Toward Novel Diagnostics for Primary Open-Angle Glaucoma? An Association Study of Polymorphic Variation in Ras Homolog Family Member (A, B, C, D) Genes RHOA, RHOB, RHOC, and RHOD.

The annual economic burden of visual disorders in the United States was estimated as $139 billion. The World Health Organization has listed glaucoma in the top 10 priority eye diseases. Primary open-angle glaucoma (POAG) is a common subtype, with a lack of clinical tools for early diagnosis. The Rho GTPases belong to the Ras superfamily of proteins; the RhoA immunostaining in the optic nerve head in human glaucoma is reportedly increased. We investigated the association of polymorphisms in the Ras Homolog Family Member A, B, C, and D genes (RHOA, RHOB, RHOC, and RHOD, respectively). In a total sample of 361 unrelated subjects (179 patients with POAG and 182 age- and sex-matched healthy controls), RHOA (rs6784820, rs974495), RHOB (rs62121967), RHOC (rs11102522), and RHOD (rs61891303, rs2282502) polymorphisms were characterized by the BioMark HD dynamic array system with real-time polymerarse chain reaction. Among these candidate genetic markers and considering the Bonferroni correction, RHOA rs974495 polymorphism was significantly associated with POAG (p = 0.0011), with the TT genotype increasing the disease risk 4.9 times (95% CI 1.630-15.023). The allele and haplotype distributions of the above RHO candidate polymorphisms did not diplay a significant association. This is the first study, to the best of our knowledge, to identify a significant genotypic association between POAG and RHOA gene rs974495 polymorphism. These observations warrant replication in independent samples in the pursuit of precision medicine for rapid and early glaucoma diagnosis, and molecular targets for innovation in therapeutics of this common eye disease.

CYP gene expressions in obesity-associated metabolic syndrome.

PURPOSE: The contribution of cytochrome P450 (CYP) gene expressions in metabolic syndrome (MetS) has not been elucidated, and was the aim of this study. METHODS: A total of 51 MetS patients and 41 healthy controls with similar age and sex were included to this study. mRNA from blood samples was extracted, and real-time polymerase chain reaction was performed for gene expressions using a dynamic array system. RESULTS: We observed marked suppressions in CYP2A6 (p=0.0123), CYP4F2 (p=0.0005), CYP3A5 (p=0.0003), and CYP17A1 (p<0.0001) gene expressions in MetS patients. CONCLUSIONS: This is the first study to provide evidence that depressed expressions of CYP2A6, CYP4F2, CYP3A5, and CYP17A1 genes may play a role in MetS.

Evidence for elevated (LIMK2 and CFL1) and suppressed (ICAM1, EZR, MAP2K2, and NOS3) gene expressions in metabolic syndrome.

The metabolic syndrome (MetS) is a common multicomponent condition including abdominal obesity, dyslipidemia, hypertension, and hyperglycaemia. The aim of this study was to investigate the associations of the expression of a panel of signalling genes with the MetS in a Turkish population. A total of 54 MetS patients and 42 healthy controls with similar age and sex were included to this study. mRNA from blood samples was extracted, and real-time polymerase chain reaction was performed for gene expressions using a BioMark 96.96 dynamic array system. We observed marked increases in LIM kinase 2 (LIMK2) and cofilin 1 (CFL1) gene expressions in MetS patients. However, there were significant decreases in intercellular adhesion molecules 1 (ICAM1), ezrin (EZR), mitogen-activated protein kinase kinase 2 (MAP2K2), and nitric oxide synthase 3 (NOS3) gene expressions in MetS patients. Additionally, no marked changes were noted in other 15 genes studied. This is the first study to provide evidence that activation of LIMK2/CFL1 pathway may play an important role in MetS.

Relationship between soluble intercellular adhesion molecules and attention-deficit/hyperactivity disorder.

OBJECTIVE: Attention-deficit/hyperactivity disorder (ADHD) is a common childhood-oneset psychiatric disease, characterized by excessive overactivity, inattention, and impulsiveness. In recent studies, it is emphasized that inflammation may have a role in ADHD. In this study, we aimed to investigate whether there are associations between ADHD and serum levels of soluble intercellular adhesion molecules (s-ICAMs) which have important role in inflammatory diseases. We also measured the levels of these molecules after treatment with oros-methylphenidate. METHODS: Twenty-five patients diagnosed with ADHD according to Diagnostic and Statistical Manual of Mental Disorders-IV-TR criteria and 18 healthy volunteer controls were included in this study. The levels of sICAMs were measured in the serum of the patients and healthy volunteers by ELISA kit as described. RESULTS: The levels of sICAM-1 and sICAM-2 were significantly higher in patients compared with controls. The level of sICAM-2 was decreased significantly in group treated with oros-methylphenidate. CONCLUSIONS: This is the first study pointing out the relationship between sICAMs and ADHD. The changes in sICAM-2 level may have a role in the effect mechanism of oros-methylphenidate, used for the treatment of ADHD.

Evaluation of the attitudes of the nurses related to rational drug use in Gaziantep University Sahinbey Research and Practice Hospital in Turkey.

BACKGROUND AND OBJECTIVES: This study aimed to evaluate the attitude of nurses about rational drug use in Gaziantep University Sahinbey Research and Practice Hospital. There are a limited number of studies available on this issue and no studies of this scale were conducted among the nurses in our region. DESIGN AND SETTING: A questionnaire generated by the Rational Drug Use Unit of Turkish Ministry of Health General Directorate of Pharmaceuticals and Pharmacy was carried out to nurses. PARTICIPANTS: The study was carried out to 162 nurses. METHODS: The data obtained from nurses by questionnaire were determined as count, percentage and Chi-square test by SPSS statistical package program. RESULTS: The most common type of medication error was giving the medicine at the wrong time. Medication errors were least common among the 36-50-year age group and with a professional experience of 11 years or longer. Nurses had the highest level of knowledge in the areas of drug administration routes and the intended use. The number of nurses reported having good/very good knowledge was higher with 4 to 10 years of professional experience and with a university degree. The nurses aged between 26 and 35 years and those with professional experience of 4 to 10 years provided drug information to patients more often than others. Forty two percent of the nurses were found to actively report any adverse events. Reporting of adverse events and reporting more than 6 adverse events were most common among university degree holders. CONCLUSIONS: Nurses required a more comprehensive education on pharmacology both during their training years and working life since the requests for medicinal products are received by the nurses and preparation and administration of drugs are under the control of nurses.

Association of Rho-kinase 1 (ROCK1) gene polymorphisms with Behçet's disease.

BACKGROUND AND OBJECTIVE: Behçet's disease (BD) is a chronic inflammatory vasculitis presenting with flares and silent periods usually between 15 and 40 years of age. The aim of this study was to evaluate the possible association between Rho-kinase 1 (ROCK1) gene polymorphisms and patients with BD in a Turkish population. METHODS: A total of 192 BD patients and 255 healthy controls of similar age and sex were enrolled in this study. Polymorphisms were analyzed in genomic DNA using a BioMark HD dynamic array system. RESULTS: In the presence of CC genotype for rs73963110, CT genotype for rs111874856 (Val355Ile), and TC genotype for rs112130712 (Lys1054Arg) polymorphisms, the risk of BD increased 12.13-, 15.05-, and 16.28-fold, respectively (p < 0.0001). There was a lower frequency of the GA genotype of the rs112108028 (Pro1164Leu) polymorphisms in BD (10.3 %) compared with controls (39.7 %; p < 0.0001). Marked associations between these polymorphisms and the manifestations of BD were recorded. CONCLUSION: This is the first study to show that ROCK1 gene polymorphisms may have a significant impact on susceptibility to BD.

Investigation of intercellular adhesion molecules (ICAMs) gene expressions in patients with Barrett's esophagus.

The adhesion molecules play a major role in inflammation as well as in neoplastic diseases. The aim of this study is to evaluate the expressions of the adhesion molecules, intercellular adhesion molecule 1 (ICAM-1), ICAM-2, and ICAM-3, in Barrett's esophagus, recognized as a premalign lesion for esophageal cancer and related to inflammation. Eighteen patients with Barrett's esophagus according to endoscopy and 25 volunteers without Barrett's esophagus disease were included in the study. Tissue samples were supplied by biopsy and used for both gene expression and immunohistochemical analysis. The significance of the differences between the two groups was assessed by Student's t test. The ICAM-1 expression level was fivefold higher in the patient group compared with that of the control. There was an increase in the serum level of ICAM-1 in patients compared to that of the controls, but this increase was not significant. ICAM-2 levels were also increased in the patient group, but it was not significant. There was no difference between controls and patients in ICAM-3 levels. Significantly higher levels of ICAM-1 gene expression make us think that ICAM-1 may play an important role in Barrett's esophagus. We think that more studies, with larger patient groups and preferably detailed histopathological and clinical evaluations, are needed to explain the severity of ICAM-1, ICAM-2, and ICAM-3 molecules in Barrett's esophagus.

Investigation of the esophageal Rho-kinase expression in patients with Barrett's esophagus.

The mechanisms responsible for the malignant transformation in Barrett's esophagus (BE) are still poorly understood. The authors have evaluated the role of Rho-kinase (ROCK1 and ROCK2) expressions in patients with BE. All patients underwent upper gastrointestinal system endoscopy, which was confirmed histologically. Real-time PCR revealed no marked change in gene expressions of ROCK1 and ROCK2 at mRNA levels in BE when compared to controls. Immunohistochemical and western blot analyses showed no change in ROCK1 and ROCK2 protein expressions in BE. This study demonstrates that Rho-kinase gene and protein expressions are not modified in BE.

Lack of association between the C276T polymorphism of the neuronal nitric oxide synthase gene and migraine.

A migraine attack is a spectacularly complex brain event that can produce a wide array of neurological and systemic symptoms. The molecular mechanisms and genetics of migraine have not yet been clarified. The objective of this study was to analyze the genotype distributions and allele frequencies for the C276T polymorphism of the neuronal nitric oxide synthase (nNOS) gene among the patients with migraine. The diagnosis of migraine was made clinically based on questionnaires. One hundred and twenty patients with migraine were genotyped for the C276T polymorphism of the nNOS gene and compared with 185 age-matched healthy controls. Genomic DNA from migraine patients and controls was analyzed by polymerase chain reaction (PCR). A PCR-restriction fragment-length polymorphism analysis of nNOS gene polymorphism was performed, and the results were compared. Neither genotype distributions nor the allele frequencies for the C276T polymorphism showed a significant difference between the groups. Additionally, there was no marked differences in genotype distribution or allele frequencies for the migraine without aura and migraine with aura subgroups when compared to control group. These results suggested that migraine of the Turkish population seemed to develop without any alterations in nNOS C276T polymorphism. Our data showed that there is no marked association between the C276T polymorphism of the nNOS gene and migraine.

Association between Rho-kinase (ROCK2) gene polymorphisms and Behçet's disease.

Behçet's disease (BD) is a multi-systemic vasculitis. The aim of this study was to investigate the association between Rho-kinase (ROCK2) gene polymorphisms and patients with BD in a Turkish population. A total of 194 BD patients and 276 healthy controls with similar age and sex were included to this study. Polymorphisms were analyzed in genomic DNA using a BioMark 96.96 dynamic array system. mRNA from blood samples was extracted, and real-time polymerase chain reaction was performed for ROCK2 gene expression. There were marked changes in both genotype (TT, 41.8%; TA, 30.3%) and allele (T, 57%; A, 43%) frequencies for the rs35768389 (Asp601Val) polymorphism in patients compared with controls (TT, 64.6%; TA, 9.4%, P < 0.0001; T, 69.3%; A, 30.7%, P = 0.0004). Although CC genotype (52.0%) of rs1515219 polymorphism were more frequent, CT genotype (27.7%) were less frequent among the patients than controls (CC, 31.7%, CT, 44.6%, P = 0.0001). There was an increase in C allele (65.8% vs 54.0%) and decrease in T allele frequencies (34.2% vs 46.0%, P = 0.001) in patients. However, no associations were found with rs726843, rs2290156, rs965665, rs10178332, rs2230774, rs6755196, rs10929732, and rs34945852 polymorphisms. There was an increase in peripheral blood mRNA ROCK2 expressions in patients. This is the first study to examine the involvement of ROCK2 gene variation in the risk of incident BD. The results strongly suggest that ROCK2 gene polymorphisms may modify individual susceptibility to BD in the Turkish population.

DD genotype of ACE gene in boys: may it be a risk factor for minimal change nephrotic syndrome?

It has been shown that angiotensin-converting enzyme (ACE) gene insertion/deletion (I/D) polymorphism affects the circulating and cellular levels of ACE and may be a risk factor in several renal diseases. We analyzed the association of ACE gene I/D polymorphism with the clinical presentation of minimal change nephrotic syndrome (MCNS) in a Turkish child population. This study consisted of 97 children with MCNS and 144 healthy controls. Genotyping of ACE gene was performed using polymerase chain reaction (PCR). The distributions of ACE genotypes were II in 13%, ID in 49%, and DD in 38% in patient group, and 9%, 49%, and 42% in control group, respectively. The frequency of the D allele was 63% and that of the I allele was 37% in patients. There were no relevant differences in the allele frequencies and genotypes of ACE I/D polymorphism between patients and controls. However, DD genotype was higher in boys in children with MCNS (78.4%. vs. 50.0%, p = 0.004). The frequencies of DD genotype and D allele in boys were 7.25 and 2.56 times higher than II genotype and I allele in the patient group, respectively. We suggest that DD genotype in boys may be one of the risk factors for MCNS.

Lack of association between the Thr431Asn and Arg83Lys polymorphisms of the ROCK2 gene and diabetic retinopathy.

PURPOSE: To analyze the genotype distributions and allele frequencies for ROCK2 Thr431Asn and Arg83Lys polymorphisms among the diabetic retinopathy patients in a Turkish population. METHODS: In this case-control study, 335 patients with diabetes mellitus were recruited and divided into three groups according to non-proliferative (n = 127), proliferative (n = 85) diabetic retinopathy, and no retinopathy (n = 123, served as a diabetic control group). Genomic DNA from the patients, and the nondiabetic healthy control cases (n = 132) was analyzed by real-time PCR using a Light-Cycler. RESULTS: Neither genotype distributions nor the allele frequencies for the Thr431Asn or Arg83Lys polymorphisms showed a significant difference between the groups. The haplotypes were also not significantly associated with diabetic retinopathy. CONCLUSION: These results suggest that there were no evidence for an association of ROCK2 gene Thr431Asn and Arg83Lys polymorphisms with diabetes or diabetic retinopathy in the Turkish population.

Nitric oxide synthase gene polymorphisms in children with minimal change nephrotic syndrome.

AIMS: Nitric oxide (NO) attenuates many functions within the kidney, and all NO synthase (NOS) isoforms are constitutively expressed in the kidney. But the exact role of NO in renal diseases is still debatable. The aim of the present study was to investigate endothelial (eNOS), and neuronal (nNOS) NOS gene polymorphisms in children with minimal change nephrotic syndrome (MCNS). MATERIALS AND METHODS: Eighty-six Turkish children with clinical MCNS, ranging in age from 2 to 10 years, were compared with 114 healthy age- and sex-matched controls. The glu 298 Asp (G/T) polymorphism of the eNOS, and C276T (C/T) polymorphism of nNOS genes were genotyped using polymerase chain reaction. RESULTS: The distribution of GG, TG, and TT genotypes for eNOS was 52%, 33% and 15% in MCNS compared with 61%, 26% and 13% in the controls (P > 0.05). The distribution of CC, TC, and TT genotypes for nNOS was 16%, 66% and 18% in MCNS compared with 10%, 43% and 47% in the controls. TT genotype distribution of nNOS was found to be lower in patients (P = 0.003). The eNOS and nNOS gene polymorphisms were not associated with gender, positive family history, frequency of relapses, or response to steroid. CONCLUSIONS: The present study is the first to investigate eNOS and nNOS gene polymorphisms in children with MCNS. The nNOS gene polymorphism may be associated with MCNS in children, but further studies in a larger population with different glomerular diseases are needed to confirm the results.

Which genotype of MAO gene that the patients have are likely to be most susceptible to the symptoms of fibromyalgia?

The objective of this study was to analyze the genotype distributions and allele frequencies for the MAO-A and MAO-B polymorphism of the MAO gene among the patients with fibromyalgia syndrome (FS). One hundred and seven fibromyalgia patients and 90 unrelated healthy subjects were included into the study. Genomic DNA of 107 FS patients and 90 healthy control subjects were analyzed by polymerase chain reaction. Polymorphism of the MAO gene was: 1-1, 1-3, 3-3, 3-4. The "allele 3" had a 2.7 to 4.8-fold increased transcription activity than the "allele 1". The frequencies of the genotypes of the patients with FS and healthy controls were compared. Although no significant difference was found in genotypes of patients and controls (P = 0.0559), it is likely that "allele 3" could be a more riskful factor for FS than "allele 1" (P = 0.033). Fibromyalgia impact questionnaire was administered to FS group as well as control group. One of our findings is that, the patients whose genotype 3-3 may be mostly affected by the symptoms of FS. In conclusion, it seems plausible to say that MAOA-dependent metabolism of the biological amines may be partly related to high-activated MAO-A, allele 3, in the occurrence of FS among Turkish population.

Nitric oxide synthase gene polymorphisms in children with primary nocturnal enuresis: a preliminary study.

AIMS: Recent studies demonstrated some differences in urinary electrolytes of enuretic children. Intrarenal nitric oxide (NO) serves as a major regulator of renal sodium and water excretion like an endogenous diuretic. This study aimed to investigate endothelial (eNOS), and neuronal (nNOS) NO synthase gene polymorphisms in children with primary nocturnal enuresis (PNE). MATERIALS AND METHODS: The eNOS gene polymorphism was investigated in 171 Turkish children (57 PNE cases and 114 healthy, non-enuretic controls), and nNOS gene polymorphism was determined in 158 Turkish children (83 PNE cases and 75 healthy, non-enuretic controls). The glu298asp (G/T) polymorphism of the eNOS and C276T (C/T) polymorphism of nNOS genes were genotyped using PCR. RESULTS: The distribution of GG, TG, and TT genotypes for eNOS gene was 48%, 33%, and 19% in PNE, compared with 61%, 26%, and 13% in the controls (p > 0.05). The distribution of CC, TC, TT and genotypes for nNOS gene was 31%, 29%, and 40% in PNE compared with 10%, 43%, and 47% in the controls. CC genotype was found higher in enuretic children (p = 0.002). The eNOS and nNOS gene polymorphisms were not associated with positive family history, frequency of enuresis, and clinical response to desmopressin. CONCLUSIONS: This study is the first to search the NOS gene polymorphisms in children with PNE. It was determined that eNOS gene polymorphism may not be associated with PNE, while nNOS gene polymorphism, a predominantly CC genotype, may be associated with PNE in Turkish children. Further studies with larger samples together with the detection of enuresis gene may help determine the exact role of nNOS gene polymorphism in enuresis.

No evidence for an association between the Glu298Asp polymorphism of the endothelial nitric oxide synthase gene and fibromyalgia syndrome.

The objective of this study was to analyze the genotype distributions and allele frequencies for the Glu298Asp (G894T) polymorphism of the eNOS gene and the serum nitric oxide level among the patients with fibromyalgia syndrome (FS). Ninety-six fibromyalgia patients and 79 unrelated healthy volunteer controls were included in the study. All patients and controls were females. Genomic DNA from 96 patients meeting the American College of Rheumatology 1990 criteria for FS and 79 healthy controls was analyzed by polymerase chain reaction. A polymerase chain reaction-restriction fragment-length polymorphism analysis of eNOS gene polymorphism was performed, and the results of the patients with FS and healthy controls were compared. Ozone-based chemiluminescence assay with Sievers NO Analyzer was used to measure the serum nitric oxide levels. Neither the frequencies of the Glu298Asp genotypes nor the serum nitric oxide levels showed a significant difference between the groups. These results suggested that FS of the Turkish population seemed to develop without any alterations in eNOS Glu298Asp genotype frequency and the serum nitric oxide level.

Effects of caffeine on placental total nitrite concentration: A 21-day, vehicle-controlled study in rats.

BACKGROUND: Nitric oxide (NO) is a potent vasodilator that might have an10 important role in the modulation of maternal and fetal vascular tone during pregnancy. The effects of caffeine intake on maternal and fetal hemodynamic properties during pregnancy have been investigated in several human and animal studies. However, based on a literature search, there has been no study of placental total nitrite (a stable product of NO) concentration (PTNC) in pregnant humans or rats given caffeine. OBJECTIVE: The aim of this study was to assess the effects of caffeine intake 10 on PTNC in rats. METHODS: This 21-day, vehicle-controlled study was conducted at the Department10 of Pharmacology, Gaziantep University Hospital, Gaziantep, Turkey. Female Wistar rats were randomly assigned; based on age and weight, to receive 25, 50, or 100 mg/kg QD caffeine or 50 mg/kg QD isotonic saline solution (vehicle; age-matched control group), intraperitoneally for 21 days. After euthanization of the rats and cesarean section, the numbers of fetuses and fetal deaths were counted. The lengths and weights of the fetuses in each study group were noted. PTNC in the rats was determined using the Greiss reaction. RESULTS: This study included 26 rats (7, 7, and 6 rats in the groups receiving10 25, 50, and 100 mg/kg. d caffeine, respectively; 6 rats in the control group). The mean (SD) lengths of the fetuses of the rats given 25, 50, and 100 mg/kg · d caffeine (4.90 [0.15], 4.02 [0.27], and 3.45 [0.17] mm, respectively) were significantly less compared with controls (5.10 [0.18] mm) (all, P < 0.001), as were the mean (SD) weights of the fetuses of rats given caffeine (5.86 [0.24], 4.97 [0.59], and 3.41 [0.23] g, respectively) versus controls (6.18 [0.21] g) (all, P < 0.001). The mean (SD) PTNCs in rats given 25, 50, and 100 mg/kg. d caffeine (19.82 [1.97], 29.39 [2.07], and 45.51 [7.66] nmol/g, respectively) were significantly higher compared with controls (16.10 [2.12] nmol/g) (all, P < 0.001). CONCLUSIONS: The results of this study in rats suggest that caffeine intake 10 might increase NO production in the placenta. In addition, based on our findings and those from previous studies, we suggest that this increase might be an adaptive physiologic response to prevent undesirable effects of caffeine on vascular tone during pregnancy.

Excessive maternal caffeine exposure during pregnancy is cataractogenic for neonatal crystalline lenses in rats: a biomicroscopic and histopathologic study.

PURPOSE: To investigate histologically the influence of maternal caffeine exposure during pregnancy in vivo on crystalline lenses in neonatal rats. METHODS: Experimentally naive, female Wistar-albino rats (200-220 g) were mated with adult male rats over 2 days for copulation. After confirming pregnancy with a vaginal smear method, 50 gravid rats (dams) were randomly divided into five groups (n = 10 in each), consisting of one control and four experimental groups. Groups 1, 2 and 3 experimental dams were treated with intraperitoneal (i.p.) caffeine at doses of 25, 50 and 100 mg/kg/day, respectively, during pregnancy from gestational day 9 through to day 21. Group 4 dams were treated with caffeine in distilled water in a gavage at a dose of 50 mg/kg/day. Group 5 control dams were given i.p. saline solution daily for the same period. After normal delivery, the eyes were examined by slit-lamp biomicroscopy. The neonates were then killed by decapitation at postnatal days 1 or 30 and the eyes removed for histopathologic investigation of the lenses. RESULTS: Group 1 and control eyes had normal anterior lens capsules with a single layer of anterior cuboidal epithelial cells, regularly oriented cortical and nuclear lens fibres, and a clear posterior lens capsule with no lining epithelial cells behind the equator. In the remaining groups, histopathologic findings suggesting cataractogenesis included eosinophilic degeneration, lens fibre cell swelling and liquefaction, central lens fibres with retained nuclei, and prominent epithelial cells lining the posterior lens capsule behind the equator. Moreover, some lenses in group 3 had immature cataract on slit-lamp biomicroscopic examination at postnatal day 30. CONCLUSION: Excessive maternal caffeine exposure during pregnancy had cataractogenic effects on developing crystalline lenses in newborn rat eyes, both macroscopically and histopathologically. If an appropriate dose of caffeine can be identified, caffeine-induced cataract formation may be used as a new experimental cataract model in animal studies.

Effect of gestational nicotine treatment on newborn rat retina: a histopathological and morphometric analysis.

BACKGROUND: Smoking is a significant risk factor in several debilitating and fatal diseases. It has been implicated in bilateral tobacco-toxic and Leber's hereditary optic neuropathies. Although it has been demonstrated that smoking has a cumulative effect on retinal and optic nerve functions and causes diffuse and localised retinal sensitivity decrease in healthy chronic heavy smokers, the affected retinal layer has not been identified and there is no experimental study investigating the effect of nicotine exposure during gestation on the newborn rat retina. PURPOSE: This experimental investigation evaluated histologically the influence in vivo of maternal nicotine treatment during pregnancy on the newborn rat retina. Different dosages of the test compound simulated the range of low, moderate, and heavy smokers in humans. METHODS: Experimentally naive, adult female Wistar-albino rats weighing 200-250 g were mated with adult male rats over 2 days for copulation in the proportion of two females for every male animal. After confirming pregnancy with vaginal smear method, 40 gravid rats (dams) were then randomly assigned into four equal groups (three experimental and one control; n = 10 in each). On day 9 of gestation, groups 1, 2, and 3 experimental dams were treated with intraperitoneal (i.p.) (-)-nicotine tartrate at doses of 0.5, 1, and 2 mg kg body weight-1 day-1, respectively during pregnancy from gestational day 9-21. Group 4 control dams were given i.p. saline solution daily for the same period. After normal delivery, the newborn litters were sacrificed at postnatal day 1 or day 30. The eyes were enucleated for histopathologic and morphometric analysis of the retinas. Nicotine-induced neuronal changes were measured by morphometric analyses on cell counts of ganglion cell layer (linear cell density in number per unit length of retina) and thickness of the various retinal layers. RESULTS: The litters in control group 4, and experimental groups 1 and 2 had normal retinal findings. On the other hand, morphometric analysis of retinal sections in experimental group 3 eyes demonstrated a 20.7% decrease in the number of surviving ganglion cells (40.7 +/- 2.0) compared with controls (51.3 +/- 1.1; p < 0.001). The thickness of whole retina (126.6 +/- 5.4 microm) was also reduced by 13.5% compared with controls (146.3 +/- 4.5 microm; p = 0.007). The main site of retinal atrophy was the inner plexiform layer (30.1 +/- 1.6 microm vs 43.5 +/- 1.3 microm; p < 0.001) with almost no change in the other retinal layers. CONCLUSIONS: Gestational nicotine treatment induces marked changes in the organisation of the developing retina in newborn rats histopathologically. Quantitative morphometric analysis clearly demonstrated that the two most affected structures were the retinal ganglion cells and the inner plexiform layer, both of which are supplied by central retinal artery.

High dose of caffeine administered to pregnant rats causes histopathological changes in the cornea of newborn pups.

BACKGROUND: Caffeine is frequently used during pregnancy and associated with teratogenic effects, such as low birth weight, hearth and digital defects, cleft palate and abortion with fetal loss. This study investigated histopathologically the effects of caffeine on neonatal rat cornea. MATERIAL/METHODS: Fifty pregnant Wistar-Albino rats (dams) were randomly divided into five groups, one control and four experimental. Between day 9 and 21 of gestation, group 1 dams (control, n=10) were exposed to intraperitoneal (i.p.) SF daily until delivery. Group 2 (n=10), group 3 (n=10) and group 4 (n=10) dams were treated with i.p. caffeine at doses of 25, 50 and 100 mg/kg/d, respectively, for the same period. Group 5 dams were given caffeine in distilled water in a gavage at a dose of 50 mg/kg/d during the same period. After normal delivery, the litters were killed at postnatal day 1 or 30 and the eyes were enucleated for corneal histopathologic investigation. RESULTS: Control and group 1 eyes had normal corneal epithelium, regular stromal fibers, descement membrane and monolayer inner corneal endothelium. The remaining experimental litters demonstrated changes, such as vacuolated endothelial cells with proliferation, hyperchromasia, polymorphism, endothelial cell agenesis, increased stromal mitotic activity and focal increase in corneal thickness with widely separated corneal lamellae in the injured area. These changes occurred most often in the litters treated with high doses of caffeine. CONCLUSIONS: Excessive gestational caffeine intake has been shown histopathologically to have some teratogenic effects on newborn rat cornea.