Rapamycin causes upregulation of autophagy and impairs islets function both in vitro and in vivo.

Autophagy is a lysosomal degradation process of redundant or faulty cell components in normal cells. However, certain diseases are associated with dysfunctional autophagy. Rapamycin, a major immunosuppressant used in islet transplantation, is an inhibitor of mammalian target of rapamycin and is known to cause induction of autophagy. The objective of this study was to evaluate the in vitro and in vivo effects of rapamycin on pancreatic ß cells. Rapamycin induced upregulation of autophagy in both cultured isolated islets and pancreatic ß cells of green fluorescent protein-microtubule-associated protein 1 light chain 3 transgenic mice. Rapamycin reduced the viability of isolated ß cells and down-regulated their insulin function, both in vitro and in vivo. In addition, rapamycin increased the percentages of apoptotic ß cells and dead cells in both isolated and in vivo intact islets. Treatment with 3-methyladenine, an inhibitor of autophagy, abrogated the effects of rapamycin and restored ß-cell function in both in vitro experiments and animal experiments. We conclude that rapamycin-induced islet dysfunction is mediated through upregulation of autophagy, with associated downregulation of insulin production and apoptosis of ß cells. The results also showed that the use of an autophagy inhibitor abrogated these effects and promoted islet function and survival. The study findings suggest that targeting the autophagy pathway could be beneficial in promoting islet graft survival after transplantation.

Laboratory and imaging features of kidney involvement in autoimmune pancreatitis: incidence, correlation, and steroid therapy response.

AIM: Autoimmune pancreatitis (AIP) is a rare subtype of chronic pancreatitis. AIP has been suggested to be complicated by tubulointerstitial nephritis or glomerulonephritis, implying that the kidney is involved as a phenotype of IgG4-positive multi-organ lymphoproliferative syndrome; however, the clinical significance of this novel entity is not well-defined. METHODS: We conducted a retrospective cohort analysis of 47 (male, 39; female, 8) AIP patients. RESULTS: The patients (mean age, 70.3 +/- 9.5 years) had a mean observation period of 4.1 years. Before treatment, renal dysfunction with an eGFR of 30 and 15 ml/min/1.73 m2 developed only in 10.6% (5/47) and 2.1% (1/47) of the patients, respectively. Nevertheless, urinary N-acetyl-beta-D-glucosaminidase and alpha1-microglobulin levels were elevated in 78.6% (11/14) and 30.8% (4/13) of the patients, respectively. Renal involvement in contrast-enhanced CT imaging was present in 18.2% (8/44) of the patients and was associated with proteinuria (p = 0.04) and a decrease in eGFR (p < 0.01). Furthermore, a follow-up CT study (mean, 545 days) revealed improved kidney lesions in 80.0% (4/5) of the patients after oral corticosteroid administration. In contrast, first-time kidney involvements appeared newly in 3.6% (1/28) of the patients after steroid therapy for nonrenal AIP symptoms, and in 14.3% (1/7) of the patients under no specific therapy (p = 0.02). CONCLUSION: Although severe renal failure develops rarely in AIP patients, renal abnormalities have been significantly detected by biochemical and radiological tests. Oral corticosteroid administration, even when not targeting symptomatic nephropathy, can treat and prevent kidney involvements in AIP.

Fine needle aspiration cytology of the malignant variant of ossifying fibromyxoid tumor of soft parts: a case report.

BACKGROUND: Ossifying fibromyxoid tumor (OFMT) of soft parts is a rare, recently defined, fibroosseous neoplasm, generally regarded as clinically benign; however, one-third of cases recur locally, and several malignant examples have been reported. Fine needle aspiration (FNA) cytology of the tumor is rarely described in the literature. We provide the first cytomorphologic study of the malignant variant. CASE: A 70-year-old man presented with an intramuscular mass in the right buttock. Computed tomography revealed ossification within the mass and multiple pulmonary nodules. FNA biopsy showed round and polygonal to spindled tumor cells, arrayed singly, cordlike or in small aggregates, with scattered dense stromal fragments and a slightly myxoid background. The nuclei showed significant pleomorphism accompanied by coarse chromatin with clumping, irregular contours, and one to two distinct nucleoli. The tumor cells were recognizable as sarcoma, with no evidence of high grade malignancy. The tumor was totally excised, histopathologically confirmed as the malignant variant of OFMT of soft parts, and immunohistochemically and ultrastracturally analyzed as of neural origin. CONCLUSION: The FNA specimen revealed that the cytomorphology was consistent with the histologic features of the malignant variant of OFMT, but several characteristic histologic parameters, such as multilobular proliferation and peripherally placed mature, bony trabeculae, were not reflected in the aspirates. Although FNA cytologic findings may be of limited diagnostic utility in OFMT, radiographic evidence of calcification/ossification suggests that OFMT should be subjected to differential diagnosis with fine needle aspiration biopsy of soft tissue tumors. Additional studies will be required for further clarification.

Effect of induced triploidy on fin regeneration of juvenile rainbow trout, Oncorhynchus mykiss.

Triploidy was induced in the rainbow trout in order to evaluate whether the altered numbers and sizes of triploid cells could modify fin regeneration. Amputation of one lobe of the tail fin of diploid and triploid juveniles resulted in regeneration for experimentals and controls. Nevertheless, both rate and frequency of regeneration in triploids were significantly increased as compared with diploids, a fact that can be attributed to the increased nuclear and cellular volume in a wide range of tissues, whereas the cell numbers were reduced. These data suggest that a great deal of interesting and important research could be done using triploid animals as experimental models for studying the regeneration of appendages.

[Mortality and prognostic factors in patients with community-acquired pneumonia: an analysis of 231 cases].

We prospectively analyzed the mortality and prognostic factors in 231 patients with community-acquired pneumonia. The patients were enrolled between May 1996 and April 1998. The average patient age was 67.6 +/- 17.5 years, and 68% were male. The mean hospital length of stay was 21.9 +/- 21.7 days. The mortality was 6.5% at the 30th day, 13.9% at hospital discharge, and 19.9% at 1 year later. Stepwise logistic regression analysis showed that liver cirrhosis, diastolic hypotension (< 60 mmHg), hypoxemia (< 50 Torr) were significantly correlated with death at the 30th day, and that alcoholism, malignancy, diastolic hypotension, hypoxemia, hypoalbuminemia (< 3.0 g/dl), and increased creatinine (> 1.2 mg/dl) were significantly correlated with discharge mortality. Furthermore, the prognostic factors at 1 year later were the same as those at hospital discharge, with the exception of alcoholism, which was replaced by ischemic heart disease in the 1-year mortality. We conclude that liver cirrhosis, diastolic hypotension, and hypoxemia are very important prognostic factors during the acute stage of pneumonia, and that alcoholism, malignancy, hypoalbuminemia and renal complications are significantly correlated with hospital death and 1-year mortality.

[Community-acquired pneumococcal pneumonia: a comparative study of bacteremic and nonbacteremic patients].

We analyzed the clinical and laboratory features of 37 adult patients with community-acquired pneumococcal pneumonia requiring hospitalization. Blood culture was positive in 11, and negative in 26. The average age of the 37 patients was 70.2 years, and 28 were male. Compared with the patients with negative blood culture, the patients with positive blood culture were more likely to have liver cirrhosis, less sputum production, lower body temperature, and higher respiratory rate on physical examination. In the laboratory data on admission, the values for serum total protein, albumin, and glucose were significantly lower in the patients with positive blood culture. Thirty-two percent of the pneumococcal isolates were resistant to penicillin G (MIC > or = 0.12 microgram/ml). There was no difference in the frequency of drug-resistant pneumococci in the two groups. The use of mechanical ventilation was more frequent in the patients with positive blood culture than in the patients with negative blood culture (27.3% vs 3.8%). Moreover, mortality was higher (27.3% vs 7.7%) and the duration of hospitalization was longer in the bacteremic group. However, from a univariate analytical perspective, these differences were not significant. We concluded that blood culture can be a highly valuable diagnostic aid and useful for the prognostic evaluation of patients with community-acquired pneumonia requiring hospitalization.

[Comparative study of clinical features of typical and atypical pneumonias].

We prospectively analyzed the clinical and laboratory features of 74 patients with community-acquired pneumonia who required hospitalization between May 1996 and October 1997. Typical pathogens were identified in 47, and atypical pathogens in 27. The average age was higher in patients affected by typical pathogens (73.9 years), than in patients affected by atypical pathogens (50.9 years). Univariable analysis found that atypical pneumonias were more frequent in healthy patients than typical pneumonias. Moreover, the presence of relatives with symptoms of airway infection, headache, and earache was more common among the patients with atypical pneumonias, while leukocytosis and elevated C-reactive protein levels were more frequent among patients with typical pneumonias. Typical pathogens accounted for up to 79.6% of the cases of pneumonia with in older patients (aged 60 years or more), whereas atypical pathogens accounted for up to 80% of the cases of pneumonia in younger patients (aged under 60 years). This difference was statistically significant. Of all 74 patients, 39 (52.7%) were afflicted by severe community-acquired pneumonia, as categorized by American Thoracic Society guidelines. The most common pathogen among these patients was Streptococcus pneumoniae. Legionella was one of the top four. Selection of the initial antimicrobial treatment is an important clinical decision that should be made on the basis of clinical features at admission, age, and severity of the patient's illness.

[Emergency room visits by patients with exacerbations of asthma].

We retrospectively analyzed patterns of emergency room visits by patients with exacerbations of asthma from December 1995 through November 1996. A total of 591 episodes in 198 patients were reviewed. The average age was 35.8 years, ranging from 15 to 71. The largest number of visits occurred in September. The number of visits per year ranged from 1 to 22; the mean was 2.9 per year. In patients who were followed on a regular basis at our institution, serve attacks accounted for 7.1% of the total, compared with 21.6 percent at other hospitals or outpatient clinics. We suspect that this difference was related to differences in the use of inhaled steroids. At our institution, 89% of patients were taking inhaled steroids; at other hospitals or clinics, only 21% were taking inhaled steroids. Of the 198 patients, 33 fulfilled one of the following criteria: (1) endotracheal intubation for respiratory failure or respiratory arrest, (2) respiratory acidosis (pH < 7.35) without endotracheal intubation; 27% of those patients had been given a diagnosis of mild asthma before the acute exacerbation. We conclude that patient education and standard guidelines for treatment of asthma, are very important for appropriate management of asthma, to prevent exacerbations and asthma-related deaths.

Peripancreatic fluid cytology: detection of early rejection versus graft pancreatitis after canine pancreatic transplantation.

It is particularly difficult to distinguish between early rejection and graft pancreatitis when early rejection produces an elevated serum amylase level. In this study we determined whether peripancreatic fluid cytology (PFC) can differentiate early acute rejection and graft pancreatitis as an alternative diagnostic tool to graft biopsy that has the potential of pancreatic fistula and hemorrhage. Sixty-two dogs received either a segmental pancreas allograft (n = 25) or autograft (n = 37) heterotopically in the neck. This study included five groups: allografts without immunosuppression (group A, n = 12), allografts with immunosuppression (group B, n = 13), autografts without immunosuppression (group C, n = 11), autografts with immunosuppression (group D, n = 12), and autografts treated by 45 minutes of pretransplant warm ischemia to induce acute graft pancreatitis (group E, n = 14). A closed suction drainage catheter was placed next to the graft to collect peripancreatic fluid daily after the transplant. PFC was performed using May-Gruenwald-Giemsa double-staining technique and compared to the corresponding histology through the observation period. In analyses of 50 functioning grafts, PFC performed on day 1 showed similar neutrophil accumulations in all groups. In sharp contrast, on days 3 and 6, group A had dramatically increased mononuclear cell concentrations in PFC, whereas groups B, C, and D showed significantly lower concentrations, the percent of mononuclear cells among total leukocytes being 47.3 +/- 23.4%, 11.8 +/- 4.9%, 4.3 +/- 1.8%, and 6.4 +/- 2.4% (day 3); and 32.7 +/- 9.8%, 10.5 +/- 4.8%, 7.2 +/- 4.2%, and 8.6 +/- 6.4% (day 6) in groups A, B, C, and D, respectively. On the other hand, in group E numerous degenerating neutrophils with a marked to moderate increase in necrotic tissue fragments were observed by PFC on days 3 and 6. In terms of graft histology on days 3 and 6, group A showed interstitial mononuclear cell infiltration indicating an acute rejection process, whereas groups B, C, and D had minimal inflammatory cell infiltration. In group E graft pancreatitis was histologically confirmed on days 3 and 6. These results suggest that PFC after pancreas transplantation could be a safe, simple, useful diagnostic tool for discriminating early graft rejection from graft pancreatitis.

Elevated serum interferon gamma and interleukin-6 in patients with necrotizing lymphadenitis (Kikuchi's disease).

We investigated serum levels of interferon alpha, interferon gamma, tumour necrosis factor alpha, interleukin-2 (IL-2) and interleukin-6 (IL-6) in patients with necrotizing lymphadenitis (Kikuchi's disease) (NL). Four male patients, diagnosed as having NL following biopsy of the affected lymph nodes and by the clinical course, were included in this study. All four patients had higher than normal serum interferon gamma and IL-6 levels during the acute phase, which returned to normal levels during the convalescent phase. Interferon alpha, tumour necrosis factor alpha and IL-2 were, however, within the normal ranges. Our findings indicate the possibility that interferon gamma and IL-6 may play an important role in the pathogenesis of NL.

Sensitivity of steroid enzyme immunoassays. Comparison of four label enzymes in an assay system using a monoclonal anti-steroid antibody.

The sensitivities of monoclonal antibody-based enzyme immunoassays for 11-deoxycortisol using alkaline phosphatase (AP), horseradish peroxidase (HRP), beta-galactosidase (beta-GAL) and glucose oxidase (GOD) as labels were compared. The anti-11-deoxycortisol antibody used was that produced in ascites by inoculating antibody-secreting hybridoma cells into mice. Enzyme labeling of 11-deoxycortisol was carried out by the N-succinimidyl ester method. The activated ester of 4-(2-carboxyethylthio)-11-deoxycortisol was treated with each enzyme to give a homologous enzyme-labeled antigen. In the competitive immunoassay, the bound and free enzyme-labeled antigens were separated by a double antibody method and the enzymic activity of the immune precipitate was determined by colorimetric and fluorimetric methods. The AP activity was measured in three ways, using p-nitrophenyl phosphate, nicotinamide adenine dinucleotide phosphate (NADP), and 4-methylumbelliferyl phosphate as substrates. o-Nitrophenyl beta-D-galactopyranoside and 4-methylumbelliferyl beta-D-galactopyranoside were used for beta-GAL, and 3,3',5,5'-tetramethylbenzidine (TMB) and 3-(p-hydroxyphenyl)propionic acid (HPPA) for HRP. In the case of GOD, TMB and HPPA were used in combination with HRP. A dose-response curve with a high sensitivity was obtained in each 11-deoxycortisol assay system by the use of a minimum amount of the enzyme-labeled antigen at an appropriate dilution of monoclonal anti-11-deoxycortisol antibody (Ka = 2 x 10(10) M-1). The amounts of 11-deoxycortisol needed to displace 50% of the bound label ranged from 5 to 15 pg in the colorimetric methods, and 4-9 pg in the fluorimetric methods.(ABSTRACT TRUNCATED AT 250 WORDS)

Unexpected loss of immunoreactivity of a glucose oxidase-labeled antigen in a steroid enzyme immunoassay system.

Glucose oxidase-labeled 11-deoxycortisol was tested for immunoreactivity in an enzyme immunoassay system using a monoclonal antibody. The antigen was labeled by the N-succinimidyl ester method. It was found that the binding affinity of the label to the anti-steroid antibody markedly decreased during storage; the half-life time was ca. 4.5 d.

Two cases of transient partial amnesia in the course of transient global amnesia.

In the course of transient global amnesia, two patients showed symptoms of transient partial amnesia. Close observation showed that at some time during the episode the patients had some degree of nonverbal memory, which recovered earlier than verbal memory. Careful observation of the progression of symptoms during an episode of transient global amnesia may often reveal symptoms of transient partial amnesia.