Assessment of Hand Motor Function in a Non-human Primate Model of Ischemic Stroke

Ischemic stroke results from arterial occlusion and can cause irreversible brain injury. A non-human primate (NHP) model of ischemic stroke was previously developed to investigate its pathophysiology and for efficacy testing of therapeutic candidates; however, fine motor impairment remains to be well-characterized. We evaluated hand motor function in a cynomolgus monkey model of ischemic stroke. Endovascular transient middle cerebral artery occlusion (MCAO) with an angiographic microcatheter induced cerebral infarction. In vivo magnetic resonance imaging mapped and measured the ischemia-induced infarct lesion. In vivo diffusion tensor imaging (DTI) of the stroke lesion to assess the neuroplastic changes and fiber tractography demonstrated three-dimensional patterns in the corticospinal tract 12 weeks after MCAO. The hand dexterity task (HDT) was used to evaluate fine motor movement of upper extremity digits. The HDT was modified for a home cage-based training system, instead of conventional chair restraint training. The lesion was localized in the middle cerebral artery territory, including the sensorimotor cortex. Maximum infarct volume was exhibited over the first week after MCAO, which progressively inhibited ischemic core expansion, manifested by enhanced functional recovery of the affected hand over 12 weeks after MCAO. The total performance time decreased with increasing success rate for both hands on the HDT. Compensatory strategies and retrieval failure improved in the chronic phase after stroke. Our findings demonstrate the recovery of fine motor skill after stroke, and outline the behavioral characteristics and features of functional disorder of NHP stroke model, providing a basis for assessing hand motor function after stroke.

Rapid Whole-genome Sequencing of Zika Viruses using Direct RNA Sequencing

Zika virus (ZIKV) is one of the pathogens which is transmitted world widely, but there are no effective drugs and vaccines. Whole genome sequencing (WGS) of viruses could be applied to viral pathogen characterization, diagnosis, molecular surveillance, and even finding novel pathogens. We established an improved method using direct RNA sequencing with Nanopore technology to obtain WGS of ZIKV, after adding poly (A) tails to viral RNA. This established method does not require specific primers, complimentary DNA (cDNA) synthesis, and polymerase chain reaction (PCR)-based enrichment, resulting in the reduction of biases as well as of the ability to find novel RNA viruses. Nanopore technology also allows to read long sequences. It makes WGS easier and faster with long-read assembly. In this study, we obtained WGS of two strains of ZIKV following the established protocol. The sequenced reads resulted in 99% and 100% genome coverage with 63.5X and 21,136X, for the ZIKV PRVABC59 and MR 766 strains, respectively. The sequence identities of the ZIKV PRVABC59 and MR 766 strains for each reference genomes were 98.76% and 99.72%, respectively. We also found that the maximum length of reads was 10,311 bp which is almost the whole genome size of ZIKV. These long-reads could make overall structure of whole genome easily, and WGS faster and easier. The protocol in this study could provide rapid and efficient WGS that could be applied to study the biology of RNA viruses including identification, characterization, and global surveillance.

Increased CD68/TGFβ Co-expressing Microglia/Macrophages after Transient Middle Cerebral Artery Occlusion in Rhesus Monkeys

The function of microglia/macrophages after ischemic stroke is poorly understood. This study examines the role of microglia/macrophages in the focal infarct area after transient middle cerebral artery occlusion (MCAO) in rhesus monkeys. We measured infarct volume and neurological function by magnetic resonance imaging (MRI) and non-human primate stroke scale (NHPSS), respectively, to assess temporal changes following MCAO. Activated phagocytic microglia/macrophages were examined by immunohistochemistry in post-mortem brains (n=6 MCAO, n=2 controls) at 3 and 24 hours (acute stage), 2 and 4 weeks (subacute stage), and 4, and 20 months (chronic stage) following MCAO. We found that the infarct volume progressively decreased between 1 and 4 weeks following MCAO, in parallel with the neurological recovery. Greater presence of cluster of differentiation 68 (CD68)-expressing microglia/macrophages was detected in the infarct lesion in the subacute and chronic stage, compared to the acute stage. Surprisingly, 98~99% of transforming growth factor beta (TGFβ) was found colocalized with CD68-expressing cells. CD68-expressing microglia/macrophages, rather than CD206⁺ cells, may exert anti-inflammatory effects by secreting TGFβ after the subacute stage of ischemic stroke. CD68⁺ microglia/macrophages can therefore be used as a potential therapeutic target.

Abnormal Mitochondria in a Non-human Primate Model of MPTP-induced Parkinson's Disease: Drp1 and CDK5/p25 Signaling

Mitochondria continuously fuse and divide to maintain homeostasis. An impairment in the balance between the fusion and fission processes can trigger mitochondrial dysfunction. Accumulating evidence suggests that mitochondrial dysfunction is related to neurodegenerative diseases such as Parkinson's disease (PD), with excessive mitochondrial fission in dopaminergic neurons being one of the pathological mechanisms of PD. Here, we investigated the balance between mitochondrial fusion and fission in the substantia nigra of a non-human primate model of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced PD. We found that MPTP induced shorter and abnormally distributed mitochondria. This phenomenon was accompanied by the activation of dynamin-related protein 1 (Drp1), a mitochondrial fission protein, through increased phosphorylation at S616. Thereafter, we assessed for activation of the components of the cyclin-dependent kinase 5 (CDK5) and extracellular signal-regulated kinase (ERK) signaling cascades, which are known regulators of Drp1(S616) phosphorylation. MPTP induced an increase in p25 and p35, which are required for CDK5 activation. Together, these findings suggest that the phosphorylation of Drp1(S616) by CDK5 is involved in mitochondrial fission in the substantia nigra of a non-human primate model of MPTP-induced PD.

Evaluation of fecal microbiomes associated with obesity in captive cynomolgus monkeys (Macaca fascicularis)

Microorganisms play important roles in obesity; however, the role of the gut microbiomes in obesity is controversial because of the inconsistent findings. This study investigated the gut microbiome communities in obese and lean groups of captive healthy cynomolgus monkeys reared under strict identical environmental conditions, including their diet. No significant differences in the relative abundance of Firmicutes, Bacteroidetes and Prevotella were observed between the obese and lean groups, but a significant difference in Spirochetes (p < 0.05) was noted. Microbial diversity and richness were similar, but highly variable results in microbial composition, diversity, and richness were observed in individuals, irrespective of their state of obesity. Distinct clustering between the groups was not observed by principal coordinate analysis using an unweighted pair group method. Higher sharedness values (95.81% ± 2.28% at the genus level, and 79.54% ± 5.88% at the species level) were identified among individual monkeys. This paper reports the association between the gut microbiome and obesity in captive non-human primate models reared under controlled environments. The relative proportion of Firmicutes and Bacteroidetes as well as the microbial diversity known to affect obesity were similar in the obese and lean groups of monkeys reared under identical conditions. Therefore, obesity-associated microbial changes reported previously appear to be associated directly with environmental factors, particularly diet, rather than obesity.

Reference values of hematological and biochemical parameters in young-adult cynomolgus monkey (Macaca fascicularis) and rhesus monkey (Macaca mulatta) anesthetized with ketamine hydrochloride / 한국실험동물학회지

Nonhuman primate models are valuable in biomedical research. However, reference data for clinical pathology parameters in cynomolgus and rhesus monkeys are limited. In the present study, we established hematologic and biochemical reference intervals for healthy cynomolgus and rhesus monkeys anesthetized with ketamine hydrochloride. A total of 142 cynomolgus monkeys (28 males and 114 females) and 42 rhesus monkeys (22 males and 20 females) were selected and analyzed in order to examine reference intervals of 20 hematological and 16 biochemical parameters. The effects of sex were also investigated. Reference intervals for hematological and biochemical parameters were separately established by species (cynomolgus and rhesus) and sex (male and female). No sex-related differences were determined in erythrocyte-related parameters for cynomolgus and rhesus monkey housed in indoor laboratory conditions. Alkaline phosphatase and gamma glutamyltransferase were significantly lower in females than males in both cynomolgus and rhesus monkeys aged 48–96 months. The reference values for hematological and biochemical parameters established herein might provide valuable information for researchers using cynomolgus and rhesus monkeys in experimental conditions for biomedical studies.

The current epidemiological status of infectious coryza and efficacy of PoulShot Coryza in specific pathogen-free chickens

Infectious coryza (IC) is an infectious disease caused by Avibacterium (Av.) paragallinarum. IC is known to cause economic losses in the poultry industry via decreased egg production in layers. Between 2012 and 2013, Av. paragallinarum was isolated from seven chicken farms by Chungbuk National University. We identified Av. paragallinarum, the causative pathogen of IC by polymerase chain reaction (PCR) and serovar serotype A, by multiplex PCR. Antibiotic sensitivity tests indicated that a few field-isolated strains showed susceptibility to erythromycin, gentamicin, lincomycin, neomycin, oxytetracycline, spectinomycin, and tylosin. A serological survey was conducted to evaluate the number of flocks that were positive for Av. paragallinarum by utilizing a HI test to determine the existence of serovar A. Serological surveys revealed high positivity rates of 86.4% in 2009, 78.9% in 2010, 70.0% in 2011, and 69.6% in 2012. We also challenged specific pathogen-free chickens with isolated domestic strains, ADL121286 and ADL121500, according to the measured efficacy of the commercial IC vaccine, PoulShot Coryza. We confirmed the effectiveness of the vaccine based on relief of clinical signs and a decreased re-isolation rate of ADL121500 strain. Our results indicate IC is currently prevalent in Korea, and that the commercial vaccine is effective at protecting against field strains.

Eggshell apex abnormalities associated with Mycoplasma synoviae infection in layers

Eggs exhibiting eggshell apex abnormalities (EAA) were evaluated for changes in shell characteristics such as strength, thickness, and ultrastructure. Mycoplasma synoviae (MS) infection was confirmed by serological assay along with isolation of MS from the trachea and oviduct. Changes in eggshell quality were shown to be statistically significant (p < 0.01). We also identified ultrastructural changes in the mammillary knob layer by Scanning Electron Microscopy. While eggs may seem to be structurally sound, ultrastructural evaluation showed that affected eggs do not regain their former quality. In our knowledge, this is the first report describing the occurrence of EAA in Korea.

An unusual case of concomitant infection with chicken astrovirus and group A avian rotavirus in broilers with a history of severe clinical signs

A molecular study of intestinal samples from 21 broiler flocks with a history of enteritis revealed that 23.8% and 14.3% were positive for chicken astrovirus (CAstV) and avian rotavirus (ARV), respectively. CAstV and group A ARV were simultaneously detected in only one broiler flock. Birds in this group developed the significant intestinal lesions characterized by frothy contents, paleness, and thin intestinal walls. In this report we present an unusual case of runting stunting syndrome (RSS) with a history of high mortality and growth retardation in broiler chickens. We also make the first identification of CAstV and group A ARV in broiler chickens in Korea.

Clinical efficiency of the measurement of fetal nuchal translucency and Doppler examination of ductus venosus as a screening tool for chromosomal abnormalities / 대한산부인과학회지

OBJECTIVE: To evaluate the efficiency of the measurement of fetal nuchal translucency (FNT) and ductus venosus Doppler examination (DV Doppler) as a screening tool for chromosomal abnormalities. METHODS: FNT measurement and DV Doppler wereperformed in 950 pregnancies between 11(+0)~13(+6) weeks' gestation. Chromosomal analysis was done when FNT was more than 3 mm and DV Doppler showed absent flow or reversed flow. The numbers of cases with increased FNT and abnormal DV Doppler were counted in the groups of abnormal and normal karyotype. RESULTS: Data were available in 912 pregnancies. 11 pregnancies showed abnormal karyotype (1.2%). In the 11 cases with abnormal karyotype,increased FNT was found in 8 cases with 72.7% sensitivity and abnormal DV Doppler was found in 5 cases with 45.4% sensitivity. In the 901 cases withnormal karyotype, increased FNT was found in 33 cases with 96.3% specificity and abnormal DV Doppler was found in 12 cases with 98.7% specificity. Positive predictive value was 19.5% in cases of increased FNT, 29.4% in cases of abnormal DV Doppler, and 44.4% in cases of increased FNT and abnormal DV Doppler both. CONCLUSION: There is no improvement in general screening for chromosomal abnormalities when FNT measurement and DV Doppler were performed together. But better specificity and positive predictive value for chromosomal abnormalities were found.

Prenatally diagnosed split hand malformation at 16 weeks' gestation by 3D ultrasonography / 대한산부인과학회지

Split hand malformation is a rare malformation with various presentations. The current report describes a case of split hand malformation detected using 2D and 3D ultrasonography at 16 weeks` gestation. 2D ultrasonographic findings were split hand malformation in the right hand and monodactyly in the left hand , and those findings were confirmed and further clarified using 3D imaging. Postmortem X ray findings were consistent with the ultrasonographic findings. We conclude that 3D ultrasonography can assist in clarifying 2D ultrasonography findings of hand malformations during the second trimester of pregnancy.

Clinical significance of E. coli O26 isolates on urine specimen of urinary tract infection / 대한산부인과학회지

OBJECTIVE: Escherichia coli (E. coli) O26 has been the most common type of non-O157 human isolates and it has been related with urinary tract infection and its sequelae. So we investigated the clinical significance of E. coli O26 among the cases of urinary tract infection. METHODS: From January, 2005 to December, 2007, the 22 E. coli isolates that were related with urinary tract infection were analyzed. The isolates were identified biochemically by Vitek 1. We performed antisera test by O157, O26, O111 diagnostic antisera about the 22 E. coli isolates. We reviewed clinical history of the same patients retrospectively. RESULTS: 331 E. coli isolates in the urine specimen were isolated from January, 2005 to December, 2007. 175 E. coli isolates that were related with urinary tract infection were analyzed by O157, O26, O111 antisera test. As a result, 22 isolates (13.5%) were O26 antisera positive. There were 8, 3, and 2 cases of watery diarrhea, hemolytic uremic syndrome, thrombotic thrombocytopenic purpura repectively. CONCLUSION: In our study, because E. coli O26 was pathogenic and developed major complications, we concluded that patients with urinary tract infection with E. coli. should examine the antisera test about E. coli O157 and O26.

Spontaneous uterine rupture from placenta percreta at 33 weeks' gestation after a single gynecologic surgery / 대한산부인과학회지

The current case describes a case of uterine rupture from placenta percreta in a woman who had only a single gynecologic surgery. We met the case of intrauterine fetal death (IUFD) with hemoperitoneum and found uterine rupture from placenta percreta by CT imaging. A 25-year-old woman was admitted to the emergency service for acute upper abdominal pain and severe hypotension at 33 weeks' gestation. She had undergone a single pelviscopic surgery due to cornual pregnancy 11 months previously. Ultrasonogram detected IUFD and hemoperitoneum. CT showed uterine rupture from placenta percreta. An emergency laparotomy was performed to correct the defect. The current case presents that placenta percreta can occur in a woman who had a single gynecologic surgery and clinicians should consider possible placenta percreta in diagnosing pregnant patients who present with acute abdominal pain and shock.

A case of lymphoepithelioma-like carcinoma of the uterine cervix / 대한산부인과학회지

Lymphoepithelioma-like carcinoma (LELC) of the uterine cervix is a rare tumor with 0.7% of all primary cervical malignancies. It may grossly range from no visible cervical lesion to a large exophytic cervical mass and histologically similar neoplasms occur in the nasopharynx. It has been classified as a subtype of squamous cell carcinoma. But, after reviewing the literature, this tumor is a distinct carcinoma of the cervix that differs from squamous cell carcinoma. LELC is composed of undifferentiated cells surrounded by a marked inflammatory infiltrate, characterized by plasma cells and lymphocytes in the stroma. It may represent a better prognostic group than the more common squamous cell carcinoma of the cervix because of lack of lymph node metastasis. Typically affects a younger population of women, is more prevalent in noncaucasian population (especially, those of Asian descent). However, it is not clearly understood, Epstein-Barr virus play in the pathogenesis of cervical LELC while humam papilloma virus (HPV) is a risk factor for squamous cell carcinoma. The prognosis is good only by surgical treatment. Radiation therapy also appears to be effective in eradicating localized, low-stage disease (radiosensitive). The authors have experienced one case of LELC of the uterine cervix. We report a case with a brief review.

Effect of prostaglandin- and nitric oxide-synthase inhibitors on the lipopolysaccharide-induced adrenomedullin productions in cultured murine macrophage cells / 대한산부인과학회지

OBJECTIVE: Adrenomedullin (AM), a potent vasorelaxing agent associated with the maintenance of pregnancy, is synthesized in response to inflammation, which is also associated with the biosynthesis of prostaglandin (PG) and nitric oxide (NO). To clarify the interrelationships of PG, NO and AM in the inflammatory process, we tested the effects of the PG synthase inhibitor indomethacin and the NO synthase inhibitor N omega-nitro-L-arginine methyl ester (L-NAME) on AM production in lipopolysaccharide (LPS)-stimulated cultured rat macrophages. METHODS: RAW 257.8, rat macrophages were incubated with LPS, and AM production was measured by ELISA. Cells were pretreated with indomethacin, L-NAME, or both, and the effect on LPS-induced AM production was assayed. To exclude the effect of cell death, a cell viability test on these cultures was performed. RESULTS: The largest increase of AM was seen between 1 microgram (36.33+/-2.05 pg/ml) and 10 microgram (89.33+/-6.02 pg/ml) of LPS concentration (p<0.01), making the latter the optimal LPS dose to stimulate AM production. AM secretion was proportional to time in culture (p<0.006). Addition of indomethacin, L-NAME, or both 1 hr before LPS stimulation decreased AM production 2 hr later, with the AM decrement greatest in cells pretreated with both indomethacin and L-NAME, followed by L-NAME alone and then indomethacin. CONCLUSION: These findings indicate that PG and NO increase AM synthesis in rat macrophages with the presence of LPS. These results suggest that the biosynthetic pathways of PG, NO, and AM may be linked.

A case of sirenomelia diagnosed after amnioinfusion / 대한산부인과학회지

Sirenomelia, also known as the mermaid syndrome, is a rare and usually lethal congenital malformation of uncertain etiology. It is characterized by fusion of the lower limbs and commonly associated with severe urogenital and gastrointestinal malformations. Early antenatal sonogram allows for earlier, and less traumatic termination of pregnancy, but usually it is impared by severe oligohydramnios related to bilateral renal agenesis. We diagnosed a sirenomelia with severe oligohydramnios at early second trimester after amnioinfusion and terminated. We report a case with a brief review of the literature.

Clinical significance of fetal nuchal translucency during early pregnancy / 대한산부인과학회지

OBJECTIVE: To examine the significance of fetal nuchal translucency during early pregnancy in detection of chromosomal abnormality and major cardiac defect METHODS: Between Jul 1998 and Jul 2002, ultrasound examination was performed in 1,253 unselected singleton pregnancies with a live fetus and fetal crown rump length of 24-88 mm. The fetal nuchal translucency thickness was measured successfully in all cases. We reviewed the pregnancy outcome about two parameters. Chromosomal abnormality was confirmed by antenatal karyotyping and newborn's feature suggestive of a chromosomal defect. Major cardiac defect was confirmed by newborn physical examination, targeted ultrasonography, neonatal echocardiography and autopsy. RESULTS: (1) Fetal nuchal translucency thickness increased significantly with crown-rump length. (2) Nuchal translucency was greater than 3 mm in 12 pregnancies with abnormal karyotypes, giving a detection rate of 75.0% with a false positive rate of 4.0%. With a cutoff of 2.5 mm, a detection rate of abnormal karyotype was 81.3% with a false positive rate of 8.3%. And with a gestational age specific cutoff, 95 percentile, a detection rate of abnormal karyotype was 75%, with a false positive rate of 6.4%. (3) With normal karyotype, the overall prevalence of major cardiac defects in this study population was 5/1,000 pregnancies (5/1,054). This prevalence increased from 48/1,000 (4/83) using 2.5 mm cutoff to 105/1,000 (4/38) using 3 mm cutoff. The detection rate for major cardiac defect of nuchal translucency above 3 mm is 80.0% with a false positive rate of 3.2%. CONCLUSION: Increased nuchal translucency is by far an important and efficient marker for screening of chromosomal defects and effective screening method for major cardiac defects.

Benefits of Antenatal Corticosteroid in Infants Delivered before 33 weeks of Gestation after Premature Rupture of Membranes / 대한산부인과학회잡지

OBJECTIVE: To evaluate the benefits of maternal corticosteroid therapy in infants delivered before 33 weeks of gestation after premature rupture of membranes (PROM). METHODS: This retrospective study included the pregnant women complicated by preterm delivery within 32 weeks of gestation after PROM at the Asan Medical Center between 1997 to 1999. Patients were divided into 2 groups according to the gestational age at delivery, i.e., one group who delivered within 28 weeks of gestation and the other group who delivered between 29 and 32 weeks of gestation. Within each group, we evaluated the effect of maternal dexamethasone therapy on the perinatal and neonatal outcomes based on the medical records. Data were analyzed with pearson's chi-square test, Fisher's exact test, and two sample t-test. p<0.05 was considered statistically significant. RESULTS: One hundred and fifteen pregnancies complicated by preterm delivery within 32 weeks of gestation after premature rupture of membranes were included. Preterm deliveries occurred within 28 weeks of gestation in 48 cases (41.7%) and between 28 and 32 weeks of gestations in 67 cases (58.3%). Antenatal dexamethasone therapy was done in 27 out of 48 mothers (56.3%) who delivered within 28 weeks of gestation and in 47 out of 67 mothers (70.1%) who delivered between 29 and 32 weeks of gestation. Antenatal dexamethasone therapy did not affect the selected perinatal outcome variables (gestational age at delivery, birth weight, Apgar scores, cesarean section rate, and maternal and neonatal WBC counts and serum C-reactive protein concentrations). Incidences of neonatal complications (respiratory distress syndrome, intraventricular hemorrhage, retinopathy of prematurity, and periventricular leukomalacia) between the groups who did and did not received antenatal dexamethasone were not significantly different in pregnancies who delivered within 28 weeks of gestation. However, incidences of respiratory distress syndrome and intraventricular hemorrhage were significantly lower in a group who received antenatal dexamethasone than in a group who did not in pregnancies who delivered between 29 and 32 weeks of gestation (p<0.05). CONCLUSION: Antenatal corticosteroid therapy may be beneficial to the infants delivered between 29 and 32 weeks of gestation after PROM. However, it may have no therapeutic advantage to the group who delivered within 28 weeks of gestation after PROM.

A Case of Congenital Intracranial Teratoma that prenatally Diagnosed / 대한주산의학회잡지

Congenital intracranial teratoma is rare and it's prognosis is very poor with death usually occurring after birth. Moreover it is very rare for intracranial teratoma to be diagnosed in utero. Recently, we experienced a case of fetal intracranial teratoma at 26 weeks of gestation, which is placed in the middle area of cranial cavity. On the monitor of ultrasonogram, the mass was seen as a hyperechogenic mass and its size was 20~30 mm in diameter and it was located in the region of thalamus. The finding of microscopic examination was that neuroepithelial and neuroglial nests and primitive epithelial cells were aggregated and this finding was consistent with immature teratoma. We report a case with a brief review of the literature.

A Case of Fetus in Fetu: Ultrasonic prenatal diagnosis / 대한산부인과학회잡지

Fetus in fetu is a very rare pathologic feature consisting of a parasitic twin included within the body of the other twin. Fetus in fetu within about 7X6X5 cm sized cystic mass in the fetal retroperitoneum was detected and diagnosed during antenatal ultrasound and confirmed by computed tomography scan postnatally. The mass was removed after birth. The cyst contain yellowish serous fluid with two solid masses. Two solid masses were confirmed fetus in fetu by gross and microscopic pathologic examination. The larger one showed well-developed and organized internal organ.