Kanji Uchida

Granulocyte/macrophage–colony-stimulating factor autoantibodies and myeloid cell immune functions in healthy subjects

High levels of granulocyte/macrophage-colony-stimulating factor (GM-CSF) autoantibodies are thought to cause pulmonary alveolar proteinosis (PAP), a rare syndrome characterized by myeloid dysfunction resulting in pulmonary surfactant accumulation and respiratory failure. Paradoxically, GM-CSF autoantibodies have been reported to occur rarely in healthy people and routinely in pharmaceutical intravenous immunoglobulin (IVIG) purified from serum pooled from healthy subjects. These findings suggest that either GM-CSF autoantibodies are normally present in healthy people at low levels that are difficult to detect or that serum pooled for IVIG purification may include asymptomatic persons with high levels of GM-CSF autoantibodies. Using several experimental approaches, GM-CSF autoantibodies were detected in all healthy subjects evaluated (n = 72) at low levels sufficient to rheostatically regulate multiple myeloid functions. Serum GM-CSF was more abundant than previously reported, but more than 99% was bound and neutralized by GM-CSF autoantibody. The critical threshold of GM-CSF autoantibodies associated with the development of PAP was determined. Results demonstrate that free serum GM-CSF is tightly maintained at low levels, identify a novel potential mechanism of innate immune regulation, help define the therapeutic window for potential clinical use of GM-CSF autoantibodies to treat inflammatory and autoimmune diseases, and have implications for the pathogenesis of PAP.

Pulmonary alveolar proteinosis, a primary immunodeficiency of impaired GM-CSF stimulation of macrophages

Pulmonary alveolar proteinosis (PAP) is a rare syndrome characterized by accumulation of pulmonary surfactant, respiratory insufficiency, and increased infections. It occurs in various clinical settings that disrupt surfactant catabolism in alveolar macrophages, including a relatively more common autoimmune disease caused by GM-CSF autoantibodies and a rare congenital disease caused by CSF2RA mutations. Recent results demonstrate that GM-CSF is crucial for alveolar macrophage terminal differentiation and immune functions, pulmonary surfactant homeostasis, and lung host defense. GM-CSF is also required to determine the basal functional capacity of circulating neutrophils, including adhesion, phagocytosis, and microbial killing. PAP research has illuminated the crucial role of GM-CSF in innate immunity and led to novel therapy for PAP and the potential use of anti-GM-CSF therapy in other common disorders.

Relationship of anti-GM-CSF antibody concentration, surfactant protein A and B levels, and serum LDH to pulmonary parameters and response to GM-CSF therapy in patients with idiopathic alveolar proteinosis

Background: Conventional measures of the severity of alveolar proteinosis (AP) include alveolar-arterial oxygen gradient ([A – a]Do2), vital capacity (VC), and carbon monoxide transfer factor (Tlco), but alternative serological measures have been sought. Granulocyte-macrophage colony stimulating factor (GM-CSF) neutralising autoantibody is found in patients with idiopathic acquired AP. We have investigated the interrelationships between the levels of this antibody and those of surfactant protein (SP)-A and -B, lactate dehydrogenase (LDH), and conventional measures of disease severity, and the capacity of these parameters to predict the response to rhGM-CSF treatment. Methods: Blood levels of anti-GM-CSF antibodies, SP-A, SP-B, LDH, and [A – a]do2, VC, and Tlco were measured before rhGM-CSF treatment and every 2 weeks thereafter in 14 patients with AP. Results: At baseline, high levels of anti-GM-CSF antibodies and increased SP-A and SP-B levels were seen in all patients, and LDH was raised in 83%. SP-A was highly correlated with [A – a]do2, VC, and Tlco (p≤0.02), but other markers were not. Only a normal LDH level was predictive of a response to rhGM-CSF treatment (p=0.03). During treatment a correlation between conventional and serological variables within patients was seen only between SP-A and [A – a]do2 (p=0.054), LDH levels and [A – a]do2 (p=0.010), and LDH levels and VC (p=0.019). Conclusions: Of the serological parameters studied, only SP-A and LDH levels were correlated with conventional measures of disease severity, with LDH most accurately reflecting [A – a]Do2 and vital capacity. Only a normal LDH level predicted a higher likelihood of response to treatment with GM-CSF.

Patient-derived Granulocyte/Macrophage Colony–Stimulating Factor Autoantibodies Reproduce Pulmonary Alveolar Proteinosis in Nonhuman Primates

RATIONALE Granulocyte/macrophage colony-stimulating factor (GM-CSF) autoantibodies (GMAb) are strongly associated with idiopathic pulmonary alveolar proteinosis (PAP) and are believed to be important in its pathogenesis. However, levels of GMAb do not correlate with disease severity and GMAb are also present at low levels in healthy individuals. OBJECTIVES Our primary objective was to determine whether human GMAb would reproduce PAP in healthy primates. A secondary objective was to determine the concentration of GMAb resulting in loss of GM-CSF signaling in vivo (i.e., critical threshold). METHODS Nonhuman primates (Macaca fascicularis) were injected with highly purified, PAP patient-derived GMAb in dose-ranging (2.2-50 mg) single and multiple administration studies, and after blocking antihuman immunoglobulin immune responses, in chronic administration studies maintaining serum levels greater than 40 microg/ml for up to 11 months. MEASUREMENTS AND MAIN RESULTS GMAb blocked GM-CSF signaling causing (1) a milky-appearing bronchoalveolar lavage fluid containing increased surfactant lipids and proteins; (2) enlarged, foamy, surfactant-filled alveolar macrophages with reduced PU.1 and PPARgamma mRNA, and reduced tumor necrosis factor-alpha secretion; (3) pulmonary leukocytosis; (4) increased serum surfactant protein-D; and (5) impaired neutrophil functions. GM-CSF signaling varied inversely with GMAb concentration below a critical threshold of 5 microg/ml, which was similar in lungs and blood and to the value observed in patients with PAP. CONCLUSIONS GMAb reproduced the molecular, cellular, and histopathologic features of PAP in healthy primates, demonstrating that GMAb directly cause PAP. These results have implications for therapy of PAP and help define the therapeutic window for potential use of GMAb to treat other disorders.

Standardized serum GM-CSF autoantibody testing for the routine clinical diagnosis of autoimmune pulmonary alveolar proteinosis.

Autoantibodies against granulocyte/macrophage colony-stimulating factor (GMAbs) cause autoimmune pulmonary alveolar proteinosis (PAP) and measurement of the GMAb level in serum is now commonly used to identify this disease, albeit, in a clinical research setting. The present study was undertaken to optimize and standardize serum GMAb concentration testing using a GMAb enzyme-linked immunosorbent assay (GMAb ELISA) to prepare for its introduction into routine clinical use. The GMAb ELISA was evaluated using serum specimens from autoimmune PAP patients, healthy people, and GMAb-spiked serum from healthy people. After optimizing assay components and procedures, its accuracy, precision, reliability, sensitivity, specificity, and ruggedness were evaluated. The coefficient of variation in repeated measurements was acceptable (<15%) for well-to-well, plate-to-plate, day-to-day, and inter-operator variation, and was not affected by repeated freeze-thaw cycles of serum specimens or the reference standards, or by storage of serum samples at -80°C. The lower limit of quantification (LLOQ) of the PAP patient-derived polyclonal GMAb reference standard (PCRS) was 0.78ng/ml. Receiver operating characteristic curve analysis identified a serum GMAb level of 5μg/ml (based on PCRS) as the optimal cut off value for distinguishing autoimmune PAP serum from normal serum. A pharmaceutical-grade, monoclonal GMAb reference standard (MCRS) was developed as the basis of a new unit of measure for GMAb concentration: one International Unit (IU) of GMAb is equivalent to 1μg/ml of MCRS. The median [interquartile range] serum GMAb level was markedly higher in autoimmune PAP patients than in healthy people (21.54 [12.83-36.38] versus 0.08 [0.05-0.14] IU; n=56, 38; respectively; P<0.0001). Results demonstrate that serum GMAb measurement using the GMAb ELISA was accurate, precise, reliable, had an acceptable LLOQ, and could be accurately expressed in standardized units. These findings support the use of this GMAb ELISA for the routine clinical diagnosis of autoimmune PAP and introduce a new unit of measure to enable standardized reporting of serum GMAb data from different laboratories.

Lactate is correlated with the indocyanine green elimination rate in liver resection for cirrhotic patients.

The role of lactate in liver ischemia-reperfusion injury in cirrhosis has not been clarified. Fifty patients with hepatocellular carcinoma who underwent partial liver resection under Pringle’s maneuver were included in this study. We performed the indocyanine green clearance test before the operation and three times during the surgery to calculate its elimination rate. Blood lactate and base excess were measured at the corresponding times. Systolic and diastolic systemic arterial pressure, heart rate, cardiac index, and esophageal temperature were monitored. Aminotransferase levels were recorded the day before the operation, 1 h after the operation, and on the first and third postoperative days. We calculated the increase or decrease in lactate levels during the preischemic, ischemic, and postischemic phases, and examined the correlation between these results and the changes in indocyanine green elimination rate and some clinical factors. The lactate levels increased before reperfusion and began to decrease after reperfusion. The lactate increase and decrease during the ischemic and postischemic phases correlated with the change in indocyanine green elimination rate (P < 0.0001 and P = 0.02 for the respective phases). The lactate increase during the preischemic phase correlated with the duration of the preischemic phase (P < 0.0001). In cirrhotic patients who undergo liver resection with Pringle’s maneuver and who do not show postoperative liver failure, the blood lactate profile might be a reliable indicator of liver metabolic capacity during surgery.

The effects on gastric emptying and carbohydrate loading of an oral nutritional supplement and an oral rehydration solution: a crossover study with magnetic resonance imaging.

BACKGROUND:Preoperative administration of clear fluids by mouth has recently been endorsed as a way to improve postoperative outcomes. A carbohydrate-containing beverage supplemented with electrolytes or proteins may have additional benefits for patients’ satisfaction. However, effects on gastric residual, nausea, and emesis and the effectiveness of these beverages for improving patients’ hydration status have not been well defined. METHODS:We evaluated changes in gastric volume over time by magnetic resonance imaging, as well as blood glucose levels, before and after administration of 500 mL oral rehydration solution (ORS) containing 1.8% glucose and electrolytes in 10 healthy volunteers. The same volume of an oral nutritional supplement (ONS) containing 18% glucose and supplemental arginine (545 mOsm/kg) was given to the same population using a crossover design. RESULTS:The mean (median, 95% confidence interval) gastric fluid volume at 1 hour after oral ingestion was 55.0 (55.3, 39.0–70.9) mL in the ORS group, whereas 409.2 (410.9, 371.4–447.0) mL in the ONS group (P = 0.0002). The gastric fluid volume of all participants in the ORS group returned to <1 mL/kg at 90 minutes after ingestion, whereas none reached <1 mL/kg at 120 minutes in the ONS group. The ONS group showed a sustained increase in the blood glucose level after ingestion (P < 0.0001 to baseline at 30, 60, 120 minutes), while the ORS group showed an initial increase (P < 0.0001, P = 0.01, P = 0.205 at each time point). CONCLUSIONS:ORS supplemented with a small amount of glucose showed faster gastric emptying, which may make it suitable for preoperative administration. In contrast, ONS supplemented with arginine with a relatively low osmolality was associated with a longer time for gastric emptying, although it showed a sustained increase in blood glucose level.

Impact of remifentanil introduction on practice patterns in general anesthesia

PurposeThe introduction of new medicine can change clinical practice patterns and may affect patient outcomes. In the present study, we investigated whether introduction of remifentanil in Japan affected the practice patterns of anesthesia.MethodsUsing the Japanese Diagnosis Procedure Combination database, we extracted records of 423,491 patients who underwent surgery with general anesthesia in 243 hospitals before (2006) and after (2007) the introduction of remifentanil, and identified anesthetic agents used for each patient. A hierarchical mixed-effects logistic regression analysis was performed to analyze the factors that affected selection of remifentanil. Further, we compared postoperative length of stay (LOS), in-hospital mortality, and total costs between 2006 and 2007.ResultsIn 2007, remifentanil was used for up to 41.4% of all general anesthesia, accompanied by a reduction in nitrous oxide use and an increase in total intravenous anesthesia. Female gender, increasing age, and preoperative comorbidities including diabetes mellitus, hypertension, liver cirrhosis, and chronic renal failure were positively associated with the use of remifentanil, whereas accompanying cardiac disease and co-application of epidural anesthesia were negatively associated. In 2007, a similar in-hospital death rate, similar or decreased total costs, slightly reduced duration of anesthesia, and substantially reduced postoperative LOS were seen compared to those in 2006.ConclusionsOur data revealed rapid changes in practice patterns in anesthesia after the introduction of remifentanil in Japan. Remifentanil was used more often in patients with comorbidities and without epidural anesthesia, and its introduction did not affect increase in total medical costs.

External manual compression of the abdominal aorta to control hemorrhage from a ruptured aneurysm.

fluid infusion and subsequent blood transfusion, her SBP gradually increased. She underwent radiological examinations, including abdominal computed tomography (CT), which revealed an infrarenal aortic aneurysm 10cm in diameter. Retroperitoneal hematoma, as well as fluid collection in the peritoneal cavity, suggestive of intraperitoneal spread of hemorrhage, was noted. Because of continuous volume resuscitation, her SBP exceeded 100mmHg at the end of the radiological examinations. By the time she arrived at the operating room, she had received fluid and homologous blood amounting to approximately 2100ml and 720ml, respectively. On arrival at the operating room, her SBP exceeded 130mmHg. Her heart rate (HR) was 90 beats per min (Fig. 1). After she was placed on the operating table, however, her SBP fell abruptly, to less than 50mmHg. Bradycardia immediately followed. After complaining of abdominal pain, the patient became comatose. Her abdomen showed marked swelling. An instantaneous diagnosis of re-rupture of the aneurysm was made, and an anesthesiologist compressed her epigastrium hard with his fist in an attempt to occlude the abdominal aorta between his fist and the patient’s vertebrae. Meanwhile, another anesthesiologist intubated the trachea, using topical lidocaine. Then, general anesthesia was induced with fentanyl (200μg), midazolam (5 mg), and vecuronium (8mg). Simultaneously, atropine (0.5 mg) and norepinephrine (25μg) were injected en bolus. Blood and fluid were administered aggressively. The patient’s anterial blood pressure (ABP) and HR promptly responded to these treatments. With the continuous external manual compression of the abdominal aorta, the femoral pulse was not palpable on either side, while the radial SBP exceeded 100mmHg within 5min of the circulatory collapse, and exceeded 150mmHg during the next 5 min, necessitating the cessation of volume loading and the initiation of vasodilator therapy, with nitroglycerine (0.5–3.0μg·kg 1·min 1) and prostag

Standardization of perioperative management on hepato-biliary-pancreatic surgery.

Japan-China Joint Medical Workshop (2012) on standardization of perioperative management on hepato-biliary-pancreatic surgery was held by the Center for Medical Standards Research, IRCA-BSSA Group in Japan on April 15-16, 2012. Experts in the fields of surgery, anesthesia, pharmacy, and public health from 21 health institutions from Japan and China presented their research achievements and shared their medical experience of perioperative management on hepato-biliary-pancreatic surgery, which should facilitate building of guidelines for hepatocellular carcinoma and be expected to promote standardized management of liver cancer in Asia.