Classification of lymphoscintigraphy and relevance to surgical indication for lymphaticovenous anastomosis in upper limb lymphedema.

Upper limb lymphedema that develops after breast cancer surgery causes physical discomfort and psychological distress, and it can require both conservative and surgical treatment. Lymphaticovenous anastomosis has been reported to be an effective treatment; however the disease severity criteria that define indications for this treatment remain unclear. Here, we examined lymphoscintigraphic findings in 78 patients with secondary upper limb lymphedema and classified them into 5 major types (Type I-V) and 3 subtypes (Subtype E, L, and 0). Results revealed that this classification is related to the clinical stage scale of the International Society of Lymphology. Based on intraoperative examination findings in 20 of the 78 patients, lymphatic pressure is likely to be further elevated in Type II-V cases which are characterized by the presence of dermal back flow. Therefore, lymphaticovenous anastomosis should be considered as a treatment option for lymphedema in Type II-V cases. Furthermore, there are only limited lymph vessel sites usable for lymphaticovenous anastomosis in more severe lymphedema types [Types IV and Type V (which is characterized by dermal backflow only in the hand)]. The findings in Type IV-V cases suggest that therapeutic strategies for severe upper limb lymphedema need further consideration.

Effects of OPB-9195, anti-glycation agent, on experimental diabetic neuropathy.

BACKGROUND: Nonenzymatic glycation of neural proteins and their end-products (advanced glycation end-products, AGE) have been implicated in the pathogenesis of diabetic neuropathy. We need a development of effective ant-glycation agents for future clinical use. MATERIALS AND METHODS: We examined the effects of OPB-9195 (OPB), a new inhibitor of glycation, on the peripheral nerve structure and function in diabetic rats. Eight-week-old Wistar rats were made diabetic by streptozotocin (40 mg kg(-1), i.v.) and OPB (60 mg kg(-1) day(-1)) was given by gavage for 24 weeks. Age- and sex-matched normal Wistar rats were used for comparison. RESULTS: During the experimental period, OPB treatment did not affect the reduced body weight, elevated levels of blood glucose and glycated haemoglobin in diabetic rats. At the end of the experiment, delayed tibial motor nerve conduction velocity was significantly improved (by 60%) in treated diabetic rats, with reduction of serum AGE levels. Expression of immunoreactive AGE in the sciatic nerve was reduced in treated diabetic rats compared with those in untreated rats. Sciatic nerve (Na+, K+)-ATPase activity was also restored in treated diabetic rats. On the cross-sectioned sciatic nerves, positive cells with oxidative stress-related DNA damage, as expressed by 8-hydroxy-2'-deoxyguanosine, were less in the peripheral nerve of treated diabetic rats compared with those of untreated rats. CONCLUSION: The current study suggested that OPB is beneficial for the reduction of serum AGE and the prevention of diabetic neuropathy.

[Automated analysis for determination of PCBs in fish].

An automated analytical method for determination of polychlorinated biphenyls (PCBs) in fish was developed using supercritical fluid extraction (SFE) in combination with an automated sample preparation instrument (Prep) and GC/MS. By incorporating basic alumina with the sample in the extraction process, and optimizing the amount of carbon dioxide used, fish lipid was selectively reduced. The extract was cleaned up on a Florisil cartridge with Prep. The method was evaluated using naturally contaminated tissues and by comparison of automated analytical method results with those obtained by the conventional method. Mean recovery of PCBs from 3 kinds of fish including hairtail, mackerel and yellowtail were 69.8%, 90.2% and 81.1%, respectively. This method is less laborious and requires far less organic solvent than the conventional method, but produced comparable results.

[Dynamic MRI and tumor angiogenesis of breast cancer].

The purpose of this study was to clarify the mechanism underlying early enhanced MR images of breast cancer by dynamic MR imaging from the aspect of tumor angiogenesis. The images depicted by dynamic MR imaging of breast cancer were divided into the following two groups: a marginal strong enhancement (MSE) pattern and a variable pattern without marginal strong enhancement (non-MSE). Twenty patients with invasive ductal carcinoma (maximum diameter < 2 cm) were examined by dynamic MR imaging, and the histological materials were submitted to two-dimensional computer image analysis with immunohistochemistry and histochemistry; morphological microvessel characteristics and microvessel density were examined; and the expression of vascular endothelial growth factor (VEGF) was investigated. In the MSE cases, vessel wall irregularity of capillaries and venules in the peripheral area adjacent to the tumor correlated (p < 0.001) with the enhancement pattern, and the total microvessel density (especially of arterioles with a maximum diameter less than 50 microns) of the peripheral area adjacent to the tumor was significantly higher than that of the tumor area. However, in the non-MSE cases, total microvessel density showed no significant difference between the peripheral area adjacent to the tumor and the tumor area, whereas the capillary density of the tumor area was four times greater than that of the peripheral area adjacent to the tumor. The expression of VEGF was strongly positive for the tumor nest adjacent to the capillaries. These results suggest that the enhanced images of the MSE pattern depend on abundant blood supply from arterioles and that the images of the non-MSE pattern might be reflective of angiogenic activity including variable VEGF expression of tumor cells. Thus the mechanism underlying early dynamic MR images of breast cancer was a complex result of tumor angiogenesis and the microcirculatory environment.

Three-dimensional imaging of tumor angiogenesis.

OBJECTIVE: To three-dimensionally visualize the microvessel environment of tumor angiogenesis by confocal laser scanning microscopy (CLSM). STUDY DESIGN: To reveal underlying mechanisms of tumor angiogenesis, a 7, 12-dimethylbenz(a) anthracene-induced rat cancer model was used. For demonstrating tumor vasculature, fluorescence injection method (FITC-conjugated gelatin solution) was employed. FITC gelatin was injected into the left ventricle of the rat heart. After complete perfusion, the mammary glands were resected, fixed under ice cold conditions and subjected to immunohistochemistry (IHC) for tumor cells. The LSM-410 (Carl Zeiss, Jena, Germany) was employed on thick sections (300-2,000 microns) to elucidate detailed microvessel networks (MVN) and tumor cells. RESULTS: Tumor vasculature on thick sections was clearly detected by CLSM at the maximum focus depth of 2,000 microns. Three-dimensional (3-D), reconstructed images of normal mammary glands showed regular and linear MVN. In DMBA-induced mammary cancer, vascular density of MVN was markedly increased and showed an anastomosing, irregular MVN pattern. Furthermore, focal segmentation and tortuous, branching patterns of microvessels were also seen. CONCLUSION: Application of the fluorescence injection method and IHC using CLSM was very useful for studying the 3-D relationship between tumor angiogenesis and neoplastic epithelial changes. These results suggest that application of this technique is ideal for studying 3-D imaging of tumor angiogenesis.

A new approach to three-dimensional reconstructed imaging of hormone-secreting cells and their microvessel environments in rat pituitary glands by confocal laser scanning microscopy.

There has been considerable interest in the relationship between hormone- secreting endocrine cells and their microvessels in human pituitary gland. However, microcirculatory networks have rarely been studied in three dimensions (3D). This study was designed to visualize and to reveal the relationship between hormone-secreting endocrine cells and their microvessel environment in 3D, using rat pituitary glands under various (hyper/hypo) experimental conditions by confocal laser scanning microscopy (CLSM). Female adult Wistar rats were used after bilateral adrenalectomy or ACTH administration for 2 weeks. Clear 3D reconstructed images of ACTH cells, the microvessel network and counterstained nuclei were obtained at a maximal focus depth of 1 mm by CLSM without any background noise. In the hyperfunctional state, slender cytoplasmic processes of hypertrophic stellate ACTH cells frequently extended to the microvessels. In the hypofunctional state, ACTH cells appeared atrophic and round with scanty cytoplasm, and cytoplasmic adhesions to microvessel network patterns were inconspicuous. Therefore, 3D reconstructed imaging by CLSM is a useful technique with which to investigate the microvessel environment of hormone-secreting cells and has the potential to reveal dynamic hormone-secreting pathways.

Transcription of some PHO genes in Saccharomyces cerevisiae is regulated by spt7p.

Spt7p is a new global transcription factor in Saccharomyces cerevisiae(Gansheroff et al., 1995). We report here that the activities of high affinity phosphate transport and acid phosphatase in particular were decreased in a spt7 null mutant. Northern blot experiments revealed that transcription of the PHO84 and PHO5 genes was impaired in this mutant; expression of the PHO regulatory genes, PHO4 and PHO2, was normal. Spt7p is thus linked with expression of several structural genes of the PHO regulon in yeast.

Potassium/proton antiport system is dispensable for growth of Enterococcus hirae at low pH.

An energy-dependent K+/H+ antiport system is found in Enterococcus hirae ATCC 9790 cultured in a standard complex medium (Y. Kakinuma, and K. Igarashi, J. Biol. Chem. 263:14166-14170, 1988). We have now found that the activity of this antiport system was totally missing in cells cultured in a defined medium. In this defined medium, E. hirae did not grow well at pH near 9, but grew normally at pH below 7.5. This antiport system is important at high pH but dispensable at lower pH for ion homeostasis of this bacterium.

Suppression of transforming growth factor beta and vascular endothelial growth factor in diabetic nephropathy in rats by a novel advanced glycation end product inhibitor, OPB-9195.

AIMS/HYPOTHESIS: Advanced glycation end products (AGEs) participate in the pathogenesis of diabetic nephropathy. We reported earlier that OPB-9195, a synthetic thiazolidine derivative and novel inhibitor of advanced glycation, prevented progression of diabetic glomerulosclerosis by lowering serum concentrations of advanced glycation end products and reducing their deposition in the glomeruli. Here, we examined their contribution and that of growth factors, such as transforming growth factor-beta (TGF-beta) and vascular endothelial growth factor (VEGF), to the progression of diabetic nephropathy. We also investigated the expression of type IV collagen in the kidneys of Otsuka-Long-Evans-Tokushima-Fatty (OLETF) rats, a Type II (non-insulin-dependent) diabetes mellitus model, after treatment with OPB-9195. METHODS: Using northern blots and immunohistochemical techniques, we determined the renal expression of TGF-beta and type IV collagen mRNAs and proteins in OLETF rats. We also examined OPB-9195's effects on renal expression of VEGF mRNA and protein. RESULTS: Concomitant increases in TGF-beta and type IV collagen expression were observed at each point in time in OLETF rats not given OPB-9195. In contrast, OPB-9195 treatment greatly suppressed the renal expression of TGF-beta, VEGF and type IV collagen mRNAs and proteins to that seen in non-diabetic rats. CONCLUSION/INTERPRETATION: Since OPB-9195, an AGE-inhibitor, prevented the progression of diabetic nephropathy by blocking type IV collagen production and suppressing overproduction of two growth factors, TGF-beta and VEGF, in diabetic rats, this compound warrants further investigation.

Involvement of Spt7p in vacuolar polyphosphate level of Saccharomyces cerevisiae.

Saccharomyces cerevisiae became less sensitive to nickel by a defect of the SPT7 gene encoding a transcription factor. Initial rate of nickel uptake by whole cells of a SPT7-negative mutant FY963 was nearly equal to that of the parent strain FY61, and FY963 accumulated nickel about 1.7-fold of the value of FY61 when cultured in medium containing 0.1 mM NiCl2; most of which was sequestered into vacuoles. The pH gradient-driven nickel uptake by vacuolar membrane vesicles was not altered in FY963, but the amount of polyphosphate in vacuoles was highly elevated. Involvement of Spt7p in nickel detoxification through regulation of vacuolar polyphosphate level in S. cerevisiae was discussed.

Progression of nephropathy in spontaneous diabetic rats is prevented by OPB-9195, a novel inhibitor of advanced glycation.

Levels of tissue advanced glycation end products (AGEs) that result from nonenzymatic reactions of glucose and proteins are high in both diabetic and aging people. Irreversible AGE formation is based on increases in AGE-derived protein-to-protein cross-linking and is considered to be a factor contributing to the complications of diabetes. A novel inhibitor of advanced glycation, OPB-9195, belongs to a group of thiazolidine derivatives, known as hypoglycemic drugs; however, they do not lower blood glucose levels. We did studies to determine if OPB-9195 would prevent the progression of nephropathy in spontaneous diabetic rats. In vitro inhibitory effects of OPB-9195 on AGE formation and AGE-derived cross-linking were examined by enzyme-linked immunosorbent assay (ELISA) and SDS-PAGE, respectively. Otsuka-Long-Evans-Tokushima-Fatty (OLETF) rats, a model of NIDDM, were used to evaluate the therapeutic effect of OPB-9195. Light microscopic findings by periodic acid-Schiff (PAS) staining, the extent of AGE accumulation detected by immunohistochemical staining in the kidneys, the levels of serum AGEs by AGE-specific ELISA, and urinary albumin excretion were examined. OPB-9195 effectively inhibited both AGE-derived cross-linking and the formation of AGEs, in a dose-dependent manner in vitro. In addition, the administration of OPB-9195 prevented the progression of glomerular sclerosis and AGE deposition in glomeruli. Elevation of circulating AGE levels and urinary albumin excretion were dramatically prevented in rats, even at 56 weeks of age and with persistent hyperglycemia. We concluded that a novel thiazolidine derivative, OPB-9195, prevented the progression of diabetic glomerular sclerosis in OLETF rats by lowering serum levels of AGEs and attenuating AGE deposition in the glomeruli.

Inflammatory Carcinoma of the Breast: Characteristic Findings of MR Imaging.

Inflammatory breast carcinoma (IBC) Shows a unique clinical appearance and has an extremely poor prognosis. Although immediate intensive therapy has been proposed, diagnosis of this disease tends to be delayed as its clinical features can be confused with acute mastitis. The conventional imaging modalities including mammography and ultrasonography are of limited value in the diagnosis of IBC, as it is difficult to delineate specific findings of the swollen dense breast. Recently, magnetic resonance (MR) imaging has been widely applied to breast diseases. One of the excellent features of this modality is its diagnostic ability in dense breasts. However, few trials to evaluate the capability of this new modality for IBC have been documented. In recent years, we found that a characteristic feature in 5 cases of IBC was a strong signal on T2-weighted images (T2WI) of MR imaging at the retromammary and subcutaneous area. Pathological features of the retromammary area showed marked interstitial edema and focal lymphatic involvement by tumor cells. These characteristic images obtained by MR imaging may be suggestive of inflammatroy breast carcinoma. Furthermore, subtracted dynamic MR imaging indicated the site of the tumor. Therefore, the application of MR imaging for swollen breasts would assist in the immediate diagnosis of IBC and would contribute to appropriate and timely therapy.

Dynamic MR Imaging and Tumor Angiogenesis in DMBA-Induced Rat Breast Cancer: Three Dimensional (3D) Reconstructed Image Analysis of Tumor Microvessels by Confocal Laser Scanning Microscopy.

We investigated the correlation between dynamic magnetic resonance imaging (MRI) findings in breast cancer and tumor angiogenesis in a dimethylbenz(a)anthracene(DMBA)-induced rat breast cancer model. In this study, we clearly demonstrated the three-dimensional (3D) architecture of tumor microvessel networks(MN)by confocal laser scanning microscopy (CLSM)and also investigated the hyperpermeability of tumor microvessels. Dynamic MRI findings were closely related to tumor angiogenesis. Three-dimensional reconstructed image analysis by CLSM revealed that the depicted images were dependent on microvessel density as well as microvessel permeability. It should be emphasized that dynamic MRI may have the potential to evaluate the tumor angiogenic activity in human breast carcinoma.

Synthesis and structure-activity relationships of cephalosporins, 2-isocephems, and 2-oxaisocephems with C-3' or C-7 catechol or related aromatics.

A series of cephalosporins, 2-isocephems, and 2-oxaisocephems with C-3' catechol-containing (pyridinium-4-thio)methyl groups and 2-isocephems with C-7 catechol related aromatics have been prepared and evaluated for antimicrobial activity. It turns out that these compounds have highly potent activity against Gram-negative bacteria, especially resistant pathogens such as Pseudomonas aeruginosa. The most active compound of the series was (6S,7S)-7-[2-(2-aminothiazol-4-yl)-2-[(Z)-[(1,5-dihydroxy-4-pyr idon-2-yl) methoxy]imino]acetamido]-3-[[[(4-methyl-5-carboxymethyl)thiazol-2- yl] thio]methyl]-8-oxo-1-aza-4-thiabicyclo [4.2.0] oct-2-ene-2-carboxylic acid which exhibited potent in vitro activity against clinically isolated P, aeruginosa and Acinetobacter baumanii which is also resistant to many anti-infectives, and good in vivo efficacy against clinically isolated P aeruginosa.

Alterations in macrophages after exposure to root canal filling materials.

The process of engulfment of overextended root canal filling materials was investigated in rat peritoneal macrophages in vitro. Root canal filling materials tested were Finapec APC, Sealapex, Canals-N, and Canals. The phagocytosis rate of macrophages for the Finapec APC was significantly (p < 0.05) higher than that for other three root canal filling materials. That for Canals was the lowest. About 95% of the cells exposed to Finapec APC were viable at 60 and 120 min. For Canals the percentage was 74% and 63% at 60 and 120 min, respectively. Ruffle formation in macrophages was observed by scanning electron microscopy after exposure to Finapec APC for 60 or 120 min. Many vacuoles were observed in macrophages exposed to Canals for 60 min. It was concluded that the phagocytic rate of macrophages for Canals that showed a strong cytotoxicity was lowest and that the rate for Finapec APC that showed a low cytotoxicity was the highest.

Three-dimensional structure of a DNA repair enzyme, 3-methyladenine DNA glycosylase II, from Escherichia coli.

The three-dimensional structure of Escherichia coli 3-methyladenine DNA glycosylase II, which removes numerous alkylated bases from DNA, was solved at 2.3 A resolution. The enzyme consists of three domains: one alpha + beta fold domain with a similarity to one-half of the eukaryotic TATA box-binding protein, and two all alpha-helical domains similar to those of Escherichia coli endonuclease III with combined N-glycosylase/abasic lyase activity. Mutagenesis and model-building studies suggest that the active site is located in a cleft between the two helical domains and that the enzyme flips the target base out of the DNA duplex into the active-site cleft. The structure of the active site implies broad substrate specificity and simple N-glycosylase activity.

Synthesis and biological evaluation of a series of new parenteral optically active 3-[[(N-alkylpyridinium-4'-yl)thio]methyl]-2-oxaisocephems.

The preparation and biological evaluation of a series of 7-[2-(2-aminothiazol-4-yl)-2-(Z)-[(cyclopentyloxy)imino]acetamido] optically active 2-oxaisocephems, substituted at the 3-position with [(N-alkylpyridinium-4'-yl)thio]methyl groups, are described. The resulting family of parenteral compounds displays a broad spectrum of in vitro antibacterial activity. These compounds exhibit increased activity against Gram-positive organisms including methicillin-resistant Staphylococcus aureus and Enterococcus faecalis which are resistant to most cephalosporins with a similar level of Gram-negative activity to that of the third-generation antibiotics. In vivo efficacy of new antibacterial agents in this investigation is excellent against both Gram-positive and Gram-negative bacteria as compared with reference compounds. The in vitro and in vivo antimicrobial activity and the structure-activity relationships are presented.

Synthesis and antibacterial activities of 2-oxaisocephems.

A series of 2-oxaisocephems with a thio-substituted methyl group at the 3-position and a 2-aminothiazol-4-yl moiety at the 7-position was synthesized via benzyl 3-acetyloxymethyl-7-azido-8- oxo-1-aza-4-oxabicyclo[4.2.0]oct-2-ene-2-carboxylate (2), derived from benzyl acetoacetate (1). The new 2-oxaisocephems were tested for antibacterial activities. Among them, the derivatives having a [2-(2-aminothiazol-4-yl)-2- (Z)-cyclopentyloxyimino]acetamido group at the 7-position characteristically showed potent activities against gram-positive bacteria including methicillin-resistant Staphylococcus aureus (MRSA) and Enterococcus faecalis as compared with cefuzonam and cefmenoxime, which are third-generation cephalosporins.

Retro runner with ischial tuberosity enthesopathy.

In reviewing the literature, no studies were found reporting the use of retro running on flat and hilly terrain, which elicited enthesopathy (stress reaction) at the ischial tuberosity. Therefore, this case study of an atypical enthesopathy condition warrants careful scrutiny in order to generate future research. This case study describes the clinical management of a female runner with bilateral patellofemoral pain who self-initiated a program of backward running and stationary bicycling after reading an article about retro running in a runners' magazine. She subsequently developed ischial tuberosity enthesopathy verified by scintigraphy (bone scan). Her symptoms gradually resolved with physical therapy intervention. Eventually, she was able to forward jog 2 miles on flat surfaces without complaint of pain but did not resume retro running. This case not only suggests the need for further research in retro running kinetics and kinematics but highlights the proactive role health professionals must assume in injury prevention.