Further simplification of ultrasensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) for anti-htlv-i igg using microplates and fluororeader

In a previously reported ultrasensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) for anti-HTLV-I IgG, polystyrene beads were handled with tweezers, and bound b?-D-galactosidase activity was measured with a fluorometer. The use of tweezers was causative of false-positivity by carryover, and testing many samples was difficult. Recently, these drawbacks have been minimized using microplates and a fluororeader. However, tweezers were still required in the initial and final steps. In the present study, the immune complex, comprising 2,4-dinitrophenyl-antigen, anti-HTLV-I IgG, and antigen-b?-D-galactosidase conjugate, was formed in and trapped onto microplate wells coated with (anti-2,4-dinitrophenyl group) IgG. Subsequently, the microplate wells were incubated with ?N-2,4-dinitrophenyl-L-lysine and polystyrene beads, modified by attaching to plates through cylindrical bars and coated with (antihuman IgG γgM-chain) IgG, to transfer the immune complex from the microplate wells to the modified polystyrene beads. Alternatively, modified polystyrene beads coated with (anti-2,4-dinitrophenyl group) IgG and microplate wells coated with (antihuman IgG γgM-chain) IgG were substituted for microplate wells coated with (anti-2,4-dinitrophenyl group) IgG and modified polystyrene beads coated with (antihuman IgG γgM-chain) IgG, respectively. The fluorescence intensity for bound b?-D-galactosidase activity was quickly measured with a fluororeader. Thus the modified polystyrene beads were transferred from wells to wells more quickly and easily without tweezers, eliminating false-positivity due to carryover, and it became easy to test many samples with high sensitivity and reliability, although the assay of bound b?-D-galactosidase activity became slightly more time-consuming.©1995 wiley-Liss, inc.     

Investigation into the 5-hydroxytryptamine-induced atropine-resistant neurogenic contraction of guinea-pig proximal colon.

1. The aim of this study was to characterize the receptors mediating the atropine-resistant neurogenic contraction to 5-hydroxytryptamine (5-HT) in the longitudinal muscle of the guinea-pig proximal colon and to determine the type of tachykinin receptors involved in the contractile response to 5-HT by the use of selective antagonists. 2. In the presence of atropine (0.3 microM), guanethidine (5 microM), hexamethonium (100 microM), ketanserin (0.1 microM) and indomethacin (3 microM), 5-HT (0.01-3 microM) produced concentration-dependent neurogenic contractions of colonic strips and at 0.3 microM produced a maximal effect (pEC50 = 7.39 +/- 0.09, n = 18). The 5-HT4 receptor stimulant, 5-methoxytryptamine (5-MeOT, 0.03-10 microM) also produced neurogenic contractions with similar maximum effect to those of 5-HT (pEC50 = 6.89 +/- 0.16). 3. The 5-HT4 receptor antagonist, DAU 6285 (3 microM) shifted the concentration-response curves to both 5-HT and 5-MeOT to the right without significant depression of the maximum, but the 5-HT1/5-HT2 receptor antagonist, metitepine (0.1 microM) and the 5-HT3 receptor antagonist, ondansetron (0.3 microM) had no effect on the control curves to 5-HT and 5-MeOT. 4. The selective NK1 receptor antagonist, FK 888 (1 microM) markedly attenuated the contractions to 5-HT and 5-MeOT. In contrast, the selective NK2 receptor antagonist, SR 48968 (10 nM) and the selective NK3 receptor antagonist, SR 142801 (10 nM) had no effect on the contractions to 5-HT and 5-MeOT. 5. These results indicate that the 5-HT-induced atropine-resistant neurogenic contraction of guinea-pig proximal colon is due to activation of 5-HT4 receptors, presumably located on excitatory motor neurones, innervating the longitudinal muscle. The contraction evoked by activation of the 5-HT4 receptors is mediated primarily via NK1 receptors but not NK2 or NK3, suggesting that the 5-HT4 receptor-mediated contraction is evoked indirectly via tachykinin release from tachykinin-releasing excitatory neurones.     

Ulnar entrapment neuropathy along the medial intermuscular septum in the midarm

We report a patient with primary ulnar entrapment neuropathy in the midarm. Stimulation of multiple sites along the ulnar nerve showed a motor conduction block at a distance of 7.5–10 cm proximal to the medial epicondyle, where the nerve was compressed by the medial intermuscular septum. Anatomically, the possibility of ulnar nerve entrapment in this segment has long been suggested, and stimulation at least 10 cm above the medial epicondyle may reveal the entrapment. Muscle Nerve 39: 707–710, 2009     

Achieving a natural glanular meatus for distal hypospadias with a narrow and shallow plate: Tubularized incised plate versus modified Barcat repair

Objectives:   Although the tubularized incised plate (TIP) repair has become the most popular surgical procedure for distal hypospadias, some authors suggest that this is not suitable for hypospadias with a narrow plate or shallow groove.
Methods:   The configuration and position of the reconstructed meatus were postoperatively analyzed in patients with distal hypospadias whose preoperative urethral plates were shallow or narrow. The findings were compared between six patients undergoing TIP repair and seven patients undergoing modified Barcat (BAVIS) repair.
Results:   Among those undergoing the TIP procedure, a slit-like meatus at the tip of the glans was achieved in one patient, a slit-like meatus at the mid portion of the glans in four patients and a round meatus at the mid portion of the glans in one patient. In those repaired by the BAVIS procedure, a slit-like meatus at the tip of glans was achieved in three patients, a round or irregularly shaped meatus at the tip of the glans in two patients, an irregularly shaped meatus at the mid portion of the glans in one patient and neourethral dehiscence in one patient.
Conclusions:   The present study confirms that a higher rate of achieving slit-like meatus but a lower rate of locating in the glans tip can be attained after TIP repair. On the other hand, there is a higher rate of locating the meatus in the glans tip but a lower rate of achieving a slit-like meatus after BAVIS repair.     

Muscle fiber type abnormalities in bulbospinal muscular atrophy. Comparison with amyotrophic lateral sclerosis

Bulbospinal muscular atrophy (BSA) is a distinct clinical type of chronic spinal muscular atrophy of adult onset, which was first reported by Kennedy et al. Histochemical findings of involved muscles are poorly described. In the present study, the muscle fiber type was studied on biopsied specimens by the histochemical method. The results were compared with those of amyotrophic lateral sclerosis (ALS). Muscle specimens were obtained mainly from biceps brachii muscles of 7 cases of BSA and 10 cases of ALS. Both group of patients were matched in respect to age, sex and stage of muscle weakness. The following results were obtained. Histopathological findings of BSA are classed as stage 4 abnormalities of Jennekins. Both fiber type grouping and type 2B deficiency are more frequent in BSA than in ALS. Type 2C fiber seems to be more in ALS than in BSA. Atrophy factor of Dubowitz is the same in BSA and in ALS. Hypertrophy factor is more prominent in BSA than in ALS. Significant correlation between fiber type grouping and type 2B deficiency was confirmed in muscles from other neurogenic atrophy. Since there is a significant association of the type 2B deficiency with fiber type grouping, type 2B fiber might be transformed to type 2A during the reinnervation process. The adaptation to the increased activities of the survived muscles and motoneurons might be contributory to the transformation, which is already known to occur in normal subjects during the endurance training.     

Studies on the nephrotoxicity of aminoglycoside antibiotics and protection from these effects. (1). Nephrotoxicity of gentamicin and mercuric chloride

The present study was designed to find useful markers for detection of renal damage due to gentamicin (GM). Following the administration of 80 mg/kg GM, there were significant increases in urinary protein contents and alkaline phosphatase, N-acetyl-beta-glucosaminidase, lactate dehydrogenase, gamma-glutamyl transpeptidase and lysozyme activities. Alterations of these parameters had a peak at the 7th or 10th day and values restored to near normal levels by the 15th day. Light microscopic observations of the kidney on the 10th day showed mainly the necrosis of proximal tubular epithelial cells in the renal outer cortex, and there was regeneration of epithelial cells on the 15th day. In addition, when 1 mg/kg HgCl2 was given to rats, there were increases in urinary enzyme activities and protein contents, and BUN. The kidney of rats that received HgCl2 showed the necrosis of tubular epithelial cells in the renal inner cortex. It is considered from these results that determination of the activities of various urinary enzymes may be useful markers to detect tubular damage induced by GM.