TarGo: network based target gene selection system for human disease related mouse models / 한국실험동물학회지

Genetically engineered mouse models are used in high-throughput phenotyping screens to understand genotype-phenotype associations and their relevance to human diseases. However, not all mutant mouse lines with detectable phenotypes are associated with human diseases. Here, we propose the “Target gene selection system for Genetically engineered mouse models” (TarGo). Using a combination of human disease descriptions, network topology, and genotype-phenotype correlations, novel genes that are potentially related to human diseases are suggested. We constructed a gene interaction network using protein-protein interactions, molecular pathways, and co-expression data. Several repositories for human disease signatures were used to obtain information on human disease-related genes. We calculated disease- or phenotype-specific gene ranks using network topology and disease signatures. In conclusion, TarGo provides many novel features for gene function prediction.

Identification of ERBB Pathway-Activated Cells in Triple-Negative Breast Cancer

Intratumor heterogeneity within a single tumor mass is one of the hallmarks of malignancy and has been reported in various tumor types. The molecular characterization of intratumor heterogeneity in breast cancer is a significant challenge for effective treatment. Using single-cell RNA sequencing (RNA-seq) data from a public resource, an ERBB pathway activated triple-negative cell population was identified. The differential expression of three subtyping marker genes (ERBB2, ESR1, and PGR) was not changed in the bulk RNA-seq data, but the single-cell transcriptomes showed intratumor heterogeneity. This result shows that ERBB signaling is activated using an indirect route and that the molecular subtype is changed on a single-cell level. Our data propose a different view on breast cancer subtypes, clarifying much confusion in this field and contributing to precision medicine.

Construction of a Transcriptome-Driven Network at the Early Stage of Infection with Influenza A H1N1 in Human Lung Alveolar Epithelial Cells

We aimed to understand the molecular changes in host cells that accompany infection by the seasonal influenza A H1N1 virus because the initial response rapidly changes owing to the fact that the virus has a robust initial propagation phase. Human epithelial alveolar A549 cells were infected and total RNA was extracted at 30 min, 1 h, 2 h, 4 h, 8 h, 24 h, and 48 h post infection (h.p.i.). The differentially expressed host genes were clustered into two distinct sets of genes as the infection progressed over time. The patterns of expression were significantly different at the early stages of infection. One of the responses showed roles similar to those associated with the enrichment gene sets to known ‘gp120 pathway in HIV.’ This gene set contains genes known to play roles in preventing the progress of apoptosis, which infected cells undergo as a response to viral infection. The other gene set showed enrichment of ‘Drug Metabolism Enzymes (DMEs).’ The identification of two distinct gene sets indicates that the virus regulates the cell's mechanisms to create a favorable environment for its stable replication and protection of gene metabolites within 8 h.

A Comparative Efficacy of Propacetamol and Ketorolac in Postoperative Patient Controlled Analgesia

BACKGROUND: Ketorolac has been used as a postoperative analgesia in combination with opioids. However, the use of ketorolac may produce serious side effects in vulnerable patients. Propacetamol is known to induce fewer side effects than ketorolac because it mainly affects the central nervous system. We compared the analgesic effects and patient satisfaction levels of each drug when combined with fentanyl patient-controlled analgesia (PCA). METHODS: The patients were divided into two groups, each with n = 46. The patients in each group were given 60 mg of ketorolac or 2 g of propacetamol (mixed with fentanyl) for 10 minutes. The patients were then given 180 mg of ketorolac or 8 g of propacetamol (mixed with fentanyl and ramosetron) through PCA. We assessed the visual analogue pain scale (VAS) at the time point immediately before administration (baseline) and at 15, 30, and 60 minutes, and 24 hours after administration. Also, the side effects of each regimen and each patient's degree of satisfaction were assessed. RESULTS: There was a significant decline in the VAS score in both groups (P < 0.05). However, there were no significant differences in the VAS scores between the groups at each time point. Satisfaction scores between the groups showed no significant difference. CONCLUSIONS: The efficacy of propacetamol is comparable to that of ketorolac in postoperative PCA with fentanyl.

Corrigendum: Expression of the spinal 5-HT7 receptor and p-ERK pathway in the carrageenan inflammatory pain of rats (Korean J Anesthesiol 2015 April 68(2): 170-174) / 대한마취과학회지

This article was inadvertently omitted Acknowledgments section for grant support.

Changes in the hemodynamic parameters between the prone and supine positions measured by an arterial pulse contour cardiac output monitoring system

BACKGROUND: The changes in the hemodynamic parameters when the patients assumed a prone position from a supine position were examined using the FloTrac/EV1000(TM) system, during general anesthesia. METHODS: A total of 56 adult patients with American Society of Anesthesiologists physical status I-II and undergoing elective lumbar spine surgery were enrolled in the study. The hemodynamic parameters, such as the mean arterial pressure, heart rate, cardiac index, stroke volume variation, systemic vascular resistance index, central venous pressure, and peak airway pressure, were recorded when the patients were in a supine position and then in a prone position. RESULTS: No significant differences were found in the subjects' hemodynamic parameters between when the subjects were in a supine position and when they were in a prone position, except in the central venous pressure and peak airway pressure, both of which were elevated when the subjects were in a prone position. CONCLUSIONS: There were no differences in hemodynamic parameters between supine and prone positions measured by FloTrac/EV1000(TM) system during general anesthesia.

The Effect of Stellate Ganglion Block on Intractable Lymphedema after Breast Cancer Surgery

Lymphedema of the upper limb after breast cancer surgery is a disease that carries a life-long risk and is difficult to cure once it occurs despite the various treatments which have been developed. Two patients were referred from general surgery department for intractable lymphedema. They were treated with stellate ganglion blocks (SGBs), and the circumferences of the mid-point of their each upper and lower arms were measured on every visit to the pain clinic. A decrease of the circumference in each patient was observed starting after the second injection. A series of blocks were established to maintain a prolonged effect. Both patients were satisfied with less swelling and pain. This case demonstrates the benefits of an SGB for intractable upper limb lymphedema.

Vasopressin ameliorates hypotension induced by beach chair positioning in a dose-dependent manner in patients undergoing arthroscopic shoulder surgery under general anesthesia / 대한마취과학회지

BACKGROUND: The beach chair position (BCP) is associated with hypotension that may lead to cerebral ischemia. Arginine vasopressin (AVP), a potent vasoconstrictor, has been shown to prevent hypotension in BCP. It also improves cerebral oxygenation in different animal models. The present study examined the effect of escalating doses of AVP on systemic hemodynamics and cerebral oxygenation during surgery in BCP under general anesthesia. METHODS: Sixty patients undergoing arthroscopic shoulder surgery in BCP under general anesthesia were randomly allocated to receive either saline (control, n = 15) or three different doses of AVP (0.025, 0.05, or 0.075 U/kg; n = 15 each) 2 minutes before BCP. Mean arterial pressure (MAP), heart rate (HR), regional cerebral oxygen saturation (SctO2), and jugular venous oxygen saturation (SjvO2) were measured after induction of anesthesia and before (presitting in supine position) and after BCP. RESULTS: AVP per se given before BCP increased MAP, and decreased SjvO2, SctO2, and HR in all patients (P 20% SctO2 decrease from the baseline value) with no differences in SjvO2 and the incidence of SjvO2 < 50% or SjvO2 < 40% among the groups. CONCLUSIONS: AVP ameliorates hypotension associated with BCP in a dose-dependent manner in patients undergoing shoulder surgery under general anesthesia. However, AVP may have negative effects on SctO2 before and after BCP and on SjvO2 before BCP.

Effects of intravenously administered indocyanine green on near-infrared cerebral oximetry and pulse oximetry readings / 대한마취과학회지

BACKGROUND: Intravenously administered indocyanine green (ICG) may cause misreadings of cerebral oximetry and pulse oximetry in patients undergoing carotid endarterectomy under general anesthesia. The present study determined the effects of two different doses (12.5 mg vs. 25 mg) of ICG on regional cerebral tissue oxygen saturation (SctO2) and percutaneous peripheral oxygen saturation (SpO2). METHODS: Twenty-six patients receiving ICG for videoangiography were divided into two groups according to the dosage (12.5 mg and 25 mg, n = 13 in each group). Heart rate, arterial blood pressure, SctO2, and SpO2 were measured before and after an intravenous bolus administration of ICG. RESULTS: Following the dye administration, no changes in heart rate or arterial blood pressure were noted in either group. SctO2 was increased in both groups; however, the magnitude of the increase was greater (21.6 +/- 5.8% vs. 12.6 +/- 4.1%, P < 0.0001) and more prolonged (28.4 +/- 9.6 min vs. 13.8 +/- 5.2 min, P < 0.0001) in the 25 mg group than in the 12.5 mg group. In contrast, SpO2 was decreased in both groups; the magnitude of the decrease was greater in the 25 mg group than in the 12.5 mg group (4.0 +/- 0.8% vs. 1.6 +/- 1.0%, P < 0.0001). There were no differences in the time to reach the peak SctO2 or to reach the nadir SpO2 between the two groups. CONCLUSIONS: In patients given ICG for videoangiography, a 25 mg bolus results in a greater and more prolonged increase in SctO2 and a greater reduction in SpO2 than a 12.5 mg bolus, with no differences in the time to reach the peak SctO2 or to reach the nadir SpO2.

Expression of the spinal 5-HT7 receptor and p-ERK pathway in the carrageenan inflammatory pain of rats / 대한마취과학회지

BACKGROUND: Although the inhibitory role of the 5-hydroxytrypatmine receptor 7(5-HT7R) on nociceptive processing is generally recognized, an excitatory effect associated with a reduced 5-HT7R expression has also been observed in the nerve injury model. In the carrageenan model, no significant effect is produced by the 5-HT7R activation, but the change in 5-HT7R expression has not been examined. Lesioning of the spinal serotonergic pathway enhances allodynia in the carrageenan model, but it also relieves several other pain states, including in the formalin model. While lesioning suppresses the activation of the extracellular signal-regulated kinase (ERK) of the spinal cord in the formalin model, its role in the carrageenan model has not been reported. METHODS: Following intraplantar injections of carrageenan, the spinal 5-HT7R expression was examined using Western blotting in male Sprague-Dawley rats. The effect of serotonergic pathway lesioning with intrathecal 5,7-dihydroxytryptamine (5,7-DHT) on the expression of the phospho-ERK was measured. RESULTS: The expression of the 5-HT7R in the carrageenan model was not significantly different from that of naive animals. The expression of the spinal p-ERK in the carrageenan model was significantly increased, but returned to the level of a naive rat 1 hour after the carrageenan injection. However, it remained significantly higher 1 hour after the injection in the animals treated with 5,7-DHT than in the naive and control rats. CONCLUSIONS: The expression of the spinal 5-HT7R is not altered by peripheral inflammation with carrageenan, suggesting that the lack of antinociceptive effect of the 5-HT7R activation is partly attributable to the absence of changes in the expression of the 5-HT7R in the spinal cord. The extended increase of the spinal p-ERK might be related to the enhanced pain behavior in the animals with lesions of the spinal serotonergic pathway in the carrageenan model.

Genetic diversity of Korean Bacillus anthracis isolates from soil evaluated with a single nucleotide repeat analysis

Bacillus (B.) anthracis, the etiological agent of anthrax, is one of the most genetically monomorphic bacteria species in the world. Due to the very limited genetic diversity of this species, classification of isolates of this bacterium requires methods with high discriminatory power. Single nucleotide repeat (SNR) analysis is a type of variable-number tandem repeat assay that evaluates regions with very high mutation rates. To subtype a collection of 21 isolates that were obtained during a B. anthracis outbreak in Korea, we analyzed four SNR marker loci using nucleotide sequencing analysis. These isolates were obtained from soil samples and the Korean Center for Disease Control and Prevention. The SNR analysis was able to detect 13 subgenotypes, which allowed a detailed evaluation of the Korean isolates. Our study demonstrated that the SNR analysis was able to discriminate between strains with the same multiple-locus variable-number tandem repeat analysis genotypes. In summary, we obtained SNR results for four SNR marker loci of newly acquired strains from Korea. Our findings will be helpful for creating marker systems and help identify markers that could be used for future forensic studies.

Antinociceptive Effect of Intrathecal Nefopam and Interaction with Morphine in Formalin-Induced Pain of Rats

BACKGROUND: Nefopam, a non-opiate analgesic, has been regarded as a substance that reduces the requirement for morphine, but conflicting results have also been reported. The inhibition of monoamine reuptake is a mechanism of action for the analgesia of nefopam. The spinal cord is an important site for the action of monoamines however, the antinociceptive effect of intrathecal nefopam was not clear. This study was performed to examine the antinociceptive effect of intrathecal (i.t.) nefopam and the pattern of pharmacologic interaction with i.t. morphine in the formalin test. METHODS: Male Sprague-Dawley rats were implanted with an i.t. catheter, and were randomly treated with a vehicle, nefopam, or morphine. Formalin was injected into the hind-paw 10 min. after an i.t. injection of the above experiment drugs. After obtaining antinociceptive ED50 of nefopam and morphine, the mixture of nefopam and morphine was tested for the antinociceptive effect in the formalin test at a dose of 1/8, 1/4, 1/2 of ED50, or ED50 of each drug followed by an isobolographic analysis. RESULTS: Intrathecal nefopam significantly reduced the flinching responses in both phases of the formalin test in a dose-dependent manner. Its effect, however, peaked at a dose of 30 microg in phase 1 (39.8% of control) and 10 microg during phase 2 (37.6% of control). The isobolograhic analysis indicated an additive interaction of nefopam and morphine during phase 2, and a synergy effect in antinociception during phase 1. CONCLUSIONS: This study demonstrated that i.t. nefopam produces an antinociceptive effect in formalin induced pain behavior during both phases of the formalin test, while interacting differently with i.t. morphine, synergistically during phase 1, and additively during phase 2.

Genetic populations of Bacillus anthracis isolates from Korea

Bacillus (B.) anthracis is the pathogen that causes fatal anthrax. Strain-specific detection of this bacterium using molecular approaches has enhanced our knowledge of microbial population genetics. In the present study, we employed molecular approaches including multiple-locus variable-number tandem repeat analysis (MLVA) and canonical single-nucleotide polymorphism (canSNP) analysis to perform molecular typing of B. anthracis strains isolated in Korea. According to the MLVA, 17 B. anthracis isolates were classified into A3a, A3b, and B1 clusters. The canSNP analyses subdivided the B. anthracis isolates into two of the three previously recognized major lineages (A and B). B. anthracis isolates from Korea were found to belong to four canSNP sub-groups (B.Br.001/2, A.Br.005/006, A.Br.001/002, and A.Br.Ames). The A.Br.001/002 and A.Br.Ames sub-lineages are closely related genotypes frequently found in central Asia and most isolates were. On the other hand, B. anthracis CH isolates were analyzed that belonged to the B.Br.001/002 sub-group which found in southern Africa, Europe and California (USA). B.Br.001/002 genotype is new lineage of B. anthracis in Korea that was not found before. This discovery will be helpful for the creation of marker systems and might be the result of human activity through the development of agriculture and increased international trade in Korea.

Roles of Serotonergic and Adrenergic Receptors in the Antinociception of Selective Cyclooxygenase-2 Inhibitor in the Rat Spinal Cord

BACKGROUND: The analgesic mechanisms of cyclooxygenase (COX)-2 inhibitors have been explained mainly on the basis of the inhibition of prostaglandin biosynthesis. However, several lines of evidence suggest that their analgesic effects are mediated through serotonergic or adrenergic transmissions. We investigated the roles of these neurotransmitters in the antinociception of a selective COX-2 inhibitor at the spinal level. METHODS: DUP-697, a selective COX-2 inhibitor, was delivered through an intrathecal catheter to male Sprague-Dawley rats to examine its effect on the flinching responses evoked by formalin injection into the hindpaw. Subsequently, the effects of intrathecal pretreatment with dihydroergocristine, prazosin, and yohimbine, which are serotonergic, alpha1 adrenergic and alpha2 adrenergic receptor antagonists, respectively, on the analgesia induced by DUP-697 were assessed. RESULTS: Intrathecal DUP-697 reduced the flinching response evoked by formalin injection during phase 1 and 2. But, intrathecal dihydroergocristine, prazosin, and yohimbine had little effect on the antinociception of intrathecal DUP-697 during both phases of the formalin test. CONCLUSIONS: Intrathecal DUP-697, a selective COX-2 inhibitor, effectively relieved inflammatory pain in rats. Either the serotonergic or adrenergic transmissions might not be involved in the analgesic activity of COX-2 inhibitors at the spinal level.

Efficacy of the combination of cold propofol and pretreatment with remifentail on propofol injection pain / 대한마취과학회지

BACKGROUND: Pain upon the injection of propofol is a common adverse effect. This study was conducted to evaluate the analgesic effect of remifentanil and cold propofol during propofol injection for the induction of anesthesia and to determine if a combination of cold propofol and remifentanil produced additional analgesic efficacy. METHODS: A total of 160 patients aged 20-65 years old were randomly allocated into one of four groups (n = 40, in each). Control and remifentanil group patients received 2 mg/kg propofol that had been stored at room temperature (20-23degrees C), while the cold and combination group received cold (4degrees C) propofol. The patients received remifentanil 0.5 microg/kg IV in the remifentanil and combination groups or saline in the control and cold groups. Ninety seconds after administration the patients were administered propofol over a 30 second period. The pain intensity and incidence were then evaluated using a 4-point verbal rating scale. RESULTS: The incidence of pain was significantly reduced in groups that received remifentanil in the cold and combination groups when compared with the control group (27.5%, 30%, and 2.5% vs. 70%, respectively). Moreover, the severity of pain was significantly lower in groups that received remifentanil in the cold and combination groups when compared with the control group. The incidence and severity of pain from the propofol injection in the combination group was significantly lower than that in the remifentanil and cold groups. CONCLUSIONS: The combination of cold propofol and pretreatment with remifentanil more effectively reduced the incidence of pain upon the injection of propofol than either treatment alone.

The role of group III metabotropic receptors on opioid tolerance

BACKGROUND: Prolonged exposure to morphine causes tolerance to morphine-induced antinociception, yet the mechanisms of such tolerance are not fully understood. Although group I and II metabolic glutamate receptors (mGluRs) are involved in the modulation of morphine tolerance, the role of the group III mGluRs has not been determined. Therefore, we examined the effect of a group III mGluRs agonist on the morphine tolerance in the spinal cord. METHODS: An intrathecal infusion of morphine (40 nmol/microl/h) for 5 days was done to examine the development of morphine tolerance in male Sprague-Dawley rats. Noxious radiant heat was applied to the hindpaw and we measured the thermal withdrawal latency. To clarify the role of the group III mGluRs, an intrathecal group III mGluRs agonist (ACPT-III) or saline was administered to the morphine tolerant rats and we observed the change of the thermal withdrawal latency at 15, 30, 60, 90 and 120 min after delivery of ACPT-III. RESULTS: A continuous intrathecal infusion of morphine significantly increased the thermal withdrawal latency, as compared with the saline infused rats on day 1, with a decline on day 3 and the increase of withdrawal latency totally disappeared on day 5 (tolerance). Intrathecal ACPT-III increased the thermal withdrawal latency in the morphine tolerance rats. CONCLUSIONS: These results suggest that the group III mGluRs may be involved in the suppression of tolerance to morphine-induced antinociception at the spinal level.

Immunoreactivity of Iron-Repressible Cell Wall Proteins and Exoproteins of Staphylococcus aureus / 감염과화학요법

In the present study, we tried to investigate immunoreactivity of iron-repressible cell wall proteins and exoproteins of Staphylococcus aureus when the convalescent serum obtained from the patient with septicemia was applied. On SDS-PAGE and western blot analysis, several iron-repressible cell wall proteins expressed in iron-deficient BHI showed strong immunoreactivity, whereas no or relatively weak immunoreactivity was shown in iron-sufficient BHI. Several exoproteins were expressed only in iron-deficient BHI and these exoproteins showed strong immunoreactivity. These results indicate that several iron-repressible cell wall proteins and exoproteins are expressed and are immunogenic in vivo. Since in vivo state is an relatively iron-restricted condition, we recommend the use of iron-deficient media for studies concerning pathogenicity of staphylococcal in human.

Immunoreactivity of Iron-Repressible Cell Wall Proteins and Exoproteins of Staphylococcus aureus / 감염과화학요법

In the present study, we tried to investigate immunoreactivity of iron-repressible cell wall proteins and exoproteins of Staphylococcus aureus when the convalescent serum obtained from the patient with septicemia was applied. On SDS-PAGE and western blot analysis, several iron-repressible cell wall proteins expressed in iron-deficient BHI showed strong immunoreactivity, whereas no or relatively weak immunoreactivity was shown in iron-sufficient BHI. Several exoproteins were expressed only in iron-deficient BHI and these exoproteins showed strong immunoreactivity. These results indicate that several iron-repressible cell wall proteins and exoproteins are expressed and are immunogenic in vivo. Since in vivo state is an relatively iron-restricted condition, we recommend the use of iron-deficient media for studies concerning pathogenicity of staphylococcal in human.

An Experimental Study of Fibroblast Suppression in Collagen Lattice

Fibroblasts are known as the key cells for wound contraction. In the plastic surgical field, wound contraction is one of the main concerns. Many drugs and treatment modalities have been tried to reduce the action of fibroblasts. To investigate the effects of minoxidil and griseofulvin on fibroblasts, fetal human fibroblasts were isolated from the anterior chest of normal fetus. Minoxidil, in addition to its effect on hypertension and hair growth and griseofulvin which has been used as antifungal agent were added on cultured fibroblasts. The morphological changes and numerical changes of the fibroblasts were observed by phase contrast microscopy. In low concentration, minoxidil does not inhibit the proliferation of fibroblasts but in high concentration, it inhibits the proliferation of fibroblasts and prevents the formation of proliferative cytoplasmic buds of fibroblasts. Griseofulvin inhibits the action of fibroblasts but not so significant as minoxidil. Also, they prevent the contraction of collagen lattice in high concentration. The importance of these findings is possible usage of these drugs to treat clinical fibrosis and hypertrophic scars. Further studies will be planned to learn more about a possible role of minoxidil and griseofulvin in prevention of wound contraction.

FEMALE PSEUDOHERMAPHRODITISM DUE TO ADRENOGENITAL SYNDROME

Female pseudohermaphroditism due to adrenogenital syndrome is a condition in which individuals with a 46XX karyotype, negative H-Y antigen, normal mullerian duct derivatives, and a lack of development of w lffian duct structures differentiate partially as phenotypic males. They usually manifest masculinization of the external genitalia as a result of excess endogenous androgens. Most female pseudohermaphrodities have one of the types of congenital virilizing adrenal hyperplasia. Adrenogenital syndrome is inborn errors transmitted by autosomal recessive genes and may be due to defects in any of the enzymic steps in the biosynthesis of cortisol. Most affected individuals have a failure of 21-hydroxylation which prevents the conversion of 17 alpha-hydroxyprogesterone to 11-deoxycortisol. Such a defect in 21-hydroxylase leads to excessive production of adrenal androgens causing virilization. The treatment is early endocrinologic support and surgical reconstruction. There are some considerations in surgical repairs including normal sized clitoris with adequate erogenous sensation, sufficiently wide vaginal introitus and normal aesthetic appearance of the external genitalia for her normalized life as a female. We have experienced four cases of female pseudohermaphroditism due to adrenogenital syndrome. In two cases, we performed clitoroplasty with nerve sparing technique, vulvoplasty with mons pubis augmentation, vaginoplasty with posterior perineal flap and urethral reconstruction. In the other cases, we performed clitoroplasty with nerve sparing technique, vulvoplasty and vaginoplasty There was no specific operative complication and the result of the correction was satisfactory.