Sustained-Release Tacrolimus Stabilizes Decline of Forced Expiratory Volume in 1 Second Through Decreasing Fluctuation of Its Trough Blood Level.

BACKGROUND: Calcineurin inhibitors are a commonly used immunosuppressive drug and over 80% of lung transplant (LTx) recipients use tacrolimus. Sustained-release tacrolimus (SRT) was developed as a once-daily formulation, resulting in slower release and reduction in peak concentration compared with twice-daily immediate-release tacrolimus (IRT). Previous reports indicate that SRT may carry fewer side effects than IRT; however, the impact of SRT in bronchiolitis obliterans syndrome (BOS) after LTx is unclear. OBJECTIVE: Our study objective was to evaluate the effect of SRT in BOS after LTx. MATERIALS AND METHODS: We investigated the effect of SRT for BOS among 75 LTx recipients who were alive in 2017 in our LTx program. All analyses were carried out using student t test or F test. RESULTS: Thirty-five recipients took IRT, 32 recipients used SRT, 7 recipients used cyclosporine, and 1 patient who received bone marrow and a lung graft from the same donor did not use a calcineurin inhibitor. The most frequent reason for conversion of IRT to SRT was kidney dysfunction, followed by other IRT complications. Five recipients underwent conversion of IRT to SRT because of decline of forced expiratory volume in 1 second (FEV1) with fluctuation of the tacrolimus trough level. After induction of SRT, the fluctuation of the tacrolimus trough level was significantly reduced in 4 of 5 patients (P < .05). Before drug form conversion, the FEV1 in these 5 patients was significantly decreased; however, this exacerbation of FEV1 was attenuated after SRT induction (P < .05). CONCLUSION: SRT appeared to stabilize decline of FEV1 in patients with BOS possibly due to reducing the fluctuation of tacrolimus trough blood concentration.

[Evaluation for medical management on catamenial pneumothorax].

Catamenial pneumothorax (CP) is one of the clinical manifestations of endometriosis, therefore the systemic hormonal therapy is indispensable and should be the 1st choice for CP treatment. However, it is refractory and repeats a recurrence, so that a combined or sequential adjuvant treatment becomes necessary. From 2003 to 2009, 5 patients with CP were treated at our institution. All patients had right-sided pneumothorax and the history of pelvic endometriosis. By thoracoscopic examination, diaphragmatic abnormalities, such as defect, pinhole, or brown spot, were identified in all patients. We performed hormonal therapy combined with chemical pleurodesis using OK-432, as an initial treatment. Two patients have been free of recurrence for 24 and 53 months, respectively. Three patients who refused or interrupted hormonal therapy caused a recurrence, but were successfully managed with the addition of pleurodesis and continuing hormonal therapy. At present, all patients are asymptomatic with 13 to 92 months recurrence-free period. From the long-term results, our therapeutic strategy consisted of hormonal therapy and chemical pleurodesis is considered appropriate.

[Emergency anterior approach for decortication with right pneumonectomy in a patient with chronic expanding hematoma].

The patient was a 68 year-old woman who had a history of treatment of pulmonary tuberculosis 35 years ago. She has experienced dyspnea and hemosputa since several years ago and has been followed up as having chronic empyema. She was admitted to our hospital due to recent exacerbation of symptoms. X-ray films and computed tomography scans of the chest showed the right thoracic cavity to be totally filled with a mass and the shift of mediastinum to the left side. After several days from admission, she needed mechanical ventilation support due to dyspnea exacerbation. Emergency decortication with right pneumonectomy through median sternotomy with anterolateral incision was performed. Postoperative course was uneventful. Pathlogical diagnosis was chronic expanding hematoma.

Prevalence of intestinal parasites and genotyping of Giardia intestinalis in pet shop puppies in east Japan.

The current study examined the prevalence of intestinal parasites and genotypes of Giardia intestinalis in puppies from nine pet shops in east Japan. Fresh fecal samples from 1794 puppies (≦3 months old) were collected on one occasion. Giardia spp. was examined for specific coproantigen using ELISA kit (SNAP®Giardia, IDEXX Laboratories, Inc., USA). Other intestinal parasites were detected microscopically using the formalin-ethyl acetate sedimentation technique. Genotyping was determined for the random 29 stool samples identified as Giardia spp. positive using PCR and direct sequencing of the glutamate dehydrogenase (gdh) gene. Overall prevalence of protozoan Giardia spp. and Cystoisospora spp. revealed 23.4% and 11.3%, respectively. Prevalence of ascarids, Strongyloides spp. and hookworms were recorded 1.8%, 1.1% and 0.1%, respectively. Protozoan Giardia spp. and Cystoisospora spp., thus, represent important pathogens among pet shop puppies. All genotyped G. intestinalis isolates were belonged to assemblage C or D, identified as dog-specific genotypes. Zoonotic assemblage A and B were not demonstrated. The result suggests that the risk of zoonotic transmission of G. intestinalis from pet shops puppies to humans may be quite low in Japan.

Evaluation of plasma C-terminal atrial natriuretic peptide in healthy cats and cats with heart disease.

BACKGROUND: The clinical implications of evaluating C-terminal atrial natriuretic peptide (ANP) concentration in cats are still controversial. HYPOTHESIS: The objective of this study was to investigate the relationship between plasma C-terminal ANP concentration and left atrial pressure (LAP) in healthy cats with volume overload (study 1), and to compare plasma C-terminal ANP in normal cats and cats with cardiomyopathy (study 2). ANIMALS: Five healthy adult cats were used in study 1, and clinically healthy cats (n=8) and cats with cardiomyopathy (n=14) were used in study 2. METHODS: In study 1, cats were anesthetized and given acetated Ringer's solution (100 mL/kg/h for 60 minute) via the cephalic vein. Hemodynamic measurements and blood samples, collected from the jugular vein, were performed at 10-min intervals. In study 2, blood samples from normal cats and cats with cardiomyopathy were collected from the cephalic vein. The plasma C-terminal ANP concentration was determined by radioimmunoassay for human alpha-ANP. RESULTS: In study 1, volume overload significantly increased the C-terminal ANP concentration and LAP from baseline. The C-terminal ANP concentration was strongly correlated with the mean LAP. In study 2, age, E wave velocity, and the ratios of the left atrium to aorta were significantly higher in the cats with cardiomyopathy compared with the normal cats. The C-terminal ANP concentration was significantly higher in the cats with cardiomyopathy compared with the normal cats. CONCLUSIONS AND CLINICAL IMPORTANCE: Our results suggest that the measurement of plasma C-terminal ANP in cats may provide additional information for the diagnosis of heart disease.

[Problems in donor lung evaluation for transplantation with regard to airway infection].

The shortage of donor organs has been 1 of the major obstacles to solid organ transplantation. Typical lung donor criteria include clear lung field on chest radiograph, adequate oxygenation, acceptable lung compliance, and satisfactory bronchoscopic findings. To extend usage of available donors, liberalization of donor lung selection criteria has been facilitated, however, marginal donor lungs must be used with discretion, because donor lung injury, especially that related to infection, has a potential leading to early post-operative death of the recipient. From March 2000 to December 2006, we evaluated 15 braindead donors and at least 1 of the lungs from 9 donors was judged suitable for transplantation. One of 9 recipients developed severe pneumonia cased by carbapenems-resistant Pseudomonas aeruginosa possibly originating from the donor lungs, eventually leading to death. The chest radiograph and oxygenation of the donor had been satisfactory, however, a moderate amount of mucopurulent secretions was observed by bronchoscopic inspection and the donor had been given a cefozopran for 9 days before the procurement operation. Remaining 8 recipients were free from air-way infection in the early postoperative period. We discuss the status and problems of donor lung evaluation for transplantation with regard to donor lung infection.

[Bronchoplasty without lung resection for the complete transection of the left main bronchus due to blunt trauma].

Blunt bronchial injury is rare but crucial injury. A 17-year-old female was admitted due to traumatic injury. She was diagnosed with bilateral lung contusion, multiple rib fractures, spleen damage and the suspicion about the complete transection of the left main bronchus on X-ray and computed tomography (CT). She was brought to our hospital at 30 hours later from injury. Bronchoscopy revealed the complete transection and the edema of the left main bronchus. She underwent a resection of the disrupted portion and end-to-end anastomosis of left main bronchus without lung resection. We should be an immediate and accurate diagnosis of tracheobronchial disruption by X-ray, CT and bronchoscopy whenever we evaluate patients with blunt chest trauma.

Relationship between serum TNF activity and insulin resistance in dairy cows affected with naturally occurring fatty liver.

To clarity the relationship between tumor necrosis factor (TNF) and insulin resistance in dairy cows affected with fatty liver, naturally occurring cases were investigated. The affected cows were classified into following three groups according to histopathologic findings of the liver: mild fat droplet deposition (group 1; n=11), severe fat droplet deposition (group 2; n=10), and cloudy swelling (group 3; n=8). Serum TNF activities in Group 2 (8.67 +/- 2.16 U/ml) and Group 3 (11.65 +/- 1.92 U/ml) were significantly higher than that in Group 1 (3.57 +/- 0.81 U/ml) (p<0.05). The insulin-tolerance tests showed that the insulin-stimulated glucose disposal rates (GDR) in Group 2 (27.6 +/- 7.8%) and Group 3 (15.8 +/- 9.1%) were significantly lower than that in Group 1 (41.7 +/- 9.8%). There was a significant negative correlation between serum TNF activity and GDR in affected cows (r=-0.56, p<0.01). These results indicate that serum TNF activity is correlated with insulin resistance in cows with fatty liver.

Acute phase response in naturally occurring coliform mastitis.

Changes in the activities of serum cytokines and in acute phase response were observed in dairy cows with naturally occurring coliform mastitis. Seven cows with severe mastitis showed systemic and mammary inflammatory response throughout the observation period, and 11 cows with mild mastitis recovered and were able to be milked within 3 days of onset of mastitis. Serum interleukin (IL)-I and tumor necrosis factor (TNF) activities were higher in the severe group than in the mild group at the first appearance of symptoms. Elevated IL-1 activity was evident in the severe group throughout the observation period. Serum alpha-1-acidglycoprotein (alpha1AG) concentration began to rise with the beginning of mastitis in the severe group, and peaked at 9 days. Serum haptoglobin (Hp) concentrations peaked at 3 days, and decreased gradually after 3 days in the severe group. These results showed that there are dynamic changes in serum IL-1 activity and in serum alpha1AG and Hp concentrations in cows with severe coliform mastitis.

Determination of canine beta2-microgloblin in plasma and urine by enzyme-linked immunosorbent assay.

An enzyme-linked immunosorbent assay was developed for the determination of canine beta2-microglobulin (beta2-m) in plasma and urine. The detectable sensitivity for pure canine beta2-m was 0.05 microg/l and the analytical range was 0.1 to 50 microg/l. The mean analytical recovery when pure canine beta2-m was added to normal plasma was 101.9%. The mean analytical recovery in the urine was 102.1%. The intra-day variation coefficient was 3.1% in plasma, 4.3% in serum and 1.9% in urine. No difference was found between the concentration of beta2-m in plasma and serum (n=17). The concentration of beta2-m in the plasma of normal dogs was 1.82 +/- 0.57 mg/l (n=31). The mean excretion in 24 hr urine collected from normal dogs was 17.6 +/- 9.2 microg/l, 0.22 +/- 0.12 microg/kg of body weight or 14.2 +/- 9.4 microg/g of urine creatinine. The beta2-m creatinine index of random urine samples was 23.5 +/- 16.6 microg/g (n=26). There was a close correlation between the beta2-m creatinine index of 24 hr urine samples and that of random urine samples (r=0.872).

Changes of immunological response after experimentally ozonated autohemoadministration in calves.

Changes in subsets of peripheral blood mononuclear cells and lymphocyte blastogenesis reaction were observed after ozonated autohemoadministration (OAHA) in calves. Ten healthy calves were used in this study. After OAHA, CD8+ cells, CD14+ cells and WCl+cells increased (p<0.05). The level of lymphocyte blastogenesis stimulated by PHA decreased after OAHA. A significant increase in serum IL-6 activity was observed in OAHA calves. These results suggested that OAHA induced immunological changes in calves.

Purification of bovine urinary beta2-microglobulin and its biochemical characteristics.

In this study, bovine beta2-m was purified from urine by ion-exchange chromatography and gel chromatography, and the characteristics were compared with those of colostral beta2-m by the immunological reactivity, isoelectric points, peptide map, and amino acid sequence. The characteristics of purified urinary beta2-m were consistent with those of the colostral beta2-m. The urinary and colostral beta2-m possessed the same polypeptide chain consisting of 98 amino acids, and its molecular weight is 11.8 kDa. Furthermore, four isoforms of beta2-m were found. The isoelectric points were different from each other.

Determination of bovine beta2-microglobulin and albumin in urine by a reversed-phase high-performance liquid chromatography.

Reversed-phase high-performance liquid chromatography (HPLC) was used to analyze beta2-microglobulin and albumin in bovine urine. The urine samples were chromatographed on TSK-gel ODS-120T column with an acetonitrile gradient. Urinary beta2-m and albumin were detected at 220 nm. For the pre-treatment, there were two steps proceeding injection: dialysis of urine with distilled water overnight, followed by concentration by solid-phase extraction method using a Sep-Pak cartridge. The retention times of beta2-microglobulin and albumin were 25.35 +/- 0.85 and 32.20 +/- 0.20 minutes (n=5), respectively. The mean analytical recoveries of beta2-microglobulin and albumin added to 0.1 ml of urine samples were 94.5 and 100.5%, respectively. The within-run coefficients of variation ranged from 1.5 to 5.3% for beta2-microglobulin and from 2.3 to 7.0% for albumin. The sensitivity for quantification of each protein was 0.5 microg in 100 microl injected urine samples. Urine samples from healthy cows and from cows with different types of proteinuria were analyzed by this reversed-phase HPLC. Results revealed albumin was remarkable in the urine from a cow with glomerulonephritis, and beta2-microglobulin was, in the urine from a cow with tubular dysfunction.

The complete amino acid sequence of dog beta2-microglobulin.

Dog beta2-microglobulin was purified from the urine of dogs with potassium dichromate induced tubular damage. It was purified by sequential use of anion exchange chromatography, gel filtration chromatography, and reversed-phase high performance liquid chromatography. Comparisons of the amino acid sequence of the dog protein with human, mouse, and rabbit beta2-microglobulin, indicated a high degree of similarity. The dog protein was very similar to human beta2-microglobulin in that it had a molecular weight of 11.8 kDa and contained two half-cystinyl residues. Dog and human beta2-microglobulin were demonstrably different at 24 of the 99 positions compared. The data supported the conclusion that the purified protein was dog beta2-microglobulin and that all four proteins from dog, human, mouse, and rabbit were closely related.

Purification of bovine beta 2-microglobulin from colostrum and its complete amino acid sequence.

Bovine beta 2-microglobulin (beta 2-m) was purified from colostrum milk in two chromatographic steps: anion-exchange chromatography, and gel filtration. The amino acid sequence was determined to confirm that the purified protein was beta 2-m. A molecular weight of 11.8 kDa beta 2-m was estimated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Bovine beta 2-m consists of 98 amino acid residues and contains one disulfide linkage connecting residues 25 and 79. The amino acid sequence determined by this study is different from a previously published sequence at three sites, but agrees with the amino acid sequence from cDNA. Thus, we can conclude that the amino acid sequence determined in this study is correct.

Development of Babesia gibsoni in the salivary glands of the larval tick, Rhipicephalus sanguineus.

The development of Babesia gibsoni in the salivary glands of larval Rhipicephalus sanguineus ticks was morphologically studied for 4 days of feeding of tick on rabbits. Babesia gibsoni showed two forms (sporont and sporozoite) in the budding process. Sporozoites were formed in the acinar cells of salivary glands and released from the hollow cytoplasmic area of these cells by day 4 of feeding. The morphology of B. gibsoni in the salivary glands of R. sanguineus shows a close similarity to those of B. gibsoni in Haemaphysalis longicornis.

Application to cows and horses of Spotchem, a dry-chemistry blood analyzer for use in veterinary clinics.

The usefulness of a dry-chemistry blood analyzer, Spotchem SP-4410 (SP-4410) in a veterinary clinic for analysis of bovine and equine blood chemistry was studied. We quantitated total protein (TP), albumin (Alb), total bilirubin (T-Bil), blood urea nitrogen (BUN), total cholesterol (T-Cho), glucose (Glu), calcium (Ca), aspartate aminotransferase (AST), gamma-glutamyltransferase (GGT), creatinine phosphokinase (CPK), and alkaline phosphatase (ALP) in bovine sera. Each sample was assayed with both the SP-4410 and an automated blood analyzer which served as a wet-chemistry reference system, and the data were analyzed with regression analysis. The correlation coefficient for AST was 0.997 being the highest for all the parameters, and all the correlation coefficients were 0.93 or higher. The coefficients of variation were lower than 5.0 except in the case of bovine T-Bil where it was 5,756. The ranges of normal reference values measured by SP-4410 were the same as those reported by other investigators in most cases, but those for GGT and CPK were slightly higher. The strongest interference was observed with hemoglobin. It seems that dry-chemical-analysis of blood serum using the SP-4410 is useful for analysis of bovine and equine blood.

Development of Babesia ovata in the salivary glands of the nymphal tick, Haemaphysalis longicornis.

The development of Babesia ovata in the salivary glands of nymphal Haemaphysalis longicornis ticks was morphologically studied for 4 days of feeding of tick on rabbits. Babesia ovata showed two forms (sporont and sporozoite) in the budding process. Sporozoites were formed in the acinar cells of salivary glands and released from the hollow cytoplasmic area of these cells by day 4 of feeding.