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Biomedical subjects

Y K Gupta

Publications and source records attributed to Y K Gupta.

At least 37 records · Page 2Linked to original sources
PubMed2002

Protective effect of adenosine against neuronal injury induced by middle cerebral artery occlusion in rats as evidenced by diffusion-weighted imaging.

In the present study, adenosine, an inhibitory neuromodulator, was studied in male Wistar rats subjected to 2 h of transient middle cerebral artery (MCA) occlusion. Adenosine (500 mg/kg ip) was administered twice-once at the time of MCA occlusion and again at the time of reperfusion-and evaluated for its protective effect by using diffusion-weighted imaging (DWI) (30 min after reperfusion). After the DWI experiments, one group of animals was euthanized 2 h after reperfusion for the estimation of oxidative stress markers, while in another group, neurological deficit was assessed 24 h after MCA occlusion. In the adenosine-treated group, percent hemispheric lesion area (%HLA) in DWI was significantly attenuated (11.7+/-5.2) as compared to vehicle-treated group (21.4+/-4.7). The level of malondialdehyde (MDA) (301.8+/-22 nmol/g wet tissue) in the adenosine-treated group was significantly decreased as compared to that in the vehicle-treated MCA-occluded rats (420+/-20 nmol/g wet tissue). An insignificant change was observed in the levels of glutathione in both the vehicle-treated MCA-occluded and the adenosine-treated groups. The neurological deficit was significantly improved in the adenosine-treated group (1.8+/-0.06) as compared to the vehicle-treated (2.9+/-0.38) group. This is the first study to demonstrate the effectiveness of adenosine using DWI in the MCA-occluded rats.

Adenosine↗
PubMed2002

Intracerebroventricular administration of colchicine produces cognitive impairment associated with oxidative stress in rats.

Oxidative stress has been implicated in neurodegenerative disorders including the Alzheimer's disease (AD). Central administration of colchicine is known to cause cognitive impairment in rats and is likened to sporadic AD in humans. However, it is not known whether this cognitive impairment is associated with free radical generation. Therefore, in the present study, the effect of intracerebroventricular colchicine was studied on paradigms of learning and memory behavior and the markers of oxidative stress in rats. Adult male Wistar rats (200-250 g) were injected with colchicine (intracerebroventricular) bilaterally (15 microg/rat; 7.5 microg/site) on the first day. The learning and memory behavior was assessed using passive avoidance paradigm, elevated plus maze and closed field activity test on Days 13, 14 and 21. The parameters of oxidative stress were assessed by measuring the malondialdehyde (MDA), glutathione, superoxide dismutase (SOD) and catalase levels in brain tissue on Day 21 of the colchicine injection. The rats developed significant learning and memory impairment as indicated by deficit in behavioral paradigms. There was a significant elevation in MDA levels and decrease in levels of glutathione. No significant difference was observed in SOD and catalase levels. Thus, the study demonstrates that central administration of colchicine causes impairment in learning and memory with associated increase in oxidative stress.

Animals↗
PubMed2002

Effect of different extracts of Centella asiatica on cognition and markers of oxidative stress in rats.

Centella asiatica, a plant mentioned in Indian literature has been described to possess CNS effects such as stimulatory-nervine tonic, rejuvenant, sedative, tranquilizer and intelligence promoting property. In the present study aqueous, methanolic and chloroform extracts of C. asiatica were investigated for their effect on cognitive functions in rats. Male Wistar rats of 200-250 g were used to study the effect on learning and memory by using shuttle box, step through, step down and elevated plus maze paradigms. Only the aqueous extract of whole plant (200 mg/kg for 14 days) showed an improvement in learning and memory in both shuttle box and step through paradigms. Therefore, further experiments were conducted with aqueous extract using 100, 200 and 300 mg/kg doses in different paradigms of learning and memory. All doses of aqueous extract increased the number of avoidances in shuttle box and prolonged the step through latency in step through apparatus in a dose dependent manner, while only two doses 200 and 300 mg/kg of aqueous extract showed significant increase in the step down latency in step down apparatus and transfer latency (TL) in elevated plus maze. Among doses of aqueous extract tested on oxidative stress parameters, only 200 and 300 mg/kg showed a significant decrease in the brain levels of malondialdehyde (MDA) with simultaneous significant increase in levels of glutathione. There was a significant increase in the levels of catalase at the 300 mg/kg but no significant change in superoxide dismutase (SOD) levels were observed. The present findings indicate that the aqueous extract of C. asiatica has cognitive enhancing effect and an antioxidant mechanism is involved.

Animals↗
PubMed2002

Antioxidant property of Celastrus paniculatus willd.: a possible mechanism in enhancing cognition.

In the present study aqueous, methanolic, chloroform and petroleum ether extracts of seeds of Celastrus paniculatus were investigated for their effect on cognitive functions in rats. Male Wistar rats weighing 200-250 g each were used to study effect on learning and memory through use of the shuttle-box, step-through, step-down and elevated plus maze paradigms. Only the aqueous seed extract (200 mg/kg body wt. for 14 days) showed an improvement in learning and memory in both the shuttle-box and step-through paradigms. Therefore, further experiments were conducted using the aqueous extract at 100, 200 and 300 mg/kg body wt. doses in different paradigms of cognition. All three doses of the aqueous extract increased the number of avoidances in the shuttle-box and step-through latency the in step-through apparatus, but no significant difference was observed between the doses tested. In the step-down apparatus, the 200- and 300-mg/kg body wt. doses of aqueous extract showed a significant increase in step-down latency, whereas no significant difference was observed in the elevated-plus-maze paradigm between drug-treated and vehicle-treated groups. Since the behavioral impairments are associated with oxidative stress, we investigated the effect of the aqueous extract on oxidative stress parameters. Among the three doses tested, only 200 and 300 mg/kg body wt. stimulated a significant decrease in the brain levels of malondialdehyde, with simultaneous significant increases in levels of glutathione and catalase. The present findings indicate that the aqueous extract of Celastrus paniculatus seed has cognitive-enhancing properties and an antioxidant effect might be involved.

Animals↗
PubMed2002

Enhanced protection by melatonin and meloxicam combination in a middle cerebral artery occlusion model of acute ischemic stroke in rat.

Mixed efficacy of neuroprotective drugs in clinical trials has led to the emergence of the approach of combination therapy in stroke. The present study was carried out to investigate the effect of the combination of melatonin (potent antioxidant) and meloxicam (preferential inhibitor of cyclooxygenase-2 enzyme) against a middle cerebral artery occlusion model of stroke in rats. Male Wistar rats in the weight range of 250-300 g were used. Rats were anesthetized using chloral hydrate (400 mg/kg i.p) and subjected to 2 h of transient middle cerebral artery occlusion. Melatonin was administered at a dose of 20 mg/kg i.p. four times: at the time of middle cerebral artery occlusion, 1.5 h after middle cerebral artery occlusion, at the time of reperfusion, and 1 h after reperfusion. Meloxicam (2.5 mg/kg) was administered 4 h after middle cerebral artery occlusion. Motor performance tests (grip test, foot fault test, rotarod performance test, spontaneous locomotor activity), markers of oxidative stress, and triphenyltetrazolium chloride staining were carried out 24 h after middle cerebral artery occlusion. A vehicle-treated group was run in parallel. It was observed that melatonin treatment improved the motor performance and significantly attenuated the levels of malondialdehyde (MDA) as compared with the middle cerebral artery occluded group. Meloxicam treatment at the dose used neither showed significant improvement on the motor performance nor decreased the levels of MDA significantly as compared with the middle cerebral artery occluded group. However, when the combination of the two drugs was used, better protection was observed as was evident by the significant decrease in the percent foot fault errors, the increase in the time spent on the rotarod, and the increase in the six-point neurological score and grip test score. There was also a significant decrease in the levels of MDA in the combination group. The results of the present study demonstrate that enhanced protection is observed with the use of a combination of melatonin plus meloxicam in the middle cerebral artery occlusion model of acute ischemic stroke in rats.

Animals↗
PubMed2002

Protective effect of resveratrol against pentylenetetrazole-induced seizures and its modulation by an adenosinergic system.

The effect of trans-resveratrol (resveratrol), a polyphenolic compound with potent antioxidant activity, was investigated against pentylenetetrazole (PTZ) induced seizures in rats. Resveratrol (20, 40, and 80 mg/kg i.p.) administered 20 min prior to convulsive challenge with PTZ (60 mg/kg i.p.) dose dependently reduced the percent incidence of generalized tonic-clonic convulsions. Resveratrol (40 mg/kg) also potentiated the effect of sodium valproate (150 mg/kg) and diazepam (2 mg/kg) against PTZ-induced seizures. Since adenosine, an endogenous anticonvulsant, has been demonstrated to modulate the action of various antiepileptics, experiments were also carried out to determine whether an adenosinergic mechanism is involved in the anticonvulsant action of resveratrol. When a subanticonvulsant dose of adenosine (500 mg/kg) was administered together with resveratrol, a significant reduction in the percent incidence of generalized tonic-clonic convulsions was observed. Moreover, the nonspecific adenosine receptor antagonist theophylline (50 mg/kg i.p.) significantly reversed the resveratrol-induced protection, whereas the specific adenosine A2 receptor antagonist 3,7-dimethyl-1-propargylxanthine (1 mg/kg i.p.) could not reverse the resveratrol-induced protection. The findings of the present study suggest an antiepileptic potential of resveratrol and that an adenosinergic mechanism may play a role in its anticonvulsant activity.

Adenosine↗
PubMed2002

Effect of melatonin and phenytoin on an intracortical ferric chloride model of posttraumatic seizures in rats.

The present study was carried out to investigate the effect of melatonin, a potent antioxidant, and phenytoin, a conventional antiepileptic, against FeCl3-induced posttraumatic seizures. Male Wistar rats weighing 200-250 g were implanted with epidural electrodes and allowed to recover. After the recording of baseline EEG, FeCl3 (5 ul, 100 mM) was administered intracortically over a period of 5 min and EEG was monitored for 2 h. Subsequently, rats were sacrificed to estimate oxidative stress, i.e., the amount of malondialdehyde (MDA) in whole brain tissue. A sham group was run parallel with saline (pH adjusted), and a similar protocol for EEG recording and estimation of oxidative stress was followed. FeCl3-treated animals exhibited epileptiform EEG changes (high amplitude sharp waves of increased frequency and polyspikes) within 15 min, which continued throughout the period of observation. MDA levels were found to be significantly elevated as compared to the sham group. Melatonin (50 mg/kg i.p.) administered 30 min before FeCl3 injection delayed the onset of appearance of epileptiform EEG changes, while at a 100 mg/kg dose of there was complete protection, as none of the animal exhibited epileptiform EEG discharge. Brain MDA levels were also significantly reduced in melatonin (50 and 100 mg/kg dose)-treated animals as compared to the vehicle-treated FeCl3-injected rats. In the phenytoin group, all animals showed epileptiform EEG discharge. However, phenytoin at both 50 and 100 mg/kg dose delayed the onset of epileptiform EEG discharge. There were no significant changes in brain MDA levels in the phenytoin-treated group as compared to controls. Melatonin and phenytoin at doses of 100 mg/kg did not show any sign of motor impairment as observed during the rota-rod test. These findings showed a superior protective effect of melatonin over phenytoin in an intracortical FeCl3 model of posttraumatic epilepsy.

Animals↗
PubMed2002

Pyrogallol-induced hepatotoxicity in rats: a model to evaluate antioxidant hepatoprotective agents.

Various hepatic disorders and hepatotoxic agents are associated with increased free radical generation. In the present study, the free radical generator pyrogallol (100 mg/kg i.p.) caused significant hepatic damage. The serum enzymes asparatate aminotransaminase (AST) and alanine aminotransaminase (ALT) increased to 357 +/- 30.7 IU/I and 147.8 +/- 28.4 IU/I, respectively in the pyrogallol-treated group compared with 208.4 +/- 4.1 IU/I and 84.5 +/- 19.5 IU/I, respectively in the control rats. Compared with control rats, the liver tissue in the pyrogallol-treated group showed an increased level of malondialdehyde (MDA) as well as glutathione (GSH). The infiltration of white blood cells into the liver tissue, as seen histologically, further substantiated liver damage. Pretreatment with a standard hepatoprotective drug (silymarin, 100 mg/kg i.p.) afforded significant protection against pyrogallol hepatotoxicity, as evidenced by amelioration of the raised serum markers of hepatic function, markers of oxidative stress and normal liver histology. Thus, pyrogallol-induced hepatotoxicity could be used as an appropriate model to evaluate hepatoprotective agents that have an antioxidant property.

Alanine Transaminase↗
PubMed2002

Involvement of 5-HT1A and 5-HT2 receptor in cisplatin induced emesis in dogs.

The effect of drug acting on 5-HT1A, 5-HT2 and 5-HT3 receptors were studied against cisplatin and apomorphine induced emesis in dogs. Buspirone, 5-HT1A receptor partial agonist significantly reduced the emetic episodes though it had no significant effect on emetic latency. Mianserin, 5-HT2 receptor antagonist exhibited significant reduction in emetic episodes and in latency. Buspirone prevented the apomorphine induced emesis while mianserin had no effect. The antiemetic activity of buspirone may be attributable to its agonistic activity at 5-HT1A receptor and antagonistic activity at dopamine receptors. These findings further confirm the involvement of 5-HT1A and 5-HT2 receptor in cytotoxic drug induced emesis, though the species difference in their antiemetic action can not be ruled out.

Animals↗
PubMed2001

Effect of melatonin on ischemia reperfusion injury induced by middle cerebral artery occlusion in rats.

Free radicals have been implicated in neuronal injury during ischemia reperfusion in stroke. Therefore, in the present study, melatonin, a potent antioxidant, was studied in male Wistar rats subjected to 2 h of transient middle cerebral artery occlusion. Melatonin (10, 20 and 40 mg/kg i.p.) was administered four times in an animal at the time of middle cerebral artery occlusion, 1 h after middle cerebral artery occlusion, at the time of reperfusion and 1 h after reperfusion. Two hours after reperfusion, rats were euthanized for estimation of oxidative stress markers (malondialdehyde and reduced glutathione). The doses of 20 and 40 mg/kg of melatonin significantly attenuated the raised level of malondialdehyde (287+/-28, 279+/-52 nmol/g wet tissue, respectively) as compared to the levels (420+/-61 nmol/g wet tissue) in vehicle-treated middle cerebral artery-occluded rats. There was an insignificant change in levels of reduced glutathione at these doses (95+/-42, 88.7+/-36 microg/g wet tissue, respectively) as compared to those in the vehicle-treated middle cerebral artery-occluded rats (108.21+/-21 microg/g wet tissue). However, there was an insignificant difference between 20 and 40 mg/kg treated rats. Therefore, the dose of 20 mg/kg i.p. was used to evaluate the neuroprotective effect by using diffusion-weighted imaging (30 min after reperfusion), assessing the neurological deficit (24 h after middle cerebral artery occlusion) and estimating oxidative stress markers (72 h after middle cerebral artery occlusion). In the 20 mg/kg melatonin-treated group, percent ischemic lesion volume on diffusion-weighted imaging was significantly attenuated (9.8+/-3.9) as compared to that in the vehicle-treated group (21.4+/-4.7). The neurological deficit was significantly improved in the melatonin group (1.8+/-0.06) as compared to that in the vehicle-treated (2.9+/-0.38) group. The level of malondialdehyde (321.4+/-31 nmol/g wet tissue) and reduced glutathione (142.6+/-13 microg/g wet tissue) in the melatonin-treated group was also significantly decreased as compared to the level of malondialdehyde (623+/-22 nmol/g wet tissue) and reduced glutathione (226.6+/-19 microg/wet tissue) in the vehicle-treated group. The present study indicates that melatonin has a neuroprotective action in focal ischemia, which may be attributed to its antioxidant property.

Animals↗
PubMed2001

Effects of intracerebroventricularly administered chimeric peptide of metenkephalin and FMRFa--[D-Ala2]YFa-on antinociception and its modulation in mice.

An enzymatically stable analog of YGGFMKKKFMRFamide (YFa), a chimeric peptide of metenkephalin and FMRFa, was synthesised. The antinociceptive effects of intracerebroventricular injections of this analog-[D-Ala2)]YAGFMKKKFMRFamide ([D-Ala2]YFa)-was then investigated using the mouse radiant-heat tail-flick test. [D-Ala2]YFa produced modest to good antinociception at 1, 2, and 5 microg/mouse (0.64, 1.28, and 3.22 nmol, respectively). This antinociceptive effect was completely reversed by the opioid receptor antagonist naloxone (1.5 microg/mouse: 4.12 nmol, intracerebroventricular [i.c.v.]), administered 5 min prior. Pretreatment (5 min) with either neuropeptides FF (1 microg/mouse: 0.92 nmol, i.c.v.) or FMRFa (1 microg/mouse: 1.69 nmol, i.c.v.) significantly attenuated the antinociceptive effects induced by [D-Ala2]YFa (1 microg/mouse, i.c.v.). Intracerebroventricular administration of [D-Ala2]YFa at 1 microg/mouse dose with morphine (2 microg/mouse: 5.86 nmol, i.c.v.) produced an additive antinociceptive effect, suggesting that [D-Ala2]YFa may have a modulatory effect on opioid (morphine) analgesia. These results provide further support for a role of such amphiactive sequences in antinociception and its modulation.

Analgesics, Opioid↗
PubMed2001

Lithium does not synergize the peripheral action of cholinomimetics as seen in the central nervous system.

Lithium is known to synergize the action of cholinomimetics in the CNS such that pilocarpine induces seizures in low concentration (1/13th of per se dose) in rats. The present study was undertaken to see if lithium priming also enhances the peripheral effects of acetylcholine and pilocarpine i.e. change in blood pressure in rats and contractions of the isolated guinea pig ileum. In anaesthetized rats the blood pressure was recorded from cannulated carotid artery connected through the pressure transducer to Coulbourn polygraph. The blood pressure response of pilocarpine was not different either in magnitude or in duration when administered 1, 2 and 4 h after lithium chloride (3 meq/kg) pretreatment as compared to the control. Similarly acetylcholine effect remained unchanged after lithium chloride priming. In the isolated guinea pig ileum experiments, ileum was incubated for 1 h in different concentrations of lithium chloride and effect on acetylcholine induced contractions were observed. Lithium in concentration of 2.8 x 10(-3) M had no effect on acetylcholine induced contractions while incubation with higher concentrations of 1.4 x 10(-2) M and 2.8 x 10(-2) M significant inhibition of acetylcholine contractions were observed. At this concentration, histamine induced contractions were also inhibited. The results indicate that lithium does not synergize the action of cholinomimetics in the periphery as that seen in the CNS. The inhibition of acetylcholine and histamine induced contractions in guinea pig ileum at high concentration of lithium seems to be non-specific effect.

Acetylcholine↗
PubMed2001

Intracerebroventricular injection of streptozotocin in rats produces both oxidative stress in the brain and cognitive impairment.

Recent reports suggest the involvement of free radicals in the pathophysiology of Alzheimer's disease [AD]. Streptozotocin [STZ] injection in the brain is known to cause cognitive impairment in rats and is likened to sporadic AD in humans. Though STZ is known to cause impairment in glucose and energy metabolism, it is not known whether this is associated with free radical generation. The present study was designed to investigate if the changes in learning and memory by intracerebroventricular administration of STZ are associated with changes in the markers of oxidative stress. Adult male Wistar rats [330-340 g] were injected with intracerebroventricular STZ [3 mg/kg] bilaterally stereotaxically under ketamine anesthesia [70 mg/kg]. The rats were treated with STZ twice, on day 1 and on day 3. The learning and memory behavior was analyzed using passive avoidance paradigms, elevated plus maze and the closed field activity test while the parameters of oxidative stress assessed were malondialdehyde [MDA] and glutathione. The behavioral tests were performed on day 17, 18 and 19. The rats developed significant deficits in learning, memory and cognitive behavior, indicated by deficits in passive avoidance paradigm and elevated plus maze as compared to sham rats. On day 21, the rats were sacrificed under ether anesthesia and the brains were analyzed for biochemical studies. There was a development of oxidative stress in the brain as indicated by significant elevations in malondialdehyde [MDA] levels and decreased levels of glutathione. The study demonstrates that intracerebroventricular STZ may be appropriate model for investigations of antioxidants as potential treatment in Alzheimer's dementia.

Animals↗
PubMed2001

Chemopreventive activity of Withania somnifera in experimentally induced fibrosarcoma tumours in Swiss albino mice.

The current experimental work deals with the chemopreventive studies of a hydroalcoholic extract of Withania somnifera roots, against 20-methylcholanthrene induced fibrosarcoma tumours in Swiss albino mice. A single subcutaneous injection of 200 microg 20-methylcholanthrene in 0.1 mL of dimethylsulphoxide into the thigh region of mice produced a high incidence (96%) of tumours. Oral treatment of animals with 400 mg/kg body weight of Withania somnifera extract (one week before injecting 20-methylcholanthrene and continued until 15 weeks thereafter) significantly reduced the tumour incidence, tumour volume and enhanced the survival of the mice, compared with 20-methylcholanthrene injected mice. The tumour incidence was also delayed in the treatment group when compared with 20-methylcholanthrene injected mice. Liver biochemical parameters revealed a significant modulation of reduced glutathione, lipid peroxides, glutathione-S-transferase, catalase and superoxide dismutase in extract treated mice compared with 20-methylcholanthrene injected mice. The mechanism of chemopreventive activity of Withania somnifera extract may be due to its antioxidant and detoxifying properties.

Administration, Oral↗
PubMed2001

Intranasal tooth as a complication of cleft lip and alveolus in a four year old child: case report and literature review.

Ectopic position of teeth is not rare. The presence of teeth have been reported in ovaries, testes, anterior mediastinum, and pre-sacral regions. In the maxillofacial region, teeth have been found in maxillary sinus, mandibular condyle, coronoid process, chin, nose, and even orbit. Approximately 50 cases of a tooth in the nasal cavity have been reported in literature. However, an intranasal tooth in cases of cleft lip and palate is comparatively rare. Intranasal teeth can cause problems such as nasal obstruction, chronic rhinorrhea and speech problems. Sometimes however, they are totally symptom-free. We present here an interesting case of an intranasal tooth in a four year-old-boy, who was operated on for cleft lip and alveolus at 6 months of age. The intranasal tooth did not cause any symptoms. The tooth was extracted under general anaesthesia when it was found to be very loosely attached to the nasal mucosa. The case is discussed in the light of relevant literature on intranasal teeth in cases of cleft lip and palate.

Child, Preschool↗
PubMed2001

Protective effect of resveratrol against intracortical FeCl3-induced model of posttraumatic seizures in rats.

The present study was carried out to investigate the effect of trans-resveratrol, a potent antioxidant, against FeCl3-induced posttraumatic seizures. Male Wistar rats weighing 200-250 g were implanted with epidural electrodes and allowed to recover After the recording of baseline EEG. FeCl3 (5 microl, 100 mM) was administered intracortically over a period of 5 min and EEG was monitored for 2 h. Subsequently, rats were sacrificed for the estimation of oxidative stress markers, ie., malondialdehyde (MDA) and glutathione in whole brain tissue. A sham group was run parallel with saline (pH adjusted) and similar protocol for EEG recording and estimation of oxidative stress markers was followed FeCl3-treated animals exhibited epileptiform EEG changes thigh amplitude sharp waves of increased frequency and polyspikes) within 15 min, which continued throughout the period of observation. The level of MDA was found to be significantly elevated, whereas insignificant change was observed in the level of glutathione. trans-Resveratrol (20 and 40 mg/kg ip.) administered 30 min before FeCl3 injection delayed the onset of the appearance of epileptiform EEG changes. The brain MDA levels were also significantly reduced in the trans-resveratrol-treated animals as compared to the vehicle-treated FeCl3-injected rats. The findings show the protective effect of trans-resveratrol in the FeCl3-induced seizure model of posttraumatic seizures.

Animals↗
PubMed2001

Reversal of pyrogallol-induced delay in gastric emptying in rats by ginger (Zingiber officinale).

The effects of the acetone extract of ginger (Zingiber officinale) was studied against pyrogallol-induced delay in gastric emptying in rats. Wistar rats of either sex, weighing between 200-250 g, were used. Pyrogallol, at a dose of 100 mg/kg i.p., significantly delayed the gastric emptying of a methyl cellulose meal. One-hour pretreatment with ginger acetone extracts (100, 250 and 500 mg/kg p.o.) reversed the pyrogallol-induced delay in gastric emptying. The effect was significant at doses of 250 and 500 mg/kg. When the low dose of ginger (100 mg/kg p.o.) was given with vitamin C and vitamin E (100 mg/kg p.o., each), the reversal of gastric emptying was more pronounced than when only two vitamins or ginger (100 mg/kg and 500 mg/kg) were given alone. The present study indicates the potential of ginger in improving symptoms such as abdominal discomfort and bloating, which may accompany several gastrointestinal illnesses.

Animals↗