Efficacy of Bisphosphonate for Urolithiasis Complicated by Osteogenesis Imperfecta.

A 44-year-old man with osteogenesis imperfecta presented with left renal colic. Non-contrast computed tomography revealed a stone (10×9 mm) in the left upper ureter. Ureteroscopic lithotripsy was performed twice and stone-free status was achieved. An analysis of the stone revealed a mixed composition including calcium oxalate and calcium phosphate. Postoperatively, we administered bisphosphonates to prevent recurrence of urolithiasis, as 24-hour urine collection revealed marked hypercalciuria. Eighteen months after surgery, the urinary calcium levels had normalized, and there was no recurrence of urolithiasis. Osteogenesis imperfecta can be complicated by urolithiasis, but bisphosphonates may be useful in preventing recurrence of this disease.

In vitro comparison of simulated intrapelvic pressure in an artificial kidney model during retrograde intrarenal surgery among various single-use ureteroscopes.

OBJECTIVES: This study compares intrapelvic pressure (IPP) during retrograde intrarenal surgery with various single-use flexible ureteroscopes (f-URS) in an artificial kidney model. METHODS: We created an artificial kidney model with a pressure sensor using a bladder evacuation device. The model was completely closed and the only backflow was on the side of the ureteroscope inside the ureteral access sheath (UAS). We tested five single-use f-URSs (LithoVue, Wiscope, PU3022A, PU3033A, and AXIS) with six different types of UAS (9.5/11.5-14/16 Fr). Using the automatic irrigation system, 30 s of irrigation was performed at various pressures (40-180 mmHg) and steady-state IPP was recorded. IPP was compared between the five single-use f-URSs. IPP cutoff value was determined at 30 mmHg. The diameter of the endoscope tip and the curved and shaft parts were also measured and recorded. RESULTS: The diameters of all parts were significantly different between single-use f-URSs. The maximum IPP tended to be higher in ureteroscopes with larger diameters of the proximal parts (curved part/shaft part). In LithoVue and Uscope PU3022A f-URSs, the maximum IPP did not exceed 30 mmHg when UAS ≥12/14 Fr was used. In AXIS and Wiscope f-URSs, it did not exceed the cutoff value when the UAS ≥11/13 Fr was used. In Uscope PU3033A f-URS, it did not exceed 30 mmHg when the UAS ≥10/12 Fr was used. CONCLUSIONS: Maximum IPP tended to be higher in f-URSs with larger diameters of the proximal part and the appropriate size of the UAS differed between various single-use f-URSs.

Dynamic Changes in Fluid Temperatures during Laser Irradiation Using Various Laser Modes: A Thermography-Based In Vitro Phantom Study.

The differences in dynamic thermal changes during laser lithotripsy between various laser pulse modes are unclear. We used thermography to evaluate the temporal changes in high-temperature areas during laser activation in order to compare different laser pulse modes. An unroofed artificial kidney model was used for the experiments. The laser fired for 60 s with a laser setting of 0.4 J/60 Hz in the following four different laser pulse modes without saline irrigation: short pulse mode (SPM), long pulse mode (LPM), virtual basket mode (VBM) and Moses mode (MM). Using the first 30 s of moving images, we compared the ratio of a high-temperature area of >43 °C to the total area every 5 seconds. The dynamic changes in fluid temperatures were shown to be different between the laser pulse modes. The extent of the high-temperature areas during the laser activation was large in the LPM and MM compared with the SPM and VBM. While the high-temperature areas expanded in an anterior direction in the early laser irradiation period using the LPM, they spread in a posterior direction in the early laser activation period using the MM. Although only the temperature profile in one specific plane was investigated, these results are considered useful for preventing thermal injuries during retrograde intrarenal surgeries.

Motor generated torque drives coupled yawing and orbital rotations of kinesin coated gold nanorods.

Kinesin motor domains generate impulses of force and movement that have both translational and rotational (torque) components. Here, we ask how the torque component influences function in cargo-attached teams of weakly processive kinesins. Using an assay in which kinesin-coated gold nanorods (kinesin-GNRs) translocate on suspended microtubules, we show that for both single-headed KIF1A and dimeric ZEN-4, the intensities of polarized light scattered by the kinesin-GNRs in two orthogonal directions periodically oscillate as the GNRs crawl towards microtubule plus ends, indicating that translocating kinesin-GNRs unidirectionally rotate about their short (yaw) axes whilst following an overall left-handed helical orbit around the microtubule axis. For orientations of the GNR that generate a signal, the period of this short axis rotation corresponds to two periods of the overall helical trajectory. Torque force thus drives both rolling and yawing of near-spherical cargoes carrying rigidly-attached weakly processive kinesins, with possible relevance to intracellular transport.

Comparison of stone retropulsion between Moses mode and virtual basket mode: an in vitro study using artificial stones.

This study aimed to evaluate stone retropulsion in various laser pulse modes in both Moses mode (MM) and virtual basket mode (VBM). Experiments were performed using a channel-shaped rubber rail and artificial stones. We compared short pulse mode and long pulse mode in both MM and VBM with the laser tip positioned so that it was touching and at 1 and 2 mm distances from the stone surface. Stone retropulsion was measured after the laser fired for 10 s in three different laser settings: 0.5 J/8 Hz, 0.8 J/8 Hz and 1.0 J/8 Hz. When the laser tip was touching the artificial stone, stone retropulsion in MM was significantly shorter than that in VBM in all laser settings (P < 0.01, P = 0.02 and P = 0.02, respectively). At 1-mm distance, stone retropulsion in MM was significantly shorter than that in VBM in 0.8 J/8 Hz and 1.0 J/8 Hz settings (P < 0.01 and P = 0.01, respectively). At 2-mm distance, however, there were no differences between MM and VBM in stone retropulsion in any laser settings. Stone retropulsion was not significantly different between the laser settings at 1-mm distance in MM, or when touching in VBM. In conclusion, stone retropulsion distance in MM can be shorter than that in VBM. Stone retropulsion in MM and VBM may be differently influenced by laser settings and laser tip position.

Highly Efficient Separation of Ultratrace Radioactive Copper Using a Flow Electrolysis Cell.

Preparing compounds containing the radioisotope 64Cu for use in positron emission tomography cancer diagnostics is an ongoing area of research. In this study, a highly efficient separation method to recover 64Cu generated by irradiating the target 64Ni with a proton beam was developed by employing a flow electrolysis cell (FE). This system consists of (1) applying a reduction potential for the selective adsorption of 64Cu from the target solution when dissolved in HCl and (2) recovering the 64Cu deposited onto the carbon working electrode by desorbing it from the FE during elution with 10 mmol/L HNO3, which applies an oxidation potential. The 64Cu was selectively eluted at approximately 30 min under a flow rate of 0.5 mL/min from the injection to recovery. The newly developed flow electrolysis system can separate the femtomolar level of ultratrace radioisotopes from the larger amount of target metals as an alternative to conventional column chromatography.

Rapid Flow-Based System for Separation of Radioactive Metals by Selective Complex Formation.

Short-lived radioactive metals are important tracers in clinical diagnosis. Radioactive metals for clinical use are produced from suitable target metals in cyclotrons. The trace amount of radioactive metal produced is contained in a relatively large amount of target metal. A rapid and effective method is required to isolate the radioactive metal. In the present study, selective complex formation followed by cation-exchange adsorption was performed in a continuous flow-based system. Ethylenediamine-N,N,N',N'-tetraacetic acid (EDTA) was selected as the ligand after simulation of the separation of radioactive Ga from the target (Zn). Selectively, the Ga-EDTA complex passed through the cation trap, while Zn2+ was trapped. This separation principle is opposite to that of typical solid-phase extraction, which captures the target ion. The proposed separation was performed in a flow-based system with a parallel, open-channel ion trap. The performance was optimized by altering the channel dimensions, channel-filling mesh, and flow rate. Finally, the target radioactive metal, Ga, was selectively and effectively (>99%) separated from a mixture of 50 fg Ga/L and 100 mg Zn/L. The concentration of Zn remaining in the Ga solution was 2.3 µg/L. The complexed Ga was converted to free Ga3+ by a simple UV irradiation method. The proposed method effectively and rapidly separates trace amounts of radioactive metals contained in larger amounts of target metals using a simple flow system that can be operated on site.

Efficacy of Methotrexate on Rat Knee Osteoarthritis Induced by Monosodium Iodoacetate.

OBJECTIVE: To explore whether methotrexate (MTX) prevents joint destruction and improves pain-related behaviors in the acute phase of knee osteoarthritis (OA) induced by monosodium iodoacetate (MIA) in a rat model. METHODS: Twenty of 25 male Wistar rats (10-14 weeks old) received 3 mg MIA via intra-articular injection into their right knee and were then administered a vehicle control (n=10) or 3 mg/kg MTX orally weekly (n=10). We assessed differences in pain-related behavior, spontaneous lifting behavior, micro-computed tomography (CT), histopathology, and expression of pain- and inflammatory-related genes using reverse transcription-quantitative polymerase chain reaction (RT-qPCR) between the two groups for 4 weeks. Five rats were used as untreated controls to assess pain- and inflammatory-related mRNA expression in the dorsal root ganglia (DRG) and knee joints using RT-qPCR. RESULTS: Joint destruction and mechanical hyperalgesia were observed in the vehicle group. Decreases in mechanical pain thresholds for the knee joint and calf muscles were improved after MTX administration; however, joint damage assessed by micro-CT and histopathology was not significantly inhibited by MTX administration, while upregulation levels of transient receptor potential cation channel, subfamily V, member 1 (TRPV-1) (P<0.01) and brain-derived neurotrophic factor (BDNF) (P=0.02) mRNA in the DRG and nerve growth factor NGF mRNA (P=0.03) in the affected knee joints were significantly suppressed in the MTX group compared with the vehicle group at week 4. CONCLUSION: Our results imply that MTX administration improves pain-related behaviors and suppresses expression of pain-related mRNAs in the DRG and knee joint; however, MTX is not expected to prevent cartilage degeneration in MIA-induced OA in rat knee.

Characterization of the motility of monomeric kinesin-5/Cin8.

Cin8, the Saccharomyces cerevisiae kinesin-5, has an essential role in mitosis. In in vitro motility assays, tetrameric and dimeric Cin8 constructs showed bidirectional motility in response to ionic strength or Cin8 motor density. However, whether property-switching directionality is present in a monomeric form of Cin8 is unknown. Here we engineered monomeric Cin8 constructs with and without the Cin8-specific ∼99 residues in the loop 8 domain and examined the directionality of these constructs using an in vitro polarity-marked microtubule gliding assay within the range of the motor density or ionic strength. We found that both monomeric constructs showed only plus end-directed activity over the ranges measured, which suggested that minus end-directed motility driven by Cin8 is necessary for at least dimeric forms. Using an in vitro microtubule corkscrewing assay, we also found that monomeric Cin8 corkscrewed microtubules around their longitudinal axes with a constant left-handed pitch. Overall, our results imply that plus-end-directed and left-handed motor activity comprise the intrinsic properties of the Cin8 motor domain as with other monomeric N-kinesins.

CYK4 relaxes the bias in the off-axis motion by MKLP1 kinesin-6.

Centralspindlin, a complex of the MKLP1 kinesin-6 and CYK4 GAP subunits, plays key roles in metazoan cytokinesis. CYK4-binding to the long neck region of MKLP1 restricts the configuration of the two MKLP1 motor domains in the centralspindlin. However, it is unclear how the CYK4-binding modulates the interaction of MKLP1 with a microtubule. Here, we performed three-dimensional nanometry of a microbead coated with multiple MKLP1 molecules on a freely suspended microtubule. We found that beads driven by dimeric MKLP1 exhibited persistently left-handed helical trajectories around the microtubule axis, indicating torque generation. By contrast, centralspindlin, like monomeric MKLP1, showed similarly left-handed but less persistent helical movement with occasional rightward movements. Analysis of the fluctuating helical movement indicated that the MKLP1 stochastically makes off-axis motions biased towards the protofilament on the left. CYK4-binding to the neck domains in MKLP1 enables more flexible off-axis motion of centralspindlin, which would help to avoid obstacles along crowded spindle microtubules.

Electrodialytic Handling of Radioactive Metal Ions for Preparation of Tracer Reagents.

Radioactive metals are applied in biochemistry, medical diagnosis such as positron emission tomography (PET), and cancer therapy. However, the activity of radioisotopes exponentially decreases with time; therefore, rapid and reliable probe preparation methods are strongly recommended. In the present study, electrodialytic radioactive metal ion handling is studied for counter ion conversion and in-line probe synthesis. Presently, counter ion conversion and probe synthesis are achieved by evaporative dryness and solution mixing, respectively. Evaporative dryness is time-consuming and is a possible process that can lead to loss of radioactive metal ions. Mixing of solutions for synthesis makes dilution and undesirable effects of counter ion on the synthesis. An optimized electrodialytic flow device can transfer a radioisotope, 64Cu2+, with high recovery from HCl matrices to HNO3 (∼100%). Matrices can also be transferred into acetic acid and citric acid, even though the concentration of the metal ion is at the picomolar level. The ion transfer can also be achieved with simultaneous counter ion conversion, complex synthesis, and enrichment. When the ligand was dissolved in an acceptor solution, the transferred metal ions from the donor were well mixed and formed a complex with the ligand in-line. The efficiency of the synthesis was ∼100% for 1.0 pM 64Cu. A relatively larger donor-to-acceptor flow rate can enrich the metal ion in the acceptor solution continuously. The flow rate ratio of 10 (donor/acceptor) can achieve 10 times enrichment. The present electrodialytic ion handling method can treat ultra-trace radioisotopes in a closed system. With this method, rapid, effective, and safe radioisotope treatments were achieved.