Differential interference contrast microscopy for cells using hard x-ray holography.

We propose a differential interference contrast method for cells using hard x-ray Gabor holography and knife-edge filtering in the spatial frequency domain, without relying on beam shearing. A phase object is holographically recorded and reconstructed by computer. Interference between the wavefronts of zeroth order weighted by ejπ/2 in the positive frequency region produces a dark image. Similarly, interference between the wavefronts of the zeroth order weighted by ej3π/2 in the negative frequency region produces a bright image. By adding these two intensity distributions, good quality phase-contrast images of 8-µm-diameter polystyrene beads and human HeLa cells were obtained.

Tophaceous gout in the bipartite patella with intra-osseous and intra-articular lesions: a case report.

We report a case of a 34-year-old man who had an acute gouty attack with tophi around the accessory bone of the bipartite patella with intra-osseous lesions. As the symptoms did not respond to conservative treatments, the patient was treated surgically. Arthroscopy revealed urate crystals surrounded by inflammatory synovitis on the surface of the bipartite patella. Arthrotomy exposed the abundant crystals around the accessory bone and in the intra-osseous lesion. These findings made us speculate that some pre-existing inflammatory conditions around the accessory bone induced deposition of urate crystals, and the destruction of the barrier between the deposits and the joint led to an acute arthritis. Curettage of the tophi resulted in immediate improvement of knee function. There had been no recurrence of symptoms at the 2-year follow-up.

Studies on antitumor agents, 2. Syntheses and antitumor activities of 1-(tetrahydro-2-furanyl)-5-fluorouracil and 1,3-bis(tetrahydro-2-furanyl)-5-fluorouracil.

Convenient and efficient methods were developed for preparing 1-(tetrahydro-2-furanyl)-5-fluorouracil (Thf-FU, 3) [trade name, Futraful (Ftorafur) or FT-207], which is used clinically as an antitumor agent, and 1,3-bis(tetrahydro-2-furanyl)-5-fluorouracil (Thf2-FU, 4). For the syntheses, 2,4-bis(trimethylsily)-5-fluorouracil (Me3Si-FU, 1) and 2-acetoxytetrahydrofuran (Thf-OAc, 2) were condensed in the presence of Friedel-Crafts catalysts, such as SnCl4 and BF3-Et2O in dichloromethane, or in the presence of NaI in acetonitrile to give Thf-Fu or Thf2-FU depending on the reaction conditions and workup procedure. A trace of 3-(tetrahydro-2-furanyl)-5-fluorouracil (3-Thf-FU, 5) was formed in these reactions. Thf2-FU was easily hydrolyzed to Thf-FU. 2-Methoxytetrahydrofuran can be used instead of Thf-OAc for preparation of Thf-FU under similar conditions. The optimal ratios of Me3Si-FU, Thf-OAc, and SnCl4 or NaI for preparation of Thf-FU and Thf2-FU were determined. In all cases, 2-2.5 equiv of Thf-OAc with respect to Me3Si-FU gave the best results. The yields of Thf-FU and more especially of Thf2-FU were greatly dependent on the relative amount of SnCl4, and 0.01-0.1 equiv of the catalyst with respect to Me3Si-FU gave the best results. Thf2-FU was found to be effective against murine solid tumors and it was less toxic than Thf-FU when given orally. The antitumor activity of 3-Thf-FU is also reported.

Studies of antitumor agents. 1. Resolution of racemic 1-(tetrahydro-2-furanyl)-k-fluorouracil into the R and S isomers and examination of the biological activities of the isomers.

1-(Tetrahydro-2-furanyl)-5-fluorouracil (Thf-FU), which is named Ftorafur or FT-207 and is used clinically as an antitumor agent, was conveniently synthesized by condensation of the trimethylsilyl derivative of 5-fluorouracil with 2-acetoxytetrahydrofuran using NaI as a catalyst. This optically inactive Thf-FU was resolved into optically active (R)-(+)- and (S)-(-)-Thf-FU in high optical purity and excellent yield by formation of diastereoisomers with brucine. 13C NMR data were obtained on Thf-FU and related compounds and the antibacterial activities and in vivo antitumor activities of these isomers were tested. The degradations of these isomers to 5-fluorouracil by liver microsomes were also examined. No significant differences were found in any of these properties of these isomers.

Studies on antitumor agents. 8. Antitumor activities of O-alkyl derivatives of 2'-deoxy-5-(trifluoromethyl)uridine and 2'-deoxy-5-fluorouridine.

O-Benzyl and O-ethyl derivatives of 2'-deoxy-5-(trifluoromethyl)uridine (F3Thd) and 2'-deoxy-5-fluorouridine (FUdR) were synthesized. The oral antitumor activity of the compounds against sarcoma 180 in mice was examined. The 5'-O-ethyl (3b), 3'-O-ethyl (3c), 5'-O-benzyl (3e), and 3'-O-benzyl (3f) derivatives of F3Thd were 4-fold more active than F3Thd itself. Among the substituted-benzyl derivatives of F3Thd, 3'-O-(p-chlorobenzyl)-F3Thd (3h) showed the highest activity, with an ED50 less than one-tenth of that of F3Thd. The activities of 5'-O-benzyl (7c) and 3'-O-benzyl (7d) derivatives of FUdR were equal to those of the effective O-alkyl derivatives of F3Thd.

Synthesis and antiallergic activity of dimethyl-2-(phenylcarbamoyl)ethylsulfonium p-toluenesulfonate derivatives.

The derivatives of dimethyl-2-(phenylcarbamoyl)ethylsulfonium p-toluenesulfonates were synthesized and evaluated for antiallergic activity. The 2,3-dihydroxyethoxy group was introduced to the phenyl ring from the standpoint of lipophilicity and electronic effects of substituent. The IgE-induced rat passive cutaneous anaphylaxis (PCA) was inhibited by oral administration of several substituted 2-[(4-propoxyphenyl)carbamoyl]ethyldimethylsulfonium p-toluenesulfonate derivatives. Among them (+/-)-2-[N-[4-(3-ethoxy-2-hydroxypropoxy)phenyl]carbamoyl]ethyldimeth ylsulfonium p-toluenesulfonate (1a, IPD-1151T) was found to possess considerable activity in the PCA test, and it was launched as Suplatast tosilate in Japan.

Soluble fragments of e-cadherin cell-adhesion molecule increase in urinary-excretion of cancer-patients, potentially indicating its shedding from epithelial tumor-cells.

E-cadherin (Ecad) is well known to be a calcium-ion-dependent cell-cell adhesion molecule expressed mostly in epithelial tissues. Previous immunohistochemical studies suggested that this cell adhesion molecule acts as an invasion suppressor and is negligibly detected in cancer metastatic regions. Soluble Ecad fragments derived from the proteolysed membrane-associated form were detected in culture supernatants of two cell lines, COLO 205 and A-431, with normal distribution of cell surface Ecad. Soluble Ecad levels released into culture of COLO 205 exhibiting reduced cell-cell adhesion were apparently elevated above those of A-431 with tight cell-cell adhesion. Furthermore, human circulation and urine continuously contain soluble Ecad which consists mainly of homogeneous 75-85 kDa extracellular domains. Soluble Ecad urinary level per urinary creatinine level was found to be significantly elevated in 53% of patients suffering from various types of cancers including lung, liver, stomach, colon and rectal cancers, as compared with those in the age-matched healthy subjects. These results suggest that dysfunction of cell surface Ecad is responsible for its enhanced proteolytic shedding in tumorigenesis, which may lead to the decrease of cell surface Ecads. Furthermore, excretion of high levels of soluble Ecad fragments potentially indicates the progression of epithelial tumors excessively degrading cell surface Ecad in clinical subjects.

Expression of recombinant mouse/human chimeric antibody specific to human GMP-140/P-selectin.

To construct a mouse/human chimeric antibody, we cloned the genomic DNAs for Ig from a murine hybridoma that produces PL7-6 monoclonal antibody specific to human P-selectin and expressed them in SP2/0 myelomas using a series of pSV2 vectors. Transfected cells that produce the mouse/human chimeric anti-human P-selectin antibody were geneticin-selected and screened by an immunoassay using immobilized antigen. The chimeric antibody, cPL-2R1, expressed by the resultant clone has the murine Ig variable region and the human Ig constant region. The native antibody PL7-6 and the chimeric antibody cPL-2R1 react equally with purified P-selectin and thrombin-stimulated platelets. Competitive inhibition tests demonstrated that the native antibody PL7-6 and the chimeric antibody cPL-2R1 had identical affinity for purified human P-selectin. Thus, although human IgG1 constant region was substituted for the murine counterpart in this chimeric antibody, its specificity and binding affinity for P-selectin was not altered. This chimeric antibody may prove useful when employed in combination with imaging reagents or therapeutic drugs for targeting activated platelets or endothelium in patients with thrombosis or intravascular inflammation.

Malignant lymphoma of the gingiva: MR evaluation.

PURPOSE: Our purpose was to document the MR imaging findings of malignant lymphoma of the gingiva. METHODS: Five patients with histologically proved malignant lymphoma of the gingiva were studied by MR imaging. The MR images were analyzed for tumor size, extent, and signal characteristics, bone involvement, and associated cervical lymph node enlargement. RESULTS: Clinical examination tended to underestimate the size of lymphomatous lesions. The signal intensity of the lesions was isointense to hyperintense relative to muscle on noncontrast T1-weighted images and showed variable contrast enhancement patterns. On T2-weighted images, signal intensity was isointense to hypointense relative to the oral mucosa. In one case, the mass extended to the submandibular space; in the remaining cases, the masses were limited to the gingiva and the adjacent bone. MR imaging revealed that gingival lymphomatous masses were broad-based along the mandible or maxilla and eroded through the cortex into the marrow space, but the cortex was still recognizable. No nodal involvement was noted in any of the patients with malignant lymphoma. CONCLUSION: The signal characteristics of gingival lymphoma overlap those of other tumors. The cortex separating marrow involvement from the broad-based gingival mass generally appears to be permeated with small erosions but is still recognizable.

Extracorporeal life support for patients undergoing prolonged external cardiac massage.

From November 1987 to February 1992, extracorporeal life support (ECLS) was used for four patients undergoing prolonged external cardiac massage following cardiac arrest. Their underlying diseases consisted of acute pulmonary embolism, pulmonary arterial thrombosis due to protein C deficiency, acute inferior left ventricular infarction accompanied by right ventricular infarction and thoracic contusion. After the initiation of ECLS, hemodynamic variables and metabolic acidosis improved in all of the cases. The case of pulmonary embolism and the case of acute myocardial infarction were successfully weaned from ECLS without complications. They were later discharged ambulatory from the hospital. The patient with pulmonary arterial thrombosis, who was comatose, became alert after the initiation of ECLS. However the patient finally died due to diffuse and massive pulmonary arterial thrombosis, which was probably related to protein C deficiency. The patient with thoracic contusion was also comatose before ECLS. He did not recover from the coma and died soon after the disconnection of ECLS. The latter two cases were suspected to have had irreversible organ failures not responsive to mechanical support of both circulation and respiration. We conclude that ECLS is a very useful method for patients requiring prolonged cardiac massage following cardiac arrest.

External and interstitial radiotherapy in the treatment of oropharyngeal squamous cell carcinoma.

The records of 165 patients with squamous cell carcinoma of the oropharynx treated at our hospital with external and/or interstitial radiotherapy between 1971 and 1990 were reviewed to evaluate the treatment results, focusing on primary control and complications. All cancers were restaged according to the UICC 1987 TNM staging system. Of these 165 patients, 11% were in Stage I, 55% in Stage II, 24% in Stage III and 8.5% in Stage IV. Local control and complications were analysed in 140 patients. The remaining 25 patients died of intercurrent or metastatic disease during the first 2 years following treatment and were excluded from the analysis. 70 patients were treated by interstitial implant using permanent implant seeds (Rn-222 in eight patients, Au-198 in 62 patients) with or without external radiation. 56 other patients received external irradiation alone and another 14 patients received pre-operative external irradiation plus surgery. 2-year recurrence-free rates were 73% (100% for T1, 76% for T2, 36% for T3 tumours) in the patients who received interstitial implant with or without external radiation and 36% (67% for T1, 48% for T2, 13% for T3 tumours) in the patients treated by external irradiation alone. The incidence of soft-tissue or bone complications requiring long-term treatment was 14%. Based on these findings, it is believed that an interstitial implant using Au-198 grains combined with external beam irradiation is a useful treatment modality for small lesions of the oropharynx.

Reconstruction of the medial patellofemoral ligament for the treatment of habitual or recurrent dislocation of the patella in children.

We investigated the clinical outcome of a reconstructive procedure of the medial patellofemoral ligament for the treatment of habitual or recurrent dislocation of the patella in four children (6 knees), with a minimum follow-up of four years. The technique involves transfer of the tendon of semitendinosus to the patella using the posterior one-third of the femoral insertion of the medial collateral ligament as a pulley. There was no recurrence of dislocation after surgery. The mean Kujala score at follow-up was 96.3 points. Radiological assessment showed that the congruence angle, the tilt angle and the lateral shift radio were restored to normal. The lateral and medial stress shift ratios and the Insall-Salvati ratio remained abnormal. We conclude that this technique can be recommended for the treatment of habitual or recurrent patellar dislocation in children, although hypermobility and patella alta are not fully corrected.

The variation of trace element concentration in human hair: the trace element profile in human long hair by sectional analysis using neutron activation analysis.

Neutron activation analysis was used to determine the distribution of trace elements in human hair. Hair samples were obtained from five infants (two to seven years of age) with hair length ranging from 15 to 40 cm. The hairs were divided into segments, each of 2.5 cm, starting from the scalp end and trace elements were analysed in each segment. Concentrations of I, Mg, Ca and Cu increased from the scalp end to the tip. Concentrations of Cl and Br decreased inversely. Different profiles of the concentrations of Hg, Se, Ca and Mn were seen in each sample. These results were discussed with reference to the indication of environmental pollution.

Primary saphenous venous aneurysms presenting in a child.

Venous aneurysms are dilatations of localized venous segments having no connection with varicosities. They are most often found post-traumatically in adults. The case presented here was a rare case of primary saphenous venous aneurysms in an 8-year-old child which were well demonstrated by Tc-99m human serum albumin (HSA) angiography.

Increase in memory (CD4+CD29+ and CD4+CD45RO+) T and naive (CD4+CD45RA+) T-cell subpopulations in smokers.

To examine the effects of smoking on lymphocyte subpopulations, we measured the following cell subpopulations: CD4+ T-cell subpopulations (i.e., CD4+CD29+, CD4+ CD45RO+, and CD4+CD45RA+ cells); CD8+ T-cell subpopulations (i.e., CD8+CD11a+ and CD8+CD11b+ cells); and natural killer cell subpopulations (i.e., CD16+CD57-, CD16+CD57+, and CD16-CD57+ cells). We measured these subpopulations, together with total CD4+ T, total CD8+ T, total CD3+ T, B (CD19+), and total lymphocytes, in 10 male heavy smokers, 38 male light-to-moderate smokers, and 33 male nonsmokers. The mean ages were 30 y, 31 y, and 32 y, respectively, and ages did not vary significantly among the smokers. CD4+CD29+ and CD4+CD45RO+ (memory T) cells in heavy smokers were significantly more numerous than those in light-to-moderate smokers and nonsmokers. Also, these memory T-cell subpopulations were significantly more numerous in light-to-moderate smokers than in nonsmokers. The number of CD4+CD45RA+ (naive T) cells was significantly larger in heavy smokers than nonsmokers; numbers of CD4+CD45RO+ T and CD4+CD29+ T cells (memory T cells) were significantly correlated with daily cigarette consumption. Numbers of CD3+ T, CD4+ T, CD19+ B, and total lymphocytes in heavy smokers were significantly larger than in nonsmokers. There were significantly more CD3+ T, CD4+ T, and total lymphocytes in light-to-moderate smokers than in nonsmokers. The numbers of CD4+ T lymphocytes in heavy smokers were significantly larger than in light-to-moderate smokers. Perhaps CD4+ T cell subpopulations, especially memory T cells, are most susceptible to the effects of smoking on lymphocyte subpopulations.

Distribution of trace elements in the human body determined by neutron activation analysis.

Neutron activation analysis and instrumental semiconductor gamma-ray spectrometry were used for analysis of 20 trace elements in 10 autopsied human organs and tissues (liver, kidney, cerebrum, cerebellum, heart, muscle, pancreas, spleen, lung, and aorta) from 63 Japanese persons, whose ages ranged from 15 days to 85 yr. Distributions of aluminum, bromine, magnesium, manganese, rubidium, selenium, and vanadium in human body were almost uniform. High concentrations of cadmium were found in kidney and liver samples. There was a high mercury concentration in the liver, kidney, and brain samples. Concentrations of other elements (arsenic, gold, cobalt, chromium, copper, iron, indium, antimony, selenium, titanium, and zinc) in each organ or tissue are also presented in this paper.

[Anesthetic management of a patient with tuberous sclerosis].

Tuberous sclerosis is characterized by facial angiofibromatosis, epilepsy, and mental retardation. There have been only a few reports of anesthetic management of patients with tuberous sclerosis. We managed a 22-year-old patient with tuberous sclerosis using nitrous oxide (67%) in oxygen supplemented with fentanyl and midazolam. There were no untoward events related to anesthesia and surgery. Problems in managing patients with tuberous sclerosis are also discussed.

[Pediatric one-lung anesthesia with a Fogarty catheter].

One-lung anesthesia using a Fogarty catheter in pediatric patients is presented. Case 1 is an 8 year old male with coarctation of the aorta scheduled for coarctectomy. Case 2 is a 13 year old female with metastatic lung tumor scheduled for partial resection of the left upper lobe under thoracoscopic surgery. One-lung anesthesia was successfully established with a Fogarty catheter in both patients and no hypoxic episode was encountered during the one-lung anesthesia. Postoperatively, there was no evidence of atelectasis of the lung or barotrauma to the trachea or bronchus. Thus, the Fogarty catheter appears useful for pediatric one-lung anesthesia, especially when the double lumen endobronchial tubes or endobronchial blockers of smaller sizes are not available.

[Subarachnoid anesthesia with hyperbaric bupivacaine].

The effect, duration and complications of subarachnoid anesthesia with hyperbaric bupivacaine solution, adjusted at a gravity of 1.044 and 1.033 by adding glucose, were compared with those of isobaric 0.5% bupivacaine anesthesia. Thirty patients undergoing surgical operations on lower abdomen or lower extremities were divided into three groups and given 2.0 to 4.0 ml of bupivacaine at L3-4 according to the body height. The upper margin of analgesic zone at 15min and 60min after the subarachnoid injection of bupivacaine was higher in the hyperbaric groups, but it receded thereafter and significant differences were found at 120min compared with the isobaric group. Hypotension occurred less frequently in the hyperbaric groups. The short duration of analgesia in the hyperbaric groups may be attributable to the lower concentrations of bupivacaine, 0.40 and 0.45%, but the low frequency of hypotension might be due to the younger mean ages of these two groups. No notable complications were found in any group. Bupivacaine solution of 0.5% with a higher gravity is desired in the market.

[Sympathetic skin response as an objective indicator of sympathetic activity].

Sympathetic excitement is not rarely accompanied by sweating over the palm or sole, and this sudatory reaction is often depressed by atropine. But, there was no simple way for quantitative expression of sympathetic activity. As an objective indicator of sympathetic activity, sympathetic skin response (SSR) was studied in 17 volunteers and 5 patients in the pain clinic. SSR was evoked by square wave electric stimulation through a pair of surface electrodes placed on the unilateral forearm. SSR recorded with a surface electrode was much larger than that was taken with a needle electrode inserted deep under the skin, and was markedly reduced by an intravenous administration of atropine sulfate 0.5mg for about 10 min. These findings support the hypothesis that the SSR is developed by an action potential of eccrine glands of a cholinergic nature. Unilateral stellate ganglion block abolished SSR in the ipsilateral palm indicating that the efferent impulse of palmal SSR is transmitted via cervical sympathetic ganglion. Inhalation of 0.1MAC halothane or enflurane in air reduced the height of SSR within 10 minutes. They obtunded SSR dose-dependently, and almost completely abolished at a concentration of 0.3 MAC. SSR may be useful to evaluate sympathetic activity under various conditions, such as under the effect of sedatives, analgesics and general anesthetics, and to evaluate the effects of a nerve block or other drugs which were used to obtund the nociceptive afferent stimuli to the sympathetic center of the brain.