Efficacy and safety of multitarget therapy with cyclophosphamide and tacrolimus for lupus nephritis: a prospective, single-arm, single-centre, open label pilot study in Japan.

Background Pulsed cyclophosphamide or mycophenolate mofetil for lupus nephritis has limited efficacy. We previously reported a case of mixed-class IV + V lupus nephritis successfully treated with cyclophosphamide and tacrolimus. This study assessed the efficacy and safety of multitarget therapy with cyclophosphamide and tacrolimus for the treatment of lupus nephritis. Methods In a prospective, single-arm, open label pilot study, we recruited 15 patients aged 18-64 years with active lupus nephritis who met the American College of Rheumatology criteria for a diagnosis of systemic lupus erythematosus (1997). The treatment protocol was a starting dose of prednisolone of 0.6-1.0 mg/kg/day for 2 weeks and then tapered to a maintenance dose, intravenous cyclophosphamide (500 mg biweekly for 3 months) and tacrolimus (3.0 mg/day). Tacrolimus was continued as maintenance therapy. Complete remission was defined as a spot urine protein/creatinine ratio of < 0.5 g/gCr with no active urine casts and a serum creatinine level that was either normal or within 30% of a previously abnormal baseline level. We retrospectively compared results for the study patients with those of 18 historical controls conventionally treated with cyclophosphamide and prednisolone. Results At baseline, the mean patient age was 41.5 ± 14.6 years (male:female ratio 2:13), urine protein/creatinine ratio 3.9 ± 2.3 g/gCr and serum creatinine 84.6 ± 34.6 µmol/L. Lupus nephritis classifications included classes IV ( n = 8), III + V ( n = 1), IV + V ( n = 5) and unclassified ( n = 1). Eleven patients completed the treatment protocol and four withdrew. At 6 months, 12 of 15 (80.0%) had achieved complete remission using intention-to-treat analysis, significantly more than historical controls (seven of 18 patients, 38.9%). A transient increase in serum creatinine and gastric symptoms occurred in three cases. One patient withdrew due to cytomegalovirus antigenemia and severe diabetes, and one patient died of thrombotic microangiopathy. Conclusions Multitarget therapy with cyclophosphamide and tacrolimus can be a therapeutic option for lupus nephritis. Clinical trials registration Combination therapy of tacrolimus and intravenous cyclophosphamide for remission induction of lupus nephritis, UMIN: 000004893, URL: https://upload.umin.ac.jp/cgi-open-bin/ctr/ctr.cgi?function=brows&action=brows&type=summary&recptno=R000005830&language=E . Date of registration: 18 January 2011.

Investigation of exercise intensity during a freestyle wrestling match.

AIM: This study investigated changes in exercise intensity during a freestyle wrestling match. METHODS: Wrestling matches that consisted of three periods of 2 min were performed by nine elite collegiate male wrestlers. Exercise intensity was measured using heart rate (HR), blood lactate concentration ([BLa-]), and rating of perceived exertion (RPE). HR was continuously recorded during the match, and mean HR during each period was obtained. [BLa-] and RPE according to the 6-20-point Borg Perceived Exertion Scale were measured immediately after each period. RESULTS: HR during the wrestling match increased as a whole, but a continuous decrease in HR was observed during the first half of the second and third periods. This was likely caused by a cautious strategy involving reduced aggressive actions. HR significantly increased (P<0.05) from the first period (81 ± 6% of maximal HR [HRmax]) to the second period (88 ± 5% of HR(max)), and from the second to the third periods (92 ± 5% of HR(max)). [BLa-] significantly increased (P<0.05) from the first period (7.6 ± 2.0 mmol.L⁻¹) to the second period (10.4 ± 4.2 mmol.L⁻¹), but not from the second to the third periods (11.5 ± 3.1 mmol.L⁻¹). RPE significantly increased across the three periods (first, 13 ± 1; second, 15 ± 2; and third periods, 17 ± 2, P<0.05). CONCLUSION: Our results on changes in exercise intensity during each period and between periods would be helpful for making strategic decisions during wrestling matches and for planning daily training.

Towards a fibrous composite with dynamically controlled stiffness: lessons from echinoderms.

Sea urchins and sea cucumbers, like other echinoderms, control the tensile properties of their connective tissues by regulating stress transfer between collagen fibrils. The collagen fibrils are spindle-shaped and up to 1 mm long with a constant aspect ratio of approx. 2000. They are organized into a tissue by an elastomeric network of fibrillin microfibrils. Interactions between the fibrils are regulated by soluble macromolecules that are secreted by local, neurally controlled, effector cells. We are characterizing the non-linear viscoelastic properties of sea cucumber dermis under different conditions, as well as the structures, molecules and molecular interactions that determine its properties. In addition, we are developing reagents that will bind covalently to fibril surfaces and reversibly form cross-links with other reagents, resulting in a chemically controlled stress-transfer capacity. The information being developed will lead to the design and construction of a synthetic analogue composed of fibres in an elastomeric matrix that contains photo- or electro-sensitive reagents that reversibly form interfibrillar cross-links.

Collagen fibril aggregation-inhibitor from sea cucumber dermis.

Collagen fibrils from the dermis of the sea cucumber Cucumaria frondosa are aggregated in vitro by the dermal glycoprotein stiparin (Trotter et al., 1996). Under physiological ionic conditions stiparin appears to be both necessary and sufficient to cause fibrils to aggregate (Trotter et al., 1997). We report here the initial biochemical and biophysical characterization of a sulfated glycoprotein from C. frondosa dermis that binds stiparin and inhibits its fibril-aggregating activity. This inhibitory glycoprotein, which has been named 'stiparin-inhibitor,' has the highest negative charge density of all the macromolecules extracted from the dermis. SDS-PAGE reveals three approximately 31-kDa bands that stain with alcian blue but not with Coomassie blue. Analytical ultracentrifugation indicates a native molecular weight of 62 kDa. Transmission electron microscopy of rotary-shadowed molecules shows curved rods about 22 nm long. The glycoprotein does not bind collagen fibrils, but does bind stiparin with a 1:1 stoichiometry. The binding of stiparin-inhibitor to stiparin prevents the binding of stiparin to collagen fibrils. The carbohydrate moiety produced by papain-digestion of the glycoprotein retains all of its inhibitory activity. The carbohydrate moiety of the inhibitor is dominated by galactose and sulfate.

Characteristic loss of heterozygosity in chromosome 3P and low frequency of replication errors in sporadic renal cell carcinoma.

PURPOSE: A high frequency of genetic loss at 4 loci on chromosome 3p has been shown in human sporadic renal cell carcinomas (RCCs), but the relative contribution of each locus is not well known, and the involvement of DNA replication errors (RERs) in carcinogenesis of RCCs remains unclear. We report the simultaneous comparison of genetic loss at the 4 chromosome 3p loci and RERs in sporadic RCCs. MATERIALS AND METHODS: DNA was extracted from 33 Japanese sporadic RCC samples, and examined for loss of heterozygosity (LOH) and RERs by amplification of 14 microsatellite regions. LOH of the von Hippel Lindau (VHL) gene was analyzed by a polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) method. The target sequences of RER, transforming growth factor beta type II receptor (TGFbetaRII) and Bcl-2-associated X protein (BAX) genes were amplified and analyzed for mutations by sequencing. RESULTS: LOH of the VHL gene was observed in 53.3% of RCCs, a higher frequency than that of the 4 regions 3p12-p13 (18.8%), 3p14.2 (17.4%), 3p21 (21.2%) and 3p25-p26 except for VHL (31.3%). There were no RERs in 14 microsatellite regions, including the mononucleotide (A)10 repeats of the TGFbetaRII gene and (G)8 repeats of the BAX gene. CONCLUSION: Japanese sporadic RCCs were characterized by predominant loss of VHL gene and low contribution of the other 3 candidate RCC tumor suppressor genes. RERs, mostly caused by a defect of DNA mismatch repair, might only rarely be involved in the carcinogenesis of sporadic RCCs.

[Immunological effect of recombinant interferon-gamma on tumor infiltrating lymphocytes of renal cell carcinoma--relationship with clinical stage].

We studied the relationship between clinical stage and tumor infiltrating lymphocytes (TIL) in 26 cases of renal cell carcinoma. In 10 patients, interferon-gamma (IFN-gamma) was administered preoperatively (administration group); 5 patients had low stage (Robson < or = I), and the remaining 5 patients had high stage (Robson > or = II) tumors. The other 16 patients underwent nephrectomy alone without preoperative IFN administration (control group); 11 patients had low stage, and the remaining 5 patients had high stage tumors. Immunohistochemical studies of tumor infiltrating lymphocytes in renal cell carcinoma showed a significantly high incidence of CD3, CD8, CD11b and ICAM-1 in the administration group, while CD4, LFA-1 and Ber-MAC3 were increased without significance (p < 0.05). Concerning clinical stage (Robson), a significant increase in CD3, CD8, CD11b and ICAM-1 was observed in the patients with high-stage tumors in the administration group compared to those in the control group (p < 0.05). This suggested that TIL could be changed by preoperative administration of IFN-gamma.

[Conservative treatment of postoperative chylothorax in a patient with chronic traumatic aneurysm of thoracic aorta].

A 66-year-old male was admitted to our hospital because of a chronic traumatic aneurysm of the thoracic aorta caused by a traffic accident 5 years earlier. Although he had been treated successfully by patch closure under extracorporeal circulation, chylothorax gradually manifested 9 days after the operation. The fact that our conservative treatment resulted in complete recovery from chylothorax, strongly suggested that progress of nutriology was a contributing factor.

Preoperative diagnosis of malignant melanoma using the touch-fluorescence method.

We have developed a touch-fluorescence method using preparations from the outer surface of elevated and ulcerative malignant melanoma lesions. This method allows the demonstration of the melanogenic activity of melanoma cells within 30 min and has made it possible to definitively diagnose a lesion as malignant melanoma at the initial examination. In 21 cases clinically diagnosed as typical melanoma, 17 were definitively diagnosed as melanoma from the touch-fluorescence microscopic findings. Of the four cases in which no fluorescent tumor cells were found, two proved not to be melanoma. There was some correlation between the morbid types of primary melanoma and the configuration of fluorescent melanoma cells, the main cellular configuration of superficial spreading melanoma being round whereas that of nodular melanoma and acral lentigenous melanoma was pleomorphic. Based on the above results, this method was concluded to be extremely reliable for the preoperative diagnosis of elevated and ulcerative lesions of malignant melanomas, which are sometimes difficult to diagnose.

5-S-cysteinyldopa in urine and tumors.

1) The urinary 5-S-CD contents in malignant melanoma subjects (n = 135) and non-melanoma subjects (n = 204) were measured by HPLC. These results suggest that, as a biochemical marker, periodic measurement of urinary 5-S-CD is quite useful for evaluating the determinations of stage classification (UICC, 1987), and the detection of metastases, the therapeutic efficacy of operation or immunochemotherapy against malignant melanoma. 2) Quantitative analyses of 5-S-CD values in tissues from primary malignant melanoma lesions (n = 24) and pigmentary tumors other than melanomas (n = 136) showed 80.6-821.4 ng/mg and N.D.-55.0 ng/mg respectively. In view of the above findings, it was suggested that the pigmentary tumors can be diagnosed as malignant melanoma if the 5-S-CD value in the tissues is higher than 100 ng/mg.

New polyenic antibiotics active against gram-positive and gram-negative bacteria. VIII. Construction of synthetic medium for production of mono-chloro-congeners of enacyloxins.

New antibiotics enacyloxins (ENXs) are a family of non-lactonic polyene antibiotics produced by Frateuria sp. W-315. For the production of antibiotics, we had to employ two-step fermentations, the first is the production of spent medium of Neurospora crassa and the second is the production of antibiotics by Frateuria. To simplify the production of antibiotics, systematic analyses have been done on the spent medium, and factors which affect the production of antibiotics characterized. From the above results, we constructed a new medium for antibiotic production. Moreover, we could get a new antibiotic named enacyloxin IIIa (1), C33H48O11NCl (m/z 669). 1 was deduced to be one of the congeners of enacyloxins because it was similar to ENX IIa or ENX IVa both in biological and physico-chemical properties. Chlorine of 1 could be replaced by bromine, biosynthetically, and the resultant bromine-containing antibiotic also showed an antibacterial activity comparable to 1.

Time allocations of various activities under multiple schedules in pigeons.

Time allocations of three pigeons' various activities (pecking, turning, preening, wing flapping, etc.) were measured under usual multiple schedules in which only key-peck response was reinforced. Positive behavioral contrast was found for both the number and the time allocation of key-peck response. Some time allocations of the other activities were inversely varied to that of key-pecks, as stated in the competition theory of behavioral contrast. However, the amount of reallocation of interim activities was not enough to complement the amount of increases or decreases of key-pecks in the unchanged component. Most of the amount of increases or decreases of key-pecks in the unchanged component was complemented with the inactive time. This result suggests that behavioral competition is not the primary mechanism for key-peck behavioral contrast in multiple schedules in pigeons. It is important to consider inactive time, which is emerged in the usual experimental setting, to clearly attribute the behavioral contrast to reallocation of time for other activities.

Moving grating and dc external field in photorefractive GaP at 633 nm.

We demonstrate that the photorefractive effect in GaP crystal at 633 nm can be enhanced using an externally applied dc field and a moving grating. A two-beam coupling gain coefficient of 2.5 cm(-1) and a steady-state phase-conjugate reflectivity of 1.9% were obtained.

New wire suture for sternal fixation.

A new sternal fixation wire, 60 cm in length, was tested clinically. The center portion of the wire is 30 cm in length and 0.8 mm in diameter, while the two end portions of the wire are 15 cm in length and 0.3 mm in diameter. A drill or gimlet with a small hole in the tip was employed to bore a hole in the sternum. The thin ends of the wire were inserted into the hole of the drill, making it easier to pass the wire through the sternal bone. We have found wire penetration through the sternum to be simpler and less traumatic using this technique.

Krypton and argon laser photocoagulation effects in subretinal hemorrhage.

Previous studies suggested that krypton laser photocoagulation was more effective in the treatment of macular diseases than argon laser. Furthermore, it could perform photocoagulation more effectively in some lesions with subretinal hemorrhage, because the krypton laser beam was poorly absorbed by hemoglobin. In the present experiment, hemorrhagic retinal detachment was produced in monkey eyes with Q-switched Nd-YAG laser, and 4 weeks later photocoagulation was performed with krypton and argon lasers to compare the differences in the effects of these two lasers. When the subretinal hemorrhage and a heavy coagulation effect was produced in the detached retina, but no coagulation effects were observed in the choroid. Krypton laser beam could go through the hemorrhage and certain coagulation effects were observed in the choroid and the detached retina. It is suggested that krypton laser photocoagulation is more effective in the lesions behind subretinal hemorrhages than photocoagulation with argon laser.

Wide field fluorescein angiography by use of contact lens.

A system has been developed to obtain fluorescein angiographs with a field measuring to 90 degrees. This involves a contact lens and a new fundus camera with a basic field of 60 degrees. The contact lens, however, can be used with any fundus camera and will result in an increase in the area of the photographs of over 2X. There are numerous clinical applications for high resolution, high contrast, wide field fluorescein angiography.