疗组（DT组）及正常对照组（NC组），4周后测定坐骨神经传导速度（NCV），血清超氧化物歧化酶（SOD）活性和丙二醛（MDA）含量以及坐骨神经内ATP、磷酸肌酸（Pcr）、肌酸（Cr）含量。结果：和DC组相比．3-AB明显提高了DT组大鼠血清SOD活性和坐骨神经Pcr含量（P〈0．05），阻止了NCV的下降（P〈0．05），降低了MDA含量（P〈0．01），而坐骨神经ATP、Cr含量差异无统计学意义（P〉0．05）。结论：短期应用3-AB改善了糖尿病早期神经病变的神经能量状态，提高了神经传导速度，为糖尿病神经病变（DPN）的防治提供了一个新的思路。     

Inhibition of poly(ADP-ribose) polymerase (PARP) influences the mode of sulfur mustard (SM)-induced cell death in HaCaT cells

Sulfur mustard (SM) is a bifunctional alkylating agent. Its primary toxic consequence is severe skin damage with blisters, occurring after skin contact. These vesicant properties of SM have been linked to cell death of proliferating keratinocytes in the basal layer of the skin. Catalytic activation of the nuclear enzyme poly(ADP-ribose) polymerase (PARP-1) has been demonstrated to be a major event in response to high levels of DNA damage, and PARP-1 activation may be part of apoptotic signaling. In other contexts, overstimulation of PARP-1 triggers necrotic cell death because of rapid consumption of its substrate, beta-nicotinamide adenine dinucleotide (NAD+) and the consequent depletion of ATP. These findings prompted us to evaluate whether SM induces apoptosis in keratinocytes like HaCaT cells and to determine whether blocking of PARP enzyme activity with 3-aminobenzamide (3AB) can influence the mode of cell death. HaCaT cells were exposed to SM (10-1,000 microM; 30 min) and then cultivated in SM-free medium with or without 3AB for up to 48 h. This treatment resulted in a time and SM dose-dependent increase of apoptotic cell death characterized by PARP-1 cleavage and DNA fragmentation during the experimental period. After just 45 min of exposure to 1 mM SM, we observed a significant increase in PARP-1 activity in HaCaT cells. About 6 h after exposure, intracellular ATP levels were diminished by 22%, which seemed to be completely prevented by the addition of 3AB directly after exposure. However, 18 h later, this 3AB effect on the SM concentration-dependent loss of ATP was no longer detectable. Interestingly, the effect of SM on total cell viability was not changed by 3AB. However, the mode of cell death was influenced by 3AB exhibiting an increase of apoptotic cells and a concomitant decrease of necrotic HaCaT cells during the first 24 h after SM exposure. Our results indicate that SM concentrations of 1 mM or higher induce a prominent PARP activation leading to ATP depletion and necrosis. In contrast, lower concentrations of SM cause minor PARP activation and, especially, PARP-1 cleavage by caspase 3 without ATP depletion. Because ATP is required for apoptosis, we suggest that ATP acts as an early molecular switch from apoptotic to necrotic modes of SM-induced cell death, at least at high concentrations (> or =1 mM). Thus, the observed early proapoptotic effect of 3AB at lower SM concentrations may point to the influence of ATP-independent cell-death regulating mechanisms.     

New inhibitors of poly(ADP-ribose) polymerase (PARP)

Poly(ADP-ribose) polymerase-1 (PARP-1), the most prominent member of the PARP family, is a DNA-binding protein that is activated by nicks in DNA occurring during inflammation, ischaemia, neurodegeneration or cancer therapy. Activated PARP-1 consumes NAD⁺ that is cleaved into nicotinamide and ADP-ribose and polymerises the latter onto nuclear acceptor proteins. This highly energy consuming process is pivotal for the maintenance of genomic stability although over-activation can culminate in cell dysfunction and necrosis. Therefore, PARP-1 is regarded as a promising target for the development of drugs useful in various forms of inflammation, ischaemia–reperfusion injury and as an adjunct in cancer therapy. This review summarises the structural classes of known PARP-1 inhibitors, with a focus on new inhibitors published for this target, between 2002 and July 2004. The chemistry and biological data disclosed in these patent applications are discussed in light of new structural knowledge of the catalytic domain of the PARP family and recent work with potent inhibitors demonstrating the effects of PARP inhibition in various animal disease models.     

鲨鱼软骨粉对S_(180)荷瘤小鼠免疫细胞调节功能的影响

目的研究鲨鱼软骨粉(SCP)对S180荷瘤小鼠免疫细胞功能的影响。方法鲨鱼软骨粉300、200、100mg·kg-1小鼠灌胃给药10 d。MTT法检测其对S180荷瘤小鼠脾淋巴细胞增殖功能的影响;流式细胞仪检测T淋巴细胞亚群的变化;ELISA法测定荷瘤小鼠血清中细胞因子TNF-α、IFN-γ含量。结果 SCP能提高小鼠脾淋巴细胞的增殖活性;不同程度地提高小鼠脾细胞CD4+细胞数,增加CD4+/CD8+细胞比值;明显提高小鼠巨噬细胞的吞噬活性;促进外周血清中TNF-α和IFN-γ的分泌水平。结论 SCP能提高T细胞介导的细胞免疫应答,提高巨噬细胞调节免疫应答的功能,促进TNF-α、IFN-γ等细胞因子的分泌,从而发挥其抗肿瘤作用。     

Effects of palmitoylation of Cys(415) in helix 8 of the CB(1) cannabinoid receptor on membrane localization and signalling

BACKGROUND AND PURPOSE

The CB₁ cannabinoid receptor is regulated by its association with membrane microdomains such as lipid rafts. Here, we investigated the role of palmitoylation of the CB₁ receptor by analysing the functional consequences of site-specific mutation of Cys⁴¹⁵, the likely site of palmitoylation at the end of helix 8, in terms of membrane association, raft targeting and signalling.

EXPERIMENTAL APPROACH

The palmitoylation state of CB₁ receptors in rat forebrain was assessed by depalmitoylation/repalmitoylation experiments. Cys⁴¹⁵ was replaced with alanine by site-directed mutagenesis. Green fluorescence protein chimeras of both wild-type and mutant receptors were transiently expressed and functionally characterized in SH-SY5Y cells and HEK-293 cells by means of confocal microscopy, cytofluorimetry and competitive binding assays. Confocal fluorescence recovery after photobleaching was used to assess receptor membrane dynamics, whereas signalling activity was assessed by [³⁵S]GTPγS, cAMP and co-immunoprecipitation assays.

KEY RESULTS

Endogenous CB₁ receptors in rat brain were palmitoylated. Mutation of Cys⁴¹⁵ prevented the palmitoylation of the receptor in transfected cells and reduced its recruitment to plasma membrane and lipid rafts; it also increased protein diffusional mobility. The same mutation markedly reduced the functional coupling of CB₁ receptors with G-proteins and adenylyl cyclase, whereas depalmitoylation abolished receptor association with a specific subset of G-proteins.

CONCLUSIONS AND IMPLICATIONS

CB₁ receptors were post-translationally modified by palmitoylation. Mutation of Cys⁴¹⁵ provides a receptor that is functionally impaired in terms of membrane targeting and signalling.

LINKED ARTICLES

This article is part of a themed section on Cannabinoids in Biology and Medicine. To view the other articles in this section visit http://dx.doi.org/10.1111/bph.2012.165.issue-8. To view Part I of Cannabinoids in Biology and Medicine visit http://dx.doi.org/10.1111/bph.2011.163.issue-7     

Biased signalling and proteinase-activated receptors (PARs): targeting inflammatory disease

Although it has been known since the 1960s that trypsin and chymotrypsin can mimic hormone action in tissues, it took until the 1990s to discover that serine proteinases can regulate cells by cleaving and activating a unique four-member family of GPCRs known as proteinase-activated receptors (PARs). PAR activation involves the proteolytic exposure of its N-terminal receptor sequence that folds back to function as a ‘tethered’ receptor-activating ligand (TL). A key N-terminal arginine in each of PARs 1 to 4 has been singled out as a target for cleavage by thrombin (PARs 1, 3 and 4), trypsin (PARs 2 and 4) or other proteases to unmask the TL that activates signalling via G_q, G_i or G_12/13. Similarly, synthetic receptor-activating peptides, corresponding to the exposed ‘TL sequences’ (e.g. SFLLRN—, for PAR1 or SLIGRL— for PAR2) can, like proteinase activation, also drive signalling via G_q, G_i and G_12/13, without requiring receptor cleavage. Recent data show, however, that distinct proteinase-revealed ‘non-canonical’ PAR tethered-ligand sequences and PAR-activating agonist and antagonist peptide analogues can induce ‘biased’ PAR signalling, for example, via G_12/13-MAPKinase instead of G_q-calcium. This overview summarizes implications of this ‘biased’ signalling by PAR agonists and antagonists for the recognized roles the PARs play in inflammatory settings.Linked ArticlesThis article is part of a themed section on Molecular Pharmacology of GPCRs. To view the other articles in this section visit http://dx.doi.org/10.1111/bph.2014.171.issue-5     

Inhibition of NMDA-gated ion channels by bis(7)-tacrine: whole-cell and single-channel studies

Bis(7)-tacrine is a novel dimeric acetylcholinesterase inhibitor derived from tacrine, and has been proposed as a promising agent to treat Alzheimer's disease. We have recently reported that bis(7)-tacrine prevents glutamate-induced neuronal apoptosis by antagonizing NMDA receptors. The purpose of this study was to characterize bis(7)-tacrine inhibition of NMDA-activated current by using patch-clamp recording techniques. In cultured rat hippocampal neurons, bis(7)-tacrine inhibited NMDA-activated whole-cell current in a concentration-dependent manner with an IC₅₀ of 0.66 ± 0.07 μM. Bis(7)-tacrine produced a gradual decline of NMDA-activated current to a steady-state, but this was not an indication of use-dependence. Also, the slow onset of inhibition by bis(7)-tacrine was not apparently due to an action at an intracellular site. Bis(7)-tacrine, 0.5 μM, decreased the maximal response to NMDA by 40% without changing its EC₅₀. Bis(7)-tacrine inhibition of NMDA-activated current was not voltage-dependent, and was independent of glycine concentration. Results of single-channel experiments obtained from cells expressing NR1 and NR2A subunits revealed that bis(7)-tacrine decreased the open probability and frequency of channel opening, but did not significantly alter the mean open time or introduce rapid closures. These results suggest that bis(7)-tacrine can inhibit NMDA receptor function in a manner that is slow in onset and offset and noncompetitive with respect to both NMDA and glycine. The noncompetitive inhibition of NMDA receptors by bis(7)-tacrine could contribute to its protective effect against glutamate-induced neurotoxicity.     

Poly (ADP-ribose) polymerase (PARP) is essential for sulfur mustard-induced DNA damage repair, but has no role in DNA ligase activation

Concurrent activation of poly (ADP-ribose) polymerase (PARP) and DNA ligase was observed in cultured human epidermal keratinocytes (HEK) exposed to the DNA alkylating compound sulfur mustard (SM), suggesting that DNA ligase activation could be due to its modification by PARP. Using HEK, intracellular 3H-labeled NAD+ (3H-adenine) was metabolically generated and then these cells were exposed to SM (1 mM). DNA ligase I isolated from these cells was not 3H-labeled, indicating that DNA ligase I is not a substrate for (ADP-ribosyl)ation by PARP. In HEK, when PARP was inhibited by 3-amino benzamide (3-AB, 2 mM), SM-activated DNA ligase had a half-life that was four-fold higher than that observed in the absence of 3-AB. These results suggest that DNA repair requires PARP, and that DNA ligase remains activated until DNA damage repair is complete. The results show that in SM-exposed HEK, DNA ligase I is activated by phosphorylation catalysed by DNA-dependent protein kinase (DNA-PK). Therefore, the role of PARP in DNA repair is other than that of DNA ligase I activation. By using the DNA ligase I phosphorylation assay and decreasing PARP chemically as well as by PARP anti-sense mRNA expression in the cells, it was confirmed that PARP does not modify DNA ligase I. In conclusion, it is proposed that PARP is essential for efficient DNA repair; however, PARP participates in DNA repair by altering the chromosomal structure to make the DNA damage site(s) accessible to the repair enzymes.     

PARP抑制剂WZ-8体内外抗肿瘤作用研究

目的 研究PARP抑制剂WZ-8对人源性肿瘤细胞增殖及裸鼠移植人结肠癌（HCT116）生长的抑制作用。方法 SRB染色法和裸小鼠移植瘤模型研究WZ-8在体内外抗肿瘤作用。结果 体外WZ-8对多种人肿瘤细胞增殖有明显抑制作用,其中对结肠癌细胞HCT116的半数抑制浓度IC50为（0.21±0.01）mg·mL^-1。体内研究表明WZ-8（20 mg·kg-1）对小鼠移植HCT116肿瘤生长有一定抑制效果,其体内外抗肿瘤活性均高于同类化合物Iniparib。结论 WZ-8在体外对人源性肿瘤细胞增殖有明显抑制作用,在体内可抑制裸鼠移植人结肠癌细胞HCT116的生长。     

Dose and Load Studies for Subcutaneous and Oral Delivery of Poly(lactide-co-glycolide) Microspheres Containing Ovalbumin

Poly(lactide-co-glycolide) microspheres containing different loads of OVA (0.05, 0.1, 0.5 and 1.0% w/w) were manufactured by a w/o/w emulsion/solvent evaporation method. Low load efficiencies of less than 20% were observed. Normal size distributions with mean volume diameters ranging from 3.7 to 4.7 µm were obtained for different batches. The in vitro release of OVA from different loaded microspheres showed an expected burst release with all batches. The in vivo dose study (1, 10, 25, 50 µg of OVA) was performed by subcutaneous and oral inoculation in mice by single (0 week) or double (0 and 3 weeks) administration of PLGA 50/50 microspheres containing 0.1% OVA. Subcutaneous administration showed an immune response (serum Ig levels by ELISA) statistically (Fishers paired t-test; P < 0.05) above OVA saline negative controls at 3, 6 and 12 weeks after administration. Oral administration of microspheres produced statistically higher systemic immune responses at the higher doses. Single and double inoculation orally and subcutaneously produced similar serum antibody levels. The in vivo load study was performed by subcutaneous and oral administration to mice of 25 µg OVA contained in various loaded (0.05, 0.1, 0.5 and 1.0% w/w) microspheres. Serum immune responses at 3, 6, and 12 weeks after inoculation were statistically above OVA saline controls and were inversely proportional to the OVA load using either route. This observation suggested a relationship between the number of microspheres delivered and the in vivo serum response. Single subcutaneous administration of 0.05 or 0.1% OVA loaded PLGA 50/50 microspheres induced larger immune responses compared with complete Freunds adjuvant.     

Pharmacology of 5HT(2C) receptor-mediated ERK1/2 phosphorylation: agonist-specific activation pathways and the impact of RNA editing

We have previously characterized a mechanism of 5HT-stimulated extracellular signal-regulated kinases 1 and 2 (ERK1/2) activation via the non-RNA-edited isoform of the serotonin 5HT(2C) receptor (5HT(2C)R-INI) in a CHO cell line. We have now used CV1 cells, which endogenously express epidermal growth factor receptors (EGFRs), to investigate whether the mechanisms underlying ERK1/2 activation by the 5HT(2C)R change in a time-, agonist-, and cell background-dependent manner. Interrogation of the CV1 5HT(2C)R-INI ERK1/2 signaling pathway, using a variety of pathway-selective inhibitors, revealed a clear time-dependence in the involvement of specific pathway components such as phosphatidylinositol 3-kinase, EGFR, matrix metalloproteases and protein kinase C. The contribution of these components to the overall response also varied with the agonist used to stimulate the receptor, providing further evidence for the ability of 5HT(2C)R-INI to signal in an agonist-specific manner. We also investigated the impact of 5HT(2C)R RNA editing on this phenomenon. Although we found no alteration in antagonist pharmacology, the partially edited VSV and fully edited VGV isoforms of the 5HT(2C)R exhibited altered temporal and pharmacological characteristics, including the degree of dependence on specific effectors, in signaling to ERK1/2 in comparison to the 5HT(2C)R-INI. In conclusion, we provide evidence for remarkable flexibility in 5HT(2C)R-mediated ERK1/2 signaling that can be pharmacologically and mechanistically distinct depending on the agonist or edited isoform involved and on the duration of receptor activation.     

Inhibition of neuroleptic-induced dopamine receptor supersensitivity by cyclo (Leu-Gly)

Behavioral supersensitivity of dopamine receptors was induced in mice by chronic administration of haloperidol (1 mg/kg/day for 21 days) and its subsequent withdrawal for 48 hr. This was evidenced by enhanced spontaneous locomotor activity and hypothermic responses to a dopamine agonist, apomorphine. Concurrent administration of cyclo (Leu-Gly), the enzymatically resistant diketopiperazine, an analog of melantropin release inhibiting factor, blocked haloperidol-induced dopamine receptor supersensitivity as evidenced by the blockade of apomorphine induced responses. Since many studies have linked the development of neuroleptic induced tardive dyskinesias with enhanced sensitivity of brain dopamine receptors, and the latter was blocked by cyclo (Leu-Gly), this agent may be of value in preventing the development of symptoms of neuroleptic-induced tardive dyskinesias.