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Biomedical subjects

A Sano

Publications and source records attributed to A Sano.

At least 91 records · Page 5Linked to original sources

Fungistatic and fungicidal activities of murine polymorphonuclear leucocytes against yeast cells of Paracoccidioides brasiliensis.

Recently, a novel culture medium for detecting live yeast cells of Paracoccidioides brasiliensis was developed by Kurita et al. Using this culture medium, murine peritoneal polymorphonuclear leucocytes (PMN) were examined for fungistatic and fungicidal activities against P. brasiliensis yeast cells. The magnitude of the antifungal effect of PMN varied depending upon the fungal isolates used. PMN exhibited a killing effect on P. brasiliensis isolate Bt-4 in 2 h of coculture. In contrast, the other three fungal isolates employed were resistant to killing by PMN. However, PMN considerably suppressed the growth of isolates Tatu and Recife in a long-term assay (approximately 72 h). The growth of isolate Bt-9 was also suppressed by PMN during the first 24 h, but was found to be considerably promoted at 72 h of coculture. Interferon-gamma (IFN-gamma), but not tumour-necrosis factor-alpha, significantly augmented the antifungal activity of PMN. IFN-gamma-treated PMN exhibited a killing effect on isolates Tatu, Recife and Bt-9 after 24 h of coculture, and showed an enhanced killing effect on isolate Bt-4. Contact between PMN and fungal cells was required for PMN to exert the antifungal effect. Our results suggest that PMN, whether activated with cytokines or not, might play a critical role in host resistance in early infection with this fungus by buying time for development of more effective immunologic responses.

Animals↗

Protein release from collagen matrices.

The effective delivery of protein drugs is an important research subject in the field of pharmacology, and to prolong the effect of protein drugs, many studies are being conducted to control the release of proteins from various carrier materials. Collagen is one of the most useful candidates for this purpose, and many studies have been reported; pharmaceutical formulations containing collagen in gel, film and sponge form are used to incorporate low-molecular-weight compounds such as antibiotics and carcinostatics, and the release of these compounds is controlled by the concentration of the gel as well as the shape and degree of crosslinking of the matrix. However, it is still difficult to retain protein drugs in the collagen. In this article, we report on the controlled release of protein drugs using collagen which exhibits good biocompatibility as a carrier, focusing on a new drug delivery system, the Minipellet, which we have developed.

Journal Article↗

Local application of basic fibroblast growth factor minipellet induces the healing of segmental bony defects in rabbits.

Fibroblast growth factor (FGF) has been reported to increase the volume of callus in a fracture model of rats. There are, however, no reports of successful repair of segmental bony defects by application of an FGF solution. In this study, the effects of basic FGF on the repair of segmental bony defects in the rabbit femur were examined. Minipellet, a new drug delivery system using atelocollagen, was employed to ensure effective delivery of FGF. Segmental bony defects (10 mm in length) were created in the right femurs of 19 rabbits. In pilot studies, no defects of this size healed spontaneously within 6 weeks. Bones were stabilized with miniexternal fixators. Minipellets containing basic FGF were implanted between fragments so as to bridge the two fragments. The healing processes were monitored radiographically and studied histologically. In rabbits in which FGF was added to the defect site at doses of 1.4 microgram or higher, approximately 90% of the defects were filled with new bone and cartilage within 6 weeks after minipellet implantation. In rabbits receiving placebo minipellets, however, approximately 15% of the defects were filled by callus within 6 weeks. Furthermore, this callus did not change into mature bone. An injection of 2 microgram of FGF solution to bony defects had no effect on the repair of segmental bony defects. These findings suggest that FGF plays a role in the production of adequate volumes of callus particularly in the initial stages of fracture healing and that sustained local release enables FGF to be effective at a low dose. In summary, large segmental bony defects healed after insertion of low-dose FGF minipellets. An adequate dose of FGF and an appropriate delivery system are required for successful healing of large bony defects. These findings imply the potential value of FGF minipellets in clinical practice.

Animals↗

Humeral attachment of the supraspinatus and infraspinatus tendons: an anatomic study.

We investigated the anatomic relationship of the supraspinatus (SSP) and infraspinatus (ISP) tendons to the three facets of the greater tuberosity. After removing the superficial layer of the cuff to expose the tendon fibers in 10 embalmed shoulders, the cuff tendon attachment to the facets was examined, and the location of attachment was measured in reference to (1) the anterior margin of the greater tuberosity and (2) the superior margin of the sulcus (anatomic neck without cartilage). The SSP tendon attached to the superior facet and the superior half of the middle facet. The ISP tendon attached to the entire middle facet, covering a portion of the SSP tendon. Thus, the anterior half of the superior cuff tendon (12.6 +/- 1.1 mm) was composed of only the SSP tendon, whereas the posterior half (9.8 +/- 3.2 mm) was composed of both the SSP and ISP tendons. The sulcus was located not at the SSP-ISP interval but slightly posterior to the posterior margin of the SSP tendon (4.3 +/- 2.4 mm). We conclude that (1) there is an overlap between the SSP and ISP tendons identifiable by the facets or the distance from the anterior greater tuberosity and (2) the sulcus is located slightly posterior to the posterior margin of the SSP tendon.

Aged↗

Distribution of nerve growth factor in cat brains following topical application of solution or Minipellet.

The distribution of nerve growth factor (NGF) was studied after topical application of NGF solution or NGF Minipellet into the caudate nucleus of cat brains. Each aliquot of NGF solution or a piece of NGF Minipellet contains 200 micrograms of mouse 2.5S beta-NGF. The concentrations of NGF in various areas of the brain were determined by enzyme-linked immunosolvent assay (ELISA). The tissue concentrations of NGF were very high at 6 h after injection of NGF solution (124.0 +/- 2.46 to 2144.0 +/- 16.03 ng g-1), but quickly decreased on day 1. The tissue concentrations of NGF were low at 6 h after implantation of NGF Minipellet, but were considerably increased on day 1 (11.22 +/- 3.36 to 72.04 +/- 20.45 ng g-1), and slowly decreased during the following 6 to 14 days. The present study demonstrates the temporal and spatial profiles of NGF distribution in the brain of middle-sized animals after topical application. Injection of NGF solution resulted in abrupt, but very transient, elevation of tissue NGF. Implantation of NGF Minipellet maintained the tissue NGF at biologically effective levels for 6 to 14 days.

Administration, Topical↗

Influence of cedar essence on spontaneous activity and sleep of rats and human daytime nap.

We investigated whether exposure to the odor of extracted cedar essence (CE) has (i) an influence on spontaneous activity and sleep-wake states of rats and (ii) a sleep-promoting effect on human daytime nap after taking an ordinary night's sleep. In rats exposed to CE, spontaneous activities and amount of wake were significantly decreased, while the amount of non-rapid eye movement (NREM) sleep was significantly increased. In human daytime nap, NREM sleep stage 2 latency was significantly shortened after exposure to CE.

Animals↗

Continuous administration of basic fibroblast growth factor (FGF-2) accelerates bone induction on rat calvaria--an application of a new drug delivery system.

Some studies have shown that locally applied basic fibroblast growth factor (FGF-2) enhances bone regeneration at a fracture site, while others have not been in agreement. We developed a new continuous FGF-2 delivery system designed to accelerate cytokine-induced new bone formation. A subperiosteal pocket was surgically formed in 36 eight-week-old male Wistar rats. The rats were administered 0, 1, 10, or 100 ng of FGF-2 contained in a collagen minipellet, mixed with allogeneic demineralized bone matrix in a dome-shaped Millipore filter and then placed into the pocket. New bone formation in the dome was evaluated at 2, 4, and 8 wks after placement. Soft x-ray radiographs disclosed an apparently larger radiopaque region in the 1-ng group at 4 wks compared with those in the other groups. Morphometrical analysis revealed that the new bone area in the 1-g group was significantly larger than that in the 0-g group (p<0.01). In the 100-ng FGF-2 group, new bone formation seemed suppressed. We concluded that continuous slow administration of a small amount of FGF-2 accelerates bone-derived osteogenic cytokine-induced new bone formation.

Animals↗

Nicotine given intracerebroventricularly does not inhibit the preovulatory surge of LH and PRL secretion in female rats.

To determine the effect of nicotine on LH and PRL secretion, nicotine bitartrate (nicotine) dissolved in saline was administered at 1400 h, just before the critical period for the preovulatory surge of LH and PRL secretion, either intracerebroventricularly (icv) or intravenously (iv) in female rats in proestrus. Nicotine neither at a dose of 5 microg nor at a dose of 10 microg injected icv at 1400 h caused significant changes in the surge of LH and PRL secretion. When nicotine was given iv at a dose of 100 microg, a significant decrease in LH and PRL concentrations occurred immediately, lasting for 2 h. After 1700 h, LH and PRL concentrations as high as that observed after 1700 h in saline-injected control rats were recovered, just as if nicotine caused a transient deficit of the surge secretion of these hormones. The results indicate that nicotine does not inhibit the preovulatory surge of LH and PRL secretion by acting at the hypothalamic level accessible via the third ventricle, but inhibits it by acting at certain other site(s).

Animals↗

Cystic changes of the humeral head on MR imaging. Relation to age and cuff-tears.

We obtained MR images of 140 painful shoulders in 134 patients to determine the relationship between cystic changes of the humeral head and integrity of the rotator cuff. Cystic changes were observed in 49 shoulders (35%) and the commonest site was in the bare bone area of the anatomical neck, and the second commonest site was at the attachment of the supraspinatus tendon. Cystic changes in the bare bone area were observed equally often in shoulders with or without rotator cuff tears (27% and 18%, respectively) and were more frequently observed in the elderly. Cystic changes at the attachment of the supraspinatus and subscapularis tendons were specific to rotator cuff tears: they were observed in 28% of rotator cuff tears, but in none of those with an intact cuff. We conclude that there are two distinct types of cystic changes: one at the attachment of the supraspinatus and subscapularis tendons, which is closely related to tears of these tendons, and the other in the bare bone area of the anatomical neck, which is related to aging.

Adolescent↗

Electromyographic activities of the biceps during arm elevation in shoulders with rotator cuff tears.

We investigated electromyographic activities of the biceps in 40 shoulders with full-thickness tears of the rotator cuff and 40 asymptomatic shoulders, with a normal rotator cuff on MRI, to determine the role of the biceps in cuff-deficient shoulders. Using surface electrodes, biceps activities were recorded during arm elevation in the scapular plane with and without a 1-kg load. The percentages of integrated electromyograms to the maximum voluntary contraction (%MVC) were obtained at 30 degrees, 60 degrees, 90 degrees, and 120 degrees of elevation. In the normal shoulders, %MVC of the biceps was always less than 10% through the arc of elevation both with and without load. Among 40 shoulders with rotator cuff tears, 14 showed increased activities of the biceps more than 10% in %MVC (p < 0.0001), whereas the remaining 26 shoulders had activities similar to the normal shoulders. The biceps activities in these 14 shoulders increased with load application and at higher angles of elevation. The muscle strength tended to be weaker in shoulders with increased biceps activities than in those without. Our findings suggest a potential supplemental function of the biceps in shoulders with rotator cuff tears.

Adult↗

Isolation of Paracoccidioides brasiliensis from armadillos (Dasypus noveminctus) captured in an endemic area of paracoccidioidomycosis.

Paracoccidioides brasiliensis, the causative agent of paracoccidioidomycosis (PCM), was first isolated from armadillos from the Amazonian region where the mycosis is uncommon. In the present study, we report on the high incidence of PCM infection in armadillos from a hyperendemic region of the disease. Four nine-banded armadillos (Dasypus novemcinctus) were captured in the endemic area of Botucatu, Sao Paulo, Brazil, killed by manual cervical dislocation and autopsied under sterile conditions. Fragments of lung, spleen, liver, and mesenteric lymph nodes were processed for histology, cultured on Mycosel agar at 37 degrees C, and homogenized for inoculation into the testis and peritoneum of hamsters. The animals were killed from week 6 to week 20 postinoculation and fragments of liver, lung, spleen, testis, and lymph nodes were cultured on brain heart infusion agar at 37 degrees C. Paracoccidioides brasiliensis was isolated from three armadillos both by direct organ culture and from the liver, spleen, lung, and mesenteric lymph nodes of hamsters. In addition, one positive armadillo presented histologically proven PCM disease in a mesenteric lymph node. The three armadillos isolates (Pb-A1, Pb-A2, and Pb-A4) presented thermodependent dimorphism, urease activity, and casein assimilation, showed amplification of the gp43 gene, and were highly virulent in intratesticularly inoculated hamsters. The isolates expressed the gp43 glycoprotein, the immunodominant antigen of the fungus, and reacted with a pool of sera from PCM patients. Taken together, the present data confirm that armadillos are a natural reservoir of P. brasiliensis and demonstrate that the animal is a sylvan host to the fungus.

Animals↗

Expression of basic fibroblast growth factor-like immunoreactivity in the nuclei of regenerating hepatocytes.

This study, utilizing rats subjected to two-thirds partial hepatectomy or sham operation, was designed (1) to investigate the content of basic fibroblast growth factor (bFGF) in the subcellular fractions of regenerating and sham-operated rat livers by immunoblot experiments and enzyme-linked immunosorbent assay (ELISA), (2) to show that bFGF immunoreactivity and proliferating cell nuclear antigen (PCNA) immunoreactivity are markers for hepatocellular mitosis before and after partial hepatectomy, and (3) to observe the location and fine structure of the bFGF immunoreaction within the regenerating liver with special attention to bFGF immunoreactivity in the nuclei of regenerating hepatocytes. Immunoblot experiments and ELISA showed a transient increase in high-molecular-weight forms of bFGF in the nuclear subcellular fraction of regenerating liver 48 h after partial hepatectomy. By light microscopy, bFGF and PCNA immunoreactivities were detected in the nuclei of regenerating hepatocytes. Electron microscopy demonstrated bFGF-like immunoreactivity mainly in the nuclear euchromatin and rarely in the heterochromatin or nucleoli of regenerating hepatocytes. The transient increase in high-molecular-weight forms of bFGF in the nuclear euchromatin of regenerating hepatocytes, together with the concomitant expression of PCNA in the regenerating liver, suggests an important role of the high-molecular-weight forms of bFGF in hepatocyte proliferation and/or mitosis, although authentic bFGF with a molecular form of 18 kDa is not considered to be involved in hepatic regeneration.

Animals↗

Effect of vitamin B12 on the sleep-wake rhythm following an 8-hour advance of the light-dark cycle in the rat.

We have studied the effect of vitamin B12 (VB12) on the sleep-wake rhythm following an 8-h advance of the light-dark (LD) cycle in the rat. The electroencephalogram and electromyogram were recorded on chart paper via a two-channel telemetry system. Infusion of VB12 (Mecobalamin; 0.25 microgram/h) or saline into the abdominal cavity was performed using an implanted miniosmotic pump. Before the phase shift, there was no significant difference between the two groups in terms of the mean daily amount of each vigilance stage. Following the phase advance, in the saline group, wake gradually decreased and nonrapid-eye-movement (NREM) sleep and REM sleep increased. In the VB12 group, no significant change was seen in any of the vigilance states, except for REM sleep on day 3. The results suggest that VB12 may maintain homeostasis of sleep and/or wake amount following an 8-h advance of the LD cycle in the rat.

Animals↗

Antigenic similarities to Paracoccidioides brasiliensis in thermo-dependent dimorphic fungi isolated from soil in Botucatu, SP, Brazil.

We compared the antigenic characteristics of two thermo-dependent dimorphic fungi isolated from soil in Botucatu, an endemic area of paracoccidioidomycosis (PCM) and Paracoccidioides brasiliensis. The soil isolates grew as cerebriform colonies at 37 degrees C (yeast form) and as cottonous colonies at 25 degrees C (mycelial form). No pathogenicity for ddY mice or hamsters were observed. In immunodiffusion test, there were precipitation bands between the 2 soil isolates and pooled PCM patient sera. There were also common precipitation bands at 21, 50 and 58 kDa between the soil isolates antigens and PCM patient sera by Western-blotting, but no gp43 kDa band. No gene for gp 43 kDa protein was detected in the soil isolates by PCR. The fact that these isolates were obtained from an endemic area of PCM and there were some antigenic similarities between the soil isolates and P. brasiliensis in immunodiffusion test and Western-blotting may have some importance in epidemiological surveys done with paracoccidioidin as well interfering with the immune response of the exposed population.

Animals↗