Preclinical studies for pharmacokinetics and biodistribution of Ad-stTRAIL, an adenovirus delivering secretable trimeric TRAIL for gene therapy

Malignant glioma is the most frequent type in brain tumors. The prognosis of this tumor has not been significantly improved for the past decades and the average survival of patients is less than one year. Thus, an effective novel therapy is urgently needed. TNF-related apoptosis inducing ligand (TRAIL), known to have tumor cell-specific killing activity, has been investigated as a novel therapeutic for cancers. We have developed Ad-stTRAIL, an adenovirus delivering secretable trimeric TRAIL for gene therapy and demonstrated the potential to treat malignant gliomas. Currently, this Ad-stTRAIL gene therapy is under phase I clinical trial for malignant gliomas. Here, we report preclinical studies for Ad-stTRAIL carried out using rats. We delivered Ad-stTRAIL intracranially and determined its pharmacokinetics and biodistribution. Most Ad-stTRAIL remained in the delivered site and the relatively low number of viral genomes was detected in the opposite site of brain and cerebrospinal fluid. Similarly, only small portion of the viral particles injected was found in the blood plasma and major organs and tissues, probably due to the brain-blood barrier. Multiple administrations did not lead to accumulation of Ad-stTRAIL at the injection site and organs. Repeated delivery of Ad-stTRAIL did not show any serious side effects. Our data indicate that intracranially delivered Ad-stTRAIL is a safe approach, demonstrating the potential as a novel therapy for treating gliomas.

Preclinical studies for pharmacokinetics and biodistribution of Ad-stTRAIL, an adenovirus delivering secretable trimeric TRAIL for gene therapy

Malignant glioma is the most frequent type in brain tumors. The prognosis of this tumor has not been significantly improved for the past decades and the average survival of patients is less than one year. Thus, an effective novel therapy is urgently needed. TNF-related apoptosis inducing ligand (TRAIL), known to have tumor cell-specific killing activity, has been investigated as a novel therapeutic for cancers. We have developed Ad-stTRAIL, an adenovirus delivering secretable trimeric TRAIL for gene therapy and demonstrated the potential to treat malignant gliomas. Currently, this Ad-stTRAIL gene therapy is under phase I clinical trial for malignant gliomas. Here, we report preclinical studies for Ad-stTRAIL carried out using rats. We delivered Ad-stTRAIL intracranially and determined its pharmacokinetics and biodistribution. Most Ad-stTRAIL remained in the delivered site and the relatively low number of viral genomes was detected in the opposite site of brain and cerebrospinal fluid. Similarly, only small portion of the viral particles injected was found in the blood plasma and major organs and tissues, probably due to the brain-blood barrier. Multiple administrations did not lead to accumulation of Ad-stTRAIL at the injection site and organs. Repeated delivery of Ad-stTRAIL did not show any serious side effects. Our data indicate that intracranially delivered Ad-stTRAIL is a safe approach, demonstrating the potential as a novel therapy for treating gliomas.

Dilated Cardiomyopathy in a 2 Month-Old Infant: A Severe Form of Hypocalcemia With Vitamin D Deficient Rickets

Dilated cardiomyopathy, which mostly has an idiopathic etiology or is caused by genetic inheritance or infection, can cause irreversible congestive heart failure. Hypocalcemia is a rare etiology of reversible dilated cardiomyopathy. Here we report the case of a two-month-old girl with congestive heart failure who was diagnosed as having dilated cardiomyopathy secondary to hypocalcemia. After calcium and vitamin D replacement therapy, the patient showed a rapid reduction in hypocalcemic tetany and a rapid recovery of left ventricular function. The cause of the hypocalcemia was vitamin D deficient rickets. She was exclusively breast-fed as an infant, and her mother had a vitamin D deficiency and was diagnosed with osteomalacia.

Limitations of 99mTc-DMSA scan in diagnosing acute pyelonephritis in children / 소아과

PURPOSE: We aimed to prove the relative limitation of 99mTc-DMSA scintigraphy (DMSA) compared to computed tomography (CT) in diagnosing acute pyelonephritis (APN) in children. METHODS: Since September 2006, after a 64-channel CT was imported, 10 DMSA false-negative patients have been identified: these patients underwent a CT scan for acute abdomen or acute febrile symptoms and were diagnosed as having APN; however, their DMSA scans were clear. We focused on these 10 DMSA false-negative patients and analyzed their clinical findings and CT results. We used Philips Brilliance Power 64-channel CT scanner for the CT scan and Siemens Orbitor Nuclear Camera 60 Hz for the DMSA scan. RESULTS: The 10 DMSA false-negative patients were mostly males (80%) and infants (80%). They had fever for a mean of 1.1-day duration before admission and showed increase in acute reactants: leukocyte, erythrocyte sedimentation rate, and C-reactive protein. The CT findings of renal lesions were focal in 6 (60%) cases and diffuse in 4 (40%) cases, and most of the lesions were unilateral in 80% of patients. CT proved that 22 renal lesions were neglected by DMSA. Differential renal function test by DMSA was also of no use in the evaluation of renal lesions. CONCLUSION: In this study, DMSA scan showed limitation in finding renal cortical lesions of CT-proven APN patients. DMSA false-negative results seem to occur at early-phase disease of infantile age, but more prospective studies are needed to determine the reasons and their prevalence.

Evolutionary Signature of Information Transfer Complexity in Cellular Membrane Proteomes

Cell membrane proteins play crucial roles in the cell's molecular interaction with its environment and within itself. They consist of membrane-bound proteins and many types of transmembrane (TM) proteins such as receptors, transporters, channel proteins, and enzymes. Membrane proteomes of cellular organisms reveal some characteristics in their global topological distribution according to their evolutionary positions, and show their own information transfer complexity. Predicted transmembrane segments (TMSs) in membrane proteomes with HMMTOP showed near power-law distribution and frequency characteristics in 6-TMS and 7-TMS proteins in prokaryotes and eukaryotes, respectively. This reaffirms the important roles of membrane receptors in cellular communication and biological evolutionary history.

Increased in vivo immunological potency of HB-110, a novel therapeutic HBV DNA vaccine, by electroporation

Pulse-induced permeabilization of cellular membranes, generally referred to as electroporation (EP), has been used for years as a tool to increase macromolecule uptake in tissues, including nucleic acids, for gene therapeutic applications, and this technique has been shown to result in improved immunogenicity. In this study, we assessed the utility of EP as a tool to improve the efficacy of HB-110, a novel therapeutic DNA vaccine against chronic hepatitis B, now in phase 1 of clinical study in South Korea. The potency of HB-110 in mice was shown to be improved by EP. The rapid onset of antigen expression and higher magnitude of humoral and cellular responses in electric pulse-treated mice revealed that EP may enable a substantial reduction in the dosage of DNA vaccine required to elicit a response similar in magnitude to that achievable via conventional administration. This study also showed that EP-based vaccination at 4-week-intervals elicited a cellular immune response which was about two-fold higher than the response elicited by conventional vaccination at 2-week intervals. These results may provide a rationale to reduce the clinical dose and increase the interval between the doses in the multidose vaccination schedule. Electric pulsing also elicited a more balanced immune response against four antigens expressed by HB-110: S, preS, Core, and Pol.

Transcriptional profiling in human HaCaT keratinocytes in response to kaempferol and identification of potential transcription factors for regulating differential gene expression

Kaempferol is the major flavonol in green tea and exhibits many biomedically useful properties such as antioxidative, cytoprotective and anti-apoptotic activities. To elucidate its effects on the skin, we investigated the transcriptional profiles of kaempferol-treated HaCaT cells using cDNA microarray analysis and identified 147 transcripts that exhibited significant changes in expression. Of these, 18 were up-regulated and 129 were down-regulated. These transcripts were then classified into 12 categories according to their functional roles: cell adhesion/cytoskeleton, cell cycle, redox homeostasis, immune/defense responses, metabolism, protein biosynthesis/modification, intracellular transport, RNA processing, DNA modification/ replication, regulation of transcription, signal transduction and transport. We then analyzed the promoter sequences of differentially-regulated genes and identified over-represented regulatory sites and candidate transcription factors (TFs) for gene regulation by kaempferol. These included c-REL, SAP-1, Ahr-ARNT, Nrf-2, Elk-1, SPI-B, NF-kappaB and p65. In addition, we validated the microarray results and promoter analyses using conventional methods such as real-time PCR and ELISA-based transcription factor assay. Our microarray analysis has provided useful information for determining the genetic regulatory network affected by kaempferol, and this approach will be useful for elucidating gene-phytochemical interactions.

A Clinical Profiles of 7 Patients with Glassy Cell Carcinoma of Uterine Cervix / 대한산부인과학회잡지

No abstract available.

A Case of Ovarian Squamous Cell Carcinoma in a Patient with Microinvasive Squamous Cell Carcinoma of Cervix / 대한산부인과학회잡지

Squamous cell carcinoma of the ovary is rare and usually arise from preexisting cystic teratomas, endometriosis or Brenner tumors which undergo carcinomatous degeneration in their epithelial elements. Some of squamous cell carcinoma of the ovary is known to be associated with cervical neoplasm, but the pathogenesis is still unknown. Recently we experienced one case of ovarian squamous cell carcinoma in patients with microinvasive squamous cell carcinoma of uterine cervix. We report this case with possible pathogenesis and brief review of literatures.

Expression of Matrix Metalloproteinase-1,2,3 and Type IV Collagen in Gastric Adenocarcinoma: Influence on Lymph Node Metastasis and Prognosis

Matrix metalloproteinases are believed to play an important role in tumor invasion and metastasis. But little is known about the role of them in the gastric adenocarcinoma. We investigated the expression of matrix metalloproteinase-1,2,3 in eighty paraffin blocks of the primary gastric adenocarcinoma tissues with immunohistochemistry and analysed their correlation with lymph node metastasis and survival. MMP-1,2,3 were expressed most intensely in the fibroblasts around the tumor stroma. In our study the increased immunoreactivity of MMP-2 only showed statistically significant correlation with lymph node metastasis (P=0.0517, Odd's ratio=2.274). But MMP-1,2,3 all were correlated with survival. Type IV collagen was observed in the vascular basement membranes and tumor basement membranes and showed statistically significant correlation with lymph node metastasis (P=0.0002, Odd's ratio=0.194) and prognosis (P=0.0001). The immunoreactivity of MMP-2 and type IV collagen was inversely correlated (Kendall's Tau-b correlation = 0.37482, P=0.0001). Our results suggest that in human gastric adenocarcinoma the increased immunoreactivity of MMP-2 and the decreased immunoreactivity of type IV collagen has an important role in lymph node metastasis and prognosis. MMP-1,3 are not correlated with lymph node metastasis but correlated with survival. The mechanism responsible for the production of MMP by the host fibroblasts remains obscure and requires further investigation.

Sarcomatoid Carcinoma with Heterologous Osteosarcomatous Component of the Prostate: A case report

Sarcomatoid carcinoma of prostate has been rarely reported and occasionally difficult to distinguish from a true sarcoma or carcinosarcoma. A case of sarcomatoid carcinoma of the prostate, which has been occured in 61-year-old male patient is presented. The tumor consists of carcinomatous areas with epithelioid cells, sarcomatoid areas with spindle cells and foci of heterologous osteosarcoma component. The phenotypic nature of the tumor was confirmed immunohistochemically by positive reaction for cytokeratin, epithelial membrane antigen, vimentin and prostate specific acid phosphatase in both sarcomatous and carcinomatous components.

The Effect of Configuration of the Needle on the Postspinal Headache in Young Males of the Third Decade / 대한마취과학회지

The incidence of postspinal headache is one of the well known complications of spinal anesthesia. The development of postspinal headache is related to age and sex of patients, size and configuration of the needle, pregnancy, direction of needle bevel, number of dural puncture. I studied the effect of configuration and size of the needle (22 gauge 25 gauge Quinke needle and 22 gauge,25 gauge Whitacre needle ) on the incidence, onset, duration and severity of postspinal headache in the 200 males of the third decade after spinal anesthesia. The following results were observed: 1) The incidence of postspinal headache in 22 gauge, 25 gauge Whitacre needle groups(4%. 2%) is lower than that of 22 gauge, 25 gauge Quinke needle groups(10%, 8%). 2) The severity of headache was mild and moderate level in 7 cases, severe level in 2 cases of all the 9 cases with postspinal headache of Quinke needle groups but mild and moderate level in 3cases of all the 3 cases with postspinal headache of Whitacre needle groups. 3) The onset of headache was within 24 hours in 55.6% in Quinke needle groups, 100% in Whitacre needle groups and the duration of headache was within 5 days in 77.8% in Quinke needle groups, 100% in Whitacre needle groups postoperatively. 4) The sites of headache were 55.6% in frontal, 33.3% in occipital region in Quinke needle groups and 66.7% in frontal, 33.3% in occipital region in Whitacre needle groups. In conclusion, the headaches which occur following use of Whitacre needle have been lower in and of less severity than those seen after use of Quinke needle in young males of the third decade.

The Effects of Continuous Epidural Fentanyl / Bupivacaine Mixtures on Analgesia and Pulmonary Function after Thoracotomy / 대한마취과학회지

Patients undergoing thoracotomy experience severe postoperative pain and marked respiratory impairment. Analgesics(narcotics or loeal anesthetics) administered via epidural catheter in epidural space have been shown to provide postoperative analgesia and improve respiratory mechanics after thoracotomy. Several different methods have been utilized in an attempt to reduce pain and pulmonary mechanics after thoracotomy. These include epidural blocks using local anesthetics, epidural narcotics, ketamine, steroid, and clonidine. These methods have been shown to provide pain relief with relative preservation of lung volumes in the postoperative period, but have disadvantages. Especially epidural local anesthetics may cause hypotension and motor blockade of lower extremities, and epidural narcotics may cause pruritus, nausea and vomiting, urinary retension and respiratory depression. In an attempt to provide excellent analgesia and improve pulmonary mechanics after thoracotomy and to decrease the side effects associated with the intermittent bolus administration of epidural narcotics or local anesthetics, we performed a study of continuously administered epidural infusion of small concentration of fentanyl combined with low concentration of bupivacaine. Twenty eight patients undergoing thoracotomy were randomized into groups based upon a postoperative pain regimen as indicated: Group I: intermittent intramuscualr injection of nalbuphine 0.2 mg/kg(n=13), Group II: continuous epidural injection of mixtures of 0.2/ bupivacaine and fentanyl 3 ug/ml(n= 15). Two, 8, 24 and 48 hours postoperative, the following indices were measured: visual analogue pain scale, vital capacity, tidal volume, arterial blood gas analysis(pH, PaCo2, PaO2), side effects, and 24 hour urine 17-ketosteroids. The results were as follows: 1) Pain score was evaluated by visual analogue pain scale postoperatively and the pain scores significantly decresed in group II as compaired with those in group L 2) Vital capacity and tidal volume in group II were more improved than group I. 3) There was no difference in arterial blood gas analysis except for decreased PaO2 at 2 hour and 24 hour compared with preoperative value in group L 4) Major complications in group II were two cases of nausea and vomiting, one case of urinary retension, whereas only I patient in group I complained of nausea and vomiting. 5) No significant difference occurred in 24 hour urine l7-ketosteroid at 24 hour and 48 hour postoperatively in group I and group II, which were within normal limits.

The Effect of Neostigmine Pretreatment on Vecuronium Induced Neuromuscular Blockade / 대한마취과학회지

Prolonged neuromuscular blockade following succinylcholine may be seen when anticho- linesterase had been administered prior to reverse nondepolarizing muscle relaxant-induced paralysis, possibly anticholinesterase has been reported to inhibit serum cholinesterase activity. Our study was undertaken in order to understand the effect of a nondepolarizing muscle relaxant (vecuronium) following neostigmine pretreatment. In this study, we assessed the effect of vecuronium induced neuromuscular blockade using a train of four, 2 Hz stimulations on ulnar nerve. Patients admitted to our hospital for elective operations were divided into two groups, each group consisting of 16 patients. In group I, vecuronium 0.1 mg/kg was .administered according to the priming principle following normal saline, in group II, vecuronium 1.0mg/kg was administered according to the priming principle following neostigmine 0.05 mg/kg and glycopyrrolate 0.003 mg/kg pretreatment. Then the time for Tl to reach 5% or less(second), and the time for Tl to reach from 25% to 75%(recovery index) were measured in both groups. The time for Tl to reach 5% or less in group I(92.8+/-14.72 second) and group II(97.5+/-16.43 seconds) were not siginificant. However the recovery indexof group I(10.2 5+/-1.93 minute) and group II(8.5+/-1.80 minute) showed significant shortening in group II(p<0. 05)

Clinical Evaluation of Supraclavicular Block with a Mixture of Local Anesthetics ; Comparison of the Effect on Onset and Duration / 대한마취과학회지

In the case of brachial plexus block, mixtures of local anesthetics can combine better features of both components, rapid onset and long duration. Combining effects may influence the onset and duration of neural blockade. Our study was undertaken in order to compare the onset time (time of injection to time of loss of pain on pin prick) and duration of analgesia (time of return of sense of pain on pin prick minus time required for onset of analgesia) of a lidocaine and bupivacaine mixture with 5 minutes interval injection of lidocaine and bupivacaine. The patients admitted to our hospital for hand or forearm operations were divided into three groups. In Group 1, 9 patients were injected with 0.5% bupivacaine 150 mg only, in Group 2, 11 patients were injected with a mixture of 29: lidocaine 200 mg and 0.5% bupivacaine 100 mg, in Group 3, 10 patients were injected with 2% lidocaine 200 mg and 5 minutes later, 0.5% bupivacaine 100 mg was injected through the same needle. Group 3 had the shortest onset time (7.2+/-0.2 minutes) with moderately long duration (9.4+/-2.4hours). Group 2 had a moderately rapid onset time (9.4+/-2.3 mintes) with the shortest duration (8.6+/-1.6 hours). Group 1 had the slowest onset time (14.8+/-4.3 minutes) with the longest duration (11.3+/-2. 4 hours). The time for analgesia to reach the C7 dermatome was the slowest in group 1 and Group 2, but in Group 3, there was no difference in the time needed to achieve analgesia in all dermatomes.