Immunohistochemical, ultrastructural and molecular study of well differentiated adenocarcinomas of the lung predominantly composed of goblet cells.

In order to clarify the morphological and biological characteristics of well differentiated adenocarcinoma of the lung predominantly composed of goblet cells (WDAG), histopathological examinations, including some molecular biological procedures, were carried out using 42 surgical specimens of primary lung carcinoma which were predominantly (>50% of the total cell population) or totally composed of goblet cells. The subjects included 19 men and 21 women, ranging in age from 41 to 81 (mean 60 years old) with predominantly nodular, peripherally located lesions. Ultrastructural examination revealed characteristic apical microvillous filamentous core rootless (AFCR) in some, but not all, cases. Histologically, these AFCR corresponded well with structures stained by phosphotungstic acid hematoxylin (PTAH). The goblet cells of WDAG were divided into PTAH-positive (26 cases) and -negative (16 cases) groups. The PTAH-positive group had larger tumor size, greater number of intrapulmonary and extrapulmonary metastases and shorter disease-free interval. The immunoexpression of p53 protein (60%) and rate of K-ras point mutation (84%) were also higher in the PTAH-positive group. Therefore the goblet cell population of WDAG, though it may appear morphologically homogeneous under light microscopy, is actually composed of heterogeneous groups of cells with different histopathological characteristics and biological behavior.

Effect of donor age on morphologic variation of cultured human corneal endothelial cells.

PURPOSE: To examine the effect of donor age on the morphologic variation of cultured human corneal endothelial cells (HCEC). METHODS: HCEC were obtained from the remaining corneoscleral rims of seven human corneas used for penetrating keratoplasty. The donor age ranged from 2 to 75 years. Primary cultures were established from explants of the endothelial cell layer, including the Descemet's membrane, and were propagated on culture dishes coated with bovine corneal endothelial extracellular matrix. At the fourth passage, frequency distribution of cell area in the confluent monolayer was calculated and the effect of donor age on cell area was analyzed. RESULTS: The percentage of HCEC with cell area over 2000 microm2 significantly increased with donor age (r = 0.935, p = 0.0007). CONCLUSION: Cultured HCEC established from older donor tissue display greater heterogeneity. The use of HCEC from younger donors may be preferable to maximize the benefits of HCEC transplantation.

[Two cases of pericardial rupture due to blunt chest trauma].

We treated two cases of pericardial rupture from blunt chest trauma. Case 1: A 55-year-old male was injured in an automobile accident. He recovered from the left hemothorax by tube drainage. One year and two months after the trauma, a left pericardial rupture was found during an operation for a left diaphragmatic hernia. As the epicardium adhered firmly to the pericardium, the ruptured pericardium was not sutured. Case 2: A 46-year-old man fell from a tree five meters in height. Chest radiography showed multiple right rib fractures, a pelvic fracture, pneumopericardium, and right hemopneumothorax. After four days, we performed a thoracoscopic examination under local anesthesia. The thoracoscopy revealed a rupture as large as an egg in the right pericardium anterior to the phrenic nerve. Judging from the size of the rupture, cardiac luxation was suspected not to have occurred. So the defect was not repaired. These two patients were discharged uneventfully without cardiac luxation. In the future, the accumulation of thoracoscopic findings may provide adequate information for judging the indication of operative repair in pericardial ruptures.

[Utility of the bronchoalveolar lavage procedure for the diagnosis of fat emboli syndrome (FES)].

In this study, we divided patients who were admitted to the Life Saving Center of our hospital with traumatic fractures into 3 groups: group F, including patients who were given diagnoses of FES according to Tsuruta's criteria; group K, including patients whose fractures resolved without any complications; and group KH, consisting of patients with fractures and pulmonary contusions. Broncho-alveolar lavage (BAL) was found to be useful for detecting FES in these groups. The subjects studied were 45 patients with traumatic fractures who were admitted to the Life Saving Center of our hospital and who underwent BAL procedures between April 1989 and March 1997 (group F : 14, group K : 17, group KH : 14) After noninvasive treatment, all patients received their first BAL by bronchoscopy within 48 hours of admission. Significant differences distinguished group F from groups K and KH in terms of features of BAL washout solution, total cell count, and values for lymphatic subsets, P-III-P, LTB 4, and granulocyte elastase. Based on these results, it was concluded that BAL is useful for detecting FES and differentiating patients in group S from those in groups K and KH.

[A case of advanced rectal cancer with unresectable liver metastasis for which chemotherapy was markedly effective].

We report a case of advanced rectal cancer accompanied by unresectable liver metastasis in which remission has been achieved for 7 years. The patient was a 64-year-old woman, and her chief complaints were a feeling of abdominal distention and melena. After low anterior resection (D3), oral administration of 600 mg/day of UFT-E and chemotherapy against the liver metastasis by hepatic arterial injection via a reservoir were started. Since the arterial injection via a reservoir became impossible due to infection, her clinical course was observed solely with oral administration of UFT-E. As a result, complete remission was achieved 7 months postoperatively, and the patient is still alive in good health without recurrences at present (7 years postoperatively). The reason why a favorable outcome has been achieved is probably that, although the initial operation resulted is non-curative resection, measures sufficient to enable the patient to lead an ordinary social life were taken at the beginning, and efforts to achieve a long-term survival were made while respecting the quality of life of the patient so that the patient would be able to coexist with the cancer.

[A comparison of asthma deaths and near-fatal asthma attacks].

This study compares the characteristics of fatal asthma patients and those of near-fatal asthma patients who had experienced severe life threatening attacks. Data from 67 fatal asthma patients and 80 near-fatal asthma patients were analyzed. The mean age of fatal asthma cases were older than near-fatal asthma cases, 51.9 yrs and 44.3 yrs, respectively. Similarities of the two cases were seen in sex, type of asthma and severity of asthma. The experiences of previous life-threatening asthma did not differ in both groups (43.3% of fatal vs 40.0% of near-fatal). From the viewpoint of type of development of severe exacerbation, rapid exacerbation was more frequent in fatal cases than near-fatal cases, but acute exacerbation after unstable asthma was more in near-fatal asthma. Seventy-six percent of all asthma deaths occurred at home or on the way to the emergency department. The rate of delay of receiving medical care was significantly higher in fatal cases (72%) than near-fatal cases (26%). There was no significant difference in the frequency of medical management before fatal or near-fatal episode such as oxygen therapy, mechanical ventilation. Use of ambulance, visiting emergency department, previous hospitalization between the two groups. The profile of fatal cases was almost same to that of near-fatal cases except delay of receiving medical care. In conclusion, the analysis of near-fatal cases was very important to make clear the causes of asthma death.

The endo-mirror, a new instrument for ophthalmic surgery requiring visualization of the ciliary sulcus.

Safely suturing a posterior chamber intraocular lens (PC-IOL) to the ciliary sulcus requires a means of visualizing the ciliary sulcus, since it cannot be viewed directly with the operating microscope. We, therefore, created several trial instruments consisting of a mirror with a built-in light source, which we call an endo-mirror, designed for observing the area behind the iris. Using these instruments, we were able satisfactorily to view the ciliary sulcus and suture a PC-IOL to it, first in a number of animal eyes, and then, using an endo-mirror incorporating the best features of the trial versions, in actual clinical cases.

[Four cases of airway infections caused by MRSA (methicillin resistant Staphylococcus aureus)].

Currently, infections caused by MRSA (methicillin-resistant Staphylococcus aureus) pose a great problem clinically. We present 4 patients with MRSA infections experienced by us. In these patients the infection was localized in the trachea and main bronchus. The first patient was a 62-year-old man. After undergoing operation for early gastric cancer, he had septic shock and was admitted to our center. The second was a 60-year-old man. After he underwent operation for advanced gastric carcinoma at another hospital septicemia developed due to suture failure and he was admitted to our center. The third was a 38-year-old woman who was admitted to our center because of grades II degrees-III degrees burns on 75 to 80% of her body surface area. The fourth was a 60-year-old man who was admitted to our center because of rupture of an aneurysm of the abdominal aorta. It is assumed that MRSA has quite different characteristics from the usual MSSA (methicillin-sensitive Staphylococcus aureus) in that it produces a new penicillin-bound protein (PBP-2') within cells. Thus, from the fiberoptic bronchoscopy findings of our own cases it is considered that there may be cases in which the observed lesion is localized in the central airway alone, without involvement of the segmental bronchi. We believe it necessary to take some prompt measures under a suspicion of airway infection caused by MRSA in the following cases: (1) compromised hosts under tracheal intubation, (2) patients who are under treatment with second or third generation cephalosporins, and (3) patients with production of bloody sputum, and (4) endotoxin-positive patients.

Characterization of the structure and determination of mRNA levels of the phenylalanine ammonia-lyase gene family from Populus kitakamiensis.

We isolated two new PAL genes, palg2b and palg4, from Populus kitakamiensis, palg2a and palg2b are clustered and palg2b encodes a polypeptide of 710 amino acids. The nucleotide sequence in the coding region of palg2b was 94.6% identical to that of palg2a. The promoter regions of palg1, palg2a and palg2b have several elements conserved among many phenylpropanoid biosynthetic genes. We measured the mRNA levels of the four PAL genes by S1 mapping using total RNA from stem tissues developing secondary xylem. Results showed that the transcript level of palg2b was higher than that of the other PAL genes.

Secondary xylem-specific expression of caffeoyl-coenzyme A 3-O-methyltransferase plays an important role in the methylation pathway associated with lignin biosynthesis in loblolly pine.

Two types of structurally distinct O-methyltransferases mediate the methylation of hydroxylated monomeric lignin precursors in angiosperms. Caffeate 3-O-methyltransferase (COMT; EC 2.1.1.68) methylates the free acids and caffeoyl CoA 3-O-methyltransferase (CCoAOMT; EC 2.1.1.104) methylates coenzyme A esters. Recently, we reported a novel hydroxycinnamic acid/hydroxycinnamoyl CoA ester O-methyltransferase (AEOMT) from loblolly pine differentiating xylem that was capable of methylating both acid and ester precursors with similar efficiency. In order to determine the possible existence and role of CCoAOMT in lignin biosynthesis in gymnosperms, a 1.3 kb CCoAOMT cDNA was isolated from loblolly pine that showed 79-82% amino acid sequence identity with many angiosperm CCoAOMTs. The recombinant CCoAOMT expressed in Escherichia coli exhibited a significant methylating activity with hydroxycinnamoyl CoA esters whereas activity with hydroxycinnamic acids was insignificant. Moreover, 3.2 times higher catalytic efficiency for methylating caffeoyl CoA over 5-hydroxyferuloyl CoA was observed which could serve as a driving force towards synthesis of guaiacyl lignin. The secondary xylem-specific expression of CCoAOMT was demonstrated using RNA blot analysis, western blot analysis, and O-methyltransferase enzyme assays. In addition, Southern blot analysis indicated that CCoAOMT may exist as a single-copy gene in loblolly pine genome. The transgenic tobacco plants carrying loblolly pine CCoAOMT promoter-GUS fusion localized the site of GUS activity at the secondary xylem tissues. These data suggest that CCoAOMT, in addition to AEOMT, plays an important role in the methylation pathway associated with lignin biosynthesis in loblolly pine.

Classic type of Kaposi's sarcoma and human herpesvirus 8 infection in Xinjiang, China.

We report 17 cases of the classic type of Kaposi's sarcoma in Xinjiang, which is located in the north-western area of China surrounded by Mongolia in the east, Russia in the north and Kazakhstan in the west. Fifteen of the patients were of the Uygur people. All patients were male and did not have acquired immunodeficiency syndrome. Most of the lesions were found in the lower and/or upper extremities, with 16 patients showing multiple lesions. Immunohistochemical examination of the lesions revealed that human herpesvirus 8 (HHV-8)-encoded latency-associated nuclear antigen was expressed in the nuclei of spindle-shaped tumor cells. HHV-8 DNA was detected by polymerase chain reaction in all seven cases examined. Phylogenetic tree analysis revealed that DNA sequences of the HHV-8-encoded K1 gene in the seven Kaposi's sarcoma cases were classified as subtype C that was common in the Mediterranean, the Middle East and East Asian countries. In addition, using immunofluorescence we investigated the seroprevalence of HHV-8 in 73 Uygur patients with diseases other than Kaposi's sarcoma. Surprisingly, the serological study revealed that 34 of the patients (46.6%) were positive for antibodies against HHV-8, suggesting that HHV-8 infection is widespread in Xinjiang area. The occurrence of the classic type of Kaposi's sarcoma with a high seropositivity rate implies that Xinjiang is the most endemic area for HHV-8 infection in the world known to date. Considering that Xinjiang is located at the middle point of the Silk Road that used to extend from Rome to China, these data imply that the virus may have been in circulation in this area due to the migration of the people via the Silk Road.

Immunocytochemical localization of phenylalanine ammonia-lyase in tissues of Populus kitakamiensis.

The polypeptide encoded by the partial fragment of cDNA of phenylalanine ammonia-lyase (PAL; EC 4.3.1.5), PALcDNA1 (Osakabe et al., 1995, Plant Sci. 105: 217-226), isolated from Populus kitakamiensis (P. sieboldii x P. grandidentata), was expressed in Escherichia coli cells. The polypeptide was purified and an antiserum raised against it. The antiserum recognized a protein of 77 kDa on nitrocellulose blots after sodium dodecyl sulfate-polyacrylamide gel electrophoresis of total protein and the partially purified PAL protein from P. kitakamiensis. Moreover, the antiserum recognized a protein on the blot after non-denaturing polyacrylamide gel electrophoresis of P. kitakamiensis proteins and this protein had PAL activity. Furthermore, the antibody inhibited PAL activity of extracts from stem tissues. These results showed that the antiserum against the partial PAL peptide recognized only the PAL subunits in extracts of P. kitakamiensis. Immunolocalization studies of P. kitakamiensis tissues revealed that the PAL protein was specifically localized in the xylem and the phloem fibers and no immunogold signal was found in the epidermis, the cortex, the pith, or the cambium of either stems or leaves.