In-situ Hybridization for the Detection of Sacbrood Virus in Infected Larvae of the Honey Bee (Apis cerana).

The aim of this study was to develop and use in-situ hybridization (ISH) for the detection and localization of the sacbrood virus (SBV) in Korean honey bee (Apis cerana) larvae that were infected naturally with SBV. A 258 base pair cDNA probe for SBV was generated by polymerase chain reaction. Cells positive for viral genome typically showed a dark brown reaction in the cytoplasm. SBV was detected consistently in trophocytes and urocytes. The ISH was successfully applied to routinely fixed and processed tissues and thus should prove helpful in the diagnosis and characterization of viral distribution in infected larvae.

Caregiving appraisal of family caregivers for older stroke patients in Korea.

PURPOSE: The purpose of this study was to investigate factors related to caregiving appraisal for family caregivers of older stroke patients in Korea. METHOD: A descriptive, cross-sectional survey of a sample of 242 family caregivers (95.7% response rate) was conducted in Korea from 1 January 2005 to 4 January 2007. Data were collected from outpatient stroke centres at three hospitals and two home health agencies located in Seoul, the capital city of Korea, its suburban area of Kyunggi-do, and the rural areas Gyeongsangbuk-do and Chungcheongbuk-do. The questionnaire was self-administered. spss 13.0 (SPSS, Inc., Chicago, IL, USA) was used to perform frequency, percentage, mean standard deviation, t-test or analysis of variance, and Pearson's correlation calculations. RESULTS: Caregiving appraisal was statistically different based on the caregiver's location of residency and the relationship between the caregiver and the recipient. The caregivers who resided in urban areas reported higher scores of caregiving appraisal compared with those who resided in rural areas. With regard to the caregiver-recipient relationship, spouses reported the highest caregiving distress status, followed by daughters-in-law, sons and, finally, daughters. In addition, caregiving distress significantly correlated with age (r = 0.151, P = 0.022), the number of caregiving hours per week (r = 0.198, P = 0.003) and filial obligations (r = 0.336, P = 0.000). DISCUSSION: The study results helped to better understand the caregiving process of Korean family caregivers. The data emphasized the importance of examining the expression of cultural roles and individual values on the caregivers' quality of life in diverse cultures. The knowledge gained from this study will be utilized to develop culture-specific interventions and social policies to directly support Korean family caregivers and to indirectly support stroke patients.

Video-based self-assessment: implementation and evaluation in an undergraduate nursing course.

This research was performed to investigate the effects of video-based self-assessment on the ability of nursing students to accurately measure vital signs, their communication skills, and their satisfaction. This research was conducted between March 2007 and June 2007 as a quasi-experimental control-group, pretest-posttest design. The study population was composed of 40 second-year student nurses who enrolled in a fundamentals of nursing course of a college of nursing, Ajou University in Korea. Results of the research indicate that there was a statistically significant difference in exam scores for assessing long-term memory video-review group demonstrating higher scores. Student satisfaction was also significantly higher in the video-review group than in the control group. These results may suggest video-based self-assessment is a beneficial and effective instructional method of training undergraduate nursing students to develop awareness of their strengths and weaknesses, and to improve their clinical and communication skills.

Cloning and characterization of cDNAs encoding the GnRH1 and GnRH2 precursors from bullfrog (Rana catesbeiana).

We have isolated the cDNAs encoding the GnRH1 and GnRH2 precursors, respectively, from bullfrog (Rana catesbeiana) brain. The first cDNA consists of 648 bp and contains an open-reading frame of 270 nucleotides, encoding the bullfrog GnRH1 precursor. The second cDNA consists of 1053 bp and contains an open-reading frame of 255 nucleotides, encoding the bullfrog GnRH2 precursor. Both types of bullfrog GnRH precursor have a similar molecular architecture as observed in other GnRH precursors, consisting of a signal peptide, followed by the GnRH decapeptide, a conserved carboxy-terminal amidation and proteolytical processing site, and a GnRH-associated peptide (GAP). In addition, we have identified a third cDNA, containing 24 additional nucleotides in its GAP-coding region. Genomic PCR and sequence analysis confirmed that this cDNA represents an alternative splice variant of the bullfrog GnRH2-precursor pre-mRNA. The bullfrog GnRH1 precursor exhibits 60% and less than 40% amino acid identity to its Xenopus and mammalian counterparts, respectively, whereas the bullfrog GnRH2 precursor displays 50% to 60% amino acid identity to that of its nonmammalian counterparts, but shares only 25% amino acid identity with its mammalian counterparts. Northern blot analysis revealed a single GnRH1-precursor mRNA species of approximately 0.75 kilobases, expressed in bullfrog forebrain, and a single GnRH2-precursor mRNA species of approximately 1.1 kilobases, expressed in bullfrog midbrain/hindbrain. Furthermore, both bullfrog GnRH-precursor mRNAs exhibited a differential spatiotemporal expression pattern. Genomic Southern blot analysis indicated that both bullfrog GnRH genes are present as single copy genes. This is the first report on the molecular cloning of a GnRH2-precursor cDNA from an amphibian species. In addition, we present data showing that alternative splicing is utilized to generate different GnRH2-precursor mRNAs. J. Exp. Zool. 289:190-201, 2001.

Molecular cloning, distribution and pharmacological characterization of a novel gonadotropin-releasing hormone ([Trp8] GnRH) in frog brain.

To date nine structural variants of GnRH have been identified in vertebrates and two additional forms have been isolated from a tunicate. In amphibians only mammalian GnRH ([Arg8] GnRH) and type II GnRH (chicken GnRH II, [His5, Trp7, Tyr8] GnRH) have been identified. In the present study, a full-length cDNA encoding a novel type of GnRH was isolated from pituitary of Rana dybowskii. The GnRH gene encodes a GnRH peptide ([Trp8] GnRH) in which tryptophan is substituted for arginine of mammalian GnRH Northern blot analysis revealed the presence of a single 500 bp transcript for the [Trp8] GnRH precursor in forebrain but its absence in testis, ovary, kidney and liver. Restriction digests of genomic DNA demonstrated a single copy of the gene. The [Trp8] GnRH immunoreactive cells were identified in the preoptic area of the frog brain. Synthetic [Trp8] GnRH was tested for its ability to stimulate inositol phosphate production by COS-1 cells transfected with the cloned Xenopus pituitary GnRH receptor and the cloned human GnRH receptor. [Trp8] GnRH had a potency of about 60% compared with mammalian GnRH ([Arg8] GnRH) for the Xenopus receptor, whereas the potency of [Trp8] GnRH was approximately 5% compared with mammalian GnRH for the human receptor. Both mammalian GnRH and [Trp8] GnRH were 1000-fold less potent than type II GnRH for the Xenopus GnRH receptor. The similar potency of [Arg8] GnRH and the novel [Trp8] GnRH for the Xenopus pituitary receptor indicates that, unlike the human receptor, the Xenopus receptor does not discriminate between these amino acids in position eight thereby allowing substitution of the arginine in the mammalian GnRH.

Retinoic acids up-regulate steroidogenic acute regulatory protein gene.

The steroidogenic acute regulatory (StAR) protein plays essential roles in the delivery of cytosolic cholesterol into the mitochondrial inner membrane, which is an acute regulated and rate-limiting step for the steroid hormone synthesis. Since retinoic acids (RAs) are known to induce the synthesis of steroid hormones in mouse Leydig cells in vitro, mouse Leydig tumour cells, K28, were used to determine the effect of RAs on the level of StAR mRNA by Northern blot analysis. The level of StAR mRNA reached the maximum in a 4-8 h treatment with all-trans-RA (atRA) or 9-cis-RA (9cRA), and the effects were dose-dependent. The effect of 9cRA on the levels of StAR mRNA was blocked by actinomycin D, which indicates that 9cRA might exert a direct effect on the transcription of the gene. Promoter/reporter constructs containing a 5'-flanking region of the mouse or rat StAR gene were prepared, and luciferase activity was assayed following transient transfection into K28 or adrenal tumour cells, Y1. The result revealed that the luciferase activity was increased by 4-5-fold in response to the treatment of 9cRA, which indicated that 9cRA participates transcriptional activation of the StAR protein gene.

Piebaldism--a case report.

We report a familial case of piebaldism affecting a 33-year-old man and his 3-month-old son. Typical skin findings, white forelock and amelanotic white macules containing hyperpigmented macules, were characteristically presented on both patients.

Partial unilateral lentiginosis associated with segmental neurofibromatosis.

Partial unilateral lentiginosis associated with segmental neurofibromatosis is rate. Therefore, we describe here a patient with partial unilateral lentiginosis associated with ipsilateral segmental neurofibromatosis who developed multiple, rice-sized, brown macules on the right side of her face, trunk, and upper arm and several bean-sized, cafe-au-lait spots on the right upper arm and right upper back. To our knowledge, partial unilateral lentiginosis associated with ipsilateral segmental neurofibromatosis has not been reported in the English literature.

An unusual combination of trisomy 21 and partial trisomy 5q.

The authors describe a male newborn with multiple congenital anomalies; craniofacial dysmorphism, bilateral cleft palate and lip, ambiguous external genitalia with absence of phallus, ventricular septal defect, agenesis of olfactory bulbs, and presence of small round cells simulating migration defect in the cerebellar white matter. Cytogenetic study demonstrated a chromosomal constitution of 47,XY, +21, +5q. Its pathological significance compared with Down's syndrome and hitherto reported partial trisomy 5q is discussed.