A Fluorescence-Labeled Heptapeptide, (FITC)KP6, as an Efficient Probe for the Specific Detection of Oxidized and Minimally Modified Low-Density Lipoprotein.

Two oxidized forms of low-density lipoprotein (LDL), oxidized LDL (ox-LDL) and minimally modified LDL (MM-LDL), are believed to play a major role in the pathogenesis of atherosclerosis. Recently, we reported that a heptapeptide (Lys-Trp-Tyr-Lys-Asp-Gly-Asp, KP6) coupled through the ε-amino group of N-terminus Lys to fluorescein isothiocyanate, (FITC)KP6, bound to ox-LDL but not to LDL. In the present study, we investigated whether (FITC)KP6 could be used as a fluorescent probe for the specific detection of MM-LDL and ox-LDL. Results from polyacrylamide gel electrophoresis and surface plasmon resonance proved that (FITC)KP6 could efficiently bind to MM-LDL as well as ox-LDL in a dose-dependent manner and with high affinity (K D = 3.16 and 3.54 ng/mL protein for MM-LDL and ox-LDL, respectively). (FITC) KP6 bound to lysophosphatidylcholine and oxidized phosphatidylcholine, both present abundantly in ox-LDL and MM-LDL, respectively. In vitro, (FITC)KP6 was detected on the surface and/or in the cytosol of human THP-1-derived macrophages incubated with ox-LDL and MM-LDL, but not LDL. These results suggest that (FITC)KP6 could be an efficient fluorescent probe for the specific detection of ox-LDL and MM-LDL and can therefore contribute to the identification, diagnosis, prevention, and treatment of atherosclerosis.

Serum γ-glutamyltransferase level is associated with periodontal disease independent of drinking habits in Japanese adults.

Background Non-alcoholic fatty liver disease is considered a hepatic manifestation of metabolic syndrome. Periodontal disease is a mild chronic inflammatory disease with systemic effects, and many studies have indicated an association between metabolic syndrome and periodontitis. In the present study, we investigated the relationship between periodontitis and liver biochemical parameters according to alcohol drinking habits through a cross-sectional study based on data from Japanese people in occupational settings. Material and Methods The subjects were 1510 employees (1218 males, 292 females, mean age 50.4 years) who underwent dental and medical checkups in 2012. Associations between the presence of periodontal pockets and serum levels of liver biochemical parameters were assessed. Results Alanine aminotransferase (ALT) and γ-glutamyltransferase (GGT) levels were higher in subjects with than without periodontal pockets. Multiple logistic regression analysis (adjusting for age, gender, cigarette smoking, and alcohol drinking habits, and components of metabolic syndrome) with GGT or ALT as the dependent variable revealed that there was a significant association between periodontal pockets and GGT (odds ratio, OR=1.48), but not ALT. Similar associations were observed when an analysis was performed according to the presence or absence of alcohol drinking habits; the OR was higher in subjects without (OR=1.84) than with drinking habits (OR=1.41). Conclusions The presence of periodontal pockets was associated with serum levels of GGT, a liver biochemical parameter, in Japanese adults with no drinking habit, suggesting that periodontal disease is associated with liver function, independent of alcohol ingestion.

Palliative surgical treatment for enterostasis caused by recurrent gastric cancer: analysis of survival and QOL based on pathological features.

To improve quality of life (QOL) and prolong survival, enterostasis caused by recurrent gastric cancer must be treated appropriately. We reviewed the current treatment retrospectively. The subjects were 43 patients with enterostasis caused by recurrent gastric cancer and treated by surgical procedures at our hospital from 1988 to 1997. Survival and QOL were analyzed in relation to the mode of recurrence, the pathological diagnosis at the initial operation, and surgical procedures. The patients treated by colostomy, ileostomy, or bypass for local occlusion caused by isolated peritoneal recurrence or lymph node recurrence had significantly better quality of life and longer survival [discharge rate: colostomy and ileostomy, 81.8% (9/11); bypass, 77.8% (14/18); survival time: colostomy and ileostomy, 223.5 +/- 171.9 days; bypass, 129.6 +/- 91.0 days] than those who underwent exploratory laparotomy, gastrostomy, or enterostomy and had diffuse disseminated lesions of peritoneal recurrence [discharge rate: 21.4% (3/14); survival time: 44.6 +/- 31.5 days; P < 0.05]. In the patients in whom the pathological diagnosis at initial surgery was differentiated type or poorly solid type, the risk of exploratory laparotomy alone was low (5.6%; 1/18; P < 0.01). Enterostasis with pathological diagnosis at initial surgery of differentiated type or poorly solid type should be treated with aggressive laparotomy and colostomy, ileostomy, or bypass to improve survival and QOL.

Sliding functional end-to-end anastomosis for colorectal surgery.

Conventional functional end-to-end anastomosis (FEEA) is not indicated for left hemicolectomy, sigmoidectomy, and anterior resection. However, our original anastomosis with stapling devices (SFEEA) can be performed at any site in the intestine. We report our novel surgical technique compared with the double stapling technique (DST). Between January 2001 and August 2003, anterior resection with stapling devices was performed in 74 patients (DST, 54; SFEEA, 20). The SEEEA group was greater than the DST group in operation time and significant intraoperative blood loss. In the DST group, two postoperative complications (3.7%) occurred (leakage and stenosis). On the other hand, no complications were noted in the SFEEA group. Our novel technique for colorectal anastomosis, SFEEA, allows safe, wide, physiological, and clean anastomosis compared with FEEA.

Implantation of rectal cancer in an anal fistula: report of a case.

A 53-year-old man who had had an anal fistula for 20 years was admitted to our hospital with a large intestinal obstruction. Barium enema and colonoscopy confirmed advanced rectal cancer and we palpated a soft tumor, 3 cm in diameter, with inflammatory induration on the right side of the rectum. After draining a perianal abscess caused by the anal fistula, we performed low anterior resection. Histological examination of the perianal necrotic tissue obtained during resection of the perianal tumor encompassing the anal fistula revealed adenocarcinoma. Since the histology of the perianal lesion was identical to that of the rectal cancer, a diagnosis of cancer implantation rather than carcinoma originating in the anal fistula was entertained. Although the recurrence of rectal cancer by mucosal implantation is not uncommon, the coincidental implantation of rectal cancer in an anal fistula is extremely rare.

Kugel herniorrhaphy: clinical results of 124 consecutive operations.

PURPOSE: Due to its recent clinical application, the results of Kugel herniorrhaphy have not yet been well documented. We analyzed our experience in performing 124 consecutive Kugel herniorrhaphies at a single institute. METHODS: Since the first introduction of Kugel herniorrhaphy in Japan in September 2001, a total of 117 patients (124 hernias) have undergone this procedure at our institute. The operative technique of Kugel herniorrhaphy was directly obtained through two hands-on training courses that were given by Dr. Kugel. The clinical results of the 124 hernias were analyzed, and to understand the feasibility of performing Kugel herniorrhaphy for various types of hernias, the operation time and the necessity for dividing the hernia sac were compared among each type based on the Nyhus classification. RESULTS: The operation times in types 2, 3A, 3B, 3C, and 4 were 48.5 +/- 19.5, 45.9 +/- 19.9, 54.2 +/- 24.8, 52.5 +/- 10.6, and 64.4 +/- 56.7 min, respectively, and the differences between each group were not statistically significant. In type 3A, no patient required a division of the hernia sac (0%), in contrast to 43% in type 3B. Peritoneal injury during operation and local hematoma/seroma were the most common complications (8.1% and 8.9%, respectively). Postoperatively the patients' quality of life factors, such as the analgesics needed, number of days before a return to normal activity, number of days before a return to work, and the recurrence rate of 0.9%, were comparable with other tension-free herniorrhaphy findings reported in the past. CONCLUSION: Our results suggest that Kugel herniorrhaphy is equally feasible for all types of Nyhus classifications, including femoral and recurrent hernias.

The activities of thymidine phosphorylase and dihydropyrimidine dehydrogenase in colorectal cancer depend on where the sample is taken.

The significance of tumor tissue thymidine phosphorylase (TP) and dihydropyrimidine dehydrogenase (DPD) levels, as well as the TP/DPD ratio have recently been reported as prognostic factors and for custom-made chemotherapy. However, there have been no distinct studies on actual tumor sampling methods. For 16 patients who had undergone resection of advanced colorectal cancer, we: i) measured TP and DPD levels in different portions of the tumor using enzyme-linked immunosorbent assay (ELISA); ii) categorized the tumor into an edge, center, and base area, and histo-pathologically calculated the ratio of cancer cell/cancer cell + stromal cell; and iii) examined the correlation between cancer and stromal cell TP expression and TP value. Variation within the same tumor was seen in each activity level and TP/DPD ratio. The ratio of cancer cell in the edge area was high, with the ratio of stromal cell in the center and base areas increasing in that order. A correlation was seen between TP expression and TP levels, and TP expression was evident in the stromal cells. It is therefore recommended to sample the edge area for tumor TP levels.

Expression of matrix metalloproteinase 7, laminin and type IV collagen-associated liver metastasis in human colorectal cancer: immunohistochemical approach.

BACKGROUND/AIMS: Matrix metalloproteinase 7 (MMP-7) plays an important role in vessel invasion and metastasis in human colorectal cancer. METHODOLOGY: The significance of MMP-7, laminin and type IV collagen expression in human colorectal cancer was investigated by immunohistochemical assay, and the correlation with liver metastasis was analyzed. RESULTS: In a synchronous metastasis group, 26 of 36 cases (72%) showed positive staining of MMP-7: There were 32 cases (89%) in the lymph channel and 28/32 cases (87%) in the vessels, and 17/34 cases (50%) showed a positive rate of laminin. In the metachronous metastasis group, 14 of 30 cases (47%) showed positive staining of MMP-7: There were 19 cases (63%) in the lymph channel and 13/19 cases (69%) in the vessels, and 17/30 cases (57%) showed a positive rate of laminin. In the control group, which was a 5-year disease-free group, despite there being no significant clinicopathological factors compared with the other two groups, 17 of 37 cases (51%) showed positive staining of MMP-7: There were 12 cases (37%) in the lymph channel and 6 cases (18%) in the vessels, and 2/31 cases (5%) showed a positive rate of laminin. The expression of type IV collagen attenuated in 19 out of 32 cases (59%) in Group S, 10 out of 19 cases (53%) in Group M, and 14 out of 37 cases (38%) in Group C, with no significant differences among the groups. Thus, the metastatic groups were significantly higher than the control group in terms of expression of laminin and MMP-7 in the lymph channel. CONCLUSIONS: These findings suggest that laminin and the expression of MMP-7 in the lymph channel is a useful parameter for predicting liver metastasis.

New approach for laparoscopic surgery of the right colon.

BACKGROUND: In patients in whom the ventral aspect of the root of the mesentery was obscured by the adherent greater omentum, laparoscopic surgery is usually abandoned. To forcefully loosen the adhesions by a laparoscopic maneuver may cause inadvertent intestinal injury. We describe a procedure, named the 'retromesen teric approach', which enables a safe laparoscopic right-sided colic operation in such circumstances. PATIENTS AND METHODS: From 2000 to 2003 in our institute, a laparoscopic right-sided colic operation was performed in 21 patients in whom the ventral aspect of the root of the mesentery was obscured by the adherent greater omentum using the retromesenteric approach (RMA; n = 16) and a conventional approach (CA; n = 5). We reviewed the medical records for the operative duration, intraoperative blood loss, conversion rates and postoperative complications. RESULTS: The duration of operation in the RMA group ranged from 75 to 120 (median 95) min, which was shorter than that in the CA group (p < 0.05). Perioperative bleeding in the RMA group ranged from 0 to 115 (median 30) ml, which was smaller than that in the CA group (p < 0.05). No conversion and no postoperative complications were noted in the both groups. CONCLUSIONS: Our new technique, in which the right colon and the ileum are dissected and lifted en bloc from the retroperitoneum, is safe and useful for laparoscopic right-sided colic operation.

A case of neonatal neuroblastoma mimicking Altman type III sacrococcygeal teratoma.

The authors report a case of neonatal neuroblastoma mimicking Altman type III sacrococcygeal teratoma. A newborn male infant delivered after a normal pregnancy was found to have an extremely large sacrococcygeal mass. Imaging studies strongly suggested Altman type III sacrococcygeal teratoma. On the 10th day after the birth, the tumor was incompletely removed at surgery. Histopathologic examination of the tumor showed neuroblastoma. The tumor disappeared completely after chemotherapy. One year after diagnosis, no local recurrence or metastasis had been detected. To the authors' knowledge, this is the first case of neonatal neuroblastoma mimicking Altman type III sacrococcygeal teratoma.

Third-line chemotherapy with paclitaxel, irinotecan hydrochloride and cisplatin for recurrent gastric cancer: a case report.

With the availability of new chemotherapeutic agents such as S-1 and paclitaxel (TXL) for advanced gastric cancer, the development of a strategy for a third-line chemotherapy is urgently needed. We treated a patient with recurrent gastric cancer using TXL, irinotecan hydrochloride (CPT-11) and cisplatin (CDDP) as a third-line chemotherapy. The patient was a 46-year-old man who had undergone total gastrectomy for advanced gastric cancer with lymph node metastases. For postoperative recurrence, he was first treated with S-1 as an outpatient; however, tumor markers increased, and para-aortic lymph node metastasis was revealed by thoracic and abdominal CT scan. A second-line therapy with weekly TXL and CDDP was then added, but resulted in PD. Therefore, combination chemotherapy with TXL, CPT-11 and CDDP was started biweekly as a third-line chemotherapy. TXL (80mg/m2) was infused over 1 hour after short premedication, followed by CPT-11 (25mg/m2) and CDDP (15mg/m2) over 30 min. After 6 courses of this therapy, the serum AFP and TPA returned to normal, and the size of the metastatic para-aortic lymph nodes reduced. The effect of this therapy was judged as PR and the toxicity of this regimen was tolerable. The patient has undergone 10 courses of this therapy and is maintaining a clinical PR. The patient was able to resume his full social activities. TXL, CPT-11 and CDDP combination chemotherapy may be useful and safe for patients with recurrent gastric cancer, even after first-or second-line therapy with S-1 or taxanes.

Implementation of a clinical pathway for laparoscopic colorectal surgery.

The purpose of this study was to assess through questionnaire the validity of our original clinical pathway for laparoscopic colorectal surgery. From 2001 to 2002, laparoscopic colorectal surgery was performed in 52 patients in our institution. All patients were out of bed within 3 days after the operation. The average postoperative hospital stay was 10.9 days. The implementation rate of the clinical pathway was 100%. As to the results of the questionnaire, 92.4% of the patients felt they could be discharged within 7 days after the operation, whereas >95% of the patients wanted to go home on a Sunday/holiday or the day before. All but 2 patients (96.2%) were satisfied with our therapy. In a clinical pathway for laparoscopic colorectomy, a high implementation rate and patients' satisfaction can be achieved by taking into account the patients' viewpoints; however, better patient education is necessary to shorten postoperative stay.

What does the public ask of the JATs? Results and feedback about insufficient public information distribution.

In order for the Japanese Association for Thoracic Surgery (JATS) to undertake appropriate internal reforms, civil receptiveness and sensitivity to the needs of the public are required. Although the JATS is conscious of the needs of physicians, the needs of the public has almost never been addressed. The JATS should place a priority on its mission to advance medicine, to contribute to the public welfare, to distribute information to the public, and to be publicly accountable over seeking the members' well-being. The web site of the "Lung Cancer Medical Treatment Network" (Haigan Chiryo Net), which was launched by one of us 5 years ago, is one example of information distribution. Visitors to the web site have gradually increased to more than 20,000 people per month, totaling 753,000 visits. More than 1200 persons have requested second opinions by e-mail or facsimile. We propose that the nature and purpose of the JATS's civic activities be specified and that the JATS consider and establish methods for public information distribution. Using a web site is an effective method for these activities, but an opinion document about medical problems and the medical system of chest surgery should be created and distributed. The establishment of a consultation service for second opinions should also be considered.

Adeno-associated viral vector-mediated expression of endostatin inhibits tumor growth and metastasis in an orthotropic pancreatic cancer model in hamsters.

We examined the feasibility of using adeno-associated virus (AAV)-mediated systemic delivery of endostatin in gene therapy to treat metastasis of pancreatic cancer. We established an animal model of orthotopic metastatic pancreatic cancer in which the pancreatic cancer cell line PGHAM-1 was inoculated into the pancreas of Syrian golden hamsters. Transplanted cells proliferated rapidly and metastasized to the liver. An AAV vector expressing endostatin (5 x 10(10) particles) was injected intramuscularly into the left quadriceps or intravenously into the portal vein. These routes of vector administration were evaluated by comparing various parameters of tumor development. Intramuscular injection of the vector modestly increased the serum endostatin level. The numbers of metastases and the incidence of hemorrhagic ascites were decreased in the treated animals. In contrast, the serum concentration of endostatin was significantly increased after intraportal injection of the vector. The antitumor effects on all parameters (including the size and microvessel density of primary pancreatic tumors, the sizes and number of liver metastases, and the incidence of hemorrhagic ascites) were significant. These results suggest that systemic delivery of endostatin represents a potentially effective treatment for pancreatic cancer and liver metastases. The route of vector administration influences the efficacy of AAV-mediated endostatin expression. Intraportal injection of the AAV vector appears to be more effective as an antiangiogenic gene therapy for pancreatic cancer.

Esophageal replacement using a reversed gastric tube for lye stricture in a child: report of a case.

A 5-year-old girl ingested lye, a detergent used in electric dishwashers. She was transferred to our hospital after balloon dilatation, performed for stenosis resulting from corrosive esophagitis, was complicated by esophageal perforation causing mediastinitis. Although the mediastinitis resolved with conservative treatment, the stenosis did not improve. Therefore, we performed esophageal replacement using a reversed gastric tube, which successfully relieved the obstruction, although she still had slight gastroesophageal reflux 6 months postoperatively.

Usefulness of radio-guided surgery using technetium-99m methoxyisobutylisonitrile for primary and secondary hyperparathyroidism.

The efficacy of technetium-99m methoxyisobutylisonitrile (99mTc-MIBI)-guided surgery for detecting abnormal parathyroid glands in patients with secondary hyperparathyroidism (2-HPT) was evaluated and compared with the results from the same examination in patients with primary hyperparathyroidism (1-HPT). The results were also compared with those found by ultrasonography (US) and preoperative 99mTc-MIBI scintigraphy was also made. At operation 99mTc-MIBI accumulated in 64 nodules of 15 cases of 2-HPT, and all of 60 parathyroid swellings were detected (true-positives 60, sensitivity 100%, accuracy 94%). In the cases of 1-HPT, 99mTc-MIBI revealed 11 hot nodules in 10 cases, and the evaluation was true-positive 10, sensitivity 100%, and accuracy 91%. US and preoperative 99mTc-MIBI scintigraphy in patients with 2-HPT had a sensitivity of 75% and 67% and an accuracy of 70% and 66%, respectively. The usefulness of 99mTc-MIBI-guided surgery for detecting abnormal parathyroid tissue in 2-HPT patients was similar to that in 1-HPT patients but was superior to US and preoperative 99mTc-MIBI scintigraphy. Intraoperative 9mTc-MIBI for patients with 2-HPT is effective and makes the surgery easier, especially when the parathyroid glands are ectopic or when a few glands are markedly enlarged but the other glands are atrophied.

[Home chemotherapy by concomitant UFT + Leucovorin (po.) in postoperative colorectal cancer patients assessed with Dukes D and curability C colorectal cancer].

UNLABELLED: Concomitant treatment with 5-fluorouracil (5-FU) and Leucovorin (LV) is positioned as the standard chemotherapy against colorectal cancer. We noted the action of LV to enhance the effect of biochemical modulation by 5-FU, and made an attempt at home chemotherapy with UFT + LV by oral administration, in consideration to the convenience of patients. SUBJECTS: The subjects of this study were 24 post-operative patients who had been assessed with Dukes D and curability C colorectal cancer with measurable metastatic lesions and who could tolerate chemotherapy. METHODS: 1 course of treatment consisted of 2 weeks of UFT at 300-400 mg/m2/day and LV at 15 mg/body/day followed by 2 weeks of drug withdrawal. The administration was conducted for 4 courses or more as the target. Unless serious adverse reaction occurred, dose increase of UFT was allowed. RESULTS: The efficacy rate in the 22 patients who were assessable was 22.7%. There were 11 NC patients, accounting for half (50%) of the subjects. This home chemotherapy is expected to become an alternative chemotherapy against colorectal cancer in the future, because the treatment does not require hospitalization and has less impact on the QOL of patients.

Fetal surgery for repair of myelomeningocele allows normal development of anal sphincter muscles in sheep.

One major problem for patients with myelomeningocele (MMC) is fecal incontinence. To prevent this problem, fetal surgery for repair of MMC has been recently undertaken. The strategy behind this surgery is to allow normal development of anal sphincter muscles. The purpose of this study was to determine whether fetal surgery for repair of MMC allows normal development of anal sphincter muscles. Myelomeningocele was surgically created in fetal sheep at 75 days of gestation. At 100 days of gestation, fetal surgery for repair of the MMC lesion was performed. Three repair methods were used: standard neurosurgical repair (4 fetal sheep), covering the MMC lesion with Alloderm (2 fetal sheep), and covering the MMC lesion with Gore-Tex (2 fetal sheep). After the sheep were delivered (140 days of gestation), external and internal anal sphincter muscles were analyzed histopathologically. In control fetal sheep (not repaired) anal sphincter muscles did not develop normally. In contrast, in fetal sheep that underwent repair of the MMC, the external and internal anal sphincter muscles developed normally. Histopathologically, in the external sphincter muscles, muscle fibers were dense. In the internal sphincter muscles, endomysial spaces were small, myofibrils were numerous, and fascicular units were larger than those in unrepaired fetal sheep. There was no difference in muscle development for the repair methods. Fetal surgery for repair of MMC allows normal development of anal sphincter muscles.

Tenascin-C regulates angiogenesis in tumor through the regulation of vascular endothelial growth factor expression.

In order to verify whether tenascin-C (TN-C) is involved in angiogenesis as an extracellular signal molecule during tumorigenesis, cancerous cell transplantation experiments and coculture experiments were carried out, focusing on the regulation of vascular endothelial growth factor (VEGF). The A375 human melanoma cells introduced the GFP gene (A375-GFP), implanted subcutaneously into BALB/cA nude (WT) and TN-C knockout BALB/cA nude (TNKO) congenic mice. Furthermore, coculture experiments between A375-GFP and embryonic mesenchyme, which was prepared from both genotypes, were carried out to investigate the molecular mechanism in the cell-cell interactions. Both the content of TN-C and that of VEGF in the tumor and the conditioned medium were analyzed by the sandwich ELISA method. Seven days after transplantation of the A375-GFP, capillary nets became far more abundant in the tumors grown in WT mice than those in TNKO mice. Interestingly, VEGF and TN-C expressions showed antithetical expression patterns between the tumors in WT mice and those in TNKO mice. This peculiar phenomenon seems to be caused by a time lag prior to the onset of the mesenchymal regulation for the TN-C expression of A375-GFP. The coculture experiments revealed that WT mesenchyme had a much stronger effect than TNKO mesenchyme on both TN-C and VEGF expression. However, the defects of TNKO mesenchyme were restored in all cases by additional TN-C. These results clearly indicated that the expressions of both TN-C and VEGF depend on the surrounding mesenchyme, and that the function of mesenchyme is regulated by its own mesenchymal TN-C. In conclusion, the present data suggest that the matrix microenvironment organized by the host mesenchyme is very important for angiogenesis in tumor development.

Successful in utero gene transfer using a gene gun in midgestational mouse fetuses.

BACKGROUND/PURPOSE: In utero gene therapy offers a number of potential advantages over postnatal gene therapy. A latest method of gene transfer to fetuses in utero uses a new tool called a gene gun. The gene gun is less invasive and simpler than other in utero methods. The current study was designed to determine whether the gene gun is an effective tool for transferring genes to mouse fetuses in utero. METHODS: Using a gene gun, we transferred plasmids that included enhanced green fluorescent protein (EGFP) genes and cytomegalo virus promoters to the abdominal skin of 40 A/J fetal mice at each of 3 gestational ages (13, 14, or 15 days). Four or 5 days after gene transfer, the number of surviving fetuses was counted, and a color image of EGFP in the skin was analyzed for gene transfer rates by fluorescence microscopy. Survival rates were analyzed using Fisher's Exact test. RESULTS: The mean survival rate was 89.2% (107 of 120) in gene transfer fetuses and 91.7% (55 of 60) in controls. There is no difference in survival rate between gene transfer fetus and control. The highest gene transfer rate was 100% (37 of 37) at the gestational age of 14 days. The rate was 97.1% (34 of 35) at gestational ages of 13 and 15 days. CONCLUSIONS: The results of this study show that in utero gene transfer by gene gun is a less-invasive technique, and the gene gun is an effective tool transferring genes to mouse fetuses in utero.