Transplantation of Schwann cells differentiated from adipose-derived stem cells modifies reactive gliosis after contusion brain injury in rats.

This study investigated whether transplantation of Schwann cells differentiated from adipose-derived stem cells (ADSC-SCs) of rats could promote functional improvement after contusion brain injury, with a focus on the effect on reactive gliosis. ADSCs were isolated and expanded from groin adipose tissue of Sprague-Dawley rats and then differentiated into Schwann cells. ADSCSCs were transplanted into the contused rat brain. Immunofluorescence and Western blotting were used to analyse reactive gliosis, and locomotor function of the rats was assessed. Hemiparalysed rats transplanted with ADSC-SCs showed significant locomotor function recovery compared with rats transplanted with undifferentiated ADSCs or control rats injected with medium alone. Transplanted ADSC-SCs significantly reduced glial scar formation and neurocan protein levels compared with transplanted undifferentiated ADSCs. In conclusion, transplantation of ADSC-SCs can effectively promote locomotor functional recovery and reduce reactive gliosis after contusion brain injury in rats.

Identification and molecular characterization of two novel chromosomal deletions associated with autism.

Autism is a childhood-onset neurodevelopmental disorder with a strong genetic basis in its etiology. Conventional karyotype analysis has revealed that chromosomal structural aberrations such as translocation, inversion, deletion, and duplication play a role in causing autism spectrum disorders (ASD). In addition, recent array-based comparative genomic hybridization (array CGH) studies discovered that submicroscopic deletion and duplication of DNA segments also contributed significantly to the genetic etiology of ASD. Together, these studies indicate that genomic rearrangement is an important genetic mechanism of ASD. Using karyotyping analysis and array CGH technology, we identified a subtelomeric deletion of approximately 6.8 Mb at 4q35.1-35.2 and a terminal deletion of approximately 2.4 Mb at 8p23.2-pter in two autistic boys, respectively. These two deletions were further validated using fluorescent in situ hybridization and real-time quantitative polymerase chain reaction, and their breakpoints were delineated using high-resolution array CGH. The 4q deletion is a rare de novo mutation, while the transmission of 8p deletion is unknown, because the father of the patient was unavailable for study. These two deletions are rare mutations and were not found in the additional 282 patients with ASD and in the 300 control subjects in our population. The identification of these two chromosomal deletions contribute to our understanding of the genetic basis of ASD, and the haploinsufficiency of several genes located at the deleted regions of chromosome 8p and 4q may contribute to the clinical phenotypes of autism.

A novel microdeletion at chromosome 2q31.1-31.2 in a three-generation family presenting duplication of great toes with clinodactyly.

HOXD gene cluster maps to chromosome 2q31 and plays a key role in embryonic limb morphogenesis. Mutations of the HOXD13 and HOXD10 genes have been found to be associated with digital and limb malformations. In addition, dysregulation of HOXD gene cluster has been proposed to account for the limb abnormalities in patients with chromosome 2q rearrangements. In this report, we investigated a three-generation family presenting clinical phenotypes of duplication of great toes, tapering fingers, and clinodactyly of the fifth finger in both hands, which were transmitted in a dominant fashion in this family. We identified and validated an interstitial microdeletion of approximately 3.4 Mb at chromosome 2q31.1-31.2 by array-based comparative genomic hybridization, fluorescence in situ hybridization, and real-time quantitative polymerase chain reaction that cosegregates with the clinical phenotypes in this family. The microdeletion removes 30 labeled genes including the entire HOXD gene cluster, suggesting that the digital abnormalities of this family may be attributed to the haploinsufficiency of the HOXD gene cluster. The delineation of the microdeletion region may contribute to the genotype-phenotype correlation study in patients with genomic rearrangements of the long arm of chromosome 2 and helps to understand the pathogenesis of haploinsufficiency of the HOXD gene cluster.

Cytogenetic and molecular characterization of a three-generation family with chromosome 5p terminal deletion.

Terminal or interstitial deletion on the short arm of chromosome 5 is associated with a genetic disorder, cri-du-chat syndrome (cat cry syndrome), which is characterized by a cat-like cry in infancy, facial dysmorphism, microcephaly, and mental retardation. There is a high degree of variation in clinical presentations of patients with cri-du-chat syndrome, which is usually associated with different sizes and locations of deletions in chromosome 5p. Most patients with a 5p deletion have de novo mutations; familial 5p deletion is rare in literature. Here, we report a three-generation family with a 5p terminal deletion. The terminal 5p deletion (5p15.2-pter) in this family was confirmed and characterized by karyotyping analysis, fluorescent in situ hybridization, array comparative genome hybridization, and quantitative polymerase chain reaction. Although the affected family members apparently share deletions of the same size, there are some variations in mental symptoms within this family. Two affected females manifest moderate mental retardation and psychotic symptoms such as delusion of persecution, auditory hallucination, self-talking, and self-laughing, which are rare in cri-du-chat syndrome. In contrast, the other three affected males express mild-to-moderate mental retardation but no psychotic symptoms. Our study suggests that other factors besides the size and location of 5p deletions may modify the mental presentations of patients with 5p deletions.

Flavonoids from Radix Scutellariae as potential stroke therapeutic agents by targeting the second postsynaptic density 95 (PSD-95)/disc large/zonula occludens-1 (PDZ) domain of PSD-95.

Excessive activation of N-methyl-D-aspartate receptors (NMDARs) and subsequent production of nitric oxide by neuronal nitric oxide synthase (nNOS) contribute to neuronal damage resulting from hypoxic and ischemic insults. NMDARs and nNOS are coupled together at the postsynaptic membrane through their interaction with postsynaptic density protein (PSD) 95 via PSD-95/disc large/zonula occludens-1 (PDZ) domains. We used NMR (nuclear magnetic resonance) spectroscopy to screen medicinal herbs used in traditional Chinese medicine (TCM) stroke therapy for compounds binding to the second PDZ domain (PDZ2) of PSD-95, the domain linking nNOS and PSD-95. Aqueous extract of Huangqin, the root of Scutellaria baicalensis Georgi (Labiatae), showed significant binding to PDZ2 of PSD-95. The binding site of the active components in the extract overlapped with the nNOS/NR2B-binding pocket of PDZ2 of PSD-95. Four flavones, baicalin, norwogonoside, oroxylin A-glucuronide (oroxyloside), and wogonoside were isolated and found to account for the PDZ-binding activity of the extract. NMR titration experiments showed that baicalin and norwogonoside displayed the highest PDZ2 binding affinity, while oroxylin A-glucuronide and wogonoside showed 4-5 fold less potency in binding to the PDZ domain. Identification of the PDZ binding activity of these compounds will allow investigating whether or not it contributes to the observed clinical effects of Radix Scutellariae. Furthermore, these molecules might provide leads for the development of drugs targeting the signaling pathways mediated by PDZ domains.

Prevalence of adults with brain arteriovenous malformations: a community based study in Scotland using capture-recapture analysis.

OBJECTIVE: To conduct a population based study of brain arteriovenous malformation (AVM) prevalence. METHODS: Multiple, overlapping sources of case ascertainment were used to establish the point prevalence of brain AVMs in the adult population of the Lothian health board of Scotland. Patients were sought retrospectively from all local general (family) practitioners, neurologists, neurosurgeons, stroke physicians, the specialist AVM clinic at the regional neuroscience centre, and routine coding of hospital discharge data. Case notes, brain imaging, and pathology reports were reviewed to validate each patient's diagnosis and to ensure that each was alive, over the age of 16 years, and resident in the geographical area of the study on the prevalence date of 30 June 1998. RESULTS: Of 148 potentially eligible people, 93 adults met the inclusion criteria. There were 40 women and 53 men. Men were significantly younger than women on the prevalence date (median age 39 years v 51 years, p = 0.003). Of those included, 25 (27%) had radiological evidence of prior therapeutic obliteration of their brain AVM and 9 (10%) had coexisting aneurysms. The minimum crude brain AVM prevalence was 15 per 100 000 adults and capture-recapture analysis gave an ascertainment adjusted prevalence of 18 (95% confidence interval 16 to 24) per 100 000 adults. CONCLUSIONS: The minimum estimate of brain AVM prevalence helps to assess its burden and comparative epidemiology and stresses the importance of brain AVMs as a cause of long term disability in adults.

Implications of sonographic identification of duplex kidney in adults.

BACKGROUND: Duplication of the renal pelvis and ureter constitutes the most common anomalies of the upper urinary tract. Most patients are asymptomatic and don't need treatment. However, the associated anomaly and acquired renal disease may cause morbidity and mortality. The aim of this study was to investigate the prevalence of duplex kidney in sonographic examinations and the predictive capability of this method. An analysis of patients with urography-confirmed duplication anomaly was also performed. METHODS: During a period of 9 year (from 1988 to 1996), sonographic examination records of 19,287 patients were reviewed. Patients with a diagnosis of duplex kidney were enrolled. Their clinical characteristics and radiological images were recorded and evaluated. RESULTS: One hundred and sixty-three patients were found by sonography to have duplex kidney, for an prevalence of 0.85%. Urography studies were done on 64 (39.2%) patients, and 42 patients were proven to have duplex kidney. The positive predictive rate of sonography was 65.6%. Eighty percent of patients with duplex kidney had associated ureteral duplication. No associated anomalies were found. Four patients had urolithiasis, and 3 patients had recurrent urinary tract infection. One patient had coexisting uroepithelial malignancy, and 1 patient had refractory gross hematuria. CONCLUSIONS: Duplex kidney is an uncommon finding during adult sonographic examinations. The sensitivity and specificity of sonography in identifying this anomaly are controversial. An experienced hand with good knowledge of the anomaly and its sonographic signs is helpful in more accurately diagnosing this anomaly.

[Study of the fluid-percussion graded model of experimental brain injury in rats].

OBJECTIVE: To study the histopathological aspects of the fluid-percussion graded model of experimental brain injury in rats and the relationship between the fluid-percussion graded model and the clinical grades of brain injury. METHODS: The graded model of rats was set up by using the device of the improved fluid-percussion model, then we observed the changes of brain tissue of rats. RESULTS: The fluid-percussion graded model of experimental brain injury could be pathologically graded. CONCLUSION: The pathological grade of the fluid-percussion graded model can be used to evaluate the degree of brain injury in clinic and in experiments.

Gonadoblastoma and chroiocarcinoma in dysgenic gonads: report of a case.

Dysgenetic gonads are well known to be associated with gonadoblastoma and various germ cell tumors, but very rarely with choriocarcinoma. We describe a 16-year-old, phenotypic female patient with a 46XY karyotype who developed gonadoblastoma on the right gonad and choriocarcinoma on the left. There was no co-existence of the two tumor cell types in the same gonad. This case emphasizes the importance of bilateral gonadectomy for "46XY female" patients, as soon as the cytogenetic evidence is known.

Patterns of immunocytochemically detected Z-DNA in the recrudescing testicular epithelium of the Turkish hamster (Mesocricetus brandti).

Z-DNA has been considered a labile but essential structural form of DNA in recombination and gene expression, two significant activities in mammalian seminiferous epithelium. The present study has utilized the recrudescing testes of Mesocricetus brandti to study in detail the potential Z-DNA sites in specific testicular cell types as detected by an immunoprobe. Testicular regression was physiologically induced by modifying environmental photoperiods and/or temperature. Partial atrophy of seminiferous epithelium occurred in all experimental groups but Sertoli cells persisted throughout regression. Recrudescence of testicular activity was marked in all experimental groups by characteristic sequences of reappearance of potential Z-DNA sites to a final positive or negative mature state of the cell type. It is suggested that Z-DNA is a functionally important from of DNA in many cell types of the active seminiferous epithelium of the Turkish hamster, and perhaps other mammals.

Some characteristics of oogenesis in the female Turkish hamster subjected to modified environments.

The hibernating female Turkish hamster (Mesocricetus brandti) was utilized for a study of possible in vivo effects of cold on oocyte maturation. Such a physiologic model offered an opportunity to analyze the ability of oocytes exposed to prolonged periods of reduced core temperature and/or light to subsequently mature to Metaphase II. Detailed observations of core temperatures, torpor/arousal, serum estradiol, and ovarian histology were made. An average incidence of 37.7% binuclearity was found in the germinal vesicle, metaphase I and II occytes of this species. Maturation to Metaphase II of total chromosome complements did not vary significantly in the experimental groups compared with the control, but aneuploidy was detected in the oocytes of animals exposed to reduced temperature or light. An effect of in vivo reduced core temperatures on oocyte chromosome complements validates many of the published in vitro studies with temperature reduction. The model presents an excellent physiologic system for perturbing and analyzing many aspects of mammalian oocyte development.

Unique state of sexual dimorphism of crossing-over in diplotene spermatocytes and oocytes of Mesocricetus brandti, a species with neonatal oogenesis.

Sexual dimorphism of recombination has been held by classic genetic theory to disfavor the heterogametic sex. Assessment of chiasma frequencies at the diplotene stage of meiosis has been used as a valid measure of this concept and in many species has revealed, as expected, an increased frequency in female vs. male germ cells. Mesocricetus brandti, a species currently used in investigations of gonadal regression, photoperiods, and hibernation, was found by this measure to be an exception to this rule, with an average of 29.81 chiasmata in spermatocytes and 23.16 in oocytes. Sites of crossing-over unique to each sex were also detected. Oogenesis occurs in this species as a postnatal phenomenon and is suggested as a possible critical factor for the exceptional recombinant behavior.

The chromomere map of the pachytene spermatocyte of the Turkish hamster (Mesocricetus brandti).

The chromomere map of the early to mid pachytene spermatocyte of the Turkish hamster (Mesocricetus brandti) is described. Each autosomal bivalent was identified and a total of 304 chromomeres was found. A sex bivalent with a despiralized Xq protruding from the sex vesicle and a small number of the polymorphic 16q bivalents were observed.

The pachytene chromomere map of the oocyte of the Turkish hamster (Mesocricetus brandti).

A study of the chromomere maps of the sex and twenty autosomal bivalents of Turkish hamster pachytene oocytes was carried out. The average total number of chromomeres in early/mid pachytene autosomes was 280 with 91 on the p (short arm) and 189 on the q (long arm). The submetacentric X1 chromosome had 20 chromomeres and the metacentric X2 had 27. Comparisons of the number and location of oocyte chromomeres are made with the pachytene spermatocyte chromomere maps of this species.

Colcemid effects on homologue pairing and crossing over during fetal mouse oogenesis.

Colcemid was administered to gestational day 13 female mice to test effects on homologue pairing, synapsis and recombination of fetal oogenesis. Pairing abnormalities were detected in pachytene oocytes by light and electron microscopy examination of bivalents and synaptonemal complexes. Reduction of total chiasmata per treated diplotene oocyte (22.74) compared to controls (31.07) was found.

The immunocytochemical localization of potential Z-DNA sites in human testicular tubule epithelium.

Z-DNA has been detected in several pro- and eukaryotic cells and possible roles in regulating transcriptional activity and meiotic recombination proposed. The present study examined the localization of reaction product to potential Z-DNA sites in human testicular tubule epithelium from three subjects using an avidin-biotin complex (ABC)-immunoperoxidase method with a specific rabbit antibody previously shown to react with rat spermatogonial nuclei. A total of 46,626 cells were scored, of which 5656 were Sertoli cells. Eighty-six percent of spermatogonia in mitotic metaphase were found to be negative. Interphase spermatogonia identified as A dark or pale were positive in 93 and 92%, respectively, of cells, and this positivity persisted through B spermatogonia into meiosis. Of 8083 leptotene/zygotene spermatocytes, 99% were positive. Pachytene spermatocytes were 98% positive in the autosomal bivalents. First meiotic metaphase nuclei were negative (92%), as were almost all cells scored of the spermiogenic series (16,195). Nuclei of Sertoli cells had reaction product over the chromatin in 81% of 5656 cells, with no reaction product on the prominent nucleolus. The presence of potential Z-DNA sites in the genome of human spermatogenesis and Sertoli cells during known active stages of transcription (pachytene) and recombination (zygotene/pachytene) suggests a role for this conformation during these stages.

Infertility in a patient with abnormal spermatogenesis and in utero DES exposure.

A young azoospermic patient is described whose spermatogenesis reflected an abnormality of meiosis. A diagnosis of asynapsis of chromosomes during early spermatogenesis was made by cytogenetic examination of a testicular biopsy. Standard histologic examination gave no indication of this abnormality. An incidental history of intrauterine diethylstilbesterol (DES) exposure of the patient was elicited. Attention is called to the dearth of cytogenetic studies of spermatogenesis in DES-treated male progeny and the usefulness in general of meiotic cytogenetic studies for obtaining accurate diagnoses in human infertility.

An analysis of the chromomere map and chiasmata characteristics of human diplotene spermatocytes.

A complete chromomere map of early/mid diplotene human spermatocytes has been developed which permits identification of each bivalent. Bivalents 9, 16, 17, and 19 demonstrated unique cytogenetic characteristics at this meiotic stage. The mean chiasma frequency per spermatocyte was 45.33 +/- 4.52 (ranging from 32 to 58) with 28% of bivalents having one chiasma, 38% having two, and 27% having three. The remaining 7% had four or more chiasmata. Fifty-eight percent of chiasmata were located distally, 31% centrally, and 11% proximally. Univalents were rare. The availability of human diplotene spermatocyte maps permits exploration of many basic questions of recombination with accuracy.

Chromosome 21 behavior during fetal oogenesis in Down's syndrome.

Oogenesis of four cases of 47,XX,+21 at gestational ages of 19 and 20 weeks was studied using pachytene cytogenetic methods. We found a variable pattern of pairing behavior of the 21 chromosomes among the cases, which included partially synapsed trivalents, a bivalent plus a univalent, and three univalents. The bivalent/univalent conformation of 21 chromosomes predominated. Pachytene chromomere maps were normal for all autosomal bivalents in 84 oocytes analyzed, except for minor variations in some 21 chromosomes. This complex system of pairing behavior of the 21 chromosomes theoretically affects subsequent disjunctional behavior and therefore may account for the observations of normal and trisomic progeny observed for Down's syndrome mothers. Further study of meiotic behavior during all stages of oogenesis in such patients would provide essential data for predicting chromosomal outcome of pregnancy in this population.