High prevalence of multilocus pathogenic variation in neurodevelopmental disorders in the Turkish population.

Neurodevelopmental disorders (NDDs) are clinically and genetically heterogenous; many such disorders are secondary to perturbation in brain development and/or function. The prevalence of NDDs is > 3%, resulting in significant sociocultural and economic challenges to society. With recent advances in family-based genomics, rare-variant analyses, and further exploration of the Clan Genomics hypothesis, there has been a logarithmic explosion in neurogenetic "disease-associated genes" molecular etiology and biology of NDDs; however, the majority of NDDs remain molecularly undiagnosed. We applied genome-wide screening technologies, including exome sequencing (ES) and whole-genome sequencing (WGS), to identify the molecular etiology of 234 newly enrolled subjects and 20 previously unsolved Turkish NDD families. In 176 of the 234 studied families (75.2%), a plausible and genetically parsimonious molecular etiology was identified. Out of 176 solved families, deleterious variants were identified in 218 distinct genes, further documenting the enormous genetic heterogeneity and diverse perturbations in human biology underlying NDDs. We propose 86 candidate disease-trait-associated genes for an NDD phenotype. Importantly, on the basis of objective and internally established variant prioritization criteria, we identified 51 families (51/176 = 28.9%) with multilocus pathogenic variation (MPV), mostly driven by runs of homozygosity (ROHs) - reflecting genomic segments/haplotypes that are identical-by-descent. Furthermore, with the use of additional bioinformatic tools and expansion of ES to additional family members, we established a molecular diagnosis in 5 out of 20 families (25%) who remained undiagnosed in our previously studied NDD cohort emanating from Turkey.

Costs of unnecessary repeated diagnostic tests in cholecystectomy.

The purpose of this study was to calculate the number, rate and cost of unnecessarily repeated tests on the patients who underwent open cholecystectomy and laparoscopic cholecystectomy and figure out the share of the unnecessarily repeated test costs in total test expenditures and total treatment expenditures. Alkaline phosphatase, alanine aminotransferase, aspartate transaminase, gamma glutamyl transferase, Total bilirubin, whole abdominal ultrasonography, upper abdominal ultrasonography, hepatobiliary ultrasonography tests, which were among the tests used in the patients who underwent cholecystectomy. The research group included 1296 patients. All records of the patients within totally 180 days of period including 90 days of pre-cholecystectomy and 90 days of post-cholecystectomy period were analysed. Necessity/Unnecessary criteria of the tests were identified in accordance with literature data, expert opinions and American Society of Anaesthesiologists Physical Status Classification scores. It was determined that unnecessary test costs consisted of 8.48% of the total test costs and 0.93% of the total treatment expenditures. This study showed that age, gender, surgical technique, and American Society of Anesthesiologists scores significantly differentiated unnecessary repeated test costs. Reducing the use of excessive health service and the related health expenditures, working to reveal its financial and medical benefits are crucial for the reimbursement agency, health service hosts and patients.

An Evidence-Based Approach to Outcome Measurement in Oral and Dental Health Services: Oral Health-Related Quality of Life and Oral Health Impact.

PURPOSE: The purpose of this study was to examine the Oral Health-Related Quality of Life (OHRQOL) and Oral Health Impact Profile (OHIP) of oral and dental health patients in terms of gender, educational status, and the reason for coming to the oral health center. Also, we investigated the relationships between OHRQOL and OHIP. METHODS: This cross-sectional study was conducted and planned for dental patients in Turkey. OHRQOL-United Kingdom (OHRQOL-UK) and OHIP-14 were used for data collection. Descriptive statistics, correlation analysis, student t-tests, and ANOVA were used for data analyses. RESULTS: Of 527 respondents, 62.8% were female, and 37.2% were male. One-hundred-forty-one (26.8%) participants were illiterate. Three-hundred-fifty-four (67.20%) dental patients had an elementary school degree. Only 32 (6.10%) participants graduated from college and bachelor programs. For dimensions of the OHIP-14 and OHRQOL-UK, we detected statistically significant differences in personal characteristics. We found that gender, marital status, age, education status, and reasons for coming to the hospital have a significant impact on OHRQOL and OHIP. LINKING EVIDENCE TO ACTION: These results are expected to provide important evidence-based information to health managers and decision-makers in health planning and reimbursement policies. Clinicians and health managers should use OHIP, quality of life (QOL), and evidence-based practice to determine individual treatments and approaches to improve oral health. QOL is an outcome indicator in healthcare services and evidence-based practice. Measurements of evidence-based health outcomes in national health systems can be made, and global comparisons and policies in oral and dental health can be developed.

IL-1RN VNTR, IL-2(-330), and IL-4 VNTR gene polymorphisms in patients with chronic rhinosinusitis with sinonasal polyposis

Background/aim: Sinonasal polyposis is a complex chronic disease displaying contributions from multiple genetic and environmental factors. In this study, we analyzed possible genetic factors that increase susceptibility to this widespread inflammatory disease. Materials and methods: A total of 176 adult patients, including 78 patients with sinonasal polyposis and 98 healthy controls, were analyzed for IL-1RN VNTR, IL-2(-330), and IL-4 VNTR gene polymorphisms using polymerase chain reaction and enzyme restriction. Results: IL-1RN and IL-4 VNTR polymorphisms were notably associated with sinonasal polyposis (P = 0.0001 and P = 0.036, respectively); however, regarding the IL-2(-330) gene polymorphism, no significant difference was shown between the patient and control groups (P = 0.235). Conclusions: Our study indicates that the RN2 allele of IL-1RN and the RP1 allele of IL-4 might be risk factors for developing sinonasal polyposis.

Rare variants in the notch signaling pathway describe a novel type of autosomal recessive Klippel-Feil syndrome.

Klippel-Feil syndrome is a rare disorder represented by a subgroup of segmentation defects of the vertebrae and characterized by fusion of the cervical vertebrae, low posterior hairline, and short neck with limited motion. Both autosomal dominant and recessive inheritance patterns were reported in families with Klippel-Feil. Mutated genes for both dominant (GDF6 and GDF3) and recessive (MEOX1) forms of Klippel-Feil syndrome have been shown to be involved in somite development via transcription regulation and signaling pathways. Heterotaxy arises from defects in proteins that function in the development of left-right asymmetry of the developing embryo. We describe a consanguineous family with a male proband who presents with classical Klippel-Feil syndrome together with heterotaxy (situs inversus totalis). The present patient also had Sprengel's deformity, deformity of the sternum, and a solitary kidney. Using exome sequencing, we identified a homozygous frameshift mutation (c.299delT; p.L100fs) in RIPPLY2, a gene shown to play a crucial role in somitogenesis and participate in the Notch signaling pathway via negatively regulating Tbx6. Our data confirm RIPPLY2 as a novel gene for autosomal recessive Klippel-Feil syndrome, and in addition-from a mechanistic standpoint-suggest the possibility that mutations in RIPPLY2 could also lead to heterotaxy. © 2015 Wiley Periodicals, Inc.

Is the extraction by Whatman FTA filter matrix technology and sequencing of large ribosomal subunit D1-D2 region sufficient for identification of clinical fungi?

Although conventional identification of pathogenic fungi is based on the combination of tests evaluating their morphological and biochemical characteristics, they can fail to identify the less common species or the differentiation of closely related species. In addition these tests are time consuming, labour-intensive and require experienced personnel. We evaluated the feasibility and sufficiency of DNA extraction by Whatman FTA filter matrix technology and DNA sequencing of D1-D2 region of the large ribosomal subunit gene for identification of clinical isolates of 21 yeast and 160 moulds in our clinical mycology laboratory. While the yeast isolates were identified at species level with 100% homology, 102 (63.75%) clinically important mould isolates were identified at species level, 56 (35%) isolates at genus level against fungal sequences existing in DNA databases and two (1.25%) isolates could not be identified. Consequently, Whatman FTA filter matrix technology was a useful method for extraction of fungal DNA; extremely rapid, practical and successful. Sequence analysis strategy of D1-D2 region of the large ribosomal subunit gene was found considerably sufficient in identification to genus level for the most clinical fungi. However, the identification to species level and especially discrimination of closely related species may require additional analysis.

Investigation of key miRNAs and target genes in bladder cancer using miRNA profiling and bioinformatic tools.

Despite the association of several miRNAs with bladder cancer, little is known about the miRNAs' regulatory networks. In this study, we aimed to construct potential networks of bladder-cancer-related miRNAs and their known target genes using miRNA expression profiling and bioinformatics tools and to investigate potential key molecules that might play roles in bladder cancer regulatory networks. Global miRNA expression profiles were obtained using microarray followed by RT-qPCR validation using two randomly selected miRNAs. Known targets of deregulated miRNAs were utilized using DIANA-TarBase database v6.0. The incorporation of deregulated miRNAs and target genes into KEGG pathways were utilized using DIANA-mirPath software. To construct potential miRNA regulatory networks, the overlapping parts of three selected KEGG pathways were visualized by Cytoscape software. We finally gained 19 deregulated miRNAs, including 5 ups- and 14 down regulated in 27 bladder-cancer tissue samples and 8 normal urothelial tissue samples. The enrichment results of deregulated miRNAs and known target genes showed that most pathways were related to cancer or cell signaling pathways. We determined the hub CDK6, BCL2, E2F3, PTEN, MYC, RB, and ERBB3 target genes and hub hsa-let-7c, hsa-miR-195-5p, hsa-miR-141-3p, hsa-miR-26a-5p, hsa-miR-23b-3p, and hsa-miR-125b-5p miRNAs of the constructed networks. These findings provide new insights into the bladder cancer regulatory networks and give us a hypothesis that hsa-let-7c, hsa-miR-195-5p, and hsa-miR-125b-5p, along with CDK4 and CDK6 genes might exist in the same bladder cancer pathway. Particularly, hub miRNAs and genes might be potential biomarkers for bladder cancer clinics.

The usefulness of DNA sequencing after extraction by Whatman FTA filter matrix technology and phenotypic tests for differentiation of Candida albicans and Candida dubliniensis.

Since C. dubliniensis is similar to C. albicans phenotypically, it can be misidentified as C. albicans. We aimed to investigate the prevalence of C. dubliniensis among isolates previously identified as C. albicans in our stocks and to compare the phenotypic methods and DNA sequencing of D1/D2 region on the ribosomal large subunit (rLSU) gene. A total of 850 isolates included in this study. Phenotypic identification was performed based on germ tube formation, chlamydospore production, colony colors on chromogenic agar, inability of growth at 45 °C and growth on hypertonic Sabouraud dextrose agar. Eighty isolates compatible with C. dubliniensis by at least one phenotypic test were included in the sequence analysis. Nested PCR amplification of D1/D2 region of the rLSU gene was performed after the fungal DNA extraction by Whatman FTA filter paper technology. The sequencing analysis of PCR products carried out by an automated capillary gel electrophoresis device. The rate of C. dubliniensis was 2.35 % (n = 20) among isolates previously described as C. albicans. Consequently, none of the phenotypic tests provided satisfactory performance alone in our study, and molecular methods required special equipment and high cost. Thus, at least two phenotypic methods can be used for identification of C. dubliniensis, and molecular methods can be used for confirmation.

Nonsense ß-thalassemia mutation at codon 37 (TGG>TGA), detected for the first time in three Turkish cases.

Thalassemias are genetically heterogeneous group of disorders with reduced or absent production of globin. ß-Thalassemia major can be caused by homozygosity or compound heterozygosity for ß-globin gene mutation. Here we report, for the first time in Turkey, three cases who carry the nonsense ß-thalassemia (ß-thal) mutation at codon 37 (TGG>TGA; Trp→Stop) causing premature stop codon.

Developmental genomics of limb malformations: Allelic series in association with gene dosage effects contribute to the clinical variability.

Genetic heterogeneity, reduced penetrance, and variable expressivity, the latter including asymmetric body axis plane presentations, have all been described in families with congenital limb malformations (CLMs). Interfamilial and intrafamilial heterogeneity highlight the complexity of the underlying genetic pathogenesis of these developmental anomalies. Family-based genomics by exome sequencing (ES) and rare variant analyses combined with whole-genome array-based comparative genomic hybridization were implemented to investigate 18 families with limb birth defects. Eleven of 18 (61%) families revealed explanatory variants, including 7 single-nucleotide variant alleles and 3 copy number variants (CNVs), at previously reported "disease trait associated loci": BHLHA9, GLI3, HOXD cluster, HOXD13, NPR2, and WNT10B. Breakpoint junction analyses for all three CNV alleles revealed mutational signatures consistent with microhomology-mediated break-induced replication, a mechanism facilitated by Alu/Alu-mediated rearrangement. Homozygous duplication of BHLHA9 was observed in one Turkish kindred and represents a novel contributory genetic mechanism to Gollop-Wolfgang Complex (MIM: 228250), where triplication of the locus has been reported in one family from Japan (i.e., 4n = 2n + 2n versus 4n = 3n + 1n allelic configurations). Genes acting on limb patterning are sensitive to a gene dosage effect and are often associated with an allelic series. We extend an allele-specific gene dosage model to potentially assist, in an adjuvant way, interpretations of interconnections among an allelic series, clinical severity, and reduced penetrance of the BHLHA9-related CLM spectrum.

Developmental expression of p97/VCP (Valosin-containing protein) and Jab1/CSN5 in the rat testis and epididymis.

BACKGROUND: The ubiquitin proteasome system (UPS) is a key player in regulating many cellular processes via proteasomal degradation of ubiquitinated proteins. Recently published data show that Jab1/CSN5 interacts with p97/VCP and controls the ubiquitination status of proteins bound to p97/VCP in mouse and human cells. However, coexpression of p97/VCP and Jab1/CSN5 in the developing rat testis and epididymis has not previously been studied. METHODS: Testicular and epididymal tissues from 5-, 15-, 30-, and 60-day-old rats were examined by immunohistochemistry and Western blotting. Colocalisation of proteins was determined by immunofluorescence microscopy. RESULTS: In the 5-day-old rat testis, p97/VCP and Jab1/CSN5 were specifically expressed in gonocytes. The expression of p97/VCP and Jab1/CSN5 significantly increased at day 15 and was found in spermatogonia, Sertoli cells and spermatocytes. In 30- and 60-day-old rat testes, p97/VCP indicated moderate to strong expression in Sertoli cells, spermatogonia, round and elongating spermatids. However, moderate to weak expression was observed in spermatocytes. Jab1/CSN5 showed strong expression in spermatogonia and spermatocytes, while relatively moderate expression was observed in round and elongating spermatids in 30- and 60-day-old rat testes. In contrast, in the epididymis, the expression of both proteins gradually increased from 5 to 60 days of age. After rats reached 2 weeks of age, the expression of both proteins was mostly restricted to the basal and principal cells of the caput epididymis. CONCLUSIONS: Our study suggests that p97/VCP and Jab1/CSN5 could be an important part of the UPS in the developing rat testis and epididymis and that both proteins may be involved in the regulation of spermatogenesis and epididymal epithelial functions.

Anterior cruciate ligament reconstruction with ToggleLoc with ZipLoop system versus transfix system: A cost-effectiveness analysis.

BACKGROUND: To evaluate the cost-effectiveness of the reconstruction of the anterior cruciate ligament tears using either ToggleLoc with ZipLoop or Transfix systems. METHODS: This study is a cost-effectiveness analysis of patients with anterior cruciate ligament reconstruction, ToggleLoc with ZipLoop and Transfix systems in our clinic between 2011 and 2016. This study was a retrospective cross-sectional study of patient's demographic, clinical and financial data. The effectiveness of surgery on patients with anterior cruciate ligament reconstruction was determined by the Lysholm Knee Score Scale. We compared two systems with the cost-effectiveness ratio. RESULTS: In this study, 103 patients were included. According to the Lysholm Knee Score Scales in both groups, the findings showed that there was no difference in effectiveness between them. The ToggleLoc with ZipLoop technique was cost-effectiveness ratio 254,57 and the Transfix technique cost-effectiveness ratio was 378,33. CONCLUSION: According to our results, ToggleLoc with ZipLoop technique was a more cost-effective method than the Transfix technique in the anterior cruciate ligament reconstruction.

Stereologic analysis of bone produced by distraction osteogenesis or autogenous bone grafting in mandible.

To our knowledge, in the literature, any other investigation that numerically compared osteoblasts retrieved from transport distraction osteogenesis and bone grafting procedures using stereological methods is not reported. The purpose of this study was to compare the total number of osteoblast cells at 3 months in bone produced by distraction osteogenesis and that in autogenous bone graft. A total of 19 growing sheep (male aged 7 or 8 mo; weighing between 21 and 28 kg) were used in this study. Mandibular discontinuity defects created in mandibles of sheep were reconstructed by transport distraction osteogenesis and iliac crest bone graft and allowed to heal for 3 months. The animals were then killed, and the jaws were resected and prepared to be decalcified. Stereological and histologic examinations were performed. Intramembranous ossification and osteoid and trabecular formations were observed in both groups. In the distraction group, the mean +/- standard deviation (SD) numerical density of the osteoblasts was found to be higher (0.0004866 +/- [0.000044])when compared with those of both the graft (0.0003458 +/- [0.000030]) and control groups (0.0002714 +/- [0.000022]). Statistically significant differences were found among the groups (P < 0.05). Stereologic evaluation of bone in the sheep model demonstrated significantly greater osteoblast density in bone formed through transport distraction osteogenesis when compared with bone grafting and the control. Therefore, further studies should be conducted to evaluate the differences in both osteoblastic and osteoclastic cellular activities at different time points in distraction osteogenesis and autogenous bone grafting to assess the healing process of bone for clinical applications.

Caffeic acid phenethyl ester protects rabbit brains against permanent focal ischemia by antioxidant action: a biochemical and planimetric study.

The present study was conducted to investigate whether caffeic acid phenethyl ester (CAPE), an active component of propolis extract, has a protective effect on brain injury after focal permanent cerebral ischemia, and to determine the possible antioxidant mechanisms. Cerebral infarction in adult male New Zealand rabbits was induced by microsurgical procedures producing right focal permanent middle cerebral artery occlusion (pMCAO). CAPE was administered to the treatment group after pMCAO at a dose of 10 micromol kg(-1) once a day intraperitoneally for 7 days. Neurological deficits were evaluated, using a modified six-point scale. Spectrophotometric assay was used to determine the contents of malondialdehyde (MDA), glutathione (GSH), catalase (CAT), nitric oxide (NO) and xanthine oxidase (XO). In the ipsilateral hemisphere, the infarct volume of the brain was assessed in brain slices stained with heamatoxylen and eosin. The results showed that treatment with CAPE significantly reduced the percentage of infarction in the ipsilateral hemisphere compared with the ischemia group. CAPE treatment significantly attenuated the elevation of plasma MDA, CAT and XO content (p<0.05), whereas it significantly increased the levels of plasma GSH and NO (p<0.05). Therefore, subacute CAPE administration plays a protective role in focal pMCAO due to attenuation of lipid peroxidation and its antioxidant activity. All of these findings suggest that CAPE provides neuroprotection against cerebral ischemia injury through its antioxidant action.

Effect of pinealectomy on the morphology of the chick cervical spinal cord: a stereological and histopathological study.

Melatonin has some effects upon morphological features of various structures in small animals and human being. In this study, the changes induced by pinealectomy procedure on morphological features of developing cervical spinal cord and their neurons in the gray matter (GM) and white matter (WM) of cervical spinal cord in the chicken were investigated. A total of 15 Hybro Broiler newly hatched chicks were randomly divided into three equal groups: unoperated control group (n=5), sham-operated group (n=5) and pinealectomy group (n=5). Pinealectomy procedure and sham operation were done in 3-day-old chicks and all animals were sacrificed at the 8th week and the 6th cervical (C6) spinal cord segment was dissected out for histopathological evaluation and subsequent stereological analysis. The volume of spinal cord segment did not show a significant difference between unoperated and sham-operated controls, but the pinealectomy group has a declined volume value compared with those of the control and sham operated groups (P<0.01). By contrast, there was no statistically significant difference between unoperated and sham-operated controls and the pinealectomy group with regard to volume fraction of the GM and WM of the cervical spinal cord. Finally, it was observed that pinealectomy procedure significantly reduces neuron number in the GM and the volume of WM of the C6 segment of cervical spinal cord in the chicken (P<0.001). The present study is the first study at all to evaluate the effects of pinealectomy on quantitative feature of the spinal cord in the chicken. Based on our findings, we suggest that pineal gland/melatonin might play an important role in morphological features of the developing spinal cord in the chicken.

Inhibitory effect of pinealectomy on the development of cerebellar granule cells in the chick: a stereological study.

Melatonin has some effects upon morphological features of various structures in small animals and human being. However, there has been no investigation concerning its physiological role on development of cerebellar granule cells. In this study, the changes induced by pinealectomy procedure on cerebellar development and their granule cell numbers in the chick were investigated using quantitative stereological methods. A total of 15 Hybro Broiler newly hatched chicks were randomly divided into three equal groups: pinealectomy group (n=5) and non-pinealectomized control group (n=5) and sham-operated group (n=5). Pinealectomy procedure and sham operation were done in 3-day-old chicks and all animals were sacrificed for histopathological evaluation and subsequent stereological analysis in the 8th week. Here, it was observed that pinealectomy significantly reduces the granular cell number in cerebellar cortex of the chicks (P<0.001). The present study is the first stereological study to evaluate the histomorphological effects of pinealectomy on the cerebellar granule cells of the chick. We suggest that the granule cell loss in the cerebellar cortex is due to developmental retardation in early postnatal period, although its exact mechanism is not clear. Based on our findings, we intimate that pineal gland/melatonin might play an important role in the development of cerebellar granule cells in the chick.

Effect of prenatal exposure to an anti-inflammatory drug on neuron number in cornu ammonis and dentate gyrus of the rat hippocampus: a stereological study.

Prenatal exposed to an anti-inflammatory drug is a major problem for the developing central nervous system. It is not well known the effect of prenatal exposed to a non-steroidal anti-inflammatory drug on the hippocampus. Total neuron number in one side of the cornu ammonis (CA) and gyrus dentatus (GD) of the hippocampal formation in control and drug-treated (diclofenac sodium, DS) groups of male rats was estimated using the optical fractionator technique. Each main group has also two subgroups that are 4 weeks old (4W-old) and 20 weeks old (20W-old). In CA, no significant difference between 4W-old DS-treated and their control was found, but a significant difference was observed between 20W-old DS-treated and their controls. A decreasing of neuron number was 12% for 20W-old DS-treated group. In GD, a decreasing of the granule cell number in 4W-old of DS-treated group was seen but an increasing of granule cell number was found in the 20W-old drug-treated rats in comparison to its control group, 7% and 9%, respectively. Although an increasing of neuron number in CA at the control group was seen with age, from 4th week to 20th week (10%), age-dependent substantial granule cell decline (17%) was observed in GD. No age effect on the total cell numbers of CA and GD of the drug-treated groups was seen in comparison to 4W-old week and 20W-old. A pronounced neuron loss observed in the drug-treated group may be attributed to the neurotoxicity of diclofenac sodium (DS) on the developing hippocampal formation. Age-dependent neuron increase in the CA of 20W-old and neuron decline in GD of 20W-old control groups may be a result of a dual effect of saline injection during the fetal life, since these animals were exposed to a stress of 15-day-period of saline injection, prenatal stress. The reason of no age effect on CA and GD cell number in the drug-treated groups may be attributed to the depletion of the progenitor cells due to neurotoxicity of DS in the fetal life of these animals.

Effect of prenatal exposure to diclofenac sodium on Purkinje cell numbers in rat cerebellum: a stereological study.

Diclofenac sodium (DS) is commonly used as a non-steroidal anti-inflammatory drug. Although several adverse effects are clearly established, it is still unknown whether prenatal exposure to DS has an effect on the development of the cerebellum. In this study, we investigated the total number of Purkinje cells of the cerebellum in a control group and in a DS-treated group of male rats using a stereological method. The DS in a dose of 1 mg/kg daily was intraperitoneally injected to the drug-treated group of pregnant rats beginning from the 5th day after mating for a period of 15 days during pregnancy. Physiological serum at 1 ml dose was intraperitoneally injected to the control group of pregnant rats at the same period. After delivery, male offspring were obtained and each main group was divided into two subgroups that were 4-week-old (4W-old) and 20-week-old (20W-old). Our results showed that the total number of Purkinje cells in offspring of drug-treated rats was significantly lower than in the offspring of control animals. These results suggest that the Purkinje cells of a developing cerebellum may be affected by administration of DS during the prenatal period.

Effects of postnatal formaldehyde exposure on pyramidal cell number, volume of cell layer in hippocampus and hemisphere in the rat: a stereological study.

The purpose of the present study was to determine whether exposure of neonatal rats to formaldehyde (FA) had either early or delayed effects on the numbers of pyramidal cells in the cornu ammonis (CA) of the hippocampus. Neonatal Wistar rats were exposed to 0 ppm (control group), 6 ppm and 12 ppm (high concentration group) of FA concentrations throughout the 30-day period following the birth by placing them for 6 h/day in a glass chamber containing FA vapor. Then, some of the animals from each FA-treated group were anesthetized and decapitated at the day 30, and the remaining ones were killed at the day 90. The brains were removed immediately and fixed in 10% neutral-buffered FA solution. The Cavalieri principle was used to determine the volumes of the CA and the entire cerebral hemisphere. The optical fractionator counting method was used to estimate the total number of pyramidal cells in the CA. The appearance of pyramidal cells was normal under light microscopy at both postnatal day (PND) 30 and PND 90 in all groups. There were concentration-related volume changes of CA at PND 30 and PND 90; low concentration of FA significantly increased, whereas high concentration decreased the volume of CA in comparison of the control at PND 30. Importantly, high concentration of FA at PND 90 increased the volume of CA in comparison of the low concentration but not with the control. Furthermore, low and high concentrations of FA decreased the volume of hemisphere at PND 30, whereas a reverse effect of these concentrations was observed at the hemisphere of PND 90 in comparison of the control. In both CA and cerebral hemisphere, an age-related volume decrease in both control and low/high concentration groups were found. On the other hand, there were significant age-related reductions in the total number of pyramidal cells at 90 days of age irrespective of the groups examined. Rats treated with high concentration FA were seen to have significantly fewer pyramidal cell neurons than either the animals treated with low concentration FA or control groups (p<0.01). These observations indicate that pyramidal cells in the hippocampus may be vulnerable to FA exposure during the early period of life.

A preliminary study of the levels of testis oxidative stress parameters after MK-801-induced experimental psychosis model: protective effects of CAPE.

We evaluated the effects of caffeic acid phenethyl ester (CAPE) on antioxidant enzyme levels and histopathologic changes in dizocilpine (MK-801) induced schizophrenic rat testis. A total of 30 adult male Wistar-Albino rats were divided into three groups. Group-I was used as control. Rats in the Group-II were intraperitoneally injected with MK-801, whereas those in Group-III were intraperitoneally injected with CAPE in addition to MK-801. The testes were collected for biochemical and histopathological examinations. Antioxidant enzyme activities, malondialdehyde, protein carbonyl and nitric oxide levels in testicular tissues were analyzed with spectrophotometric methods. Induction of schizophrenia resulted in a significant oxidative stress by increasing the levels of antioxidant enzymes. Tissue malondialdehyde and protein carbonyl levels were also increased. Treatment with CAPE led to significant decrease in oxidative injury. Administration of CAPE reduced the detrimental histopathologic changes caused by MK-801. The results showed that experimentally induced schizophrenia caused oxidative stress in testes of rats and treatment with CAPE reduced these harmful effects.