Identification of Potential mRNA Biomarkers in Milk Small Extracellular Vesicles of Enzootic Bovine Leukosis Cattle

Enzootic bovine leukosis (EBL) is a disease caused by bovine leukemia virus (BLV); only a small percentage of BLV-infected cattle develop EBL and present with B-cell lymphosarcoma. There is no vaccine against BLV, treatment for EBL, or method for predicting the possibility of EBL onset, thus making EBL control difficult. Herein, to explore biomarkers for EBL in milk, we examined the mRNA profiles of small extracellular vesicles (sEVs) in milk from four BLV-uninfected and four EBL cattle by microarray analysis. It was revealed that 14 mRNAs were encapsulated in significantly higher quantities, and these mRNAs were therefore selected as biomarker candidates. Primers for these mRNAs were designed, and nine primer sets were available for quantitative real-time PCR. Nine mRNAs were evaluated for their availability as biomarkers for EBL using sEVs from newly-collected milk of 7 uninfected and 10 EBL cattle. The quantities of eight mRNAs (TMEM156, SRGN, CXCL8, DEFB4A, FABP5, LAPTM5, LGALS1, and VIM) were significantly higher in milk sEVs of EBL cattle than in those of uninfected cattle. Therefore, our findings indicate that these eight mRNAs in milk sEVs can be used as potential EBL biomarkers with combination use, although single mRNA use is not enough. Consequently, cattle at risk of EBL onset can be identified by monitoring the fluctuation in quantities of these mRNAs in milk before they develop EBL.     

Age estimation by quantitative features of pubic symphysis using multidetector computed tomography

Macroscopic assessment of the pubic symphysis is commonly used for age estimation because its surface changes over time. However, postmortem computed tomography (PMCT), a method several forensic medical departments and institutes have begun to adopt, has the potential to simplify the information gathering process from the pelvic bone without requiring soft tissue removal. Some studies have previously evaluated the use of three-dimensional images of the pubic symphysis, but because of variance in the graphics processing among image analysis software packages, certain differences have been observed between these studies. Therefore, in this study, the PMCT findings of 199 subjects of known age and sex were retrospectively reviewed to examine the feasibility of age estimation using planar images of the pubic bones and soft tissue. The coronal and axial sectional images were observed at the center of the symphyseal surface, and the pubic bone length and thickness of the connective tissue of the pubic symphysis were measured at each slice. Our results revealed a significant positive correlation between the length of the pubic bone of the coronal section and age, suggesting that the use of a cutoff value for pubic bone length might be feasible for age estimations. In addition, the thickness of the connective tissue tended to narrow over time. Although the prediction interval range of planar images obtained by PMCT was major and is not usable in practice at this moment, it may still be a useful tool if used in conjunction with other findings obtained by PMCT.     

Bevacizumab for non‐small‐cell lung cancer: A nested case control study of risk factors for hemoptysis

Potentially life‐threatening, serious hemoptysis is an adverse event associated with bevacizumab in non‐squamous non‐small‐cell lung cancer (NSCLC) trials. Suggested risk factors include central tumor location and cavitation; however, the profile of hemoptysis occurrence in clinical practice is still unclear. A nested case‐control study was conducted to assess the onset profile and risk factors for hemoptysis in bevacizumab‐treated patients in a real‐world setting in Japan. After bevacizumab was approved for NSCLC, physicians registered all NSCLC patients scheduled for bevacizumab therapy, from November 2009 to August 2011. Patients developing grade 2 hemoptysis requiring an injectable hemostatic agent or grade ≥3 hemoptysis were selected as case subjects, matched with four control subjects each. Case report forms were collected after an observation period of 24 weeks. Radiologists assessed blinded thoracic images. Risk factors for hemoptysis were assessed by univariate and stepwise multivariate analysis. Of 6774 patients registered, 23 (0.3%) experienced grade ≥2 drug‐related hemoptysis. A total of 104 patients (21 cases, 83 controls) were analyzed by central reviewers for risk factors of hemoptysis occurrence. In the univariate analysis seven factors were associated with hemoptysis. In the step‐wise multivariate analysis, prior thoracic radiotherapy (P = 0.1844), presence of tumor exposure in the central airway (P = 0.0256) and concomitant radiotherapy (P = 0.1169) were identified as risk factors for hemoptysis. While the incidence of hemoptysis was low in the real‐world setting in Japan, the three risk factors identified, prior thoracic radiotherapy, presence of tumor exposure in the central airway and concomitant radiotherapy, should be considered when selecting patients for bevacizumab treatment. Although technically classed as a clinical trial, a nested case‐control study was a non‐interventional surveillance study analyzing all NSCLC patients receiving bevacizumab in Japan, therefore it was not registered as a phase II/III clinical trial would be.     

Stepped care medical treatment for cirrhotic ascites: Analysis of factors influencing the response to treatment

Abstract Thirty-two patients with liver cirrhosis and ascites were treated by stepped care diuretic treatment as follows: step 1, placed on a 35 mEq sodium diet; step 2, given 400 mg/day of potassium canrenoate in addition to step 1 treatment; step 3, given 40-80 mg/day of furosemide in addition to step 2 treatment. Eleven out of 32 patients (34.4%, group 1) and 12 of 21 patients (57.1%, group 2) lost their ascites at step 1 and step 2, respectively. The remaining nine patients (group 3) required step 3 treatment. Basal urinary sodium excretion and creatinine clearance were significantly lower and β₂-microglobulin was significantly higher in group 3 than those in groups 1 and 2. Elevation of basal plasma renin activity and norepinephrine was evident only in group 3. In group 1, urinary sodium excretion decreased after the treatment. In group 2, plasma α-atrial natriuretic polypeptide was lowered and plasma renin activity and norepinephrine were elevated after the treatment. These results suggest that basal renal function and plasma renin activity and norepinephrine levels are useful indices to predict the effect of ascites treatment and that responders to sodium restriction or potassium canrenoate may be in the state of vascular overflow, while non-responders to potassium canrenoate may be in the state of vascular underfilling. In summary, this stepped care treatment is safe without any side effects, although the diuretics themselves may lead to relative vascular underfilling.     

Retroperitoneal haematoma due to ruptured microaneurysm of the pancreaticoduodenal artery

A case of retroperitoneal haematoma due to a ruptured microaneurysm of the posterior superior pancreaticoduodenal artery in a 61 year old man is described. Ultrasonography and computed tomography revealed cystic masses near the gall-bladder. Selective coeliac angiography disclosed a microaneurysm of the posterior superior pancreaticoduodenal artery. Surgical extirpation of the cystic masses was performed, and the histological finding was an encapsulated old haematoma.     

Exanthem subitum (roseola infantum) with vesicular lesions

Summary We report a 7-month-old boy who developed vesicular lesions during the course of exanthem subitum. Human herpesvirus-6 (HHV-6) DNA was detected both in the skin lesions and in the throat, by polymerase chain reaction. IgG and IgM antibodies against HHV-6 were 1:10 and <1:10, respectively on the tenth day of the illness, and >1:640 and 1:10 on the twenty-second day, respectively. These results suggest that this was primary HHV-6 infection.     

The Colon Mucus Test in Comparison with the Fecal Occult Blood Test in the Detection of Gastrointestinal Disease

Abstract: This study was undertaken to assess the diagnostic significance of the colon mucus test (CMT) by which the cancer-associated carbohydrate antigen, β-D-Gal (1 - 3)-D-GalNAc (T antigen) is measured in rectal mucus, chiefly in patients with various alimentary tract diseases. The incidence of a positive CMT was relatively high in colorectal cancer (69%) and gastric cancer (40%), and was similarly high in some benign colorectal diseases such as colonic diverticulum (42%) and colonic polyps (39%), and in gastric polyps (75%). On the other hand, the incidence of a positive CMT in other benign diseases or in healthy controls was less than 10%. A positive CMT was significantly more common in colorectal disease and in gastric tumors (cancer and polyps) than in the other diseases (p < 0.001). Of 28 colonic polyp patients, 20 (71%) had negative fecal occult blood test results (FOBT), and nine of them (45%) had positive CMT results. These results suggest that the CMT may be a useful screening test for alimentary tract disease, particularly colorectal disease and gastric tumors, although its specificity for colorectal cancer is only moderate. Furthermore, a combination assay using FOBT plus CMT may help to improve the diagnostic rate.