In vivo imaging of axonal transport of mitochondria in the diseased and aged mammalian CNS

The lack of intravital imaging of axonal transport of mitochondria in the mammalian CNS precludes characterization of the dynamics of axonal transport of mitochondria in the diseased and aged mammalian CNS. Glaucoma, the most common neurodegenerative eye disease, is characterized by axon degeneration and the death of retinal ganglion cells (RGCs) and by an age-related increase in incidence. RGC death is hypothesized to result from disturbances in axonal transport and in mitochondrial function. Here we report minimally invasive intravital multiphoton imaging of anesthetized mouse RGCs through the sclera that provides sequential time-lapse images of mitochondria transported in a single axon with submicrometer resolution. Unlike findings from explants, we show that the axonal transport of mitochondria is highly dynamic in the mammalian CNS in vivo under physiological conditions. Furthermore, in the early stage of glaucoma modeled in adult (4-mo-old) mice, the number of transported mitochondria decreases before RGC death, although transport does not shorten. However, with increasing age up to 23–25 mo, mitochondrial transport (duration, distance, and duty cycle) shortens. In axons, mitochondria-free regions increase and lengths of transported mitochondria decrease with aging, although totally organized transport patterns are preserved in old (23- to 25-mo-old) mice. Moreover, axonal transport of mitochondria is more vulnerable to glaucomatous insults in old mice than in adult mice. These mitochondrial changes with aging may underlie the age-related increase in glaucoma incidence. Our method is useful for characterizing the dynamics of axonal transport of mitochondria and may be applied to other submicrometer structures in the diseased and aged mammalian CNS in vivo.Globally, longevity is increasing, and the cohort aged 60 y or over is the fastest growing portion of the population. These trends place neurodegenerative diseases among the greatest clinical threats. Glaucoma, the most common progressive neurodegenerative eye disease (1), globally affects an estimated 60.5 million people, of whom 8.4 million are bilaterally blind (2). Similar to other neurodegenerative diseases of the CNS, including Alzheimer’s disease (3) and Parkinson’s disease (4), the incidence of glaucoma (5) increases with aging. Glaucoma is characterized by axon degeneration and the death of retinal ganglion cells (RGCs) (6). Histological studies in glaucoma models have suggested disturbances in the axonal transport of mitochondria in RGCs (7, 8).Axonal transport is essential for delivering the organelles and proteins that are required for axonal function and maintenance. Mitochondria are organelles that must be transported in axons and distributed appropriately to function (9, 10), because mitochondria play a pivotal role in the function and survival of neurons by generating ATP, maintaining Ca²⁺ and reduction-oxidation (redox) homeostasis, and signaling in apoptosis. Disturbances in mitochondrial dynamics are suggested to be involved in neurodegenerative diseases and CNS aging (11–14). In vivo imaging of axonal transport of mitochondria has been reported using explant imaging of the Drosophila nervous system (15) and rat cerebellar slice cultures (16) and intravital imaging (direct in vivo imaging of living animals at subcellular resolution) of the mouse peripheral nervous system (17) and zebrafish nervous system (18). However, intravital imaging of axonal transport of mitochondria has not been achieved in the mammalian CNS. Importantly, the lack of intravital imaging of axonal transport of mitochondria in the mammalian CNS under physiological oxygen levels and metabolism and with intact blood flow has precluded the characterization of the dynamics of axonal transport of mitochondria in the CNS of diseased and aged mammals in vivo.To perform intravital imaging of axonal transport of mitochondria in the mammalian CNS, we developed the technique we call “MIMIR” (for “minimally invasive intravital imaging of mitochondrial axonal transport in RGCs”). MIMIR does not involve thinning or opening of the sclera or produce changes in the intraocular humor. MIMIR directly showed, at submicrometer resolution, that axonal transport of mitochondria is highly dynamic in the mammalian CNS in vivo under physiological conditions. It enabled us to characterize disturbances of mitochondrial transport in a mouse model of glaucoma and age-related changes of mitochondrial transport in old (23- to 25-mo-old) mice.     

Pancreatic arteriovenous malformation with duodenal ulcer

Summary We report the color Doppler ultrasonography features of arteriovenous malformation (AVM) of the pancreas, a very rare disease. The patient was a 52-year-old man with congenital AVM of the pancreas and a duodenal ulcer that had been resistant to medication. Endoscopic color Doppler ultrasonography (color Doppler EUS) revealed many abnormal color signals showing pulsatile wave form at the portion of the duodenal wall involving the duodenal ulcer. Extracorporeal color Doppler ultrasonography revealed a mosaic-like color signal, caused by turbulent flow, in the portal trunk. Angiography demonstrated a vascular network with extensive proliferation at the pancreatic head and early portal filling. It is possible that the pancreatic AVM had caused the duodenal ulcer. Color Doppler EUS can be a useful modality for detection of vessel abnormalities of the gastrointestinal tract.     

Reactivity of recombinant treponema pallidum (r-Tp) antigens with anti-Tp antibodies in human syphilitic sera evaluated by ELISA

We evaluated the immunoreactivity of recombinant Treponema pallidum (r-Tp) antigens with human sera by indirect enzyme-linked immunoabsorbant assay (ELISA). We expressed antigens with a molecular weight (MW) of 17KDa, 15KDa, 47KDa, and 42KDa, which are believed to be major immunoreactive membrane proteins of Tp cells. The expressed proteins were described by adding the prefix M, S, and G to the corresponding Tp antigens, namely, mature antigens, signal sequence containing antigens, and glutathione s-transferase (GST)-fused antigen in this report. A rather high expression occurred for M47 and S42 proteins in the Escherichia coli system, whereas for M15 and M17 proteins, a poor expression was observed. However, a fairly high expression occurred for G15 and G17. Thus expressed proteins were purified by means of chromatographies to a level of > 95%, and the purified proteins were found to be reactive with TPHA positive serum by Western blotting (WB). An ELISA performed with a serum of 1/1000 dilution using these purified antigens for coating on the solid phase showed that G17 antigen was more effective in detecting syphilis antibodies in human serum than M47, S42, and G15. There was a good consistency between ELISA and TPHA, whereby the cutoff indexes (CI) on ELISA showed a correlation coefficient of 0.7276 in logarithmic TPHA titers. J. Clin. Lab. Anal. 11:315–322, 1997. © 1997 Wiley-Liss, Inc.     

The patterns of beta-cell regeneration in untreated diabetic and insulin-treated diabetic Syrian hamsters after streptozotocin treatment.

Our previous study showed that Syrian hamsters recover from streptozotocin (SZ)-induced diabetes spontaneously and that insulin therapy adversely affects diabetes. The recovery and lack of recovery was associated with beta-cell regeneration and lack of regeneration. In this study, we examined the patterns of beta-cell regeneration in hamsters by using an immunohistochemical-autoradiographical method. We used tritiated thymidine to examine the DNA synthesis in islet cells and evaluated the number of labeled cells that were or were not immunoreactive with antiinsulin, antiglucagon, and anatisomatostatin. The results showed that, in untreated control hamsters, beta-cell regeneration took place both from preexisting beta cells and from undifferentiated cells (labeled cells unstained with any of the three antibodies) in equal proportions. In the SZ-treated hamsters, however, most regenerating cells seemed to derive from undifferentiated cells within the islets. Insulin therapy inhibited beta-cell regeneration from both the preexisting cells, but more so from the undifferentiated cells. The number of extrainsular islet cell foci was the same in all treatment groups and paralleled the number of islets in each animal during the entire experiment. We concluded the following about diabetic hamsters: 1. beta-Cell regeneration occurs primarily from undifferentiated cells within the islet; 2. Exogenous insulin inhibits beta-cell differentiation from the precursor cells; and 3. The response of intrainsular and extrainsular islet cells to SZ and insulin therapy is similar and the number of extrainsular islet cell foci per pancreas parallel the changes in the number of islets.     

Acute pancreatitis associated with anomalous union of the pancreaticobiliary ductal system.

Between 1978 and 1989, 13 of 48 patients with anomalous union of the pancreaticobiliary ductal system (AUPBD) were diagnosed as having acute pancreatitis. We have studied the clinical, radiologic, and surgical features of these 13 patients. A transient rise in the intraductal pressure of the pancreatic duct during an episode of abdominal pain is responsible for pancreatitis in patients with AUPBD. This rise in the intraductal pressure must be due to bile reflux into the pancreatic duct when an abnormally long common channel is blocked by cholelithiasis, protein plug, or dysfunction of the sphincter of Oddi. The pancreatitis resolves when the common channel obstruction is removed, and bile and pancreatic juice flow easily into the duodenum. We believe that this phenomenon is responsible for acute relapsing pancreatitis. It is our belief that the pancreas appears almost normal during symptom-free intervals.     

The impact of anti-inflammatory cytokines provoked by CD163 positive macrophages on ventricular functional recovery after myocardial infarction

Present study aimed to investigate the impact of anti-inflammatory cytokines provoked by the hemoglobin scavenger receptor, CD163, on left ventricular (LV) functional recovery after successful reperfusion in patients with acute myocardial infarction (AMI). Intraplaque hemorrhage accelerates plaque destabilization. Extracellular hemoglobin is cleared by CD163, a macrophage scavenger receptor. This process provokes secretion of anti-inflammatory atheroprotective cytokine, interleukin (IL)-10. In 40 patients with the first AMI, coronary atherothrombotic debris was retrieved during percutaneous coronary intervention (PCI), stained with antibodies to CD163 and IL-10. LV function was determined by echocardiography before PCI and 6 months after PCI. %CD163 was defined as ratio of CD163 (+)-cells to whole cells. %IL-10 was expressed as the ratio of positively stained areas per total tissue. Patients were divided into two groups depending on the amount of CD163 (+)-cells: CD163 > 10 % (CD_163high, n = 20) and CD163 ≤ 10 % (CD_163low, n = 20). CD_163high group had significantly higher %IL-10. Final thrombolysis in myocardial infarction (TIMI) flow grade was significantly lower in CD_163high group. In subgroups with the final TIMI-3 flow (CD_163high-Reflow, n = 15 and CD_163low-Reflow, n = 20), the time to reperfusion, infarct size, LV dimensions and fractional shortening (%FS) before PCI were similar. Significant correlation was observed between %IL10 and changes in LV dimensions (diastole, r = ?0.49, P = 0.01; systole, r = ?0.65, P < 0.01) or %FS (r = 0.51, P < 0.01) at 6 months after PCI. Plaque with CD163(+)-macrophages could impair distal flow after primary PCI. However, CD163(+)-macrophages enhance the anti-inflammatory cytokine expression that aids in ventricular functional recovery if distal flow can be achieved by successful reperfusion.     

Impact of the perioperative neutrophil-to-lymphocyte ratio on the long-term survival following an elective resection of colorectal carcinoma

Purposes

The present study was designed to evaluate the prognostic value of the perioperative neutrophil-to-lymphocyte ratio (NLR) for the long-term survival in patients with colorectal cancer.

Methods

This was a retrospective study of 524 patients with histologically proven stage II or III colorectal cancer who underwent curative colorectal resection. We classified patients into a low NLR group or high NLR group base on their NLR values at three time points (before surgery (Pre), on the first postoperative day (POD1), and on the third or fourth postoperative day (POD3)) and evaluated the survival according to the group.

Results

The cancer-specific survival was significantly longer in the groups with a low NLR on POD3. The disease-free survival was significantly longer in the group with a low NLR on Pre. We subsequently allocated a score of 1 to patients with a high NLR at each point and reclassified patients into those with a low perioperative NLR group (score of 0 or 1) and high perioperative NLR group (score of 2 or 3). Both the cancer-specific survival and disease-free survival rates were significantly different between the two perioperative NLR groups. Both univariate and multivariate analyses demonstrated that being in the high perioperative NLR group was an independent risk factor for both the cancer-specific survival and disease-free survival.

Conclusions

Not only the preoperative but also the postoperative NTR is thus considered to be a predictor of the long-time survival in patients with colorectal cancer.     

Role of prostaglandin D2 in the hypothermia of rats caused by bacterial lipopolysaccharide.

The intraperitoneal administration of lipopolysaccharide from Salmonella typhimurium (1 mg/kg) caused a fall in the rat colonic temperature of about 2 degrees C at an ambient temperature of 22 +/- 3 degrees C. The hypothermia induced by the lipopolysaccharide was abated in a dose-dependent manner by the administration of indomethacin. Other inhibitors of prostaglandin synthetase such as aspirin, flufenamic acid, and phenylbutazone had effects similar to those of indomethacin. When various prostaglandins were injected intracerebroventricularly, only prostaglandin D2 caused a dose-dependent fall in the colonic temperature at doses between 1.2 and 6 nmol/kg. Microinjection of prostaglandin D2 into the preoptic area caused hypothermia of about 1 degree C. However, injection of prostaglandin D2 into the posterior hypothalamus had little effect on the colonic temperature. The hypothermia caused by prostaglandin D2 was not abated by the administration of indomethacin. The amount of prostaglandin D2 increased significantly in the preoptic/hypothalamic region of rat brain 1 hr after the intraperitoneal administration of the lipopolysaccharide, whereas such increase was not observed in rats pretreated with indomethacin. The in vitro incubation of the preoptic/hypothalamic slices with the lipopolysaccharide also increased the amount of prostaglandin D2. These results suggest that the intraperitoneal administration of the lipopolysaccharide induces the release of prostaglandin D2 in the preoptic/hypothalamic area of rat brain and that the latter compound is involved in the hypothermic response of rats to the lipopolysaccharide.     

Detection of intracardiac thrombi in a case of cardiomyopathy by two dimensional echocardiography.

A case of cardiomyopathy with systemic emboli is reported. Two dimensional and M-mode echocardiography showed abnormal intracavitary echoes. After further systemic emboli, echocardiograms showed the complete disappearance of the abnormal echoes found previously in the apex, which suggested that the emboli resulted from detachment of the left ventricular mural thrombi.     

Clinical investigation on acute respiratory failure in patients with severe hematologic malignancy

Ten patients with severe hematologic malignancies (four with acute leukemia, three with multiple myeloma, one with prolymphocytic leukemia, one with malignant lymphoma and one with blastic crisis of chronic myelogenous leukemia) developed respiratory failure during the period between April 1986 and May 1990. Clinically, the patients manifested high-fever, dyspnea refractory to oxygen therapy, diffuse pulmonary rales and severe hypoxemia without evidence of cardiogenic pulmonary edema. Chest roentgenograms displayed diffuse alveolar infiltrates. Respiratory failure occurred as early as 48 hours and as late as 66 days after the administration of intensive anti-neoplastic chemotherapy. At that time leukocyte count was between 100/microliters and 54,900/microliters. Marked leukocytosis was observed in two patients with AML and PLL. Respiratory failure was preceded by sepsis in one patient with AML and by pneumonia in nine patients. DIC was diagnosed in four patients. All patients treated with high dose methyl prednisolone (mPSL) within 12 hours after the onset of respiratory failure. Only one patient required assisted ventilation. High dose mPSL had significant effect on seven of ten patients. But three patients died from progressive respiratory failure, sepsis, pneumonia and multi-organ failure.