Multiplexed analysis of post-PCR fluorescence-labeled microsatellite alleles and statistical evaluation of their imbalance in brain tumors.

Detection of the loss of chromosomal regions in cancerous tissues has diagnostic and prognostic relevance, and the development of a reliable and cost-effective technique for this is clinically important. Here we present an efficient technique for quantitative detection of microsatellite alleles, using a post-PCR fluorescence-labeling procedure and multiplexed analysis. We also present a new statistical method for the interpretation of the data that permits reliable and sensitive evaluation of the allelic status of sampled DNA. A high-resolution analysis of allelic imbalance on chromosomes 1p, 10 and 19q in 28 glioma samples of various types using this method revealed that allelic imbalances are more frequent than have been reported, suggesting the diagnostic value of this method in examining the genetic profiles of gliomas.     

Dorsal horn lesion resulting from spinal root avulsion leads to the accumulation of stress-responsive proteins

The aim of this study was to demonstrate acute to subacute molecular episodes in the dorsal horn following root avulsion using immunohistochemical methods with the markers for synapses, astrocytes and such stress-responsive molecules as heat shock proteins (Hsps) and p38 MAP kinase (p38). Among them, Hsp27 was accumulated selectively in the injured substantia gelatinosa 24 h after avulsion injury. The localization of Hsp27 in astrocytes within the substantia gelatinosa was confirmed by the double immunofluorescence method using anti-Hsp27 antibody and either anti-synaptophysin antibody or anti-glutamine synthetase antibody and by immunoelectron microscopy for Hsp27. The pattern of Hsp27 expression subsequently changed from glial pattern to punctate pattern by 7 days. Immunoelectron microscopy revealed that the punctate pattern in the subacute stage corresponded to distal parts of the astrocytic processes. Hsp27 immunoreaction was decreased 21 days after root avulsion. In the distal axotomy model, Hsp27 was accumulated later in the ipsilateral dorsal horn in a punctate pattern from 7 days after the axotomy. Phosphorylation of p38 was detected in microglia in the dorsal horn following both avulsion and axotomy. Substance P was slightly decreased in the injured substantia gelatinosa in both the avulsion and axotomy models around 14-21 days. We conclude that Hsp27 is a useful marker for demonstrating dorsal horn lesions following avulsion injury and that avulsion injury may induce Hsp27 in the dorsal horn more rapidly than distal axotomy.     

Percutaneous penetration of ozagrel and the enhancement produced by saturated fatty acids

The effects of a series of fatty acids on the percutaneous penetration of ozagrel (OZ), a selective thromboxane A2 synthetase inhibitor, through rat skin and the mechanism by which fatty acids enhance the skin penetration of OZ were examined in vitro. Lauric acid, at the fatty acid: OZ molar ratio of 2 : 1, was the most potent agent as far as increasing the skin penetration was concerned, with a flux 24-fold higher than that without fatty acid. A molar ratio of 3 : 1 also produced a large enhancing effect, comparable with that of a molar ratio of 2 : 1. When the gel formulation with lauric acid (molar ratio of 2 : 1) was applied to the skin for 6 h, the amount of drug penetrating into the skin was significantly increased compared with that after the formulations without lauric acid and with capric and palmitic acids. However, lauric acid did not change the apparent partition coefficient of OZ between n-heptane and phosphate buffer (pH 7.4). The 13C-NMR spectra of OZ was also unaffected by the addition of lauric acid, indicating that a complex or ion pair with lauric acid was not formed. A possible mechanism for the enhancing effect is the increased incorporation of lauric acid with OZ into the bulk lipid phase of the stratum corneum, where the fatty acid would act as a co-penetrant enhancing passage through the stratum corneum.     

Influence of cefotiam (SCE-963, CTM) on bowel flora (author's transl)

The influence on bowel flora of CTM was studied in 5 children who were taking normal diet. 1) In the cases following no diarrhea, administration of CTM caused no significant changes in bowel flora. In the cases following diarrhea, administration of CTM caused a fall in coliform, BEP group, Lactobacillus and Peptostreptococcus. However, after the administration was discontinued, the reduced bowel flora was returned to the normal range within a few days. 2) No overgrowth of bowel flora by Pseudomonas, Clostridium difficile or Candida was observed. 3) One strain of nontoxigenic Clostridium difficile was observed in the case of following no diarrhea. 4) As for the side effects, diarrhea was observed in 2 cases. BEP group: Nonspore making anaerobic Gram positive rods of Bifidobacterium, Eubacterium, Propionibacterium.     

Cosmetic reconstruction after resection of breast cancer: use of the ELD-MC flap and EVRAM flap

Breast reconstruction following the resection of breast cancer with inadequate residual chest-wall tissue may be performed with an implant or a myocutaneous flap, such as the latissimus dorsi or a rectus abdominis. Among a variety of operative procedures, each method has advantages and disadvantages. The insertion of a silicone-bag prosthesis is the easiest method, but the prosthetic implant sometimes has complications, such as unfavorable capsular contracture formation around the implant, rupture, infection, or exposure. We therefore use an extended latissimus dorsi myocutaneous (ELD-MC) flap with some amount of surrounding subcutaneous fat from the lumbar area, and avoid the use of any implant with an MC flap. Also, for the reconstruction and correction of infraclavicular and axillary depression, we use the extended vertical rectus abdominis myocutaneous (EVRAM) flap. This method uses the skin and fat on both sides of the umbilicus as a lenticular flap vascularized by only one of the rectus abdominis muscles. The patients are satisfied with the outcome because symmetry and good breast volume can be obtained. There have been no functional or anatomical defects of the donor area. No abdominal hernia after an EVRAM flap has resulted to date. Both the ELD-MC and EVRAM flaps can be successfully used for cosmetic breast reconstruction after the resection of breast cancer.     

Usefulness of the nurse-assisted screening and psychiatric referral program

BACKGROUND: Major depression and adjustment disorders are common psychiatric disorders in patients with cancer, but they are often overlooked in clinical oncology settings. The nurse-assisted screening and psychiatric referral program (NASPRP) has been introduced in clinical practice to facilitate psychiatric treatment for major depression and adjustment disorders. This study assessed the usefulness of the NASPRP and compared it with usual practice. METHODS: The program consists of two stages. In the first stage, consecutive patients newly admitted to the Oncology/Hematology Unit are administered the Distress and Impact Thermometer (DIT) by nurses as a brief screening tool for major depression and adjustment disorders. In the second stage, the nurses recommend psychiatric referral to patients with scores above the cutoff point. Patients' records were reviewed for a 3-month period before the start of the program and during the 3-month period after the start of the program. These records were then compared. RESULTS: Of 157 patients newly admitted during the program period, 86.0% (135/157) completed the DIT and results were positive in 49.6% (67/135), but only 28.2% (19/67) accepted psychiatric referral. Ultimately, 11.5% (18/157) of patients newly admitted were diagnosed with major depression or adjustment disorders and treated by psychiatric service, a significantly higher proportion than during the preceding 3-month period, before the program was begun (2.5%; P = 0.001). CONCLUSION: The NASPRP enabled identification of major depression and adjustment disorders in patients with cancer and introduced them to psychiatric treatment. Nevertheless, there is room for improvement in the program.     

Anti-inflammatory activity of benzo(c) phenanthridine derivatives and possible mechanisms of action (author's transl)]

Of five newly synthesized benzo[c]phenanthridine derivatives tested, the two compounds, BPD-I and BPD-II were found to have potent anti-edematous activity with intraperitoneal administration to S.D. rats. BPD-I showed a marked inhibitory effect against acute inflammation such as induced rat paw edema and leucocyte emigration and protein exudation by means of CMC pouch method and capillary permeability enhancement induced by various phlogists. This compound also inhibited subacute and chronic inflammatory responses such as granuloma formation induced by croton oil or cotton pellet. The anti-inflammatory activities of this compound resembled those of hydrocortisone. The inhibitory effects of carragenan edema and capillary permeability enhancement by ATP were strikingly reduced in adrenalectomized rats suggesting involvement of the hypophysis-adrenal systems. Rat serum corticosterone level and hepatic tyrosine aminotransferase activity (TAT) were then measured after BPD-I injection. The serum corticosterone level was increased and shortly after the elevation of corticosterone, hepatic TAT levels also increased. Thus it is concluded that the corticosterone release from adrenal gland plays a role in the anti-inflammatory action of BPD-I.     

G protein-coupled receptor signaling through Gq and JNK negatively regulates neural progenitor cell migration

In the early development of the central nervous system, neural progenitor cells divide in an asymmetric manner and migrate along the radial glia cells. The radial migration is an important process for the proper lamination of the cerebral cortex. Recently, a new mode of the radial migration was found at the intermediate zone where the neural progenitor cells become multipolar and reduce the migration rate. However, the regulatory signals for the radial migration are unknown. Using the migration assay in vitro, we examined how neural progenitor cell migration is regulated. Neural progenitor cells derived from embryonic mouse telencephalon migrated on laminin-coated dishes. Endothelin (ET)-1 inhibited the neural progenitor cell migration. This ET-1 effect was blocked by BQ788, a specific inhibitor of the ETB receptor, and by the expression of a carboxyl-terminal peptide of Galpha q but not Galpha i. The expression of constitutively active mutant of Galpha q, Galpha qR183C, inhibited the migration of neural progenitor cells. Moreover, the inhibitory effect of ET-1 was suppressed by the c-Jun N-terminal kinase (JNK) inhibitor SP600125 and the expression of the JNK-binding domain of JNK-interacting protein-1, a specific inhibitor of the JNK pathway. Using the slice culture system of embryonic brain, we demonstrated that ET-1 and the constitutively active mutant of Galpha q caused the retention of the neural progenitor cells in the intermediate zone and JNK-binding domain of JNK-interacting protein-1 abrogated the effect of ET-1. These results indicated that G protein-coupled receptor signaling negatively regulates neural progenitor cell migration through Gq and JNK.