Measurements of Strong-Interaction Effects in Kaonic-Helium Isotopes at Sub-eV Precision with X-Ray Microcalorimeters.

We have measured the 3d→2p transition x rays of kaonic ^{3}He and ^{4}He atoms using superconducting transition-edge-sensor microcalorimeters with an energy resolution better than 6 eV (FWHM). We determined the energies to be 6224.5±0.4(stat)±0.2(syst) eV and 6463.7±0.3(stat)±0.1(syst) eV, and widths to be 2.5±1.0(stat)±0.4(syst) eV and 1.0±0.6(stat)±0.3(stat) eV, for kaonic ^{3}He and ^{4}He, respectively. These values are nearly 10 times more precise than in previous measurements. Our results exclude the large strong-interaction shifts and widths that are suggested by a coupled-channel approach and agree with calculations based on optical-potential models.

Observation of Coulomb-Assisted Nuclear Bound State of Ξ

In an emulsion-counter hybrid experiment performed at J-PARC, a Ξ^{-} absorption event was observed which decayed into twin single-Λ hypernuclei. Kinematic calculations enabled a unique identification of the reaction process as Ξ^{-}+^{14}N→_{Λ}^{10}Be+_{Λ}^{5}He. For the binding energy of the Ξ^{-} hyperon in the Ξ^{-}-^{14}N system a value of 1.27±0.21 MeV was deduced. The energy level of Ξ^{-} is likely a nuclear 1p state which indicates a weak ΞN-ΛΛ coupling.

[Acute respiratory failure due to inhalation of aerosol water proof agent].

A 65-year-old laundryman who suffered from cough, fever, and dyspnea was admitted to our hospital. Laboratory finding disclosed leukocytosis of peripheral blood, elevated CRP, and hypoxemia. Chest X-ray films showed diffuse interstitial nodular shadows in both lungs. The patient was given a diagnosis of pneumonia and treated with oxygen inhalation, antibiotics, and steroids. The abnormal findings disappeared rapidly, and the patient was discharged after 11 days. However, he was readmitted 3 days later with the same chest X-ray film findings as observed before. Examination of bronchoalveolar lavage fluid disclosed an increased neutrophil count. Transbronchial lung biopsy specimens revealed alveolar collapse, which was considered to have contributed to hypoxemia and ventilation perfusion mismatch. However, no signs of infiltration of inflammatory cells into alveolar walls, or of acute-phase interstitial edema, were observed. Environmental studies supported a strong relationship between the patient's symptoms and clothes waterproofing agent. From these findings, we reasoned that the inhalation of aerosol waterproofing agent containing fluoroplastics can induce respiratory failure.

Morphine augments excitatory synaptic transmission in the dentate gyrus through GABAergic disinhibition.

The present study investigated the effect of morphine on synaptic transmission and long-term potentiation (LTP) in the dentate gyrus using rat hippocampal slice preparations. Field excitatory postsynaptic potential (fEPSP) and population spike (PS), evoked by stimulation of the perforant path, were recorded from the dentate molecular layer and the stratum granulosum, respectively. Following application of 10 microM morphine, PS amplitude increased gradually in 10 min and was eventually potentiated by approximately 50%. The phenomenon showed a concentration-dependent manner and was completely canceled by naloxone, a mu opioid receptor antagonist. Furthermore, morphine-induced PS augmentation was not detected in disinhibited hippocampal slices, which suggests that the inhibitory input to the dentate granule cells was required for the facilitatory effect of morphine. Neither fEPSP nor tetanus-induced LTP of PS was altered by morphine application. The data support the hypothesis that mu opioid receptor activity modulates inhibitory recurrent circuits in the dentate gyrus and thereby, indirectly plays a regulatory role for hippocampal excitatory neurotransmission.

Cell cycle inhibitory protein p27 in esophageal squamous cell carcinoma.

BACKGROUND: p27 protein is one of the cdk inhibitors which regulates the progression from G1 to S phase of the cell cycle. Reduced expression of p27 protein has been reported to be correlated with poor clinical outcome in patients with various cancers. MATERIALS AND METHODS: We performed immunohistochemistry of both p27 and Ki67 in 136 cases of resected human esophageal squamous cell carcinoma, and evaluated the association between p27 immunoreactivity and clinicopathological features including clinical outcome in these patients. We also examined the correlation between labeling index or the percentage of positive tumor cells for p27 and Ki67 in serial tissue sections in these cases. RESULTS: Cases with invasion of the muscularis propria or adventitia had significantly (p < 0.05) higher p27 LI (65.0 +/- 23.7) than those with invasion limited to mucosa or submucosa and those with carcinoma in situ (58.9 +/- 18.3). There were no significant correlations between p27 and other clinicopathological factors such as sex, age, tumor size, differentiation type, nodal status and histological stage. The cases with p27 LI below 40% tended to have a worse prognosis than those with p27 LI above 40%. There was no significant correlation between Ki67 and p27 LIs. CONCLUSIONS: Reduced expression of p27 may be correlated with the biological behavior of esophageal squamous cell carcinoma and may play an important role in the early stages of cancer.

Topoisomerase II alpha expression in esophageal squamous cell carcinoma.

BACKGROUND: DNA topoisomerase II alpha (topo II alpha) is associated with active cell proliferation of mammalian cells. Topo II alpha overexpression has been reported in a number of human malignancies and is considered to be related to their biological behaviors. MATERIALS AND METHODS: We examined the expression of topo II alpha immunohistochemically in 136 cases of human esophageal squamous cell carcinoma, 10 foci of squamous dysplasia and 10 non-pathologic squamous epithelium. We calculated the labeling index (LI) or the percentage of immunopositive cells for Topo II alpha and Ki67, and Topo II alpha LI/Ki67 LI (T/K ratio). These findings were then correlated with clinicopathological features of the patients including their clinical outcome. RESULTS: Both topo II alpha and Ki67 immunoreactivity were detected in the nuclei. A significant positive correlation was obtained between Topo II alpha and Ki67 LIs in all the specimens examined. Topo II alpha LI and T/K ratio were 24.5 +/- 8.0% and 1.04 +/- 0.64 for carcinoma, 19.1 +/- 15.2% and 0.68 +/- 0.29 for dysplasia and 14.0 +/- 14.1% and 0.55 +/- 0.17 for non-pathologic epithelium, respectively. Topo II alpha LI and T/K ratio in carcinoma cases were significantly higher than those of normal epithelium. Topo II alpha LI alone did not correlate with any of clinicopathological parameters examined but among carcinoma cases, cases with lymph nodes metastasis or higher histological stages had significantly higher T/K ratio than those without lymph node metastasis or lower histological stages. In addition, carcinoma cases with T/K ratio of greater than 0.8 demonstrated significantly worse prognosis than those with T/K ratio of smaller than 0.8. CONCLUSIONS: The relative overexpression of topo II alpha as compared with Ki67, i.e., increased T/K ratio was detected in esophageal squamous cell carcinoma and is considered to represent a dysregulation or qualitative alteration in topo II alpha, possibly associated with malignances, as reported in other human cancers. In addition, topo II alpha overexpression may also be correlated with the aggressive biological behavior of the patients with esophageal squamous cell carcinoma.

Automation system of X-ray filter in digital subtraction angiography.

A newly developed automation system for making X-ray filter in DSA was applied to clinical study. A low-dose test exposure image of the patient was input to a computer as a template. The image was scaled down and used to engrave the X-ray filter shape on a polystyrene foam block with a modeling machine according to the image data. The carved part was filled with barium paste, and the block was placed on the tube as an X-ray filter. In this study, the system contributed to make a patient-specific filter of sufficient amount of attenuation in a short time. Also, it could avoid the unreasonable exposure of the patient during the preparation of the filter.

The chimeric protein, PEBP2 beta/CBF beta-SMMHC, disorganizes cytoplasmic stress fibers and inhibits transcriptional activation.

The chromosomal inversion 16(p13;q22) associated with human acute myeloid leukemia generates the chimeric PEBP2 beta/CBF beta-SMMHC gene. The PEBP2 beta/CBF beta portion of the chimeric polypeptide harbors most of the amino acid sequence of the PEBP2 beta/CBF beta protein, the non-DNA binding subunit of the heterodimeric transcription factor, PEBP2/CBF, whereas the SMMHC portion of the chimera consists of the rod domain of the smooth muscle myosin heavy chain molecule. In this study we examined the subcellular localization of the chimeric protein and its effect both on stress fibers and transcriptional activation by transfecting cDNA into tissue culture cells. The localization of the chimera was investigated by immunocytochemical staining of cells and was found to be both cytoplasmic and nuclear. One aspect of the effect of expression of the chimera was a drastic alteration of cell morphology. The cells appeared elongated and possessed long cytoplasmic processes. Double fluorescent labeling revealed disorganization of the stress fibers and an altered F-actin staining pattern in the transfected cells. Studies using a deletion mutant showed that both the PEBP2 beta/CBF beta and SMMHC domains are necessary for the induction of the morphological alteration. A significant proportion of the chimeric protein was retained in the cytoskeleton after detergent extraction of the cells and could be recuperated as a membrane fraction, suggesting that this is one of the probable sites of action of the PEBP2 beta/CBF beta-SMMHC protein. Another effect of the chimeric protein was inhibition of transcriptional activation dependent on the PEBP2/CBF binding DNA sequence. However, deregulation of PEBP2/CBF site dependent transcription by itself was not sufficient to induce cell morphological changes. Taken together, these results indicate that the PEBP2 beta/CBF beta-SMMHC chimeric protein acts at two levels, at the level of stress fiber organization and at the level of transcriptional activation. We suggest that the action of PEBP2 beta/CBF beta-SMMHC depends to a great extent on whether it is located in the cytoplasm or in the nucleus.

Difference between PA700-like proteasome activator complex and the regulatory complex dissociated from the 26S proteasome implies the involvement of modulating factors in the 26S proteasome assembly.

The PA700-like proteasome activator complex was highly purified from porcine erythrocytes, and its properties were compared with those of the regulatory complex disassembled from the purified 26S proteasome. The molecular mass of the PA700-like complex, which comprises 25-110-kDa subunits, was estimated to be 800 kDa by Superose 6 gel filtration. This complex showed neither ATPase activity nor peptidase activity toward Suc-Leu-Leu-Val-Tyr-MCA. Nevertheless, it was possible to make a high molecular mass complex from the purified PA700-like complex by incubating with the 20S proteasome in the presence of ATP. In contrast, the regulatory complex dissociated from the 26S proteasome did not reconstitute a larger complex under the same conditions. The subunit composition of the PA700-like complex was similar but not identical to that of the regulator complex dissociated from the 26S proteasome: the former complex had a 25-kDa subunit which is absent in the latter, whereas the latter had two or three 43-kDa subunits lacking in the former. These results indicate that the purified PA700-like proteasome activator complex is structurally and functionally distinct from the regulatory complex dissociated from the 26S proteasome, implying the involvement of modulating factors in the 26S proteasome assembly.

M3 muscarinic receptor mediates regulation of protein secretion in rabbit lacrimal gland.

PURPOSE: To identify which muscarinic receptor subtypes mediate protein secretion in isolated rabbit lacrimal glands. To compare protein secretory profiles in vitro with those in rabbit tears. METHODS: Rabbit lacrimal gland slices were incubated with carbachol in the presence and absence of different muscarinic antagonists. Protein secretion into the incubation medium was characterized with a Bio-Rad protein assay dye reagent in conjunction with Laemmli's method of SDS-PAGE. The medium protein profile was compared to that in tear samples. RESULTS: Dose dependent increases in protein secretion were elicited by carbachol between 10(-7) and 10(-4) M. During the first 20 min period, a maximal increase of 64% above the basal level was seen at the highest concentration. With 10(-4) M carbachol, the response was transient because after 100 min it decreased to its basal level. The increases in protein secretion caused by 10(-4) M carbachol were completely suppressed in the presence of 10(-5) M atropine. On the other hand, the relatively selective M1 antagonist, pirenzepine, at concentrations from 10(-6) M to 10(-4) M, had no effect on either the basal levels or the stimulatory effects of carbachol. Similarly, the relatively selective M2 antagonist, gallamine, at concentrations from 10(-6) M to 10(-4) M, had no effect on either of these levels. In contrast, the relatively selective M3 antagonist, 4-DAMP, at concentrations from 10(-6) M to 10(-4) M, progressively suppressed the stimulated level of protein secretion elicited by 10(-4) M carbachol without affecting the basal level. The protein profiles found in the tears and in the incubation medium were similar to one another. CONCLUSION: In vitro rabbit lacrimal gland protein secretion is comparable to that in vivo. Cholinergic-mediated control of rabbit lacrimal gland protein secretion occurs through stimulation of the M3 muscarinic receptor subtype.

GnRH inhibits neuronal activity in the ventral tegmental area of the estrogen-primed ovariectomized rat.

In urethane-anesthetized ovariectomized rats, estrogen-sensitive descending neurons were identified in the midbrain ventral tegmental area (VTA), based on estrogen-induced changes in the excitability in antidromic responses to midbrain central gray stimulation. Estrogen increased the threshold and decreased the firing rate of the identified neurons. Responses of the identified neurons to the microiontophoresis of gonadotropin-releasing hormone (GnRH) or D-Phe2, D-Ala6-GnRH, a behaviorally active analog, but not to glutamate or gamma-aminobutyric acid (GABA), depended on estrogen. In the ovariectomized rat, GnRH excited a few neurons; the analog had no effect. GnRH suppressed spontaneous or glutamate-induced firing in almost all neurons in the estrogen-primed rat. The analog had mixed effects. The facilitation of female rat sexual behavior induced by infusion of GnRH in the VTA is due to the inhibition of VTA neurons.

Detection of cell adhesion molecules in lens epithelial cells of human cataracts.

PURPOSE: To detect the expression of cell adhesion molecules (CAMs), including beta integrins, intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1, leukocyte-endothelial cell adhesion molecule-2, E-cadherin, and CD44, in cultured lens epithelial cells (LECs) of human cataracts. To show that LECs attach to cells or extracellular matrix components by the detected adhesion molecules. METHODS: A circular section of the anterior capsule with attached LECs obtained by anterior capsulotomy during cataract surgery was cultured in a well of an eight-chamber slide. The LEC immediately after surgery and the cell outgrowth beyond the capsular margin at 2 weeks of culture were observed after the culture was stained immunohistochemically. Functional assays of LEC growth on collagen- or laminin-coated plates were performed in the presence and absence of the antibody blocking the detected adhesion molecules. RESULTS: beta 1 integrin, ICAM-1, and CD44 were detected in both the original specimens and the cultured cells. When the antihuman anti-beta 1 integrin monoclonal antibody (mAb), anti-ICAM-1 mAb, or anti-CD44 mAb was added at 10 micrograms/ml to the incubation medium, LEC migration and proliferation were inhibited significantly on the collagen- or laminin-coated plates. When the mAb blocking these three CAMs were added each at 1 microgram/ml, LEC proliferation also were inhibited. CONCLUSIONS: beta 1 integrin, ICAM-1 and CD44 are all involved in LEC attachment and growth on collagen and laminin in vitro. It can be assumed that these CAMs are involved in adhesion of LECs to extracellular matrix components of the lens capsule. Understanding the characteristics of the adhesion molecules in LEC may lead to the development of a new approach to inhibit secondary cataract formation.

Anatomic segmental resection of the lung by thoracoscopy: an experimental study.

In patients who are unable to undergo a lobectomy for a small peripheral lung cancer, a partial thoracoscopic resection appears to be one viable alternative. However, since the regional lymphatics are disrupted in an anatomical fashion with a segmentectomy, it appears superior to a wedge resection. This experimental study was conducted to determine whether or not an anatomical segmental resection is feasible by thoracoscopy. A segmental resection of porcine lungs was performed using thoracoscopy. The segmental vessels were divided between ligatures. The segmental bronchus was divided by an endoscopic stapler. The intersegmental lung parenchyma was divided using a cotton dissector and a contact neodymium-yttrium aluminum garnet laser. Forty-three pigs were divided into seven groups as follows. Group 1: S1 + 2; group 2: S3; group 3: upper division; group 4: lower division; group 5: S6; group 6:S8; and group 7: S9 + 10. The operating times ranged from 145 +/- 15 min to 191 +/- 47 min. Blood loss ranged from 36 +/- 35 ml to 151 +/- 48 ml in all groups. The blood loss in the group with a resection of S6 and S9 + 10 was significantly greater than that of the other five groups. Most of the blood loss occurred during the division between the intersegmental planes. In conclusion, a thoracoscopic segmentectomy is considered to be technically feasible; however, further refinements in this technique are warranted before beginning clinical trials.

Thoracoscopic en bloc total esophagectomy with radical mediastinal lymphadenectomy.

OBJECTIVE: Total esophagectomy with en bloc mediastinal lymphadenectomy for cancer carries a substantial morbidity and mortality rate. To investigate the feasibility of thoracoscopic technique, we carried out an extensive laboratory study. Encouraged by our excellent results, we conducted a clinical trial. METHODS: From September 1994 to September 1995, 39 patients thoracic esophageal cancer lesions not invading surrounding organs underwent total esophagectomy with mediastinal lymphadenectomy by means of thoracoscopy. Ages ranged from 47 to 86 years. The procedures were conventional except for the thoracic portion, which was performed as a thoracoscopic procedure with six trocar holes instead of thoracotomy. All harvested lymph nodes were counted for each station. Spirometric data and plethysmographically determined vital capacity were measured before and after operation for all patients. RESULTS: All procedures were accomplished as scheduled, and none was converted to open thoracotomy. The operating time was 200 +/- 41 minutes (mean +/- standard deviation). Estimated blood loss was 270 +/- 157 ml. The harvested lymph nodes numbered 19.7 +/- 11.1 per patient. Seventeen patients (45%) had positive lymph nodes. There were no in-hospital deaths within 30 days. Twenty-two patients did not require postoperative ventilatory support. Vital capacity decreased to 85% +/- 11% of the preoperative values, and forced expiratory volume in 1 second decreased to 82% +/- 16%. CONCLUSIONS: Thoracoscopic mediastinal lymphadenectomy is technically feasible, and its completeness is comparable to that of the open technique. The decline in pulmonary function is significantly less than that seen in our previous experience with the open technique.

Estrogen modifies the baroreceptor-induced responses of supraoptic vasopressin neurons in the ovariectomized rat.

Effects of estrogen (E2) and estrogen plus progesterone (EP) on the baroreceptor-induced response of the spontaneous discharge activity in the supraoptic (SON) vasopressin (AVP) neurons were examined in ovariectomized (OVX) female rats. An increase or a decrease in arterial blood pressure induced by intravenous injection of phenylephrine or nitroprusside, resulted in an inhibition or an acceleration of the discharge activity in the identified AVP neurons. The slopes that were derived from a least squares regression line for the arterial blood pressure and baroreceptor-induced responses of AVP neurons in both OVX + E2 and OVX + EP rats were statistically smaller (P < 0.05) than that in the control OVX rat. These results suggest a functional relationship between ovarian endocrine and autonomic functions in the female rat.