Length of laparotomy incision and surgical stress assessed by serum IL-6 level

BACKGROUND: Because surgical stress is thought to have an effect on morbidity, mortality, and remnant tumour progression after surgery, diminishing surgical stress is important. The purpose of this study was to assess in a murine model whether the length and type of laparotomy incision influence surgical stress. METHODS: Serum IL-6 concentrations were measured sequentionally in 220 male BALB/c mice who were assigned to different basic laparotomies, (1-cm versus 2-cm versus 3-cm laparotomy with or without caecal resection), other types of laparotomy (3-cm, 1-cm x 3, 3-cm transverse, 3-cm laparotomy with rapid closure), or 3-cm skin incision with or without laparotomy. The serum level of IL-6 was measured by ELISA. RESULTS: Serum IL-6 levels at 3 and 6h after surgery were significantly higher in the 3-cm laparotomy group (1,680+/-802pg/ml and 1,066+/-507pg/ml, respectively), than in the 1-cm laparotomy group (797+/-427pg/ml and 515+/-212pg/ml, respectively). When caecal resection was added, the serum IL-6 level at 6h was significantly higher in the 3-cm laparotomy group (2,844+/-134pg/ml) than in the 1-cm laparotomy group (2,200+/-379pg/ml). Although the type of laparotomy incision was not associated with the serum level of IL-6, the serum IL-6 level after midline skin incision without laparotomy (245+/-142pg/ml) was significantly lower than that after 3-cm laparotomy (1,680+/-802pg/ml). CONCLUSIONS: The length of laparotomy incision was correlated with the serum level of IL-6 in a murine model. The surgical stress related to abdominal procedures might be decreased when laparotomy wounds are kept as small as possible.     

Laparoscopic surgery for gynecologic malignancy

The gynecologic laparoscopic surgery that has developed in the reproductive field is performed for most cases of benign disease. Introduction of laparoscopic surgery for a gynecologic malignancy is relatively late in comparison with other surgical regions and does not qualify for insurance. An operation for a gynecologic malignancy is classified roughly into a surgery of internal pelvic organs and lymph nodes. The techniques which extensive technology is pursued involve radical hysterectomy and paraaortic lymph node excision. In the surgeons who performed many laparoscopic surgeries, it seems that a surgical procedure other than these can be performed at present. The experienced laparoscopic techniques and the particular knowledge of pelvic anatomy were necessary to perform laparoscopic surgery for gynecologic malignancy.     

Dendritic cell immunotherapy with poly(D,L-2,4-diaminobutyric acid)-mediated intratumoral delivery of the interleukin-12 gene suppresses tumor growth significantly

A conventional DC-based immunotherapy has been tested clinically for treatment of patients with advanced cancer but requires modification to further improve the clinical results. In this study, we evaluated the in vivo antitumor effects of DC therapy, non-viral-mediated IL-12 gene therapy, and a combination of the two in a murine bilateral subcutaneous tumor model. DC therapy alone and IL-12 gene therapy alone suppressed tumor growth at the injected sites. However, the antitumor effect on the distant contralateral tumor was insufficient. When DC therapy and IL-12 gene therapy were carried out simultaneously, tumor growth was significantly suppressed bilaterally (P < 0.001). Cytolytic activity was augmented significantly in mice given the combination treatment compared to in mice treated with either DC or IL-12 gene therapy alone (P < 0.05). Microvessel density of both tumors was significantly lower in mice subjected to the combination therapy than in mice treated otherwise (P < 0.05). Furthermore, no side-effects were observed in the treated mice. DC therapy combined with non-viral-mediated intratumoral IL-12 gene delivery has a synergistic antitumor effect not only on targeted tumors but also on contralateral distant tumors and may be of great potential as a therapeutic treatment for patients with advanced cancer.     

The epidemiology of developmental dysplasia of the hip in Japan: Findings from a nationwide multi-center survey

Background

It has been reported that the national incidence of developmental dysplasia of the hip (DDH) has decreased in Japan. This is because of prevention activities after birth since around 1970. However, cases of late-diagnosed DDH have still been noted in some children's hospitals. There has been no recent survey of DDH in Japan. The purpose of this study was to investigate the current epidemiology of DDH using a comprehensive nationwide survey.

Expression of monocyte chemoattractant protein 1 (MCP-1) in adult periodontal disease: increased monocyte chemotactic activity in crevicular fluids and induction of MCP-1 expression in gingival tissues.

The present study shows that monocyte chemotactic activity in crevicular fluids increases with severity of the disease and that a monocyte chemoattractant, monocyte chemoattractant protein 1 (MCP-1), is expressed as the predominant cytokine of gingival tissues and their fibroblasts treated with Porphyromonas (Bacteroides) gingivalis lipopolysaccharide (P-LPS). High monocyte chemotactic activity in the crevicular fluids was neutralized significantly by antiserum specific for the JE/MCP-1 protein. Marked expression of the MCP-1 gene was observed in the gingival tissues of all adult periodontal patients tested, but not in those of healthy subjects. Monocyte chemotactic activity was observed in culture supernatants of human normal gingival tissues treated with P-LPS, and the chemotactic activity increased in a dose-related manner. Expression of MCP-1 in P-LPS-treated human gingival fibroblasts was further examined. P-LPS induced the MCP-1 gene expression in a dose- and treatment time-dependent manner. The MCP-1 gene product in the culture supernatant was detected as two forms with molecular masses of 11,000 and 15,000 Da by immunoprecipitation with the specific antiserum. The MCP-1 gene expression was induced in the fibroblasts treated with interleukin-1 beta and tumor necrosis factor alpha, but not with interleukin-6. These results suggest that gingival fibroblasts can participate in monocyte recruitment in gingival tissues of adult periodontal patients via the MCP-1 gene product and that MCP-1 plays an important role in the inflammatory reaction in the disease.     

Effective production of a human monoclonal antibody against tetanus toxoid by selection of high productivity clones of a heterohybridoma

A mouse.human-human heterohybridoma, N12-16.63, has been described which produces an anti-tetanus toxoid human monoclonal antibody (MoAb). A clone, N12-16.63.49.19, which produces eight times as much MoAb as that produced by the original cell line, was selected by repeating the recloning and selection twice. Two clones, N12-16.63.49.19.69 and N12-16.63.49.19.127, further selected from this clone produced almost 20 times more than that produced by the original cell line. Though the production of MoAb by these clones gradually decreased with repeating transfers, they still produced a large amount of human MoAb even after 3 months of transfer. Human MoAb (IgM) was isolated from the culture supernatants of the original and high productivity clones and the products were confirmed to be identical. Human MoAb was effectively produced by batch culture on the 20 liter scale or a perfusion culture on the 1 liter scale using these high productivity clones.     

Ca(2+)-dependent inhibition of inwardly rectifying K(+) channel in opossum kidney cells

The effect of intracellular Ca(2+) on the activity of the inwardly rectifying ATP-regulated K(+) channel with an inward conductance of about 90 pS was examined by using the patch-clamp technique in opossum kidney proximal tubule (OKP) cells. The activity of the inwardly rectifying K(+) channel rapidly declined with an application of ionomycin (1 microM) in the presence of 10(-6) M Ca(2+) in cell-attached patches. The application of 10 microM phorbor-12-myristate-acetate (PMA) with 10(-6) M Ca(2+) reduced the K(+) channel activity. Although the channel activity was not influenced by an increase of bath Ca(2+) from 10(-7.5) to 10(-6) M, the activity was inhibited by protein kinase C (PKC, 1 U/ml) with 10(-6) M Ca(2+) in inside-out patches. The inhibitory effect of Ca(2+) with ionomycin on the channel activity was diminished by the pretreatment with a specific PKC inhibitor, GF 109203X (5 microM), in cell-attached patches. By contrast, the application of Ca(2+)/calmodulin kinase II (CaMK II, 300 pM) dramatically increased this channel activity in inside-out patches. In cell-attached patches, the addition of both GF 109203X and cyclospolin A (5 microM), a potent inhibitor of protein phosphatase 2B (calcineurin), instead stimulated the K(+) channel activity with ionomycin and 10(-6) M Ca(2+). The addition of protein phosphatase 2B (calcineurin) (2 U/ml) to the bath with calmodulin (1 microM) and Ni(2+) (10 microM) to stimulate calcineurin inhibited the channel activity in inside-out patches. Furthermore, the inhibitory effect of PKC or calcineurin on this channel activity was abolished by a removal of Ca(2+) from bath solution. These results suggest that Ca(2+)-dependent inhibitory effect on the inwardly rectifying K(+) channel in OKP cells was mainly mediated by Ca(2+)-PKC-mediated phosphorylation, and that the Ca(2+)-calmodulin-dependent phosphorylation process may be counterbalanced by the Ca(2+)-calmodulin-dependent dephosphorylation process.     

Astroglial expression of ceramide in Alzheimer's disease brains: a role during neuronal apoptosis

Accumulating evidences indicate that ceramide is closely involved in apoptotic cell death in neurodegenerative disorders and aging. We examined ceramide levels in the cerebrospinal fluid (CSF) or brain tissues from patients with neurodegenerative disorders and the mechanism of how intra- and extracellular ceramide was regulated during neuronal apoptosis. We screened the ceramide levels in the CSF of patients with neurodegenerative disorders, and found that ceramide was significantly increased in patients with Alzheimer's disease (AD) than in patients with age-matched amyotrophic lateral sclerosis (ALS) and other neurological controls. With immunohistochemistry in AD brains, ceramide was aberrantly expressed in astroglia in the frontal cortices, but not detected in ALS and control brains. To explore for the regulation of ceramide in astroglia in Alzheimer's disease brains, we examined the metabolism of ceramide during neuronal apoptosis. In retinoic acid (RA)-induced neuronal apoptosis, RA slightly increased de novo synthesis of ceramide, but interestingly, RA dramatically inhibited conversion of [14C] ceramide to glucosylceramide (GlcCer), suggesting that the increase of ceramide mass is mainly due to inhibition of the ceramide-metabolizing enzyme GlcCer synthase. In addition, a significant increase of the [14C] ceramide level in the culture medium was detected by chasing and turnover experiments without alteration of extracellular [14C] sphingomyelin levels. A 2.5-fold increase of ceramide mass in the supernatant was also detected after 48 h of treatment with RA. These results suggest a regulatory mechanism of intracellular ceramide through inhibition of GlcCer synthase and a possible role of ceramide as an extracellular/intercellular mediator for neuronal apoptosis. The increased ceramide level in the CSF from AD patients, which may be derived from astroglia, raises a possibility of neuronal apoptosis by the response to intercellular ceramide in AD.     

Catalytic effects of cationic polymeric liposomes on the alkaline hydrolysis of aniomic phenyl esters

Alkaline hydrolyses of anionic phenyl esters such as 4-acetoxy-3-nitrobenzoic acid and 4-butyryloxy-3-nitrobenzoic acid were examined in the presence of cationic and polymeric liposomes, liposomes of low molecular weight compounds, and micelles. All the additives accelerate the reaction due to the hydrophobic interaction between substrates and additives and the electrostatic interaction both between substrates and additives and between OH^? and additives. In the Arrhenius plots of the reactions catalyzed by the liposomes, discontinuous regions were observed due to the phase transition of liposomes from the gel state to the liquid crystal state. Activation parameters ΔH^≠, ΔS^≠ and ΔV^≠ for these reaction systems were evaluated. Both ΔS^≠ and ΔV^≠ values increase upon the addition of cationic liposomes and micelles. These results were attributed to the desolvation of the activated complex by the strong electrostatic affinity to the cationic colloidal additives.