Novel strategy of gene therapy in cardiovascular disease with HVJ-liposome method.
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Cardiovascular Diseases↗
Biomedical subjects
M Aoki
Publications and source records attributed to M Aoki.
Role of transcriptional cis-elements, angiotensinogen gene-activating elements, of angiotensinogen gene in blood pressure regulation.
Results of recent genetic studies suggest that the angiotensinogen gene is a possible determinant of hypertension. Using antisense technology, we demonstrated that generation of circulating angiotensinogen is a rate-limiting step in blood pressure regulation. In the present study, we examined how the angiotensinogen gene is regulated in vivo. The transcriptional cis-elements, angiotensinogen gene-activating elements (AGE) 2 and 3, have been reported to regulate angiotensinogen production in human hepatocytes in vitro. To determine the critical transcriptional regulator of angiotensinogen production in vivo, we used synthetic double-stranded oligodeoxynucleotides (ODN) as "decoy" cis-elements to block the binding of nuclear factors to promoter regions of the targeted gene, resulting in the inhibition of gene transactivation. Here we examined whether AGE 2 and AGE 3 in the promoter region of the angiotensinogen gene have a pivotal role in hepatic angiotensinogen production in vivo. Hepatic angiotensinogen mRNA was decreased by the transfection of AGE 2 but not mismatched decoy ODN. Transfection of decoy but not mismatched ODN against AGE 2 resulted in a transient decrease in blood pressure of spontaneously hypertensive rats (SHR), accompanied by a reduction in plasma angiotensinogen and angiotensin II levels. In contrast, transfection of AGE 3 decoy ODN had little effect on blood pressure. Overall, our results demonstrate that transfection of decoy ODN against AGE 2, but not against AGE 3, of the angiotensinogen gene resulted in a transient decrease in high blood pressure of SHR, suggesting that the transcriptional cis-element AGE 2, rather than AGE 3, has an important role in blood pressure regulation through the control of circulating angiotensinogen.
Angiotensinogen↗
A vascular modulator, hepatocyte growth factor, is associated with systolic pressure.
Endothelial cells are known to secrete various antiproliferative and vasodilating factors, such as nitric oxide and natriuretic peptides. The presence of endothelial dysfunction, well known in hypertensive individuals, potentially results in the development and progression of atherosclerosis. Therefore, it is important to know the factors that might influence endothelial cell growth. We examined the mitogenic actions of hepatocyte growth factor (HGF) on human endothelial and vascular smooth muscle cells. Exogenously added human recombinant HGF stimulated endothelial but not vascular smooth muscle cell growth in a dose-dependent manner. We also compared the mitogenic action of HGF with that of basic fibroblast growth factor and vascular endothelial growth factor. Interestingly, the mitogenic action of HGF on endothelial cells was greater than the actions of basic fibroblast growth factor and vascular endothelial growth factor, whereas basic fibroblast growth factor but not HGF and vascular endothelial growth factor stimulated vascular smooth muscle cell growth. Given the characteristics of HGF as an endothelium-specific growth factor, we evaluated the relationship of circulating HGF and blood pressure in normotensive and hypertensive subjects. Serum HGF concentration has been reported to be elevated in response to organ damage, such as in hepatitis and nephritis, and recent findings show that HGF may play an important role in tissue regeneration. We hypothesized that HGF might contribute to the protection or repair of vascular endothelial cells. If so, serum HGF level might be elevated in response to endothelial cell damage induced by hypertension. To test this hypothesis, we measured serum levels of HGF, lipoprotein(a), plasminogen activator inhibitor-1, tissue plasminogen activator, total cholesterol, and blood pressure in 41 normotensive and hypertensive subjects without liver, kidney, or lung damage. Serum HGF concentration was significantly correlated with systolic pressure (P < .01, r = .43) but not diastolic pressure. Serum HGF concentration in hypertensive subjects was significantly higher than in normotensive subjects. None of the other factors showed any correlation with blood pressure. We have demonstrated that HGF is an endothelium-specific growth factor whose serum concentration is significantly associated with systolic pressure. These results suggest that HGF secretion might be elevated in response to high blood pressure as a counterregulatory system against endothelial dysfunction.
Aged↗
Operative treatment of intramedullary hematoma associated with congenital deficiency of alpha 2-plasmin inhibitor, A report of three cases.
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Adolescent↗
Identification of trinucleotide repeat-containing genes in human pancreatic islets.
In the search for diabetes genes, the combined approaches of positional cloning with random markers and subsequent evaluation of candidate genes mapping to areas of interest will be increasingly used. For islet candidate genes of unknown function, expressed trinucleotide (triplet) repeats represent a unique subset. It is unlikely that abnormal expansion of expressed islet triplet repeats would be a major cause of diabetes, yet the triplet repeats are frequently polymorphic and can thus be used to map the genes in the human genome. In this study, a human islet cDNA library was screened with (CGG)7 and (CAG)7, and 23 triplet repeats were isolated. Sequencing revealed four known and six novel islet genes containing 4-15 triplet repeats. The four known cDNAs included ferritin, the major iron-binding protein in cells; HSGSA2R, a full-length clone of the alpha-subunit of the G-regulatory protein; HUMSATB1A, a DNA-binding protein expressed predominantly in thymus; and HUMPPA-PRO, a ribosomal protein. The triplet repeats in ferritin and HUMPPAPRO were found to be monomorphic. Characterization of the six unique novel expressed islet triplet cDNAs revealed that they were 0.6-1.5 kb in size, contained 4-15 triplet repeats, and were expressed in islets and all other tissues examined. Four of the novel clones, CGG-isl 10, CGG-isl 11, CAG-isl 6, and CAG-isl 7, were mapped to human chromosomes 19, 16, 12, and 3, respectively, via somatic cell hybrids. One islet cDNA, CAG-isl 7, contained a repeat that was highly polymorphic, with 14 alleles (4-18 triplets) in African-Americans (heterozygosity = 0.86) and 6 alleles (heterozygosity = 0.77) in whites. Northern analysis indicated that the mRNA was abundant in pancreatic islets. A putative full-length clone contained an open reading frame encoding 213 amino acids with a variable number of alanines (4-18) within the COOH-terminal. The gene was uniquely mapped with odds > 1,000:1 on chromosome 3p in Centre d'Etude du Polymorphisme Humain pedigrees. There were no differences in CAG-isl 7 allele frequencies between African-American patients with NIDDM (n = 108) and control subjects (n = 116), nor was expansion above 18 repeats noted. Linkage analysis in 14 nonglucokinase maturity-onset diabetes of the young pedigrees showed a cumulative logarithm of odds score of -33.19 at theta = 0.00. Abnormal expansion was not observed in 20 IDDM patients with one NIDDM parent. While these data suggest no major role for CAG-isl 7 in diabetes, at least four of the six novel islet triplet genes are coexpressed in pancreatic islets and neural tissue, and these genes can now be considered as candidates for diabetes and/or neuropsychiatric diseases.
Amino Acid Sequence↗
Mitochondrial glycerol-3-phosphate dehydrogenase. Cloning of an alternatively spliced human islet-cell cDNA, tissue distribution, physical mapping, and identification of a polymorphic genetic marker.
Pancreatic beta-cell mitochondrial glycerol-3-phosphate dehydrogenase (mGPDH) plays a major role in glucose-induced insulin secretion. Decreased activity of this enzyme has thus been proposed to play a role in the pathogenesis of NIDDM. Cloning of human insulinoma mGPDH cDNAs disclosed the existence of two variant transcripts with different 5' ends. Reverse transcription polymerase chain reaction (PCR) confirmed the presence of both mGPDH mRNAs in purified native human pancreatic islets and other tissues. A major 6.5-Kb mGPDH transcript was detected by Northern blot analysis in RNA from human and rat pancreatic islets, with distinctly lower levels in other human tissues, indicating that previously reported high mGPDH enzymatic activity in beta-cells is determined by high transcript levels. The mGPDH gene was mapped to chromosome 2 by PCR analysis of genomic DNA from human/rodent somatic cell hybrids, and five independent overlapping yeast artificial chromosome (YAC) clones containing the mGPDH sequence were identified from the Centre d'Etude du Polymorphisme Humain YAC library. Analysis of these YAC clones identified a highly polymorphic chromosome 2q21-q33 dinucleotide repeat genetic marker (D2S141) physically linked to the mGPDH gene. These studies provide the means to investigate the role of the human mGPDH gene in the pathogenesis of NIDDM and illustrate the value of a novel strategy to identify genetic markers for diabetes candidate genes.
Alternative Splicing↗
[Clinical experience with laparoscopy-assisted live donor nephrectomy].
PURPOSE: Laparoscopic nephrectomy has become widely performed because of its minimally invasive nature. We have performed laparoscopic nephrectomies for non-functioning kidneys and laparoscopy-assisted radical nephrectomies for renal carcinomas. Recently, we have successfully performed laparoscopy-assisted transperitoneal living related donor nephrectomy for kidney transplantation. To determine the efficacy of laparoscopy-assisted live donor nephrectomy, we investigated the clinical results of 2 patients operated upon with this new operation. PATIENTS: Case 1 was a 64-year-old healthy male and the left kidney was removed for his son who had a history of 4 months hemodialysis. Case 2 was 67-year-old healthy female who decided to donate her left kidney for her son with the history of 3 years and 2 months hemodialysis. METHODS: An upper abdominal midline incision of approximately 10 cm in length was made. Two retractors were attached to either side of the midline incision. The abdominal wall was raised by suspending these retractors from a special hanger. Then three trocars were introduced. The Gerota's fascia was incised and the kidney was suspended by grasping the perirenal fatty tissues. The renal artery and vein were carefully isolated without any manipulations of the kidney (non-touch method). After the transection of the ureter, the renal artery was ligated with a free tie of 1-0 silk and a suture ligature of 3-0 silk and the renal vain was clamped with a Satinski forceps. Immediately after the transection of the renal pedicle, the kidney was irrigated and transplanted to the right iliac fossa of the recipient in the usual fashion. A pen-rose drain was placed in the retroperitoneal space and the posterior peritoneal membrane was completely closed with 3-0 silk interrupted sutures. RESULTS: The mean operating time was 298 minutes and the mean blood loss was minimal. The average time of warm ischemia and cold ischemia of the graft were 4.5 minute and 37 minutes, respectively. There were no complications during either the operation or the postoperative period in both patients. The donors began oral intake and ambulation within 48 hours and resumed their normal daily activities by postoperative day 6.5 on average. Postoperative recovery of the patients were far faster than that of the patients receiving open donor nephrectomy. The graft functions were also excellent. The serum creatinine concentration of the recipients fell down to 0.82 mg/dl and 1.02 mg/dl at the third postoperative day. CONCLUSION: Laparoscopy-assisted live donor nephrectomy might be advantageous for kidney transplantation because of its minimally invasive procedure.
Female↗
[Extraperitoneal laparoscopic adrenalectomy: the initial 9 cases].
(PURPOSE). To determine the efficacy of extraperitoneal laparoscopic adrenalectomy for patients with adrenal tumor, the clinical results of 9 patients treated with this procedure were analysed. (PATIENT AND METHOD). Between July 1994 and March 1995, we have performed extraperitoneal laparoscopic adrenalectomy using the modified Gaur technique of balloon dilatation of the retroperitoneum in 4 men and 5 women with unilaterla small adrenal tumor who were 30 to 79 years old (mean age 56 years). Preoperative diagnosis was primary aldosteronism in 2, 18-hydroxycorticosterone producing adnoma in 1, pre-Cushing syndrome in 5, and nonfunctioning adrenal tumor in 1 patient. The operations were performed with the patients on lateral position and 4 torocars were positioned. The retroperitoneal space was first dissected bluntly by the index finger and a balloon dissector through a small skin incision, and the retroperitoneal space was insufflated with carbon dioxide at the pressure of 10 to 12 mmHg. After the dissection of the adrenal gland, adrenal vein was clipped and transected. The completely freed adrenal gland was enclosed in an entrapment sack and removed en bloc through the open laparoscopy wound. (RESULTS). All 9 procedures were successfully performed. The mean estimated blood loss and the mean operating time were 53 ml and 168 min respectively. There was no intraoperative complication. A postoperative retroperitoneal hematoma was observed in one patient, however it spontaneously resolved without surgical management. (CONCLUSION). In open adrenalectomy, the benefit of the extraperitoneal approach (excluding patients with pheochromocytoma or bilateral lesions) have already been established elsewhere. Although the working space is smaller than that of peritoneal approach, and the surgical techniques are slightly more difficult, extraperitoneal laparoscopic adrenalectomy promises to be safe and a minimally invasive treatment for patients with unilateral small adrenal tumors except for pheochromocytoma.
Adrenal Gland Neoplasms↗
[Laparoscopic adrenalectomy for Cushing's syndrome: a study of comparisons between laparoscopic and open adrenalectomy, and laparoscopic adrenalectomy for differing adrenal tumors].
In order to evaluate the utility of laparoscopic adrenalectomy for Cushing's syndrome, the results of 6 laparoscopic adrenalectomies for Cushing's syndrome were compared with those of 34 other laparoscopic adrenalectomies, consisting of 7 pre-Cushing's syndromes, 13 primary aldosteronisms, and 14 non-functioning adrenal tumors. The results were also compared with 5 open adrenalectomies for Cushing's syndrome. The results of the laparoscopic adrenalectomies for Cushing's syndrome were as follows: the mean operating time was 216 +/- 46 min, and the mean estimated blood loss was 180 +/- 194 ml. From the third patient, an ultrasonic surgical system was used and the estimated blood loss decreased significantly. The ultrasonic surgical system, together with a more experienced surgical technique, also cut down the operating times. During surgical intervention, 2 complications occurred; a hemorrhage of more than 500 ml in one patient, and splenic injury in another, which was treated by compression. Postoperative complications occurred in 2 patients; paralytic ileus in one, and abdominal pain due to the pneumoperitoneum in the other. All patients except 2 patients with vertebral fracture began oral intake and ambulation 1 to 4 days postoperatively, and resumed normal daily activity on postoperative day 5 to 7. Compared with the laparoscopic adrenalectomies for the other adrenal tumors, the operating time and estimated blood loss in the Cushing's syndrome patients was not substantially different, though postoperative recovery was slightly longer. When compared with the open adrenalectomies, the operating time was longer, but the postoperative recovery period was significantly shorter. We conclude that with careful surgical intervention, experience of technique, and the introduction of proper equipment, a laparoscopic adrenalectomy for Cushing's syndrome can be performed as less-invasively as a laparoscopic adrenalectomy is for the other adrenal tumors. Furthermore, our findings suggest that laparoscopic adrenalectomy for Cushing's syndrome is likelier to have better postoperative results than conventional procedures, including a more rapid recovery to normal daily activity.
Adrenal Gland Neoplasms↗
[A device for rapid elevation of plasma methotrexate (MTX) concentration and its maintenance in high-dose MTX therapy].
For 5 osteosarcomas and one synovial sarcoma, 24 courses of high-dose methotrexate (MTX) therapy were performed. At MTX doses of 300 mg/kg with several infusion patterns, the plasma MTX concentrations were measured by fluorescence polarization immunoassay. In the various types of infusions, five hours oblique and five hours bolus infusions were well maintained at 1,000 micromol/l for several hours. In particular, five hours bolus infusion of MTX needed only 20 minutes to reach 822 micromol/l and maintained 1,000 micromol/l during almost the entire infusion period. Optimization of chemotherapy for sarcomas by MTX requires individual adaptation of the infusion pattern.
Adolescent↗
[Retroperitoneal laparoscopic pelvic lymphadenectomy for patients with prostate cancer].
Between February 1994 and February 1995, we performed retroperitoneal laparoscopic pelvic lymphadenectomy on 7 patients with prostate cancer between 63 and 76 years old, consisting of 6 patients with clinical stage B2, and 1 patient with stage C. Retroperitoneal laparoscopic procedures were performed under general anesthesia. First, the prevesical space was separated by a balloon dissector to create the working space. Then, three trocars were inserted into the retroperitoneal space at the lower abdominal midline. Except for the first patient, only two surgeons were needed to dissect the obturator lymph nodes, under 5 mmHg CO2 insufflation. The mean operating time was 133 minutes for bilateral lymphadenectomy. The mean number of removed lymph nodes was 4.8 on the left side and 4.2 on the right side. The estimated blood loss ranged from 10 to 66 ml. As none of the patients showed node involvement on the microscopic examination of the frozen section, they immediately underwent total prostatectomy. Additionally removed lymph nodes were 0.8 on the left side and 1.5 on the right side. As for intraoperative complications, endotidal CO2 was elevated in the first patient, but the other patients had no complications. Retroperitoneal laparoscopic pelvic lymphadenectomy is a safe and useful procedure, and may facilitate a lymphadenectomy better than a peritoneal procedure.
Adenocarcinoma↗
[Usefulness of endoscopic ultrasonography in deciding indication for high dose rate intracavitary brachytherapy for esophageal carcinoma].
We retrospectively analyzed 20 patients with stage I-IV esophageal carcinoma treated by limited external radiotherapy (EXR) combined with high dose rate intracavitary brachytherapy (ICBT) using 60Co as boost therapy. In 10 of 20 patients treated by combined therapy, we used endoscopic ultrasonography (EUS) before ICBT and measured the thickness of the residual tumor following external beam therapy. Of the patients treated by combined therapy, the local control rate of esophageal carcinoma with a thickness under 10 mm before ICBT was significantly better than that with a thickness over 10 mm (confidence limit 95%). Therefore, when administering high dose rate intracavitary brachytherapy after EXR, we should measure the thickness of esophageal carcinoma with millimeter unit precision. One of the most precise methods currently available for measuring thickness is EUS. Our results indicate that esophageal carcinoma under 10 mm in thickness after EXR is the most favorable indication for ICBT.
Aged↗
[Continuous systemic venous oxygen saturation monitoring immediately after Fontan procedure].
Changes in cardiac output (CO), systemic venous oxygen saturation (SvO2), systemic oxygen consumption, and urinary output immediately after Fontan procedure were measured in 10 patients at the intensive unit (ICU) to assess the effects of aorusal from anesthesia, hypothermic management, and respiratory condition. The measurements were taken at the following phases; phase A in deep sedation under hypothermia (33-35 degrees C rectal temperature) and controlled ventilation; phase B in mild sedation under normothermia and controlled ventilation; phase C when awake under normothermia and assisted ventilation; phase D when awake under assisted ventilation; phase D when awake under normothermia immediately after extubation; and phase E 24 hours after extubation. Oxygen delivery (O2 Del.) and fractional oxygen fractions were calculated in each phase. Two patients whose SvO2 values were below 55% during the postoperative course needed reoperation for atrioventricular valve regurgitation in one case for PV stenosis in the other case. CO increased significantly (p < 0.05) after extubation (phase D), compared with that of controlled ventilation (phase B). Under hypothermia (phase A), urinary output was relatively higher with lower CO. There was a significant correlation between SvO2 and CO (R = 0.61) in phase A, however there was no correlation in phase E. Fractional oxygen extraction in phase A was significantly lower than in phase B. In conclusion, the continuous SvO2 measurements reflected real-time changes in cardiac output in the immediate post-Fontan patients. Induced hypothermia was beneficial in increasing urinary output presumably through the correction of maldistribution of cardiac output in post-Fontant patients. Arousal from anesthesia and spontaneous ventilation seemed advantageous for increasing cardiac output, hence, early extubation should be encouraged in the management of post-Fontan patients.
Blood Gas Monitoring, Transcutaneous↗
In vivo gene transfer of insulin gene into neonatal rats by the HVJ-liposome method resulted in sustained transgene expression.
In Many areas of disease such as homozygous familial hypercholesterolemia, no known effective therapy exists. The time is ripe for the introduction of gene therapy for the management of incurable disorders. However, for clinical gene therapy it is necessary to establish an efficient and non toxic gene delivery system. Recently, we have developed a gene delivery system mediated by Sendai virus as a potential means of gene therapy. However, this HVJ-liposome method has some disadvantages: (1) transient transgene expression; and (2) no integration. In gene therapy, it is desirable for transgenes to be expressed for as long as possible. To deal with this issue, we hypothesized that gene transfer into newborn animals would result in sustained expression of transgene. Our results demonstrated that transgene expression in liver was subsequently sustained for at least up to 8 weeks, as detected by reverse transcriptase PCR and radioimmunoassay, while our previous studies documented that transgene expression in adult animals disappeared within 2 weeks. Interestingly, we detected the presence of transgene in the host genomic DNA extracted from the nuclei of hepatic cells assessed by PCR. These findings provide new important information for gene therapy, although the mechanisms of sustained transgene expression are not clear.
Aging↗
"Tuberculosis".
Tuberculosis has decreased so rapidly and has changed its epidemiological feature so markedly in these several decades in Japan, that tuberculosis epidemiologists had done and have to do numerous studies to understand the situations and to adjust tuberculosis control programme of high prevalent country to that of low prevalent country. Main themes of epidemiological research are being focused on 1) the causes of stagnation of the decrease of tuberculosis incidence and estimation of future trend in Japan, 2) risk factors of development of disease at present when more than 95% of diseases are the exacerbation of the remote infection, 3) the efficacy and efficiency of each control measure to improve the existing programme in Japan, 4) the mode of tuberculosis infection in the community using RFLP analysis of tubercle bacilli, 5) epidemiology of tuberculosis in the developing countries where tuberculosis is increasing at present, and so on. Because of the increase of tuberculosis in the world, epidemiological research in tuberculosis is becoming more and more active to-day. It is being expected that epidemiological research on tuberculosis will have more and more fruitful results in near future.
Humans↗
Transfer of latissimus dorsi for irreparable rotator-cuff tears.
We treated 12 shoulders in ten patients with irreparable rotator-cuff tears by transfer of the latissimus dorsi. There were nine men and one woman. Their average age was 64.0 years and the average follow-up was 35.6 months (26 to 42). The results were excellent in four shoulders, good in four, fair in one, and poor in three. Active forward flexion improved from a preoperative average of 99 degrees to a postoperative average of 135 degrees. Osteoarthritic changes appeared in five shoulders and proximal migration of the humeral head progressed in six. EMG revealed that nine of the 12 transferred muscles showed activity which was synergistic with the supraspinatus on external rotation with abduction. We conclude that latissimus dorsi transfer can be effective in restoring shoulder function after massive irreparable tears of the rotator cuff.
Activities of Daily Living↗
[A study on prevalence of resistance to antituberculosis drugs in Japan: comparison of results in the local facilities and in the reference laboratory].
During five years since the last survey of drug-resistant tuberculosis in Japan, features of tuberculosis in Japan have been changed. A nationwide survey was conducted by the Tuberculosis Research Committee of Japan. A total of 38 hospitals in various districts of Japan participated in the cooperative study. Each collaborating laboratory sent all mycobacterial cultures isolated during 1 June to 30 November, 1992 to the reference laboratory of the Committee, where species of the isolates were identified and drug susceptibility of Mycobacterium tuberculosis isolates were reexamined. The reference laboratory received a total of 1,236 cultures. Among them, 290 cultures were excluded from further examination by various reasons, such as contamination (52 cultures), non-viability (53), growth of nontuberculous mycobacteria (182) and other reasons (3). Thus, drug susceptibility test results were available for 946 cultures, including 26 cultures from non-Japanese persons. In the local laboratories, two methods, the absolute concentration method using 1% Ogawa egg slant (standard method, 26 hospitals) and its modified method using a microwell plate (microtiter method, 12 hospitals), were used for drug susceptibility testing, and the standard method was used in the reference laboratory. The results in the local laboratories were compared with those in the reference laboratory. The overall coincidence rate between drug susceptibility results reported from the local laboratories and those from the reference laboratory was 92.5%. A high coincidence rate (94.3%) was seen when the standard method was used in both local and reference laboratories. On the other hand, the coincidence rates between the results with the microtiter method in the local laboratory and those with the standard method in the reference laboratory were lower (standard method vs microtiter method; P < 0.01). Out of 19 hospitals, when the isolates were tested by the standard method 17 (89.5%) showed high coincidence rates (over 85%). Three hospitals using the microtiter method showed the coincidence rate over 90%, while other three showed lower rate (less than 80%) with high overestimation rates (over 19%), indicating that there are variations among facilities in performing the microtiter test. A part of the results concerning the resistance patterns to five antituberculosis drugs will be reported elsewhere.
Adolescent↗
In vivo transfer of gene and oligodeoxynucleotides into skin of fetal rats by incubation in amniotic fluid.
One of the tools to test an in vitro hypothesis in vivo is transgenic/gene targeting technology. Transgenic technology provides many advantages such as: (1) the study of specific gene function as systemic and developmental effects; and (2) testing of specific gene function chronically. nevertheless, one disadvantage of this technology is the difficulty in tissue-specific targeting of the transgenic expression. Another useful tool is the in vivo gene transfer approach. Therefore, we tested a potential novel model for the study of transgene expression and knock-out using antisense technology. Here, we have demonstrated that incubation of hemagglutinating virus of Japan (HVJ)-liposome complex containing FITC-labeled oligonucleotides in the amniotic fluid of fetal rats resulted in the nuclear localization of fluorescence in the skin (epidermis and dermis). Similarly, transfection of beta-galactosidase gene resulted in positive staining in several surface layers of the skin. Thus, local gene or antisense oligonucleotide transfer approach into the skin may be useful for studying the role of autocrine/paracrine mediators and treating diseases.
Amniotic Fluid↗