Laparoscopic salvage lateral pelvic lymph node dissection for locally recurrent rectal cancer.

AIM: The lateral pelvic lymph nodes are one of the major sites and sources of local recurrence (LR) after surgery for rectal cancer. Salvage lateral pelvic lymph node dissection (LPLD) is potentially curative, but the value of laparoscopic surgery in such cases is unknown. Our aim was to report the technical details of laparoscopic salvage LPLD for LR at these nodes after rectal cancer surgery. METHOD: The study was based on nine patients who underwent laparoscopic salvage LPLD for LR at the lateral pelvic lymph nodes after surgery for rectal cancer. The safety and feasibility of this procedure were determined. RESULTS: The median operation time was 381 min and the median estimated blood loss was 130 ml. There were no conversions. Adjacent structures removed en bloc were the pelvic plexus in four patients, the internal iliac artery in seven patients and the seminal vesicle in one patient. The median number of metastatic lymph nodes was 1 (range 1-11). CONCLUSION: Our novel technique of laparoscopic salvage LPLD for LR at the lateral pelvic lymph nodes is safe and feasible.

Serum procalcitonin (PCT) as a negative screening test for colonic ischemia after open abdominal aortic surgery.

BACKGROUND AND AIM: We assessed serum procalcitonin (PCT) as a screening test for early detection of ischemic colitis. PATIENTS AND METHODS: Ninety-three patients (81 men and 12 women) undergoing elective aortic surgery were enrolled in this study. Their mean age was 70.3+/-8.1 years old. Serum procalcitonin was measured postoperatively. RESULTS: Four patients suffered from colon ischemia. Based on a cut off value of serum PCT>/=2.0 ng/ml, fourteen patients had a false positive but none had a false negative result. Sensitivity was 100%, and specificity was 83.9% in detecting ischemic colitis. Negative predictive vale was 100%. CONCLUSION: Serum PCT is a non-invasive test that has a high negative predictive vale in ruling out colon ischemia after aortic surgery.

Influence of forward head posture on condylar position.

There are several reports suggesting that forward head posture is associated with temporomandibular disorders and restraint of mandibular growth, possibly due to mandibular displacement posteriorly. However, there have been few reports in which the condylar position was examined in forward head posture. The purpose of this study was to test the hypothesis that the condyle moves posteriorly in the forward head posture. The condylar position and electromyography from the masseter, temporal and digastric muscles were recorded on 15 healthy male adults at mandibular rest position in the natural head posture and deliberate forward head posture. The condylar position in the deliberate forward head posture was significantly more posterior than that in the natural head posture. The activity of the masseter and digastric muscles in the deliberate forward head posture was slightly increased. These results suggest that the condyle moves posteriorly in subjects with forward head posture.

Consistent disruption of the AML1 gene occurs within a single intron in the t(8;21) chromosomal translocation.

The AML1 gene on chromosome 21 was rearranged by the t(8;21) chromosomal translocation in acute myeloid leukemia (AML). Southern blot analysis of 21 AML patients with t(8;21), including three with complex translocations, t(8;V;21), demonstrated that all the breakpoints occurred at random within a single intron between two coding exons of AML1. Clustering of the breakpoints in the restricted intron suggests the formation of a unique fusion gene between the AML1 gene and a presumable counterpart gene on chromosome 8. Nucleotide sequencing of the breakpoint region revealed that the translocation event was accompanied by deletion of a short stretch of nucleotides.

Precursor role of branched-chain amino acids in the biosynthesis of iso and anteiso fatty acids in rat skin.

Monoester fraction of rat skin surface lipid has been shown to contain more than trace amounts of branched-long-chain fatty acids (BCFAs) of the iso and anteiso series. These BCFAs are biosynthesized using either branched-chain amino acids (BCAAs) or branched-chain alpha-keto acids (BCKAs), or using both of them as precursor. This study has been carried out to address which precursor, BCAAs or BCKAs in the circulation, are mainly utilized for biosynthesis of BCFAs. Dietary supplement of [14C]-valine and isoleucine-induced sharp rise of serum concentration of these two amino acids and their respective alpha-keto acids, and elevated the levels of related BCFAs and branched-chain fatty alcohols in the monoester fraction. A larger proportion of label in the total skin surface lipid was found in the monoester fraction in which fatty acid and alcohol accounted for approx. 80% of total radioactivity. Incorporation of intravenously administered [14C]-BCAAs and BCKAs into the monoester fraction revealed that BCAAs were far better as precursors than BCKAs for BCFA biosynthesis in rat skin. Among three BCAAs, leucine differed from valine or isoleucine in that this amino acid was primarily utilized for production of straight-chain fatty acids rather than for production of related BCFA.

Accumulation of 1-o-alkyl-2,3-diacylglycerols in cultured rat keratinocytes.

The present study was undertaken to identify the chemical structure of neutral lipid accumulated in cultured rat keratinocytes and to address their metabolism. Neutral lipid of similar mobility with alkyldiacylglycerol was isolated from cultured rat keratinocytes by thin layer chromatography. The long-chain diols derived from the neutral lipids were identified as 1-alkylglycerol based on the mass spectra of their nicotinylidene derivatives. Thus these neutral lipids were identified as 1-o-alkyl-2,3-diacylglycerols (ADAG). Addition of rat serum elevated the level of ADAG with increasing trend of linoleic acid concentration in this fraction. [14C]Acetate added to the confluent plates was incorporated into alkyl- and acyl-chains of ADAG with incubation in 24 h, and remained un-metabolized up to 72 h. This, however, is not the case for the label incorporation into phospholipid and triacylglycerol. Radioactivities of these two lipid fractions appeared to reach the maximum in 24 h, and thereafter decreased to 72 h with a similar decay curve. Incorporation of [14C]acetate into phospholipid and ADAG was significantly depressed, and that into triacylglycerol and free cholesterol was increased by the supplementation of the medium with rat serum. In concomitance with the accumulation of ADAG, the concentration of ethanolamine-plasmalogen increased in the cultured keratinocytes. The results of the present study first showed the elevated level of ether lipid synthesis in the proliferating primary culture of rat keratinocytes.

Lipoprotein and apoprotein profile of Japanese quail.

The present study delineated the lipoprotein and apoprotein distribution in Japanese quail. Quail lipoprotein was composed of three fractions: VLDL, d < 1.020; LDL, 1.020 < d < 1.081; HDL, 1.081 < d < 1.210. When animals were fed the cholesterol-free diet, HDL was the predominant form, LDL intermediate, VLDL and chylomicron were smallest in amount. Feeding of cholesterol induced a marked change in the lipoprotein profile: VLDL or chylomicron predominated over HDL and LDL. An apoprotein of 26 kD (molecular weight) was the major protein moiety comprising more than 50% of total apoprotein in the entire density range of lipoprotein class. Amino acid composition of 26 kD protein was similar to hen, rat and human apo A-I. N-Terminal 36 amino acid sequence of 26 kD protein showed 92% homology to chicken apo A-I and 11% homology to human apo A-1. The 26 kD protein did not react with the antibody raised against human apo A-I. These observation showed that the 26 kD protein was partially identical to apo A-I.

Significant correlation between interleukin 10 expression and vascularization through angiopoietin/TIE2 networks in non-small cell lung cancer.

The expression of interleukin 10 (IL-10) is correlated with clinical prognosis in non-small cell lung cancer [NSCLC (H. Hatanaka et al., ANN: ONCOL:, 11: 815--819, 2000)]. However, the effects of IL-10 expression on vascularization in NSCLC are not apparent. We examined the gene expression of IL-10/IL-10 receptor and various angiogenic/angioinhibitory factors in 95 NSCLC samples to determine the correlation between IL-10 production and vascularization. Vascular endothelial growth factor, angiopoietin [Ang (Ang-1 and Ang-2)], thrombospondin, brain-specific angiogenesis inhibitor 1, vascular endothelial growth factor receptors (KDR and flt-1), and Ang receptor (TIE2) gene expression were evaluated by reverse transcription-PCR. The cellular localization of these factors and vascularity in the cancer stroma were examined immunohistochemically. Seventy-eight (82.1%) and 93 (97.9%) of these 95 NSCLCs were positive for IL-10 and IL-10 receptor, respectively. Ang-1, Ang-2, and TIE2 gene expression was seen in 76 (97.4%), 73 (93.6%), and 78 (100%) of 78 IL-10-positive NSCLCs, respectively, and was significantly correlated with IL-10 gene expression (P < 0.0088, <0.0008, and 0.0305, respectively; Fisher's exact method). The localizations of Ang-1, Ang-2, and TIE2 were confirmed within tumor cells immunohistochemically. Vascular number and measurement area were significantly higher in the IL-10-positive NSCLCs (33.500 +/- 9.299/microm(2) and 4.742 +/- 1.287%) as compared with IL-10-negative NSCLCs (10.611 +/- 2.839/microm(2) and 0.718 +/- 0.331%; Mann-Whitney U test, P = 0.0039). The IL-10 expression did not show any significant correlation with the expression of other factors. These results suggested that tumor-produced IL-10 promotes stromal vascularization through expression of Ang-1, Ang-2, and TIE2.

High dose of Garcinia cambogia is effective in suppressing fat accumulation in developing male Zucker obese rats, but highly toxic to the testis.

We investigated the ability of Garcinia cambogia extract containing (-)-hydroxycitric acid (HCA) to suppress body fat accumulation in developing male Zucker obese (fa/fa) rats. We also examined histopathologically the safety of its high doses. Diets containing different levels of HCA (0, 10, 51, 102 and 154 mmol/kg diet) were fed to 6-week-old rats for 92 or 93 days. Each diet group was pair-fed to the 154 mmol HCA/kg diet group. Epididymal fat accumulation and histopathological changes in tissues were observed. The highest dose of HCA-containing Garcinia cambogia (154 mmol HCA/kg diet) showed significant suppression of epididymal fat accumulation in developing male Zucker obese rats, compared with the other groups. However, the diets containing 102 mmol HCA/kg diet and higher (778 and 1244 mg HCA/kg BW/d, respectively) caused potent testicular atrophy and toxicity, whereas diets containing 51 mmol HCA/kg diet (389 mg HCA/kg BW/d) or less did not. Accordingly, 51 mmol HCA/kg diet (389 mg HCA/kg BW/d) was deemed to be the no observed adverse effect level (NOAEL).

Pulp revascularization for immature replanted teeth: a case report.

Immature avulsed teeth are not usually treated with pulp revascularization because of the possibility of complications. However, this therapy has shown success in the treatment of immature teeth with periapical lesions. This report describes the case of an immature replanted tooth that was successfully treated by pulp revascularization. An 8-year-old boy suffered avulsion on his maxillary left lateral incisor. The tooth showed incomplete root development and was replanted after 30 minutes. After diagnosis, revascularization therapy was performed by irrigating the root canal and applying a calcium hydroxide paste and 2% chlorhexidine gel for 21 days. In the second session, the intracanal dressing was removed and a blood clot was stimulated up to the cervical third of the root canal. Mineral trioxide aggregate was placed as a cervical barrier at the entrance of the root canal and the crown was restored. During the follow-up period, periapical repair, apical closure and calcification in the apical 4 mm of the root canal was observed. An avulsed immature tooth replanted after a brief extra-alveolar period and maintained in a viable storage medium may be treated with revascularization.

Reestablishing Biology, Function, and Esthetics for Fractured, Immature Incisors.

A seven-year-old boy with enamel-dentin fractures on both maxillary central incisors presented to the Piracicaba Dental School-UNICAMP seven days after the trauma. At the clinical evaluation, there were no clinical signs of pulp exposure, neither tooth was mobile, and both affected teeth presented a positive response to sensitivity tests and a negative response for percussion and palpation. The radiographic examination showed an undeveloped root and opened apex for both teeth. Indirect pulp capping was performed on the left maxillary central incisor, followed by a direct restoration. After one month, the patient complained of pain in the left central incisor, which responded negatively to sensitivity testing. Pulp revascularization was performed only on this tooth and was followed for 18 months. During this period, the left maxillary central incisor did not recover sensitivity, although radiographic examination showed apical closure, a slight increase in root length, and the formation of a mineralized barrier between the root canal and sealing material. The technique achieved its goal of restoring biological aspects, function, and esthetics of traumatized teeth when using this multidisciplinary approach.

Hammerhead ribozyme specifically inhibits vascular endothelial growth factor gene expression in a human hepatocellular carcinoma cell line.

Hammerhead-type ribozymes are often utilized to suppress the expression of target genes. We evaluated the efficacy of an anti-vascular endothelial growth factor (VEGF) hammerhead-type ribozyme against GUC at exon 1 of the VEGF gene in a cell-free system (in vitro) as well as in the hepatocellular carcinoma cell line HLF (in vivo). The anti-VEGF ribozyme (alphaVRz) specifically cleaved synthetic VEGF RNA substrate, but not other triplet sequences of VEGF RNA substrate in vitro. When the alphaVRz was introduced into HLF cells, the ribozyme suppressed not only VEGF mRNA level but also that of VEGF protein. These results suggest that this ribozyme selectively inhibits VEGF gene expression in human hepatocellular carcinoma cells.

Interleukin 10 expression is correlated with thrombospondin expression and decreased vascular involvement in colon cancer.

Interleukin 10 (IL-10) is an immuno-suppressive cytokine produced by T-lymphocytes, and a regulatory molecule for angiogenesis in various cancers. We examined IL-10 gene expression in 53 colon cancer patients who underwent surgical resection. IL-10 gene expression was correlated with TSP1 and TSP2 gene expression (P=0.0049, P=0.0285). Colon cancer with IL-10 gene expression (19/53) showed significantly decreased venous involvement (P=0.0433). The mean vessel counts in the colon cancers with IL-10 gene expression were significantly lower than those without IL-10 gene expression (P<0.001). These results suggested that IL-10 stimulates angiostatic factor gene expression, and results in suppression of venous involvement.

High single-dose alternate-day corticosteroid regimens in treatment of polymyositis.

High single-dose alternate-day prednisolone therapy (ADT) was compared with daily-dose prednisolone therapy (DDT) for treatment of polymyositis. Thirty patients with polymyositis were treated with ADT for an average of 33.9 months. The combined number of improvements was 21 out of 30, a response rate of 70%. Side-effects were very rare and mild. On the other hand, 9 of 17 patients treated with DDT for an average 18.5 months improved, a response rate of 53%. The incidence of side-effects was strikingly higher than with ADT. ADT is therefore strongly advocated for treatment of polymyositis to avoid infectious complications and lessen cushingoid side-effects.

Progress of cell proliferation in striated muscle tissues during development of the mouse tongue.

The developmental stages of and places for the proliferation of tongue muscle cells have not yet been determined. To determine the stages of and places for proliferation between embryonic day (E) 9 and birth, we analyzed the expression of cyclin D1 mRNA and the immunolocalization for proliferating cell nuclear antigen (PCNA). The ratio of PCNA-positive nuclei to total nuclei (PCNA-labeling index) was obtained in the anterior, middle, and posterior regions. Cyclin D1 mRNA was highly expressed between E11 and E13, but decreased thereafter until birth. The distribution of PCNA-positive cell nuclei was consistent with that of myogenic cells in the occipital somites at E9. The PCNA-labeling index was highest at E11, then decreased until birth without a significant difference among the 3 regions. These findings suggest that some tongue muscle progenitor cells begin proliferation in the occipital somites at E9, and that the proliferation in the whole tongue region occurred most actively between E11 and E13, then decreased until birth without regional differences.

Role of surgery in hypertensive intracerebral hematoma. A comparative study of 305 nonsurgical and 154 surgical cases.

The indications for surgery in hypertensive intracerebral hematoma are still controversial. The reason for this may be: 1) lack of adequate and comparable data in conservative and surgical therapy from the same institution; 2) lack of adequate close follow-up monitoring over an extended period of time; or 3) lack of proper classification of hematomas for comparison of results from different institutions. The authors have treated 459 cases of hypertensive intracerebral hematoma between October, 1975, and July, 1983. The hematomas have been classified according to their mode of extension on computerized tomography. The long-term outcome was assessed on the basis of activity of daily living. Putaminal hematomas were classified as mild, moderate, severe, and very severe. In general, there was no significant difference in outcome between the surgical and nonsurgical cases; however, the outcome in the moderate and severe hematomas was found to be a little better for the surgical cases in some restricted areas. Thalamic and pontine hemorrhages were classified as mild, moderate, or severe. If the hematoma is localized to the thalamus or pons, and if it extends to the midbrain, there is no indication for surgery; however, in patients with moderate hematomas, the prognosis showed a variable outcome, and the indications for surgery were questionable. In cerebellar hematomas, the authors propose that even a hematoma with a diameter greater than 3 cm might show a good outcome with nonsurgical therapy.

Modulation of multidrug resistance in a cancer cell line by anti-multidrug resistance-associated protein (MRP) ribozyme.

Multidrug resistance-associated protein (MRP) is the major candidate molecule responsible for non-P-glycoprotein (PGp)-mediated multidrug resistance. We used a hammerhead anti-MRP ribozyme (alpha MRP-Rz) to inactivate MRP function in a multidrug resistant cancer cell line, KB8-5. The beta-actin promoter-driven alpha MRP-Rz sequence (pH beta/alpha MRP-Rz) was introduced into KB8-5 cells (KB8-5/alpha MRP-Rz) and we evaluated growth of the cell line. The gene expression of multidrug resistance-related molecules was estimated. Drug sensitivity was estimated by MTT assay in vitro. MRP mRNA expression was decreased in KB8-5/alpha MRP-Rz cells. The MTT assay showed increased IC50 values or resistance to doxorubicin (DOX), etoposide (VP-16) and cisplatin (CDDP) in KB8-5/alpha MRP-Rz cells. No significant differences were observed in expression of multidrug resistance gene (MDR1), thymidylate synthase, glutathione S-transferase pi or topoisomerase II alpha. The hammerhead ribozyme-mediated simple suppression of MRP mRNA expression was not sufficient to reverse multidrug resistance in the cancer cell line KB8-5.

Adenocarcinoma and multiple adenomas of the large intestine, associated with Cronkhite-Canada syndrome.

The Cronkhite-Canada syndrome is a rare non-hereditary disorder with generalised gastrointestinal polyposis, associated with ectodermal changes. We report here a case of adenocarcinoma and multiple adenomas of the large intestine associated with Cronkhite-Canada syndrome in a 61-year-old Japanese man. Histologically, the rectal tumour was composed of well-differentiated tubular adenocarcinoma, admixed with foci of adenomatous components, and associated with hyperplastic mucosa of Cronkhite-Canada syndrome. Multiple polyps, >20 polyps < or = 2.0 cm in diameter, were found near the carcinoma of the resected rectum. Histologically, superficial parts of the polyps were composed of tubular adenomas, and basal parts of the polyps were hyperplastic dilated glands. It was speculated that, in the present case, the rectal adenocarcinoma arose from mucosal hyperplasia (Cronkhite-Canada polyp)-adenoma-carcinoma pathway. This suggested that Cronkhite-Canada syndrome has definite malignant potential, although the frequency of malignant transformation is thought to be low in Cronkhite-Canada syndrome.

Pathological study on sibling autopsy cases of the late infantile form of neuronal ceroid lipofuscinosis.

We report autopsy cases of two brothers with the late infantile form of neuronal ceroid lipofuscinosis (LINCL) and examine apoptotic cell death in autopsied brains. Both patients showed psychomotor developmental delay, cerebellar ataxia, convulsions, visual disturbance and myoclonus, and they became bedridden around the age of 6-7 years. Macular changes, mimicking cherry-red spots, were observed on funduscopy, but conjunctival biopsy failed to disclose storage materials. In these cases, the autopsies demonstrated severe atrophy with neuronal loss and gliosis throughout the brain and spinal cord, except the hypothalamic neurons and motor neurons in the brain-stem and spinal cord, and autofluorescent lipofuscin-like materials of two types, fine granular deposits and coarse round bodies, were stored in the remaining neurons and glial cells, and in the epithelial cells of various visceral organs. Immunostaining for mitochondrial subunit C visualized the fine granular deposits but not the coarse round bodies. The nuclei of neurons and glia cells were stained by in situ nick end labeling, which was more pronounced in the younger case, although the expression of both bcl-2 and bcl-x was not significantly altered in these cases. It is suggested that immunohistochemistry for subunit C may be useful for diagnosis of NCL, and further investigations are necessary to clarify the relationship between LINCL and apoptosis, especially in severely affected cases.

Serum-free culture of rat keratinocytes.

Procedures for the serum-free culture of rat keratinocytes have been established. Basal cells prepared from epidermis of newborn rat were stored in liquid nitrogen and used for primary culture. Among the available media, MCDB 153, developed originally for human keratinocyte (HK) culture, was the best for the development of serum-free formulation. To grow rat keratinocytes, bovine serum albumin was arbitrarily substituted for the macromolecule supplements needed for HK culture, i.e. fetal bovine serum protein or bovine pituitary extract. Qualitative and quantitative adjustment of supplements was thereafter made to support rapid cell growth. Satisfactory cell growth was achieved in the optimized medium of MCDB 153 supplemented with growth factors and amino acids: insulin (10 micrograms/ml), hydrocortisone (0.1 microgram/ml), epidermal growth factor (25 ng/ml), calcium chloride (0.2 mM), histidine (0.23 mM), isoleucine (0.05 mM), tryptophane (0.015 mM), threonine (1.25 mM), tyrosine (0.031 mM), alanine (4.08 mM), and albumin (2 mg/ml). This optimized culture system was superior to the original HK culture condition for rapid growth of rat keratinocytes. Under our condition, cells grew as a monolayer, becoming confluent, but without stratification, and were passaged 2 to 3 times without any changes in morphology. The serum-free formulation allows us to control more accurately the concentrations of biomolecules in the medium including lipids and hormones, and therefore will be suitable for the study focusing on lipid metabolism or hormonal regulation of rat keratinocytes.