Clinical impact of hyperglycemia on days 0-7 after allogeneic stem cell transplantation.

In order to clarify the association between hyperglycemia during the early period after allogeneic stem cell transplantation (allo-SCT) and adverse outcomes, we retrospectively analyzed 563 consecutive patients who underwent allo-SCT at our institute between 2008 and 2015. Patients were categorized into three groups according to mean fasting blood glucose levels on days 0-7 (normoglycemia group<110 mg/dL, n=347; mild hyperglycemia group 110-149 mg/dL, n=192 and moderate/severe hyperglycemia group≥150 mg/dL, n=24). The median follow-up was 2.7 years. Patients in the moderate/severe hyperglycemia group had significantly worse characteristics. The cumulative incidences of 2-year non-relapse mortality (NRM) and the probabilities of 2-year overall survival (OS) in the normoglycemia, mild hyperglycemia and moderate/severe hyperglycemia groups were 7.5%, 19% and 29%, respectively (P<0.01), and 69%, 53% and 33%, respectively (P<0.01). In multivariate analyses, hyperglycemia was an independent predictor of high NRM (vs normoglycemia; mild hyperglycemia, hazard ratio (HR) 2.56, 95% confidence interval (CI) 1.56-4.18; moderate/severe hyperglycemia, HR 4.46, 95% CI 1.92-10.3) and poor OS (vs normoglycemia; mild hyperglycemia, HR 1.54, 95% CI 1.14-2.07; moderate/severe hyperglycemia, HR 1.61, 95% CI 0.89-2.91). In conclusion, hyperglycemia on days 0-7 after allo-SCT was associated with inferior outcomes.

Historical changes in epidemiology of diffuse panbronchiolitis.

BACKGROUND AND OBJECTIVE: Japanese pulmonologists, experienced in treating patients with diffuse panbronchiolitis (DPB) prior to the 1980s, have uniformly observed that new incidences of DPB are now a rare event in Japan. However, there is no epidemiological data to support this observation. We examined epidemiological trends of the number of patients with DPB in a large company. DESIGN: The computerized health records of JR East Company employees were used to identify patients with DPB and then these were followed up using the assessments of these patients in JR Tokyo General Hospital and two other JR hospitals. The whole study period was 27 years (1976-2003), although detailed analyses were carried out for three specific periods; the first was 1976-1980, the second was 1989-1993, and the third was 1999-2003. RESULTS: In the first period, 11 DPB cases (four incidence, and seven prevalence) were detected among a total of 355,572 workers. In the second period, three DPB cases (one incidence, and two prevalence) were identified from a total of 180,359 workers. In the third period, no case was found in a total of 144,485 workers. CONCLUSION: This epidemiological trend suggests that both the incidence and prevalence of DPB may have decreased.

FUS immunoreactivity of neuronal and glial intranuclear inclusions in intranuclear inclusion body disease.

AIMS: Recent studies have shown that fused-in-sarcoma (FUS) protein is a component of 'neuronal' intranuclear inclusion bodies (INIBs) in the brains of patients with intranuclear inclusion body disease (INIBD). However, the extent and frequency of FUS-immunoreactive structures in INIBD are uncertain. METHODS: We immunohistochemically examined the brain, spinal cord and peripheral ganglia from five patients with INIBD and five control subjects, using anti-FUS antibodies. RESULTS: In controls, the nuclei of both neurones and glial cells were intensely immunolabelled with anti-FUS and neuronal cytoplasm was weakly positive for FUS. In INIBD, neuronal and glial INIBs in the brain and spinal cord were positive for FUS. FUS-positive INIBs were also found in the peripheral ganglia. The proportion of FUS-positive neuronal INIBs relative to the total number of inclusion-bearing neurones ranged from 55.6% to 83.3% (average 73.2%) and that of FUS-positive glial INIBs ranged from 45.9% to 85.7% (average 62.7%). The nucleus and cytoplasm of inclusion-bearing neurones and glial cells showed no FUS immunoreactivity. CONCLUSIONS: These findings suggest that FUS is incorporated into INIBs in both neurones and glial cells and that loss of normal FUS immunoreactivity may result from reduced protein expression and/or sequestration within inclusions.

Evidence for intermolecular interaction between sulfonium and sulfide sulfur atoms and its application to synthesis of cyclic bis(disulfide) dimer.

On the basis of the remote Pummerer reaction of p-bis(alkylthio)-aromatic S-oxides, the intermolecular interaction between the sulfonium and sulfide sulfur atoms is described. (1) In marked contrast to the Pummerer reaction of 1b-d(3) with (CF(3)CO)(2)O (J. Org. Chem. 1999, 64, 3190-3195), the reaction of 3,3',5,5'-tetramesityl-4-(trideuteriomethylsulfinyl)-4'-(methylthio)biphenyl (1a-d(3)) as a sterically hindered analogue of 1b gave only 2a-d(2). (2) Both reactions of the two unsymmetrical regioisomers of 1-(ethylthio)-4-(methylthio)benzene S-oxide (5a and 5b) with (CF(3)CO)(2)O afforded a mixture of the mono-Pummerer products 6a and 6b, the bis-Pummerer product 7, and the bis-sulfide 8 in a similar ratio. The quenching at the initial stage of both reactions produced 5a, 5b, 8, and the bis-sulfoxide 10 in a similar ratio. These results indicate the equilibrium in the intermolecular interaction between the sulfur atoms. (3) The reaction of the p-bis(benzylthio)-aromatic S-oxide 16 with (CF(3)SO(2))(2)O gave the cyclic bis(disulfide) dimer 17 for the diphenyl sulfide and diphenylmethane spacers or the cyclic tetrakis(disulfide) tetramer 19 for the benzene and biphenyl spacers via the debenzylation of an intermolecular dithia dication. The cyclic bis(dithia dication) dimer A resulting from the intermolecular interaction between the sulfonium and sulfide sulfur atoms is proposed as an intermediate throughout the present reactions.

Intramedullary hemorrhage caused by arteriovenous malformation: a case of mixed lateral and medial medullary syndrome.

A 48-year-old man with no known risk factor for cerebrovascular disease, other than cigarette smoking, experienced the sudden onset of a mixed lateral and medial medullary syndrome. Computed tomography scan failed to show any definite abnormality. Magnetic resonance imaging scans revealed hemorrhage restricted to the left dorsolateral medulla. Angiography showed abnormal arteries originating from the left vertebral artery with small niduses located on the surface of the medulla and contralateral cerebellum. Small brain-stem hemorrhages are a contraindication to thrombolytic or anticoagulant therapy, and therefore must be recognized in the acute stage.

Dysphagia in children with oculo-auriculo-vertebral spectrum.

Feeding difficulty necessitating tube feeding after the infantile period was seen in 3 children with oculo-auriculo-vertebral spectrum. Videofluorographic imaging showed impaired pharyngeal function, which was thought to result from dysplasia of the pharyngeal muscles. Note should be made of feeding difficulty in patients with oculo-auriculo-vertebral spectrum.

Subsequent epilepsy in very-low-birthweight infants: a long-term follow-up study from birth.

We studied 197 survivors of 290 very-low-birthweight (VLBW, < 1,500 g) infants admitted to our neonatal intensive care unit from 1977 through 1982. The children were all followed until at least age 6 years (mean 10 years 6 months). Eight children (4.1%) had epilepsy: 5 had generalized, 2 had unilateral, and 1 had partial seizures. Two (1.0%) had active and poorly controlled epilepsy. Three had a history of epileptic seizures, but none for > or = 6 years, and 3 were no longer receiving antiepileptic drug (AED) treatment. Most (5 of 8) were severely multiply handicapped. As compared with VLBW children without epileptic seizures and neurodevelopmental abnormalities, VLBW children with epileptic seizures had a gestational age < 27 weeks, a weight < 1,000 g, severe perinatal complications as indicated by an Apgar score of < 4 at 5 min, and the need for long-term oxygen administration.

Inherent bias in correlation averaged images.

The correlation averaging algorithm frequently used to enhance micrographs of repeating structures contains an inherent bias that favours images with larger pixel values or positive noise levels. This bias not only skews the composite image toward higher pixel values, but also distorts the image by increasing the value of high-valued pixels more than that of low-valued pixels. These errors are especially important in scanning probe microscopy images where the pixel value reflects a distinct height. A similar algorithm that uses a structure function in place of the correlation function eliminates this bias.

Ultrastructural and biochemical observations on antineutrophil antibody- and complement-induced immuno-injury to equine neutrophils.

Antibody-induced damage to neutrophils was studied to elucidate processes associated with destruction of neutrophils in immune-mediated neutropenias. Cytomorphological changes and release of certain cellular constituents were determined for neutrophils treated with an antineutrophil serum in the presence or absence of rabbit complement. Neutrophils exposed to the antineutrophil serum alone showed endocytotic vacuoles and degranulation. In contrast, neutrophils exposed to the antineutrophil serum and complement showed marked morphologic changes. The plasma membrane developed numerous vesicles, villous processes and minute areas of bilayer discontinuity. Highly damaged cells exhibited cellular and nuclear swellings, disruption of cytoplasmic integrity and disordered distribution of lysosomal granules. Cytoplasmic constituents (K+ and lactate dehydrogenase) were released extracellularly from neutrophils exposed to the antineutrophil serum with or without complement. Cytological changes induced by the antineutrophil serum and complement were analogous to those reported for leucocytes exposed to the activated complement components C5b-9 (the membrane attack complex) and bacterial toxins. It was concluded that the cytological abnormalities observed were most probably associated with immune-mediated damage to the cell membrane, leading to leakage of cytoplasmic constituents like K+, colloidal osmotic swelling, and disruption of the cytoskeletal system.

Methods for detection of immune-mediated neutropenia in horses, using antineutrophil serum of rabbit origin.

Equine neutrophil antibody was raised in rabbits inoculated with equine neutrophils isolated to purity greater than 99.0%, using Percoll density-gradient sedimentation. Neutrophil antibody was detected by use of agar gel diffusion, leukoagglutination, indirect immunofluorescence, staphylococcal protein A and streptococcal protein G binding, and phagocytic inhibition techniques. Precipitin lines and leukoagglutination were seen in antiserum dilutions of 1:4 and 1:64, respectively. The specific nature of leukoagglutination was characterized by the formation of rosette-like clumps of neutrophils. Specific bright membranous fluorescence was seen in neutrophils treated with the antiserum and exposed to fluorescein-conjugated goat anti-rabbit immunoglobulin, and staphylococcal protein A and streptococcal protein G. Whereas the indirect immunofluorescence and protein G-binding tests were equally sensitive and resulted in titer of 1:256, the protein A-binding test was less sensitive and resulted in titer of only 1:32. Nonspecific binding of protein A and protein G was noticed as uniform or patchy cellular fluorescence in a small number of neutrophils. Treatment of neutrophils with antiserum up to dilution of 1:8 resulted in a significant (P less than 0.05) suppression of phagocytosis of opsonized zymosan particles. Thus, protein G-binding and indirect immunofluorescence tests are highly sensitive to detect neutrophil antibody and may be used to diagnose immune-mediated neutropenias in horses and, possibly, in other animal species.

Cytochemical studies of normal feline blood and bone marrow cells.

Blood and bone marrow cells of ten clinically healthy cats were stained for alkaline phosphatase (ALP), peroxidase (PO), chloroacetate esterase (CAE), alpha-naphthyl butyrate esterase (NBE), sudanophilia, and periodic acid-Schiff (PAS) reaction. Mature neutrophils in blood and bone marrow were devoid of ALP and NBE, but exhibited modest to strong PO, CAE, sudanophilia, and PAS reaction. In bone marrow, sudanophilia, PO, and CAE were prominent at the promyelocyte stage and diminished with cellular differentiation and maturation, while PAS reactivity increased with cell maturation usually from the myelocyte stage onwards. Myeloblasts were negative for all cytochemical reactions, but some large unidentifiable cells reacted strongly for ALP. Eosinophils were slightly reactive for ALP, CAE, and PAS, but not for PO, sudanophilia, and NBE. Basophil granules stained strongly for CAE, revealed PAS positivity, and stained negatively for PO, NBE, ALP, and sudanophilia. Slight ALP activity was detected in the intergranular cytoplasm of basophils. Lymphocytes and monocytes, with few exceptions, stained negatively. An occasional lymphocyte revealed slight globular NBE activity (NaF-resistant) and diffuse PAS reaction, while an occasional monocyte contained a few PO-positive and sudanophilic granules. Monocytes reacted modestly, whereas bone marrow macrophages reacted strongly for NBE (NaF-sensitive). Cells of the erythroid series stained negatively for all cytochemical reactions, megakaryocytes were PAS-positive, and platelets gave positive reactions for PAS and CAE.

Shape changes in caprine erythrocytes exposed to chlorpromazine and lysolecithin.

Stomatocytic and echinocytic transformations of caprine erythrocytes were studied in vitro using chlorpromazine as a stomatocyyic agent and lysolecithin as an echinocytic agent. Morphologic changes in erythrocytes generally varied with the cell shape and the concentration of the substances used. Discoytic, triangular, and pear-shaped red cells common to normal goats, exhibited classical stomatocytic and echinocytic changes with formation of spherostomatocytes, sphero-echinocytes, and spherocytes. In comparison, fusiform and spindle-shaped red cells found in certain Angora goats seemed less prone to shape changes and required greater concentrations of the inducing agents to effect such changes. Chlorpromazine at higher concentrations also inflicted localized membrane damage in form of tiny pits and lysolecithin likewise induced formation of fragile smooth or beaded filaments.

In vitro platelet release by rat megakaryocytes: effect of metabolic inhibitors and cytoskeletal disrupting agents.

Development of an in vitro visual assay facilitated the study of large numbers of megakaryocytes undergoing proplatelet formation in short-term cultures. Approximately 9% of megakaryocytes formed platelets during a 24-hour period. In the presence of an inhibitor of anaerobic glycolysis (NaF), proplatelet formation was inhibited, whereas inhibitors of respiration (NaCN) did not significantly (P greater than 0.05) decrease proplatelet formation. Presence of the microtubule-disrupting agents colchicine and vincristine sulfate in culture medium inhibited proplatelet formation, whereas the microfilament-disrupting agent cytochalasin B had a less pronounced inhibition.

In vitro platelet release by rat megakaryocytes: effect of heterologous antiplatelet serum.

A visual assay to study megakaryocyte platelet release via proplatelet formation in vitro was established. Samples of megakaryocyte-enriched rat bone marrow were incubated (37 C) in RPMI-1640 medium with 15% autologous serum in specially prepared chambers. In the culture system, approximately 6% of megakaryocytes formed proplatelet processes within 24 hours. Inclusion of a heterologous antiplatelet antibody in the culture system inhibited proplatelet formation, compared with that in controls.

Scanning electron microscope study of platelet release by canine megakaryocytes in vitro.

Megakaryocytes were isolated from bone marrow from healthy dogs, using a combination of density-gradient centrifugation and polysucrose-velocity sedimentation techniques. The 2-step separation technique resulted in a preparation comprising 30% to 35% megakaryocytes of total nucleated cells. Accessibility to large numbers of viable canine megakaryocytes allowed investigation of platelet release by these cells in short-term cultures. Megakaryocytes were observed to form long cytoplasmic processes that gradually developed segmental constrictions and subsequently fragmented into platelet-sized pieces. Some platelet-sized cytoplasmic pieces of megakaryocytes presumably underwent discoid transformation.

Circulating proplatelets: isolation and quantitation in healthy rats and in rats with induced acute blood loss.

A technique to isolate megakaryocyte proplatelet processes from blood of rats' hearts, using colloidal silica coated with polyvinylpyrrolidone density gradient, was developed. The proplatelet concentration in blood from right ventricles was significantly higher (P less than 0.001) than that in blood from left ventricles in healthy rats, as well as in rats with induced acute blood loss. The proplatelet concentration of blood from the heart, 24 hours after acute blood loss was induced was significantly (P less than 0.001) increased, indicating that platelet production was accelerated. The demonstration of proplatelets entering the pulmonary circulation indicates platelet release via proplatelet formation. Seemingly, proplatelets are fragmented in the lungs at predesignated locations along the proplatelet process.

Scanning electron microscopic studies of megakaryocytes and platelet formation in the dog and rat.

Megakaryocyte morphology and platelet formation in canine and murine bone marrows were studied by scanning electron microscopy. In situ-fixed bone marrow preparations and cell suspensions of bone marrow provided complementary information for the 2 species (dogs and rats). Cylindrical processes (proplatelets) of variable length and thickness, originating from the megakaryocyte surface, were in the larger marrow sinusoids and the central vein. Regional constrictions along the length of proplatelets, particularly near their apical region, and the presence of fragments of such processes supported the concept of platelet formation through segmentation of proplatelets. Megakaryocytes presented varied morphology. Surface features resembling platelets were observed on megakaryocytes, indicating that platelets may have been released through surface budding. In conclusion, megakaryocytes formed long proplatelet processes that actively migrated to venous sinusoids to release platelets by fragmentation. Scanning electron microscopy analysis revealed a complex and variable megakaryocyte surface topography. The platelet-like structures on megakaryocyte surfaces may represent platelet release by a budding mechanism. The similarity between murine platelet release and canine platelet release demonstrates that data from rodent models may be applicable to nonrodents.