Quantification and visualization of cellular NAD(P)H in young and aged female facial skin with in vivo two-photon tomography.

BACKGROUND: In vivo two-photon tomography is a novel noninvasive three-dimensional optical skin imaging technology with subcellular resolution which enables the sensitive detection of endogenous fluorophores. One of these fluorophores, NAD(P)H (a coenzyme which plays an important role in the release of free energy during glycolysis, and influences filaggrin and lipid synthesis), can be selectively detected in keratinocytes (granular cells) with two-photon tomography. OBJECTIVES: To quantify NAD(P)H levels in subsurface human facial skin in vivo as a measure to determine if there are changes with age. METHODS: A total of 80 healthy Asian females were enrolled in this study, aged 21-68 years. Measurements were performed on facial skin using in vivo two-photon tomography (DermaInspect/MPTflex™, JenLab GmbH, Jena, Germany). The laser beam scans a skin field of interest in pulses, focused at a depth to reach the granular layer. The near-infrared laser pulses excite the endogenous fluorophores NAD(P)H. Image processing was performed to obtain high-resolution autofluorescence images (optical biopsies) and to quantify the fluorescent grey scale to determine NAD(P)H levels. Additional skin surface measures taken were hydration (corneometer), elasticity (cutometer) and wrinkles (image capture and analysis). RESULTS: Statistically significant changes in all measured parameters as a function of age were observed. Most importantly, the mean fluorescent grey scale values for NAD(P)H in the youngest group studied (women in their 20s) was 38.8 (SD ± 12.39), while that of the oldest group studied (women in their 60s) was 32.7 (SD ± 12.47). These NAD(P)H levels are statistically significantly different (P = 0.0078). CONCLUSIONS: The level of NAD(P)H in the epidermis is significantly greater in younger vs. older skin in vivo. This likely reflects decreased production and/or increased degradation of NAD(P)H in older skin, possibly as a result of chronological ageing and environmental damage (e.g. photodamage). NAD(P)H levels in epidermal skin may be a useful biomarker of skin ageing in vivo. It is also likely that maintaining NAD(P)H production is a useful approach to maintaining good skin condition and caring for ageing skin.

A novel branching pattern of the superior mesenteric artery found in the bullfrog (Rana catesbeiana) amphibian.

The branching and distribution patterns of the superior mesenteric artery were studied in 10 adult bullfrogs (Rana catesbeiana) after injection of coloured latex solution into the vasculature. The abdominal digestive organs in the bullfrog were mainly supplied by the coeliac artery and the superior mesenteric artery, both of which arose as a common trunk, the coeliacomesenteric artery, from the abdominal aorta. The coeliac artery supplied the stomach, liver, gallbladder and the pancreas, whereas the first branch of the superior mesenteric artery was the splenic artery with other branches supplying the greater part of intestine. The apex of the intestinal loop was defined as the region supplied by the trunk of the superior mesenteric artery, and its intestinal branches constituted a 'nested formation' which had the following characteristics. (1) The branches of the trunk were distributed to both sides of the apex, and the distribution regions of younger branches were located more distant from the apex than those of older branches. (2) Two branches directed towards both sides of the trunk frequently made a common stem arising from the trunk. The second branch of the superior mesenteric artery constituted a secondary trunk and its distribution region could be defined as a secondary apex, since 1 of its branches also constituted a nested formation which was distributed to both sides of the primary and secondary apices. The intestinal branches of the superior mesenteric artery were divided into 4 types on the basis of their pattern of branching and course. It is suggested that the nested formation of the superior mesenteric artery in the bullfrog is a remnant of the vascular pattern of the tadpole, which possesses a double spiral mode of intestinal convolution, probably supplied by arteries with the nested formation in a latent form.

A genetic analysis of hydrologically dispersed seeds of Hibiscus moscheutos (Malvaceae).

The dispersal of floating seeds in wetland habitats should influence the genetic characteristics of plant metapopulations. We examined gene flow of a hydrochorous wetland macrophyte, Hibiscus moscheutos L. (Malvaceae), by analyzing allozyme variation in current-year floating-seed populations. The genetic composition of floating seeds was compared to the genetic composition of established populations of H. moscheutos that had been previously analyzed in the same areas. The F statistics demonstrated that genetic structuring among floating-seed populations was weak or absent, indicating that seeds from source populations were thoroughly mixed. Floating-seed populations had an excess of homozygotes, a different situation than had previously been found in established populations. The exchange of seeds was greatest among H. moscheutos populations that were adjacent to a tidal stream. We conclude that populations adjacent to the tidal streams are part of a metapopulation that serves as a reserve of genetic variation in the system. Although established populations of H. moscheutos that are not close to the tidal stream are relatively isolated genetically, we found evidence that they also contribute to the floating-seed populations within the estuary.

Enhanced tumorigenicity caused by truncation of the extracellular domain of GP125/CD98 heavy chain.

GP125/CD98 is a heterodimeric 125-kDa glycoprotein, which consists of an 85-kDa heavy chain (hc) and a 40-kDa light chain (lc), and is strongly expressed on the cell surface of various tumor cells, irrespective of their tissue of origin. We have recently demonstrated that overexpression of the CD98hc cDNA causes malignant transformation of NIH3T3 cells. To investigate the function of the extracellular domain of CD98hc in cell proliferation and malignant transformation, we established two NIH3T3-derived clones transfected with human truncated CD98hc cDNAs, and compared their characteristics with parental NIH3T3 and clones transfected with full-length CD98hc cDNA. Truncated as well as full-length CD98hc-transfected clones grew to a higher saturation density than control cells. Efficiency of colony formation in soft agar was augmented in all CD98hc-transfected clones, and the degrees of augmented colony formation of the transfectants expressing full-length CD98hc of 529 a.a. or truncated CD98hc of 418 a.a. were reduced by anti-human CD98hc antibodies, while that of the transfectant expressing truncated CD98hc of 237 a.a. lacking the epitopes recognized by anti-human CD98hc antibodies was not affected by the addition of antibodies. CD98hc-transfected clones developed tumors in athymic mice, and tumor growth of truncated CD98hc-transfected clones was faster than that of full-length CD98hc-transfected clones.

Malignant transformation of NIH3T3 cells by overexpression of early lymphocyte activation antigen CD98.

CD98, which forms a heterodimer of relative molecular mass (M(r)) 125, 000, was originally identified as an early T cell activation antigen. It consists of a heavy chain of M(r) 85,000 that bears the CD98 epitope and a light chain of M(r) 40, 000. CD98 is strongly expressed on the surface of activated lymphocytes and various tumor cells irrespective of tissue origins. To investigate the participation of CD98 in cellular proliferation and malignant transformation, we established and characterized human CD98-transfected NIH3T3 clones. Although the doubling times of the control cells and CD98-transfected clones were almost the same, CD98-transfected clones grew to a higher saturation density than control cells. Effciency of colony formation in soft agar was augmented in CD98-transfected clones, and this augmentation was significantly reduced by anti-human CD98 mAb. Furthermore, CD98-transfected clones developed tumors in athymic mice. These results indicated that overexpression of CD98 participates in the process of malignant transformation, suggesting that CD98 has oncogenic potential.

The consistent presence of the human accessory deep peroneal nerve.

Twenty-four human legs were dissected macroscopically to study the morphological details of the accessory deep peroneal nerve. This nerve arose from the superficial peroneal nerve and descended in the lateral compartment of the leg, deep to peroneus longus along the posterior border of peroneus brevis. Approaching the ankle joint, this nerve passed through the peroneal tunnels to wind around the lateral malleolus; it then crossed beneath the peroneus brevis tendon anteriorly to reach the dorsum of the foot. The accessory deep peroneal nerve was found in every case examined and constantly gave off muscular branches to peroneus brevis and sensory branches to the ankle region. In addition, this nerve occasionally had muscular branches to peroneus longus and extensor digitorum brevis, and sensory branches to the fibula and the foot. The anomalous muscles around the lateral malleolus were also innervated by this nerve. Neither cutaneous branches nor communicating branches with other nerves were found. The present study reveals that the accessory deep peroneal nerve is consistently present and possesses a proper motor and sensory distribution in the lateral region of the leg and ankle. It is not an anomalous nerve as has previously been suggested.

[Extramedullary relapse in the external auditory canal in a patient with acute promyelocytic leukemia treated with all-trans retinoic acid and autologous peripheral blood stem cell transplantation].

A 41-year-old man was given a diagnosis with of acute promyelocytic leukemia (APL) in August 1994. A chromosome analysis showed 46, XY, t(15; 17) and 47, XY, idem, +8 at that time. Because initial induction chemotherapy (BHAC-DMP) has not been successful, the patient was given 45 mg/m2 of all-trans retinoic acid (ATRA) and achieved complete remission (CR) after 26 days on this regimen. Following intensified chemotherapy, he received an autologous peripheral blood stem cell transplant (PBSCT) with high-dose busulfan and cyclophosphamide in April 1995. Competitive RT-PCR for PML-RAR alpha mRNA did not find any of APL cells in the collected stem-cell fraction. Although the patient remained in CR without therapy, a myeloblastoma was found in his left external auditory canal in August 1996. Recurrence in bone marrow, moreover, was discovered the following month. A chromosome analysis of bone marrow cells showed 47, XY, t(15; 17), +8 at this time. Thus, the extramedullary relapse developed after autologous PBSCT. This case provides information linking ATRA to the development of extramedullary relapse in patients with APL.

Development of variable dose X-ray fluoroscopic system--aimed at reduction of X-ray exposure during interventional radiologic procedure.

Recent advancement in IVR by the catheterization technique has raised the problem of increased X-ray exposure dose. To minimize this problem, we developed an X-ray fluoroscopy system with which the operator can select the X-ray dose according to the required image quality. The optical system features an optical iris that adjusts the amount of light input to the camera tube (the optical iris is usually fixed). If a rough fluoroscopic image with low S/N is acceptable, the optical iris is opened and the fluoroscopic dose is automatically reduced. This reduces exposure dose to 30-60% of the conventional dose.

Microgeographic genetic structure and gene flow in Hibiscus moscheutos (Malvaceae) populations.

Microgeographic genetic variation in populations of a wetland macrophyte, Hibiscus moscheutos L. (Malvaceae), was investigated using allozyme polymorphism. The species is a self-compatible insect-pollinated perennial, and seeds are water dispersed (hydrochory). Six hundred plants were analyzed from eight brackish and two freshwater populations within the Rhode River watershed/estuarine system. The genetic structure of the populations was assessed by fixation indices and spatial autocorrelation analyses. The degree of genetic differentiation among sites and gene flow between all paired combinations of sites (M ) was analyzed using three hypothetical gene flow models. Fixation indices indicated almost complete panmixia within populations, and spatial autocorrelations showed that genotypes were randomly distributed within sites, most likely the result of water dispersal of seeds. Allele frequencies were significantly different among sites, and estimated FST indicated moderate genetic differentiation (_ = 0.062). Genetic differences between populations were mostly explained by a gene flow model that accounted for the location of populations relative to the tidal stream. The importance of hydrochory in affecting spatial genetic structure was thus suggested both within and among H. moscheutos populations.

[Anesthetic management of a patient with erythroderma].

We gave anesthesia to a patient with erythroderma for distal partial gastrectomy. Erythroderma is an inflammatory disorder in which generalized erythema and scaling occur, either as a specific phase of processes such as T cell lymphoma, as a manifestation of underlying malignancy or in the absence of preexisting diseases. Severe cases of erythroderma may involve disturbances in the cardiovascular, thermoregulatory and metabolic systems which make the management of general anesthesia difficult. Systemic scale also causes a difficulty in placing ECG monitor electrodes and endotracheal tube. In the present case, dyspnea and asthmatic bronchitis occurred postoperatively. Therefore, in a patient with erythroderma, we recommend careful systemic management throughout the perioperative period.

[A case of inflammatory pseudotumor of the lung with suppressed immune response in the patient with renal cell carcinoma after nephrectomy and administration of interferon].

A 60-year-old man with renal cell carcinoma was treated of right nephrectomy and high dose administration of Interferon-alpha (IFN) in 1990. Three and half years after operation, he complained of cough and hemoptysis. Chest X-ray showed a abnormal shadow (5.5 x 3.5 cm) in the upper lung field. On chest CT, a tumor mass with small cavity was located at S2, attended with the speculation and pleural indentation. A rough nodule suspected daughter tumor was pointed out near the tumor mass. Laboratory investigation revealed mild leucocytosis, an elevated ESR and C reactive protein. On lymphocyte subset, CD 4/8 ratio was 0.8 and NK cell activity was 11%. Immunosuppressive acid protein (IAP) in serum was 1,137 ng/ml. No organisms or malignant cells could be demonstrated in the biopsy materials or sputum. Right thoracotomy and upper lobectomy was carried out. Postoperative pathological diagnosis was inflammatory pseudotumor. Namely, the lesion was occupied with variable admixture of lymphocytes, plasma cells, histiocytes, phagocytes included hemosiderin, foam cells and whorled fibrosis. Daughter tumor was scar tissue. Now, he is well without recurrence and his immune response is within normal level. Some discussion of the literature was mentioned.

Ramification patterns of the nerves innervating the forearm extensors in mammals and reptiles.

This paper discusses muscle phylogeny based on muscle innervation and defends a new concept of nerve-muscle specificity. Some researchers argued strongly against this concept in the first half of this century. However, we think their arguments were partially based on insufficiently examined findings of the ramification manner of the radial nerve. We have dissected 140 limbs of 25 species in mammals and reptiles to examine the manners of ramification of nerves supplying the forearm extensors. The pattern of the radial nerve has been revealed to consist of consistent and inconsistent elements. The branches to the forearm extensors except for the supinator follow quite consistent patterns, while the branch(es) to the muscle is much less consistent. Comparing the ramification patterns of the nerves between mammals and lizards, it can be concluded that the radial nerve in mammals is formed by the phylogenetic path alteration of its partial nerve fibers from the pathway along the flexor side in lizards to the route in the extensor side.

[Neoadjuvant chemotherapy for elderly advanced gastric cancer: a case report of MTX/5-FU sequential therapy followed by radical gastrectomy].

A 70-year-old-male was diagnosed as having gastric cancer (Borr. 2, 78 mm in size on the body region; mod. dif. adenocarcinoma). CT scan revealed direct invasion of the liver. A course of MTX/5-FU sequential therapy was started (MTX 100 mg/m2, 5-FU 500 mg/m2, i.v., weekly; interval, 2 hours) on March 23, 1992. The gastric tumor showed 67% reduction on week 8 and 86% on week 10. The liver invasion had almost disappeared by CT scan (total doses, MTX 1,040 mg, 5-FU 6,750 mg). On June 25, the tumor was completely resected by radical.gastrectomy. The pathological specimen showed a "submucosal carcinoma" (10 mm in size, Stage I). Twenty-two months after the operation, the patient is alive and in good health. No cancer recurrence has been found without post-operative chemotherapy.

[Non-Hodgkin's T cell lymphoma associated with marked eosinophilia].

A case of non-Hodgkin's T cell lymphoma (diffuse lymphoma, large cell type) associated with marked eosinophilia and pleurisy in a 57-year-old male is reported. The leukocyte count was 12.5 x 10(3)/microliters and eosinophil count was 53% and the absolute count of 6.6 x 10(3)/microliters. The patient's serum and pleural effusion fluid, containing abundant lymphoma cells, showed eosinophil colony stimulating factor (Eo-CSF) activity. Conditioned medium (CM) prepared from patient's T cells (T-CM) produced Eo-CSF and this was enhanced by interleukin-2 (IL-2) stimulation. We demonstrated that the patient's serum contained a significant amount of interleukin-5 (IL-5) and the patient's T-CM, particularly after IL-2 stimulation contained a significant amount of granulocyte-macrophage colony-stimulating factor (GM-CSF). These findings suggest that Eo-CSF produced by neoplastic T cells or normal T cells activated by tumor antigen stimulated the production of eosinophils in this patient and that both IL-5 and GM-CSF might play a role in Eo-CSF activity.

Ischemic tolerance due to the induction of HSP70 in a rat ischemic recirculation model.

Various studies have demonstrated an increase in heat shock protein 70 (HSP70) synthesis in the brain following transiently induced ischemia, suggesting a protective role for HSP70 against ischemic insult. In this study, we determined the time course of HSP70 mRNA and protein induction in rat hippocampus following ischemia using Pulsinelli's four-vessel occlusion model, and suggested a protective role for HSP70 induction in limiting ischemic damage to neurons and delayed neuronal death. In Northern blotting analysis using human HSP70 DNA as a probe, the accumulation of HSP70 mRNA after 5 min ischemia became evident at 4 h, and continued until 16 h, while after 30 min ischemia, HSP70 mRNA appeared at 2 h, and continued above control level until 24 h after treatment. In immunoblot analysis using anti-HSP70 antibody, induction of HSP70 protein appeared 24 h and reached a maximum 48 h after 5 min ischemia. In immunohistochemical analysis using anti-HSP70 antibody, staining was not detected in CA1 neurons until 16 h after 5 min ischemia, but staining in CA1 gradually increased 1 day after ischemia and reached a maximum level 2 days after ischemia. Similar time profiles in the staining pattern of HSP70 protein were observed in CA3 and CA4 neuronal cells following 30 min ischemia. When rats pretreated with 5 min ischemia (non-lethal for CA1 pyramidal neurons) were exposed to a 30 min, lethal period of ischemia, 2 days after pretreatment, considerable staining of HSP70 was observed. Pretreated rats had much less neuronal damage in the CA1 sector than did rats subjected to lethal, 30 min ischemia alone.(ABSTRACT TRUNCATED AT 250 WORDS)

[Clinicopathological study of 6 cases of mantle zone lymphoma].

Clinicopathological analyses of 6 cases of mantle zone lymphoma (MZL) were carried out. The median age of the patients was 62 years with a range of 41 to 71 years and the male-to-female ratio was 1:1. Superficial lymph node (LN) swelling was present only in 2 patients. Giant LN swellings of the mesenteric or inguinal regions were present in 4, and bone marrow involvement by lymphoma cells in 5. Serum protein electrophoresis revealed a monoclonal protein of IgM kappa type in 2 patients. One of these also had polyclonal hypergammaglobulinemia. An immunohistochemical study of 6 patients revealed LN-1-(-)+, LN-2+(-)++, sIgM+, sIgD+, CALLA +/-(-)+, DRC-1+(-)++. The immunohistochemical features of the cases were similar to those of small lymphocytic lymphoma or follicular lymphoma. Only 1 patients out of 6 achieved complete remission. Two patients died, one of pneumonia after chemotherapy and the other of cancer. The others were alive 4 to 100 months after the diagnoses. Although giant LN swelling and bone marrow involvement of lymphoma cells which were refractory to treatments were frequently observed, we consider MZL to be a slowly progressive and low-grade type of non-Hodgkin's lymphoma.

Innervation of the sternalis muscle accompanied by congenital partial absence of the pectoralis major muscle.

In one case accompanied by congenital partial absence of the pectoralis major muscle the sternalis muscle was examined to confirm its innervation by means of analysis of intramuscular nerve distribution. It was proved that the sternalis muscle was supplied only by the pectoral nerves even in the case of sternalis in direct contact with the proper thoracic wall. These findings as well as the results of Ura (1937) and Morita (1944) favor the interpretation presented by Eisler (1901), in which the sternalis muscle was described as being supplied only by the pectoral nerves. However, the problem of double innervation of the sternalis requires continued discussion because the relationships between the pectoral nerves and the branches of the intercostal nerves or extramural nerves (Yamada & Mannen, 1985; Kodama et al., 1986) have not yet been resolved. The precise genesis of the sternalis muscle should be also examined though it has already been proved to be derived from the pectoralis muscle group including the subcutaneous trunci muscle.

[An autopsy case of parathyroid carcinoma with primary hyperparathyroidism].

An autopsy case of a 72-year-old woman who was determined as having had a parathyroid carcinoma with primary hyperparathyroidism is reported. On autopsy, a tumor, measuring 6 X 3 X 2 cm, was found beneath the left lobe of the thyroid gland, adherent to the left recurrent nerve and the trachea, though there was no metastasis evident. The tumor was found to be lobulated on cross section inspection. Histologically, the tumor cells were arranged in trabecular and in solid patterns with fibrous bands. A few tumor cells showed mitosis. Invasions to the capsules and the blood vessels were found present. Immunohistochemically, the parathyroid hormone was found positive in the cytoplasm of the tumor cells. Otitis fibrosa generalisata, metastatic calcification, acute pancreatitis, and core pulmonale also were observed.