Preferential localization of heat shock protein 47 in dilated endoplasmic reticulum of chicken chondrocytes.

We investigated the distribution of heat shock protein 47 (hsp47) in cultured chicken embryonic chondrocytes and epiphyseal chondrocytes of tibial bones from 1-day-old to 6-week-old chickens. Northern blot and immunoblot analyses revealed that hsp47 exists in epiphyseal cartilage and cultured chondrocytes. Confocal laser immunofluorescence microscopy showed that hsp47 was localized mainly in the many granular structures found in the cytoplasm that contain Type II collagen. Epiphyseal cartilage and cultured chondrocytes were embedded in LR White resin and hsp47 was detected by protein A-immunogold electron microscopy. Gold particles were localized exclusively in the cisternal space of the endoplasmic reticulum (ER), and the labeling density of the cisternal space of the dilated ER was always higher than that of the non-dilated ER. In all the differentiating zones of epiphyseal cartilage, the labeling density was highest in the hypertrophic cells. These findings suggest that hsp47 plays an important role(s) in the synthesis, processing, and assembly of Type II collagen.

Enhancement of chemotherapeutic effects with focused shock waves: extracorporeal shock wave chemotherapy (ESWC).

The effects of shock waves in combination with various anti-cancer agents i.e. Bleomycin (BLM), Cisplatin (CDDP) and 5-fluorouracil (5-FU) on tumor cells suspended in media containing these agents were examined. GCIY cells derived from human gastric cancer and LS 174T and SW480 cells derived from human colon cancers were used for in vitro experiments; GCIY and SW480 cells were also transplanted into nude mice for in vivo study. It was only with BLM that enhancement was evident in all three cell lines, with a degree of chemotherapeutic enhancement proportional to the amount of shock wave energy applied. Ladder formation of DNA in GCIY cells was observed only when treated with both BLM and shock waves in combination. When SW480 and GCIY cells transplanted into the backs of nude mice were treated with a combination of intravenously (i.v.) injected BLM and regional exposure to shock waves, a significant enhancement of chemotherapeutic effects was observed in terms of the tumor growth curve.

[Initial experience using a new type extracorporeal lithotripter with an anti-miss-shot control device].

The results of clinical trial using a TOSHIBA piezolectric extracorporeal lithotripter are presented. The advantages of this machine are the installation of an anti-miss-shot control device (AMCD) to prevent miss-shots to the renal tissue and the improved ultrasound stone localization by an over-head type applicator. AMCD functions so that shock waves are generated only when the stone is in the focal region. Forty patients with urinary tract stone were treated by this machine from October 1989 to March 1990 at the Department of Urology, Tohoku University School of Medicine. Almost all patients could be treated with an analgesic only. The X-ray film obtained three months after ESWL treatment showed that 23 cases (57.5%) were stone-free and 10 cases (25.0%) had stone fragments of less than 4 mm. The successful rate of ESWL using this machine was 82.5%, the result was better than that using other piezolectric machines. No serious complication was observed, except mild hematuria in all cases. Milder postoperative hematuria suggested the decrease of renal tissue injury. It is concluded that the treatment with this machine is safe and seems to be more favorable than the conventional ones.

[Ultrastructural renal damage after shock wave exposure].

This report clarified the direct damage to renal cell ultrastructure on shock wave exposure. The renal cortex of the rabbit was exposed to the shock waves (1200 bar) generated by the piezo-ceramics. The change was examined by electron microscopy immediately after 20 shots exposure and 500 shots exposure as well as one week after 500 shots exposure. In all groups, the most critical damage was observed in the proximal tubule. Immediately after 20 shots exposure, large vacuoles were found to be enclosed by infoldings in the proximal tubular cell. The damaged cell lost its brush border and jutted out into the lumen. The ruins of organelles (nuclei, mitochondria, etc.) were seen in the lumina of proximal and distal tubules. The damage progressed, in the 500 shots exposure group, to include the distal tubular cytoplasm. One week after the 500 shots exposure, a few of atrophic tubular cells were found in wide scars. The tubular cell which contained a deformed nucleus and destroyed organelles was desquamated from the basement membrane. The change in the tubular cell after shock wave exposure was apparently different from that after such intervention as one hour ischemia. It was concluded that the direct damage to renal cells by underwater shock wave exposure was really present.

[The pathogenesis of hypercalciuria from the aspect of the response to human parathyroid hormone in Ca containing stone formers].

In order to clarify the pathogenesis of hypercalciuria, the response to extrinsic human parathyroid hormone (h-PTH) was studied the 21 patients with calcium containing urinary stone(s) and 5 normal controls (NO). The stone patients were classified into 3 groups from the result of the oral calcium loading test, i.e. Non-hypercalciura (NH, n = 8) and absorptive hypercalciuria (AH, n = 8) and renal hypercalciuria (RH, n = 5). Only in the AH group, urinary excretion of calcium (u-Ca) was strongly correlated to that of sodium (u-Na) in pre-load of h-PTH, and both increments were also correlated in post-load of h-PTH. As of this fact the increase in Na excretion seems to be responsible for a cause of hypercalciuria in the AH group. There was a significant correlation between the value of %TRP in pre-load of h-PTH and the rate of urinary phosphorus (P) increment between pre-load and post-load of h-PTH in the NO and NH groups. However, this relationship was not found in the AH and RH groups. These findings indicate that there is response disorder of P to h-PTH. In addition, serum P was low, plasma 1,25 (OH)2D was high, N-c-AMP was low in the AH group, whereas both serum P and %TRP were low in the RH group in pre-load of h-PTH. These findings are compatible with the primary renal P leak.(ABSTRACT TRUNCATED AT 250 WORDS)

[Radiopacity of the cystine calculi on KUB].

To study the radiopacity of cystine calculi. We have compared them with oxalate and/or phosphate calculi on plain X-ray films (KUB). Eleven pure cystine calculi from 5 people in 4 families, and 8 oxalate and/or phosphate calculi as controls were obtained from patients by lithotomy or spontaneous discharge in Dokkyo University School of Medicine and affiliated hospitals, since 1978. The radiopacity of cystine calculi was measured by a densitometer PDA-60 (Konika Co.), comparing the density of the calculus with that of the area surrounding the calculus on KUB. All cystine calculi were easily recognized on KUB, and their densities were fairly identical with those of oxalate and/or phosphate calculi.

[Estimation of glomerular filtration rate from the urinary excreted contrast medium concentration].

In the animal experiments, glomerular filtration rate (GFR) values were estimated from the urinary response to the intravenously administrated iopamidol. By introducing some reasonable assumptions, mass balance equations about the contrast medium were transformed and GFR was defined as the ratio of logarithm of urinary concentration by time. Urinary concentrations were determined by measuring the photodensity on the X-ray film of urine specimens. Obtained values of GFR were well fitted to those obtained by the analysis of the plasma disappearance curve. The advantage of this method is that measurement of urinary volume is not necessary.

Mechanism of fragmentation of urinary stones by underwater shock wave.

The focusing of an underwater shock wave, generated by an underwater microexplosion, has been studied by several methods, such as holography, pressure measurement and pressure print. It has been shown that the shock wave could be focused within the range of a few millimeters and with an amplitude of 1 kbar. The acoustic impedances of various kinds of urinary stones were measured by the original graphical method using holographic interferrometry. The process of breaking a stone by a focused underwater shock wave was observed with high-speed cinematography. It was supposed that the main mechanism of breaking the stone is the tensile stress at the solid-water acoustic interface.

Clinical application of extracorporeal shock wave lithotripsy using microexplosions.

Extracorporeal stone disintegration using a chemical explosive (10 mg. lead azide) as an energy source of underwater shock waves was performed in 105 patients 11 to 72 years old who had stones in the upper urinary tract. We used a prototype disintegrator in this series. The over-all rate free of stones 3 months after treatment was 82 per cent. Shock wave therapy was performed alone in 77 patients (73 per cent), while the remainder required combined treatment with percutaneous and/or transurethral lithotripsy. The most common complications were colic pain (30 per cent) and fever (23 per cent). In 4 patients other complications, that is bacteremia, gastrointestinal bleeding, ureteral injury and subcapsular renal hematoma, were observed but they were treated conservatively with no serious adverse effects. Our study demonstrates the safe use of this method for clinical treatment.

Initial experience using a new type extracorporeal lithotripter with an anti-misshot control device.

Initial clinical trials using a new overhead piezoelectric extracorporeal lithotripter is presented. The advantages of this machine are the installation of an anti-misshot control device (AMCD) to prevent misshots to renal tissue, simplicity for ultrasound stone localization by the overhead type applicator, and elimination of necessity for anesthesia. Thirty patients with upper urinary tract calculi were treated by this machine with a success rate of 77% and stone destruction rate of 93%. No serious complication was observed. The AMCD functioned in such a manner that shock waves were generated only when the stone was in the focal region. Although there were AMCD nonfunctioning cases (small and deeply located stones), it is indicated that this machine is favorable for safe, extracorporeal lithotripsy.

Tenosynovial injection for carpal tunnel syndrome.

Although local steroid injection into the tenosynovium is a frequent treatment for carpal tunnel syndrome, it involves some risk and is not always effective. We simulated injection on 16 fresh cadaver forearms, instilling 1 ml (group 1) or 2 ml (group 2) of methylene blue at 1 cm or 3 cm proximal to the most distal wrist crease. Passive flexion and extension were simulated 2 minutes after injection by application of traction to the appropriate digital tendons. Specimens were dissected under loupe magnification from midpalm to midforearm, dye diffusion was quantified and photographed, sections of the carpal tunnel and contents were graded for presence of dye, and average values were determined for each of the four groups. Diffusion of dye was best in group 2B in which 2 ml was injected 3 cm proximal to the distal wrist flexion crease.

Intermittent irrigation system for dissolution of renal calculi monitored by computer.

An intermittent irrigation system was tested in vitro and in a clinical case. In vitro experiments showed that the intermittent irrigation was more efficient to dissolve calculi compared to the continuous irrigation. The pelvic irrigation was performed clinically by a cystoscopically passed angiographic catheter. The system could be operated in constant low pressure without supervision. The result shows that this system will be safe and valuable for pelvic irrigation.

Extracorporeal stone disintegration using chemical explosive pellets as an energy source of underwater shock waves.

Extracorporeal renal stone disintegration using a chemical explosive pellet (lead azide 10 mg.) as an energy source of underwater shock waves has been successfully performed in animals. The shock wave was observed by holographic interferometry. Shock wave generation was performed by a reflector whose configuration was part of a pseudoellipsoid. The explosions were conducted 10 to 100 times for each animal and the stone (extracted human renal calculus or model calculus of activated alumina) placed in the renal pelvis was disintegrated satisfactorily. Negative findings in explorative laparotomy and histological examination, except for minor bleeding in several tubular lumina of the kidney, indicated that the method was clinically applicable.

Wrist position and extensor tendon amplitude following repair.

After primary repair of severed extensor tendons, various methods are used to limit tendon adhesions and avoid rupture. Early passive digital motion with wrist extension (a "reverse Kleinert" protocol) has been advocated. However, there are no data to support an optimum wrist position or to indicate how much finger motion may safely be permitted. In this study we used eight fresh cadaver limbs to measure extensor tendon gliding in Verdan's zones 3 to 8 when active grip and passive extension were simulated at different wrist positions. We found that if the wrist is extended more than 21 degrees, the extensor tendon glides with little or no tension in zones 5 and 6 throughout full simulated grip to full passive extension, permitting "passive motion" exercises to minimize tendon adhesions without risking rupture. In addition, we found that up to 6.4 mm of tendon can be debrided safely and full grip can still be permitted postoperatively if the wrist is splinted at 45 degrees extension.