Histone methyltransferase DOT1L coordinates AR and MYC stability in prostate cancer.

The histone methyltransferase DOT1L methylates lysine 79 (K79) on histone H3 and is involved in Mixed Lineage Leukemia (MLL) fusion leukemogenesis; however, its role in prostate cancer (PCa) is undefined. Here we show that DOT1L is overexpressed in PCa and is associated with poor outcome. Genetic and chemical inhibition of DOT1L selectively impaired the viability of androgen receptor (AR)-positive PCa cells and organoids, including castration-resistant and enzalutamide-resistant cells. The sensitivity of AR-positive cells is due to a distal K79 methylation-marked enhancer in the MYC gene bound by AR and DOT1L not present in AR-negative cells. DOT1L inhibition leads to reduced MYC expression and upregulation of MYC-regulated E3 ubiquitin ligases HECTD4 and MYCBP2, which promote AR and MYC degradation. This leads to further repression of MYC in a negative feed forward manner. Thus DOT1L selectively regulates the tumorigenicity of AR-positive prostate cancer cells and is a promising therapeutic target for PCa.

Chloroplast development in Arabidopsis thaliana requires the nuclear-encoded transcription factor sigma B.

Development of plastids into chloroplasts, the organelles of photosynthesis, is triggered by light. However, little is known of the factors involved in the complex coordination of light-induced plastid gene expression, which must be directed by both nuclear and plastid genomes. We have isolated an Arabidopsis mutant, abc1, with impaired chloroplast development, which results in a pale green leaf phenotype. The mutated nuclear gene encodes a sigma factor, SigB, presumably for the eubacterial-like plastid RNA polymerase. Our results provide direct evidence that a nuclear-derived prokaryotic-like SigB protein, plays a critical role in the coordination of the two genomes for chloroplast development.

Increase in basal level of Hsp70, consisting chiefly of constitutively expressed Hsp70 (Hsc70) in aged rat brain.

Alteration of proteins in the function and higher structure has been observed in aged organisms. Stress proteins, which have a role to protect proteins from denaturation, may respond to the protein denaturation with aging. We found that the basal level of Hsp70, consisting chiefly of constitutively expressed Hsp70 (Hsc70), in 24-month-old Wistar rats was significantly higher in some parts of the brain than that in 6-month-old rats. On the other hand, the basal level of Hsp70 was significantly lower in 24-month-old rats after restricted feeding, which is known to suppress aging, than those fed the diet ad libitum. In the pons, medulla, striatum, and thalamus of 24-month-old rat brain, the level of Hsp70 increased and the denaturation of the cytosol proteins was suppressed. These results suggest that the expression of Hsp70, mostly Hsc70, increases with aging and may have a role to suppress protein denaturation.

RT-PCR suggests human skeletal muscle origin of alveolar soft-part sarcoma.

In 1952, Christopherson et al. proposed that alveolar soft-part sarcoma (ASPS) was a distinct entity with unique clinical and pathological features. Since their report, its histogenesis has not been determined. In order to clarify the histogenesis of ASPS, a study using reverse transcription polymerase chain reaction using cDNAs from MyoD1 and myogenin, and the actin filament from human-skeletal-muscle-related mRNAs has been performed in 5 cases of ASPS. The expression of MyoD1 and myogenin was determined in 5 and 2 cases of the 5 cases, respectively. Moreover, expression of the many mRNAs from the actin filament of skeletal muscle was also found in ASPS. According to these findings, it is now postulated that ASPS is of skeletal muscle origin.

Detection of urinary macrophages expressing the CD16 (Fc gamma RIII) molecule: a novel marker of acute inflammatory glomerular injury.

BACKGROUND: The CD16 antigen is the Fc gamma receptor III. CD14+CD16+ cells are proinflammatory monocytes/macrophages (Mo/M phi) that constitute a minor population in the peripheral blood of healthy individuals. Little is known about the expression of CD16 antigen on Mo/M phi in glomerulonephritis. METHODS: Flow cytometric analyses were performed on urine and blood samples obtained from 209 patients with various renal diseases. Patients variously suffered from rapidly progressive crescentic glomerulonephritis (RPGN), membranoproliferative glomerulonephritis (MPGN), postinfectious acute glomerulonephritis (AGN), Henoch-Schönlein purpura nephritis (HSPN), IgA nephropathy (IgAN), membranous nephropathy (MN), minimal change nephrotic syndrome (MCNS), lupus nephritis (LN), acute interstitial nephritis, hereditary nephropathy, idiopathic renal hematuria (IRH), and renal stone. RESULTS: The CD16+ M phi population of cells was present in the urine of hematuria-positive patients with proliferative glomerulonephritis, including AGN, IgAN, RPGN, MPGN, and LN with acute inflammatory lesions, such as endocapillary proliferation, tuft necrosis, and cellular crescents. In contrast, the urinary CD16+ M phi population was negligible in hematuria-positive patients with nonproliferative renal disease, including hereditary nephropathy, IRH, and renal stone and also in patients with proliferative glomerulonephritis lacking acute inflammatory lesions. Total urinary M phi of these patients were much less than those of patients having proliferative glomerulonephritis with acute inflammatory lesions. Transient expansion of the CD16+ M phi population in urine was observed during the acute exacerbation of urinary abnormalities, whereas the disappearance of CD16+ M phi closely preceded the amelioration of urinary abnormalities in patients with proliferative glomerulonephritis. In 38 of the 98 patients positive for CD16+ M phi population in urine, the CD16+ Mo population was negligible in peripheral blood. Immunohistochemically, CD16+ M phi were present in the glomeruli of active proliferative glomerulonephritis, whereas such cells were absent in inactive proliferative glomerulonephritis or nonproliferative glomerular diseases. CONCLUSION: CD16+ M phi may be effector cells involved in the acute inflammation common to all types of proliferative glomerulonephritis. Furthermore, the detection of CD16+ M phi in urine, as well as urinary M phi counts, may serve as a useful indicator of the active stage of proliferative glomerulonephritis.

Effect of over-expressed hsp26 on cell growth of yeast.

Mammalian small heat shock protein (s-hsp) has been suggested to participate not only in stress tolerance but also in the growth regulation and differentiation of cells. To confirm the role of s-hsp in cell growth, we investigated the relationship between the expression of hsp26 and yeast cell growth. Cells lacking constitutive hsp70, ssa1ssa2, have been known to have a poor growth rate and to over-express hsp26 and some other hsps. We obtained several cell clones of ssa1ssa2 whose doubling times were different from one another. The amount of hsp26 was closely linked to the doubling time of ssa1ssa2 cells. This result suggests that the expression of hsp26 modulates the growth rate of yeast cells lacking constitutive hsp70, similarly to mammalian cells.

[Topical treatment of onychomycosis by occlusive dressing using bifonazole cream containing 40% urea].

Topical treatment with an antifungal agent was performed in 70 cases of onychomycosis (69 cases of tinea unguium and 1 case of Candida onychomycosis). The patients were treated every night by occlusive dressing using bifonazole cream containing 40% urea, and then by simple application of 1% bifonazole solution in the morning. During the treatment period, the softened affected nails were removed as completely as possible with nail clippers and files. The nails were observed every two weeks, and the efficacy was evaluated after 12 weeks. Two cases were excluded and 28 cases dropped out leaving 40 cases for efficacy evaluation. Among the 40 patients, 20 appeared to be mycologically negative (mycological cure rate: 50.0%). Clinical response was judged by the changes in opacity and thickness of the nails. Nine patients were evaluated as showing "marked improvement" and 16 as showing "improvement" (improvement rate: 62.5%). Erosion was noted as an adverse reaction in 2 patients. In one patient, treatment was stopped and the lesion was improved by antibiotic ointment, and in another, treatment was resumed when erosion was cured by discontinuation of the treatment. These results suggest that the topical treatment of onychomycosis by occlusive dressing is a useful method for those patients who have difficulties in or do not wish to be treated with oral antifungal agents.

Role of Hsp70 subfamily, Ssa, in protein folding in yeast cells, seen in luciferase-transformed ssa mutants.

To investigate the role of constitutive hsp70 in protein folding and to probe the supplementation by other hsps in this folding, yeast cells expressing reduced constitutive hsp70 proteins, ssa1ssa2, were transformed with a plasmid expressing a bacterial luciferase protein. With several independent clone cells of transformants, the levels of luciferase activity and some hsps, such as hsp104, hsp90, hsp70 and hsp26, were examined. The luciferase activity was significantly lower in ssa1ssa2 transformants than in the wild type (wt) cell transformed with the same plasmid. Among several clone cells of ssa1ssa2, the cells with higher luciferase activities exhibited higher amounts of Ssa4 which is known to be expressed instead of lacking Ssa1 and Ssa2. The luciferase activity closely correlated with the amount of Ssa proteins, more than with the amount of other hsps. It is suggested that constitutional Ssa "chaperones" are needed for the folding of proteins and, in cells lacking Ssa1 and Ssa2, the increased Ssa4 is thought to partly compensate for their role in the folding of luciferase in vivo.

Viral behavior of paracrystalline inclusions in osteoclasts of Paget's disease of bone.

Fresh tissues from six patients with Paget's disease of bone were examined ultrastructurally to investigate whether the characteristic paracrystalline inclusions in pagetic osteoclasts revealed viral behavior. These inclusions appeared as microfilamentous aggregates in both nuclei and cytoplasm of the osteoclasts in all six cases. The filamentous elements of the inclusions with a diameter of 11-15 nm showed tubular structures with a central electron-lucent zone measuring 5-7 nm in diameter. Viral budding-like structures containing these inclusions were found at the peripheral cytoplasm or cell processes in the ruffled border of some pagetic osteoclasts in two cases. The inclusions in the budding-like structures were often arrayed in a parallel fashion on the cytoplasmic side of the cell membranes of extruded cytoplasm or cell processes. Virion-like particles were also found in the extracellular spaces of the ruffled border. Marked nuclear degeneration was often seen in pagetic osteoclasts of three cases, although other nuclei in the same osteoclasts appeared normal. The degenerated nuclei showed nuclear ring formation where destroyed nuclear membranes were seen and disappearance of nuclear matrices was noted. Since the modifications were always associated with the accumulation of abundant inclusions, they were probably caused by the inclusions. These findings suggested that the inclusions showed viral behavior in pagetic osteoclasts, and that the nuclear modifications were caused by virus infection.

A case of angiocentric T-cell lymphoma presenting as lethal midline granuloma.

A case of angiocentric T-cell lymphoma with glabellar skin being the only affected site was reported. A 43-year-old woman had a several months' history of glabellar swelling followed by progressive and destructive ulceration of the region. Histopathology of the biopsied specimen showed panniculitis with atypical lymphocytes and degenerative vessels filled with atypical cells. Most of the infiltrated lymphocytes were positive for CD2, CD3, CD4 and CD5. Antibodies to Epstein-Barr virus were detected in the patient serum. Intensive chemotherapy using a protocol of the L17M regimen was partially effective for clinical manifestations. Angiocentric T-cell lymphoma presenting as lethal midline granuloma is a rare but distinct entity with an acute fatal course.

Smooth tubular aggregates associated with plasmalemmal invagination in alveolar soft part sarcoma.

The ultrastructure of four alveolar soft part sarcomas was examined to search for ultrastructural signs of myogenic or neural origin. A new ultrastructural structure, an unusual tubular structure, was found in two of four cases. The structure appeared as a large, smooth, tubular aggregate in the cytoplasm of some tumor cells but did not show a honeycomb arrangement of tubules. The aggregate was composed of long, serpentine, branching, smooth, irregularly arranged tubules without ribosomes that ran in various directions. The aggregates intermingled with small amounts of cytoplasmic organelles. Because the aggregated tubules were at times continuous with cell membranes, it was shown that they were the complex extensions or invaginations of cell membranes. Neither myelin-axon complexes nor myofilaments, including Z band material, were seen in any case. There was a possibility that the smooth tubular aggregate was a T-tubule-like structure, suggesting that the tumors were derived from skeletal muscle cells.

The effect of cyclosporin A on Angiostrongylus cantonensis infection and eosinophilia in mice.

Male BALB/c mice were infected with Angiostrongylus cantonensis and at various times p.i. treated with cyclosporin A (CsA) either for 5 days continuously or intermittently, or for 12-16 days on alternate days. They were monitored for peripheral blood eosinophilia and at necropsy examined for CSF and bone marrow eosinophilia, and worm recovery. CsA treatment provoked a transient inhibition of peripheral blood eosinophilia in all groups examined, followed by rebounding eosinophilia. There was no significant difference in bone marrow and CSF eosinophilia between CsA- and vehicle-treated groups. Mice treated with CsA on alternate days yielded lower intracranial worm recovery with small-sized worms at days 7, 21 and 30 p.i. than vehicle-treated groups did. No significant reduction in worm recovery was noted in mice treated for 5 days either continuously or intermittently, although worms, especially female ones, harvested from groups treated with CsA from days--1 to 3 or from days 13 to 17 were significantly smaller than those from vehicle-treated groups. CsA-treatment suppressed blastogenic responses of spleen cells against Con A or worm antigen at days 7 and 21 p.i. In vitro treatment of the 3rd stage larvae with CsA did not adversely influence survival of A. cantonensis in mice. These data indicate that CsA exerts anti-parasitic effects against A. cantonensis in mice.

Morphogenesis and origin of fibrous long-spacing collagen fibers in collagenase-treated mouse skin tissues.

Morphogenesis and origin of fibrous long-spacing collagen (FLS) fibers in newborn mouse skin tissues treated with collagenase were examined using ultrastructural observation, morphometry, histochemical methods, and immunoelectron microscopy. The enzyme caused both the partial destruction of basal laminae and the formation of abundant FLS fibers in the dermal matrix. The fibers were usually distributed in the vicinity of basal laminae in the capillaries or basal layer cells. The fibers were characterized by the cross-striated dark bands with about 91 nm periodicity and longitudinally aligned filaments with a diameter of about 6.5 nm. The dark bands of FLS fibers were often continuous with the basal laminae. Histochemical results showed that the dark bands contained the similar mucopolysaccharides which were involved in the basal laminae. Immunoelectron microscopic results showed that laminin was present in the dark bands as well as in the basal laminae, and that type VI collagen was located in the filaments of FLS fibers. These results suggest that the dark bands are formed by products similar to basal laminae and that the products were precipitated on type VI collagen-contained filaments with periodic intervals of about 91 nm. Morphometric examination revealed that there was no differences in ultrastructure between FLS fibers of a collagenase-treated mouse and those of a human neural tumor.

[Studies on HLA typing in a family line with spontaneous pneumothorax].

Our patient with spontaneous pneumothorax is the oldest brother of 3 siblings. Pneumothorax had occurred in his younger sister and in 3 of his mother's 4 siblings also. Namely, pneumothorax had occurred in 5 (including our patient) of 7 persons, family line on mother's side, and thoracotomy had been performed in 3 of them. Two cases in which operative findings were obtained were introduced. Next, HLA and alpha 1-antitrypsin (alpha 1-AT) were determined in this family line. As the result, the alpha 1-AT all remained in the normal range and HLA typing was not in a special relation to the occurrence of pneumothorax in this family line.

Study on the biodistribution of deuterated biomolecules in mice aiming at new diagnostic radio-imaging agents.

Deuterated compounds (2H-compounds) labeled with 14C prepared from deuterated algae, Chlorella ellipsoidea, were examined for their time-coursed distribution in mice after intravenous administration. The 14C-2H-compounds were fractionated and isolated from algae grown in practically 100 mol% 2H2O in the presence of 14C-bicarbonate. The fractions obtained were the "basic" and "acid" fractions, composed mainly of amino acids and sugar phosphates, respectively, and glucose, galactose, and lipid fractions. All fractions were examined for their biodistribution in mice bearing Ehrlich solid tumor in comparison with the fractions isolated from ordinary Chlorella (1H-Chlorella). 2H-Compounds thus examined showed some behaviors different from 1H-compounds. The 2H- "basic" fraction distributed more slowly in heart, lung and liver than the 1H-fraction. The 2H-specific large distribution in tumor was also observed on this fraction. The 2H-dependent characteristics in the distribution of glucose and galactose differed. The 2H-glucose level was lower in blood and higher in brain, resulting in a brain/blood ratio approximately twice that of 1H-glucose, while 2H-galactose did not show such a characteristic. These findings may be useful for the application of 2H-biomolecules to functional radio-imaging agents for nuclear medicine.

[Chondrosarcoma of the ribs: report of two cases].

Chondrosarcoma of rib origin is relatively rare in Japan. Two patients with chondrosarcoma of rib origin were treated surgically. Case 1: A 79-year-old male, 4 years ago, he noticed a tumor in the left anteroinferior part of the chest. The histological examination of the specimen obtained by incisional biopsy showed osteochondroma in another hospital and he had been under observation. Since the tumor has recently increased in size, he was referred to our department. Radical resection of the chest wall was performed. As expected, the tumor was found to be a secondary chondrosarcoma by postoperative histological examination. Case 2: A 49-year-old female. In the follow up studies after radical nephrectomy, Tc-MDP scintigram revealed an accumulation in the left second rib. Under the diagnosis of rib metastasis, radical resection was performed. The histological examination showed chondrosarcoma of rib origin postoperatively. This lesion was a primary central type, and is considered to have been resected in the earliest stage of all cases we have found in the Japanese literature.

[Expanded indication of percutaneous transluminal coronary angioplasty to the infarcted zone--using thallium-201 exercise scintigram after rehabilitation].

In some cases of old myocardial infarction (OMI), left ventricular wall motion was improved after revascularization, though viability of the infarcted myocardium was not detected by left ventriculography (LVG) and exercise thallium-201 myocardial scintigraphy (EX-Tl). So to obtain more correct information of the viability, EX-Tl was performed again after ergometer rehabilitation for an OMI patient whose myocardial viability of the infarcted zone could not detected by LVG and EX-Tl. Incomplete fill in was showed in the EX-Tl after rehabilitation. So percutaneous transluminal coronary angioplasty (PTCA) was performed. Left ventricular wall motion was improved after three months. EX-Tl after rehabilitation is useful to evaluate the viability that could not detected by LVG and EX-Tl.

[Angiographic findings in a patient with spontaneous hemopneumothorax].

We experienced one patient with spontaneous hemopneumothorax in whom angiography was performed preoperatively. The patient was a 23-year-old female and her plain chest X-ray revealed an index-finger-tip-sized bulla at the left apex and a narrow restiform shadow connecting the pleural cupola with the bulla. Left subclavian arteriography revealed preoperatively that the restiform shadow consisted of aberrant vessels branched from the left costocervical trunk and distributed in and around the bulla at the apex. In the present study very rare angiographic findings and excised specimens obtained from the patient are shown, with reference to surgical indication of spontaneous pneumothorax.