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Although pituitary endocrine tumors (PitNETs) are usually benign, some are highly invasive and recurrent. Recurrent PitNETs are often treatment-resistant and there is currently no effective evidence-based treatment. Tumor-associated macrophages (TAMs) promote tumor growth in many cancers, but the effect of TAMs on PitNETs remains unclear. This study investigated the role of TAMs in the incidence of recurrent PitNETs. Immunohistochemical analysis revealed that the densities of CD163- and CD204-positive TAMs tended to increase in recurrent PitNETs. Compared to TAMs in primary lesions, those in recurrent lesions were enlarged. To clarify the cell-cell interactions between TAMs and PitNETs, in vitro experiments were performed using a mouse PitNET cell line AtT20 and the mouse macrophage cell line J774. Several cytokines related to macrophage chemotaxis and differentiation, such as macrophage colony stimulating factor (M-CSF), were elevated significantly by stimulation with macrophage conditioned medium. When M-CSF immunohistochemistry analysis was performed using human PitNET samples, M-CSF expression increased significantly in recurrent lesions compared to primary lesions. Although no M-CSF receptor (M-CSFR) expression was observed in tumor cells of primary and recurrent PitNETs, flow cytometry analysis revealed that the mouse PitNET cell line expressed M-CSFR. Cellular proliferation in mouse PitNETs was inhibited by high concentrations of M-CSFR inhibitors, suggesting that cell-to-cell communication between PitNETs and macrophages induces M-CSF expression, which in turn enhances TAM chemotaxis and maturation in the tumor microenvironment. Blocking the M-CSFR signaling pathway might be a novel therapeutic adjuvant in treating recurrent PitNETs. This article is protected by copyright. All rights reserved.

Chordoma is a rare malignant bone tumor arising from notochordal tissue. Conventional treatments, such as radical resection and high-dose irradiation, frequently fail to control the tumor, resulting in recurrence and re-growth. In this study, genetic analysis of the tumor in a 72-year-old male patient with refractory conventional chordoma of the skull base revealed a high tumor mutational burden (TMB) and mutations in the MSH6 and MLH1 genes, which are found in Lynch syndrome. The patient and his family had a dense cancer history, and subsequent germline genetic testing revealed Lynch syndrome. This is the first report of a chordoma that has been genetically proven to be Lynch syndrome. Chordomas usually have low TMB; however, this is an unusual case, because the TMB was high, and immune checkpoint inhibitors effectively controlled the tumor. This case provides a basis for determining the indications for immunotherapy of chordoma based on the genetic analysis. Therefore, further extensive genetic analysis in the future will help to stratify the treatment of chordoma.

Objective We describe a case of intracranial and extracranial multiple arterial dissecting aneurysms in rheumatoid arthritis (RA). Case Presentation A 29-year-old man with a medical history of RA since 18 years of age was admitted to our hospital for vomiting, dysarthria, and conscious disturbance. At 23, he underwent ligation of the left internal carotid artery (ICA) with superficial temporal artery to middle cerebral artery anastomosis because of acute infarct of the left hemisphere caused by arterial dissection of the left ICA. During the current admission, computed tomography (CT) revealed subarachnoid hemorrhage, and digital subtraction angiography (DSA) demonstrated dissecting aneurysms of the left intracranial vertebral artery (VA) and right extracranial VA. We diagnosed him with a ruptured dissecting aneurysm of the left intracranial VA and performed endovascular parent artery occlusion on the left VA. For the right unruptured VA aneurysm, we performed coil embolization simultaneously. At 2 weeks after the endovascular treatment, follow-up DSA revealed that multiple de novo dissecting aneurysms developed on the origin of the left VA and left and right internal thoracic arteries. Those aneurysms were treated with coil embolization. Other remaining aneurysms on the left thyrocervical trunk, right transverse cervical artery, and both common iliac arteries were treated by conservative therapy. While continuing medical treatment for RA, the patient recovered and was discharged to a rehabilitation hospital. Conclusion Considering that RA-induced vasculitis can be a potential risk of vascular complications including multiple arterial dissections, physicians should carefully perform endovascular interventional procedures for patients with long-term RA.

If genetically modified organisms (GMOs) are spread through the natural environment, it might affect the natural environment. To help prevent the spread of GMOs, we examined whether it is possible to introduce conditional lethality by excising centromeric DNA from a chromosome by site-specific recombination in Saccharomyces cerevisiae as model organism. First, we constructed haploid cells in which excision of the centromeric DNA from chromosome IV can occur due to recombinase induced by galactose. By this excision, cell death can occur. In diploid cells, cell death can also occur by excision from both homologous chromosomes IV. Furthermore, cell death can occur in the case of chromosome V. A small number of surviving cells appeared with excision of centromeric DNA, and the diploid showed greater viability than the haploid in both chromosomes IV and V. The surviving cells appeared mainly due to deletion of a recombination target site (RS) from the chromosome.

Objective Ventriculostomy from Paine's point is an effective technique to ensure that the brain is relaxed for aneurysm surgery. This study aimed to use Paine's point for other neurosurgical procedures (except for those that require a pterional approach) by delineation of surface landmarks for identification of Paine's point on the cranium and scalp. Methods Based on the anatomical knowledge and examination of 3-dimensional computed tomography images of skull, we determined novel surface landmarks to identify Paine's point on the cranium and scalp. The new method was used in patients with intraventricular hemorrhage and aneurysmal subarachnoid hemorrhage caused by ruptured aneurysm of the anterior communicating artery. Results The puncture point was determined at a point located 2.5 cm superior to the supraorbital margin on linea temporalis on the skull and 2.5 cm superior to the eyebrow along the anterior edge of temporal muscle on the skin. Ventriculostomy was performed from Paine's point in patients with intraventricular hemorrhage or aneurysmal subarachnoid hemorrhage who underwent aneurysm surgery via an interhemispheric approach. No adverse events were observed in any of the patients. Conclusions By accurate surface marking on skull and skin, the use of Paine's point for ventriculostomy performed via an interhemispheric approach or for simple burr-hole surgery was found to be safe and reliable.

We report new conditions for obtaining high-strength and high-modulus polyethylene using the swell-drawing process which was introduced by Mackley and Solbai [Polymer, 28, 1115 (1987)]. Cyclohexane-toluene mixture was used to allow polyethylene with Mw of 1.75 × 106 to swell significantly at relatively low temperature. We found that a dumbbell-shaped polyethylene specimen swelled under stress gained higher drawability in subsequent drawing processes. It was confirmed that in a polyethylene specimen swelled and elongated under an optimal stress, the fibrillar textures were oriented without recrystallization in the direction of elongation. The specimen showed highest draw ratio when the solvent in the swollen polyethylene was reduced to 30% of the polyethylene in weight. Under these conditions a maximum tensile strength of 3.5 GPa and modulus of 120 GPa were achieved. These values are greater than those for the fiber prepared by Mackley and Solbai [Polymer, 28, 1115 (1987)] who used polyethylene with molecular weight greater than 106. The present results show that the improved swell-drawing method, where polyethylene specimen swelled and elongated under appropriate stress is drawn, can be used for the production of high-strength and high-modulus fiber from ultra-high molecular weight polyethylene. The result suggests the applicability of the method for polyethylene of much larger molecular weight.

To establish the molecular bases for development of a microbiological system approaching excretive fermentation of useful lipids, a mutant strain that accumulates lipids in the medium was isolated from the laboratory yeast Saccharomyces cerevisiae. Following the mutagenesis to strain YP1, a long chain fatty acid utilizer with ethylmethane sulfonate, the mutant strain, STG1, was selected from about 80,000 colonies. The analysis of extracellular lipids and the monitoring of leakage of intracellular proteins indicated that strain STG1 secreted lipids containing triacylglycerols into the extracellular space without cell lysis. Genetic studies clarified that this mutation was recessive and was complemented by wild-type genomic DNA fragments. STG1 was considered to be a good tool for elucidation of the molecular mechanism for transmembrane lipid transport.

A single copy of a reporter gene cassette, such as PGKP-lacZ-LEU2 (promoter-reporter-marker gene) cassette, was inserted into one of 32 positions along chromosome III in Saccharomyces cerevisiae with an interval of approximately 10 kb. The amounts of translational gene product (beta-galactosidase) synthesized by the cassette-transformed cells were then determined. The region specificity in chromosome III could be demonstrated in gene expression: two higher-expressed regions (hot regions), 133 and 199 (MAT) regions, and seven lower-expressed regions (cold regions). For the steady and high production of polypeptide, foreign gene products, by yeast, we would like to state that we hope for an insertion of the artificially prepared gene cassette [(promoter)-(foreign gene)-(marker gene) ] into a hot region, such as 199 (MAT) region of chromosome III.

A cDNA copy of the carboxymethylcellulase (CMCase 1) gene, CMC1 , of the yeast Cryptococcus flavus was expressed in Saccharomyces cerevisiae under the control of the glyceraldehyde-3-phosphate dehydrogenase gene ( GAP ) promoter derived from S. cerevisiae . In glucose medium, extracellular production of a large amount of CMCase 1 was achieved by S. cerevisiae cells transformed with YEp-plasmid carrying CMC1 . Two major proteins (36 kDa and 34 kDa) secreted in the medium both had CMCase activity and immuno-responsibility to anti-CMCase 1-antibody. From an analysis of amino acid sequences, it was found that the 36 kDa protein is a pro-CMCase 1 having an N-terminal residue, Ala, the 19th amino acid of the peptide deduced from the nucleotide sequence of CMC1 on the cDNA, and that the 34 kDa protein is a mature CMCase 1. Alteration of the cell morphology of S. cerevisiae was induced by transformation with YEp-plasmid carrying CMC1 .

Saccharomyces cerevisiae transformants which secrete high levels of cellulolytic enzymes, with chromosome-integrated multicopies of heterologous DNA sequences encoding the cellulolytic enzymes were constructed. An expression construct of β-glucosidase and carboxymethyl cellulase directed by the GAP promoter was integrated into the chromosomes of the haploid S. cerevisiae using the δ sequence-mediated integration system. Southern blot analysis of the chromosomes prepared from various integrants and separated by pulse-field gel electrophoresis demonstrated that the integration occurred mainly in a particular chromosome and the copy number of the integration was variable. The amount of enzymes secreted by the transformants correlated with the copy number of integration. For each enzyme, the highest activity was about 1.4-fold that produced by the transformant harboring the same expression cassette on a YEp-type plasmid. The δ-integrated exogenous DNA was mitotically stable in rich medium. A haploid double transformant which coexpresses and secretes β-glucosidase and carboxymethyl cellulase was further constructed by genetic crossing of the haploid transformant that produces a high level of the enzyme, followed by meiotic segregation of the resulting diploid strain. The haploid double transformant, but neither of the single transformant, could grow on a plate containing carboxymethyl cellulose as a sole carbon source. It is suggested that the δ-sequence-mediated integration system is a very useful means for the genetic engineering of yeast, especially when overproduction and secretion of multiple heterologous enzymes are desired.

Eukaryotic cells are characterized by very large chromosomal DNAs efficiently packed within the nucleus. To identify the mechanism of chromosomal packaging based on the uniqueness of the centromere region in Saccharomyces cerevisiae, we isolated the HCH6 mutant, which shows 2.5-fold higher efficiency of site-specific recombination between the CEN5 and HIS3 loci than the wild-type CH53 strain. This mutant also displayed defects in cell integrity at high temperature. The SSD1 gene was perhaps responsible for this defect. The efficiency of site-specific recombination was decreased by the introduction of SSD1 in HCH6 cells and increased by disruption of SSD1 in the wild-type cells. Furthermore, the distances between the CEN5 and HIS3 loci and between the CEN5 locus and the spindle pole body (SPB) indicated that disrupting SSD1 caused a loss of the anchoring of the CEN5 locus near SPB. These results suggest Ssd1p-dependent cross-talk between chromosomal positioning within the nucleus and the positioning of cellular components within the cell.

The PHO80/TUP7 locus in Saccharomyces cerevisiae is reported to be located on the right arm of chromosome XV close to its centromere. In the present study, the locus has been reassigned to the left arm of the same chromosome by reciprocal recombination between chromosomes V and XV at URA3 (on chromosome V) and PHO80/TUP7 loci by using the site-specific recombination system of the yeast plasmid pSR1.

Recombinase encoded by the R gene of pSR1 of Zygosaccharomyces rouxii mediates reciprocal recombination between two specific recombination sites (RSs) to induce excision or inversion of the DNA segment that is flanked by the RSs. We report here that site-specific recombination mediated by this system takes place effeciently in tobacco cells. To monitor the recombination events in tobacco cells, we have constructed two types of cryptic beta-glucuronidase reporter gene in such a way that recombination such as inversion of the construct or excision of the intervening sequence results in their expression. When these cryptic reporter constructs were transiently introduced together with the R gene by electroporation into protoplasts of tobacco cells, beta-glucuronidase activity was detected. The cryptic reporter genes, when stably resident in the chromosome of tobacco cells, were also activated by the R gene. Structural analyses of the genomic DNA isolated from these tobacco cells showed that the R protein did in fact catalyze precise recombination between two copies of RSs in tobacco cells, with resultant activation of the cryptic reporter genes. This observation provides the basis for development of a DNA technology whereby large regions of DNA can be manipulated in plant chromosomes. Potential uses of this recombination system are discussed.

We reported a case of pycnodysostosis and review of 67 cases in the Japanese literature, especially on its oral and maxillofacial aspects. The following oral and maxillofacial findings were commonly reported in most cases: 1. mandibular angle loss, 2. hypoplasia of the maxilla and mandible, 3. grooved palate and 4. malpositioned teeth. Other findings reported occasionally included: osteomyelitis, anterior cross bite, mandibular fracture, permanent tooth missing, hypercementosis, microdontia, delayed eruption of permanent tooth, persistence of deciduous tooth, enamel hypoplasia, obliterated pulp chamber, macroglossia, hypoplasia of dental root and dentin hypoplasia.

A strain, L-12, which secretes triglycerides (TG) was selected from low cell-density mutants of yeast Trichosporon sp. The strain was superior to the wild type strain in the extracellular production of TG from glucose; the production is over 75% of total TG accumulated.

A simple measuring system for the frequency of recombinant clones generated by R-protein mediated recombination was constructed. By applying the system, the frequencies of recombinant clones due to site-specific recombination between two loci on one or two different nonhomologous chromosome(s) were monitored during cultivation. Subsequently, it was found that the frequencies increase nearly in proportion to the culture period and are different among the individual two selected loci. Thus, the system is thought to be a candidate for analyzing physicochemical situations, such as folding and localization, of chromosomes in the nucleus.

The R gene product (R protein) of Zygosaccharomyces rouxii plasmid pSR1 catalyzes site-specific recombination within a 58 base-pair (bp) sequence present in the 959 bp inverted repeats of this plasmid. The R protein was produced in Escherichia coli and partially purified. The partially purified protein catalyzed site-specific recombination in vitro without the supply of an energy source. Recombination resulted in intramolecular inversion or deletion, depending on whether the orientations of the two recombination sites on the substrate plasmid were the same or opposite. Presumably, R protein is the only protein required for the recombination reaction. A circular DNA molecule appears to be a better substrate than a linear molecule in R-mediated in vitro intramolecular recombination. The R protein binds to a set of six 12 bp elements within the inverted repeats of pSR1. Two of these 12 bp elements are arranged in an inverted configuration with a 7 bp spacer in the 58 bp sequence. The R protein mediates strand cleavage in vitro at the junction between the 12 bp elements and the 7 bp spacer. The cleavage sites on the top and bottom strands are staggered and flanked by polypurine tracts that form part of the 12 bp elements.

We have developed an effective method to delete or invert a chromosomal segment and to create reciprocal recombination between two nonhomologous chromosomes in Saccharomyces cerevisiae, using the site-specific recombination system of pSR1, a circular cryptic DNA plasmid resembling 2 microns DNA of S. cerevisiae but originating from another yeast, Zygosaccharomyces rouxii. A 2.1-kilobase-pair DNA fragment bearing the specific recombination site on the inverted repeats of pSR1 was inserted at target sites on a single or two different chromosomes of S. cerevisiae by using integrative vectors. The cells were then transformed with a plasmid bearing the R gene of pSR1, which encodes the site-specific recombination enzyme and is placed downstream of the GAL1 promoter. When the transformants were cultivated in galactose medium, the recombination enzyme produced by expression of the R gene created the modified chromosome(s) by recombination between two specific recombination sites inserted on the chromosome(s).

The aneurysmal bone cyst is an uncommon bone lesion in the jaws. This lesion is a solitary cystic lesion of bone and was first described as a distinct clinicopathologic entity by Jeffe and Lichtenstein in 1942.We experienced a case of aneurysmal bone cyst of the right mandible in an 11-year-old girl. In April, 1983 she received a blood transfusion to treat a massive hemorrhage that arose from the gingiva at 7 of the mandible. Her course of recovery was without incident and no abnormal bleeding was seen. On April, 1984 she was admitted to The Dental Department of The Hakodate Municipal Hospital because of massive hemorrhage of the gingiva of Ti of the mandible. Roentogenography findings revealed a pear sharped radiolucent area between roots 6 and 7. Angiographical findings showed a coincidence with this roentogenography image and aggregation of contrast medium. We suspected the lesion of blood vessels, and embolization of the maxillary artery was conducted using gelform, but haemostasis was not obtained. After ligation of the right external carotid artery, fenestration of the 7-4 buccal cortid bone of the mandible was conducted.The lesion extended widely from 76 alveolar crest to the diaphysis of the mandible, and a bone cavity of the little finger in size wase found at this site, a large artery branching from the inferior alveolar artery was seen. The extraction of 76 and curettage of the affected site was conducted. On August 3 rd she was admitted to Higashi-Nippon-Gakuen University Hospital for detailed examination and treatment.Roentogenography findings of the 5 site showed a multiloculer radiolucent area of the mesial bone of the mandible. The radiolucent of the roentogenography coincided with the angiographic image of the left externar carotid artery. On August 10, a marginal resection of the mandible was conducted, and the histopathologic findings revealed an cvidence which led to the diagnosis of anrurysmal bone cyst.At present alter a two year lapse nn signs of recuriance wrre seen.

The alveolar ridge forming operation on the edentulous case with the sub-periosteal tunnel process using hydroxyapatite (HAP) is a pre-prosthetic surgical treatment that can be adapted to the aged, is quite simple under local anesthesia, and has wide applications. HAP is also attracting clinical attention for its utility as a prosthetic and compensatory compound for osseus tissue, with excellent histoaffinity and osseus induction capability.In the alveolar ridge forming operation on the edentulous case with the sub-periosteal tunnel process using HAP, the mucous membrane of the canine region is incised longitudinally under local anesthesia. Centering about the alveolar crest and up to front of the retromolar region, heperiosteum is elevated blindly, and HAP is injected in this region with an injection cylinder and the shape of the alveolar ridge is fixed with a plate splint and circumferential wiring of the mandible with the alveolar ridge forming operation, X-ray measurements showed that the height increased about 1.4 times in the anterior region and about 1.8 times in the molar region. There was a remarkable alveolar ridge lifting by the operation, with the buccal shelf extensively flattened to a form that is advantageous to improvements in maintenance and stability of dentures and support of masticatory pressure. A 20% enlargement of the denture-bearing area was observed after the operation.

A circular DNA plasmid, pSR1, isolated from Zygosaccharomyces rouxii has a pair of inverted repeats consisting of completely homologous 959-base pair (bp) sequences. Intramolecular recombination occurs frequently at the inverted repeats in cells of Saccharomyces cerevisiae, as well as in Z. rouxii, and is catalyzed by a protein encoded by the R gene of its own genome. The recombination is, however, independent of the RAD52 gene of the host genome. A site for initiation of the intramolecular recombination in the S. cerevisiae host was delimited into, at most, a 58-bp region in the inverted repeats by using mutant plasmids created by linker insertion. The 58-bp region contains a pair with 14-bp dyad symmetry separated by a 3-bp spacer sequence. The recombination initiated at this site was accompanied by a high frequency of gene conversion (3 to 50% of the plasmid clones examined). Heterogeneity created by the linker insertion or by a deletion (at most 153 bp so far tested) at any place on the inverted repeats was converted to a homologous combination by the gene conversion, even in the rad52-1 mutant host. A mechanism implying branch migration coupled with DNA replication is discussed.

The glycogen-rich clear cell adenocarcinoma of minor salivary gland is a very rare tumor. This entity, originally reported in 1975 by Mohamed and Cherrick, was distinguished from other recognized clear cell neoplasms of salivary gland origin metastatic adenocarcinomas, and glycogen-rich clear cell adenocarcinoma under a light microscopic, and by histochemical, and ultrastructural analysis. We experienced a case of glycogen-rich clear cell adenocarcinoma originating in the minor salivary gland of the left buccal mucosa, and reported the clinical course of a patient, and histopathologic findings and ultrastructural findings.The patient, a 56-year-old man admitted to Higashi-Nippon-Gakuen University Hospital in January, 1982 complaining of a tumor lesion of the left buccal mucosa. Oral examination revealed a lesion the size of a small finger, which was elastically hard and movablelesion. A tentative clinical diagnosis of malignant neoplasm of the minor salivary glands was made. An incisional biopsy was performed and the microscopic diagnosis was clear cell tumor. the tumor of left buccal mucosa was extirpoted, left upper neck dissection was conducted, and metastasis to the cervical lymph nodes was seen.A one year follow up showed evidence of left neck metastasis. Left radical neck dissection was conducted, and treated with chemotherapy.But 6 months later metastasis to the lungs was seen, and evidence of recurrence was noted at the interior of the mandiblular angle. The patient died March, 1986. Microscopic examination revealed an encapsulated, infiltrating tumor composed of nests, cords, and sheets of clear cells. The cytoplasma of the tumor cells was PAS-positive, diastase soluble, and Mucicalmin and Alcian blue negative. Electron microscopic examination revealed a glycogen. rich clear cell adenocarcinoma which is said to consist of two types of cells. One is rich organella, it seemed to have dark cells. The other had less organella with glycogen granule and appeared to have light clear cytoplasma.