The use of ultrasound in central vascular ligation during laparoscopic right-sided colon cancer surgery: technical notes.

BACKGROUND: Complete mesocolic excision (CME) with central vascular ligation (CVL) requires the surgeon to sharply dissect the mesocolon and approach the superior mesenteric artery (SMA) and superior mesenteric vein (SMV) for ligation of the supplying vessels relating to right-sided colon cancer at their origin. Even with preoperative images, it can still be challenging to identify these structures during laparoscopic surgery because of various intraoperative conditions. The aim of this study was to assess the efficacy of intraoperative ultrasound (IOUS) for identification of blood vessels during right-sided colon cancer surgery. METHODS: We performed IOUS on 19 patients diagnosed with right-sided colon cancer at our institution, in January-October 2020. Preoperatively, a three-dimensional computed tomography (3D-CT) angiogram was obtained for the majority of patients to visualize the SMA, SMV, and their respective branches. The running position of the ileocolic artery (ICA) and right colic artery (RCA) related to the SMV and the presence of the middle colic artery were identified and compared using preoperative 3D-CT, IOUS, and intraoperative findings. RESULTS: Nineteen patients [seven men and 12 women with a mean age of 73.9 ± 8.4 years (range 58-82 years)] were studied, including some with a body mass index of > 30 kg/m2, locally advanced cancer, and severe adhesion. There were IOUSs that detected the SMA, SMV, and their tributaries in all patients. The positional relationships between the SMV and the ICA and RCA revealed by IOUS were consistent with the preoperative and intraoperative findings. CONCLUSION: IOUS is a safe, feasible, and reproducible technique that can assist in detecting the branching of the SMA and SMV during CME with CVL in laparoscopic right-sided colon cancer surgery, regardless of individual conditions.

Interleukin-6 and tumour necrosis factor-α cooperatively promote cell cycle regulators and proliferate rheumatoid arthritis fibroblast-like synovial cells.

Objective: To elucidate the roles of interleukin-6 (IL-6) and tumour necrosis factor-α (TNF-α) in cell cycle regulation and proliferation of rheumatoid arthritis fibroblast-like synovial cells (RA-FLSs). Methods: Under stimulation with IL-6/soluble interleukin-6 receptor (sIL-6R) and TNF-α, we examined the expression of cell cycle regulators [p16INK4a, p21Cip1, p27Kip1, cyclin-dependent kinase-4 (CDK4), CDK6, Cyclin D, Cyclin E, and retinoblastoma protein (pRB)] by quantitative polymerase chain reaction, Western blotting, and immunofluorescence staining. The expression of pRB, with or without 10% foetal bovine serum, was examined by Western blotting. DNA synthesis and cell viability were examined by the BrdU assay and WST-8 assay, respectively. After transfection with siRNA/p16INK4a, siRNA/p21Cip1, siRNA/p27Kip1, siRNA/CDK4, or siRNA/CDK6, RA-FLSs were successively stimulated with or without IL-6/sIL-6R or TNF-α to determine cell viability. Results: IL-6/sIL-6R significantly decreased the expression of p16INK4a, and increased p21Cip1, Cyclin E1, CYCLIN D, and pRB. TNF-α decreased the expression of CDK4, and significantly increased p27Kip1, CDK6, Cyclin E1/E2, CYCLIN D, CYCLIN E, pRB, and phosphorylated pRB (phospho-pRB). By immunofluorescence staining, CYCLIN D and phospho-pRB were simultaneously stained in the single cell. In serum-free culture, the expression of pRB was apparently decreased. DNA synthesis and cell viability were significantly increased by IL-6/sIL-6R and TNF-α. Silencing of CDK6 attenuated the cell viability induced by IL-6 and TNF-α. Conclusion: The results indicate that IL-6 and TNF-α interact with each other in regulating the cell cycle and accelerate the proliferation of RA-FLSs.

Identification of novel androgen response genes in prostate cancer cells by coupling chromatin immunoprecipitation and genomic microarray analysis.

The androgen receptor (AR) plays a key role as a transcriptional factor in prostate development and carcinogenesis. Identification of androgen-regulated genes is essential to elucidate the AR pathophysiology in prostate cancer. Here, we identified androgen target genes that are directly regulated by AR in LNCaP cells, by combining chromatin immunoprecipitation (ChIP) with tiling microarrays (ChIP-chip). ChIP-enriched or control DNAs from the cells treated with R1881 were hybridized with the ENCODE array, in which a set of regions representing approximately 1% of the whole genome. We chose 10 bona fide AR-binding sites (ARBSs) (P<1e-5) and validated their significant AR recruitment ligand dependently. Eight upregulated genes by R1881 were identified in the vicinity of the ARBSs. Among the upregulated genes, we focused on UGT1A and CDH2 as AR target genes, because the ARBSs close to these genes (in UGT1A distal promoter and CDH2 intron 1) were most significantly associated with acetylated histone H3/H4, RNA polymerase II and p160 family co-activators. Luciferase reporter constructs including those two ARBSs exhibited ligand-dependent transcriptional regulator/enhancer activities. The present study would be powerful to extend our knowledge of the diversity of androgen genetic network and steroid action in prostate cancer cells.

Peroxisome proliferator-activated receptor-gamma ligands inhibit nuclear but not cytosolic extracellular signal-regulated kinase/mitogen-activated protein kinase-regulated steps in vascular smooth muscle cell migration.

Vascular smooth muscle cell (VSMC) migration involves adhesion, locomotion, and invasion regulated by various signaling molecules, among which the extracellular signal-regulated kinase (ERK)/mitogen-activated protein kinases (MAPK) play a critical role. We have shown that the peroxisome proliferator-activated receptor-gamma (PPAR-gamma) ligands troglitazone and rosiglitazone inhibit VSMC migration downstream of ERK MAPK. The purpose of the current study was to more specifically determine which step(s) in VSMC migration are targeted by inhibition of the ERK MAPK pathway or activation of PPAR-gamma. VSMC adhesion was not affected by the ERK MAPK pathway inhibitor PD98059 or PPAR-gamma ligands. Phosphorylation and activation of myosin light chain kinase (MLCK) play important roles in cell locomotion. Platelet-derived growth factor (PDGF)-induced MLCK phosphorylation (1.7-fold) was completely blocked by PD98059 at 30 microM (p < 0.05), but not by troglitazone or rosiglitazone. PDGF-directed migration (5.8-fold) was inhibited by PD98059 (-88% at 30 microM) and the MLCK inhibitor ML9 (0.1-1 microM, -84% at 1 microM) (all p < 0.05). The transcription factor Ets-1 mediates matrix metalloproteinase induction required for tissue invasion by VSMC. PDGF (20 ng/ml) stimulated an Ets-1 protein expression (14-fold at 60 min) in VSMC, which was inhibited by PD98059 (-72% at 30 microM), troglitazone (-69% at 20 microM), and rosiglitazone (-54% at 10 microM) (all p < 0.05). Immunohistochemistry of rat aortae 2 h after balloon injury showed a dramatic upregulation of Ets-1, which was markedly inhibited in animals that had received troglitazone treatment. In contrast, phosphorylated ERK MAPK was not affected by troglitazone. These data are consistent with PPAR-gamma ligands exerting their anti-migratory effects downstream of ERK MAPK activation by blocking nuclear events, such as Ets-1 expression, required for cell invasion in response to arterial injury.

TNFalpha induces expression of transcription factors c-fos, Egr-1, and Ets-1 in vascular lesions through extracellular signal-regulated kinases 1/2.

Migration, proliferation and differentiation of vascular smooth muscle cells (VSMC) and macrophages are important pathological responses that contribute to the development and progression of vascular lesions. Cytokines such as TNFalpha are present at sites of vascular injury and regulate a variety of cellular functions of inflammatory cells and VSMC. Cell migration, proliferation and differentiation require de novo gene transcription resulting from extracellular signals being transduced to the nucleus, where multiple genes are regulated to participate in lesion formation. In VSMC and macrophages, TNFalpha induces activation of the extracellular signal-regulated kinases 1/2 (ERK 1/2), which transmit signals from the cytosol to the nucleus. Potential nuclear targets of TNFalpha-activated ERK 1/2 include the transcription factors Ets-1, Egr-1, and c-fos, which are known to regulate cellular growth, differentiation, and migration. The aim of this study was to investigate the expression of the transcription factors Ets-1, Egr-1 and c-fos in different types of vascular lesions, their regulation by TNFalpha and the role of ERK 1/2 in these signaling events. Atherosclerotic lesions from fructose-fed LDL-receptor deficient mice and neointimal lesions from rat aortae 2 weeks post balloon injury demonstrated the presence and colocalization of TNFalpha, phosphorylated and activated ERK 1/2, and transcription factors Ets-1, Egr-1 and c-fos. Neointimal lesions consisted primarily of VSMC, whereas atherosclerotic lesions predominantly contained macrophages. In cultured rat aortic VSMC, TNFalpha (100 U/ml) stimulated a rapid and transient expression of Ets-1, Egr-1 and c-fos with a maximal induction 1 h after stimulation. In cultured RAW 264.7 mouse macrophages, TNFalpha similarly induced the expression of Ets-1, Egr-1, and c-fos. Induction of these transcription factors was mediated via ERK 1/2 activation, since the ERK 1/2-pathway inhibitor PD98059 (10-30 microM) significantly inhibited their TNFalpha-induced expression. TNFalpha induced ERK 1/2 activation in both cell types. These findings underscore the importance of the ERK 1/2 pathway in the expression of TNFalpha-regulated transcription factors, which may participate in different forms of vascular lesion formation.

All kinesin superfamily protein, KIF, genes in mouse and human.

Intracellular transport is essential for morphogenesis and functioning of the cell. The kinesin superfamily proteins (KIFs) have been shown to transport membranous organelles and protein complexes in a microtubule- and ATP-dependent manner. More than 30 KIFs have been reported in mice. However, the nomenclature of KIFs has not been clearly established, resulting in various designations and redundant names for a single KIF. Here, we report the identification and classification of all KIFs in mouse and human genome transcripts. Previously unidentified murine KIFs were found by a PCR-based search. The identification of all KIFs was confirmed by a database search of the total human genome. As a result, there are a total of 45 KIFs. The nomenclature of all KIFs is presented. To understand the function of KIFs in intracellular transport in a single tissue, we focused on the brain. The expression of 38 KIFs was detected in brain tissue by Northern blotting or PCR using cDNA. The brain, mainly composed of highly differentiated and polarized cells such as neurons and glia, requires a highly complex intracellular transport system as indicated by the increased number of KIFs for their sophisticated functions. It is becoming increasingly clear that the cell uses a number of KIFs and tightly controls the direction, destination, and velocity of transportation of various important functional molecules, including mRNA. This report will set the foundation of KIF and intracellular transport research.

Teaching life-long learning skills in a fourth-year medical curriculum.

For more than twenty years, the Ruth Lilly Medical Library has been a traditional part of the Indiana University School of Medicine curriculum. Recently, following changes to the curriculum, the Library's role has evolved to include responsibility for developing and teaching a Medical Informatics rotation as part of the senior year clerkships. Heavy emphasis is placed on acquiring life-long learning skills, especially on locating and critically appraising the best clinical evidence in the medical literature. In its first four months, the rotation has been quite favorably received by both students and faculty, but will continue changing to keep pace with future curriculum alterations and new technology.

Interleukin-6 and soluble interleukin-6 receptor in the colonic mucosa of inflammatory bowel disease.

BACKGROUND: Interleukin-6 (IL-6) has multiple immunological effects on a wide variety of cells and tissues. The expression of IL-6 and IL-6 receptor (IL-6R) may be important to the pathogenesis of inflammatory bowel disease (IBD). METHODS: In the present study, we examined whether mucosal IL-6 and soluble IL-6R were associated with the pathophysiology of IBD using the colonic mucosal specimens obtained from patients with IBD. Enzyme-linked immunosorbent assay was used to measure the levels of IL-6 and sIL-6R in organ cultures of mucosal tissues and in cell cultures of fractionated mucosal cells as well as in the serum. Expression of IL-6 and IL-6R was analysed by reverse transcription-polymerase chain reaction analysis using freshly isolated lamina propria mononuclear cells (LPMC). RESULTS: The levels of IL-6 and sIL-6R in organ cultures were substantially elevated in patients with IBD, especially in those with histologically active inflammation. In contrast, considerably higher levels of sIL-6R were detected in patients with other types of colonic inflammation who were included as inflammatory controls, but elevation of IL-6 was less prominent in such patients. The positivity for expression of IL-6 and IL-6R mRNA in LPMC was in parallel with the results obtained in organ cultures. In cell cultures, mucosal macrophages were the main cell type producing both IL-6 and sIL-6R on a per cell basis and other cell fractions including colonic epithelial cells and lymphocytes produced substantially lower amounts of these molecules. The levels of IL-6 and sIL-6R in organ cultures, but not those in the serum, showed a significantly positive correlation with the degree of clinical disease activity in patients with IBD. CONCLUSIONS: Enhanced IL-6/sIL-6R-mediated immune and inflammatory responses may be implicated, at least partly, in the continuation of intestinal inflammation in patients with IBD.

Sexual selection in relation to pest-management strategies.

The application of principles derived from the sexual selection literature can assist attempts to subvert the normal mating behavior of pests. Sexual selection encompasses both intermale competition for access to females and female choice of mates. It can operate during long-range attraction and short-range courtship, as well as after copulation. We review the major aspects of sexual selection and illustrate their application to pheromonal and SIT pest-management programs. Pheromones are important both in long-range attraction and in close-range mate choice; parapheromones may be very useful in pest management because of their influence on male mating success. Sexual selection theory provides a scheme for studying the normal mating behavior of a pest species and thus determining which attributes of the mass-reared sterile males are critical to their success with wild females. We hope that our review will suggest novel ways of attacking pests as well as encourage behavioral ecologists to study pest species.

[Chemosensitivity test with endoscopic biopsy specimens].

In SDI test using tetrazolium salt, we compared MTT with XTT, which has been recently developed and is more sensitive than MTT. Further, a chemosensitivity test with endoscopic biopsy specimens using XTT was conducted. The reproducibility of the MTT assay was assessed under various conditions. The I.I of MTT assay did not vary according to type or concentration of anticancer drugs and the biopsy site of specimens. Then MTT and XTT assay were compared, revealing the results of these 2 assays were significantly correlated. Thus, XTT assay was performed with tissues from surgical specimens using biotome. Of the 16 specimens subjected to XTT assay, 6 showed an OD value of 0.100 or more, which was well within assessment. XTT assay was also performed in biopsy specimens from patients using a biotome manipulated under endoscopic guidance before operation. Three among 6 specimens showed an OD value of 0.100 or more. Our findings have demonstrated that the SDI test using XTT can be used for the chemosensitivity test, even when these specimens contain a relatively small number of cells collected using a biotome.

Change in the signal-response sequence responsible for asymmetric isolation between Drosophila planitibia and Drosophila silvestris.

Drosophila planitibia and Drosophila silvestris form a species pair that is an example of species diverged through a founder event. These species exhibit asymmetric sexual isolation, courtships between D. planitibia males and D. silvestris females being more successful than courtships between D. silvestris males and D. planitibia females. When analyzing the signal-response courtship sequence in these species, we found that D. silvestris females responded to male circling by standing or preening while D. planitibia females required further signals from the male to stop walking. The main reason for the reduced mating success rate of D. silvestris males with D. planitibia females was that the females responded to male circling by walking and the males did not proceed to the head-under-wings (HUW) position of a walking female. Another critical phase in these courtships was the HUW position in D. silvestris, where males proceeded almost immediately to wing and leg vibration. The courtships between D. planitibia male and D. silvestris female proceeded in a signal-response coordination until the male went to the HUW position, where he fanned his wings for too long a period before proceeding to wing and leg vibration. Thus, it seems that the asymmetric isolation between D. planitibia (ancestral species) and D. silvestris (derived species) is mainly due to a loss of transitions in the signal-response chain of D. silvestris. A change in the behavior of the males in the HUW position has caused further isolation between the species in both directions.

NATURAL HYBRIDIZATION BETWEEN THE SYMPATRIC HAWAIIAN SPECIES DROSOPHILA SILVESTRIS AND DROSOPHILA HETERONEURA.

Two newly formed, morphologically distinct species of Drosophila from the island of Hawaii have been found to form fertile hybrids in two areas of sympatry. Both F1 and backcross hybrids have been recognized in nature; in one case, the hybridization events extended over three years. Original hybridizations involved one or more D. silvestris females mating with D. heteroneura males. Female F1 hybrids from this cross have participated in backcrosses to D. silvestris. In any one locality, less than 2% hybrids have been found in nature. A hybrid swarm was not formed; selection appears to favor a strict maintenence of morphologies characteristic of the separate species. This result is attributed to pervasive sexual selection, which serves to preserve the syndromes of sexual characteristics that arose during past allopatric divergence. Populations of D. silvestris both within and outside the present range of D. heteroneura often display heritable variation in color patterns involving the abdomen, pleurae, legs, and wings. Genes effecting variation in these characters may be derived from genes involved in a past introgression from D. heteroneura. Independent evidence for past hybridization between these species comes from study of mitochondrial DNA. Although the inferred direction of the cross is the opposite of that observed in the recent case described here, both reciprocal crosses have been obtained experimentally in the laboratory. Accordingly, we suggest that these species may have been open to hybridization since their first sympatic encounters following their inception in allopatry. That they remain as strictly recognizable morphological entities is due both to their current partial allopatry and to the action of sexual selection in maintaining two separate major modes of efficient reproduction. There is no reason to invoke specific reinforcing selection that has imposed reproductive isolation.

[FEP and FAP combination chemotherapy in advanced gastric cancer. Research Group for Gastric Cancer].

Twenty-six patients with unresectable gastric cancer, divided into two groups, were treated with combination chemotherapy of FEP (14 patients) or FAP (12 patients). The FEP regimen performed every 4 weeks was as follows: UFT 400 mg/m2 (p.o.) everyday, etoposide 50 mg/m2 (i.v.) and CDDP 30 mg/m2 (i.v.) on days 1, 8 and 15. FAP regimen was performed every 4 weeks was: UFT 400 mg/m2 (p.o.) everyday, adriamycin 10 mg/m2 (i.v.) and CDDP 30 mg/m2 (i.v.) on days 1, 8 and 15. In FEP group, (7 males and 7 females) the average age was 64 (range 53 to 75). Two patients were in PS 1, 4 in PS 2 and 8 in PS 3. In FAP group (7 males and 5 females) the average age was 55 (range 30 to 74). One patient was in PS 1, 5 in PS 2 and 6 in PS 3. No one in either group had prior chemotherapy. Response rates in FEP and FAP groups were 28.5% (4/14) and 33.3% (4/12), respectively. The median survival periods were 4.5 months in FEP group and 6.5 months in FAP group. As for side effects, myelosuppression appeared most frequently, being followed by alopecia and nausea, although the degree of alopecia was milder in FEP than in FAP. We conclude that new combination chemotherapies of either FEP or FAP are useful for advanced gastric cancer.

Hawaiian courtship songs: evolutionary innovation in communication signals of Drosophila.

In Hawaii, flies of the genus Drosophila have undergone spectacular adaptive radiation, resulting in the evolution of more than 500 species of Drosophila that are found nowhere else on earth. This taxonomic uniqueness is reflected in behavior and morphology. Hawaiian Drosophila sing songs, as do continental Drosophila; however, the Hawaiian songs have diverged strongly in form and mechanism of production. The click-song of D. fasciculisetae's (Maui) has a carrier frequency an order of magnitude higher than those reported in familiar continental species, such as D. melanogaster (170 hertz). Drosophila fasciculisetae's song resembles a cicada's more than a fly's song. The song of D. cyrtoloma (Maui) has a complex pulse rhythm more typical of crickets than flies. The pulse song of D. silvestris (Hawaii) closely resembles that of D. melanogaster in both pulse rhythm and carrier frequency, but D. melanogaster sings by vibrating its wings, whereas D. silvestris sings through abdominal vibrations. These mechanisms are radical departures from the continental wing song mechanism and are further examples of the remarkable behavioral innovation that has occurred in the Drosophila of Hawaii during their evolutionary transit through these islands.

Sexual selection and speciation: Issues raised by Hawaiian Drosophila.

The Drosophila fauna of Hawaii is extraordinarily diverse, representing about 25% of the world's described species. The most notable characteristics which differentiate the species in Hawaii are morphological and behavioral ones used in courtship and mating. These flies are excellent model species for investigating the evolution of sexually selected traits. Hypotheses regarding the associations between species formation and mating behaviour have been formulated as a result of work on this group, leading to further empirical and theoretical research.

[An autopsy case of malignant lymphoma of the pancreas].

Case reports of malignant lymphoma of the pancreas are extremely rare. We experienced a 36-year-old man with malignant lymphoma involving the pancreas associated with obstructive jaundice, confirmed by autopsy. It is very difficult to differentiate between pancreatic carcinoma and pancreatic malignant lymphoma. Age, icterus, diffuse pancreatic enlargement, pathological findings and tumor markers may be useful in suggesting lymphoma. Trials of positive biopsy, appropriate radiotherapy and chemotherapy are necessary in patients who may have malignant lymphoma of the pancreas.

Mitochondrial DNA variability in natural populations of Hawaiian Drosophila. II. Genetic and phylogenetic relationships of natural populations of D. silvestris and D. heteroneura.

Restriction site mapping of mitochondrial DNA with 23 restriction endonucleases was used to examine the genetic and phylogenetic relationships of populations of D. silvestris and D. heteroneura from the island of Hawaii. Two morphological races of D. silvestris are known on the island of Hawaii. One has three bristle rows on the tibia of the foreleg and is found on the east side of the island. The other is found on the west side of the island and has the ancestral bristle row character of two rows on the tibia of the foreleg. All D. heteroneura have the ancestral bristle row character state. We demonstrate that mtDNA restriction site analysis can also differentiate the two D. silvestris races, and that the two bristle row D. silvestris are more closely related to D. heteroneura than they are to their three bristle row conspecifics using both distance and character state analysis. Our study (which uses six base recognition restriction endonucleases) is not sensitive enough to determine the phylogenetic relationships of populations within either of the D. silvestris lineages.