Effects of hyaluronic acid and basic fibroblast growth factor on motility of chondrocytes and synovial cells in culture

The capacity of chondrogenic precursor cells to migrate and proliferate in an injured area is considered to be essential for cartilage repair. We examined cell motility of chondrocytes and synovial cells in monolayer culture and the chemokinetic effects of hyaluronic acid (HA) and basic fibroblast growth factors (bFGF) on these cells. The velocity of chondrocyte migration was accelerated by giving bFGF and simultaneously administering of both HA and bFGF, but it was not affected by HA alone. The velocity of synovial cell migration was increased by HA, but not by bFGF. HA had a chemokinetic effect on synovial cells and bFGF had the same effect on chondrocytes. Treatment with exogenous HA and bFGF may be of value for repairing articular cartilage injury by recruiting chondrogenic cells and promoting migration of chondrocytes in the cartilage tissue.     

Influence of hypothalamically induced restlessness under fasted and fed conditions on plasma levels of glucose,free fatty acids and total cholesterol

Influence of hypothalamically induced emotional behavior on plasma levels of epinephrine (E), norepinephrine (NE), glucose, cortisol, free fatty acids (FFA) and total cholesterol (T‐C) were examined in cats under both fasted and fed conditions. Restlessness was evoked intermittently for 6 h by electrical stimulation of the anteromedial hypothalamus (AMH). Blood was sampled immediately before, 1 h after and 6 h after the start of stimulation. Both E and glucose levels in the restlessness group increased markedly after 1 h and then tended to decrease after 6 h in fasted and fed states. In the restlessness group the NE level increased after 1 h and further increased after 6 h and cortisol level increased markedly, whether cats were fasted or fed. The FFA level in the fasted restlessness group showed no change while it increased in the fasted control group after 6 h. By contrast, the FFA level in the fed restlessness group significantly increased after 6 h while no change was observed in the fed control group. The T‐C level decreased only under the fasted condition in both restlessness and control groups. These results indicated that AMH‐induced restlessness elicited marked sympatho‐adrenal activation and hyperglycemia whether cats were fasted or fed while the behavior showed a different pattern in the response of plasma FFA between fasted and fed conditions. Copyright © 2001 John Wiley & Sons, Ltd.     

Assessing liver function by magnetic resonance imaging two-dimensional phase-shift flow measurement of portal venous blood flow after oral intake of glucose

We have already reported that the ratio of portal venous flow 30 min after oral intake of glucose 75 g to that before intake (PVFR30), measured using pulsed-Doppler ultrasonography (US), correlated significantly with other indicators of liver function and that it could be used to estimate hepatic function before surgery, including liver resection. In this study, to assess the disadvantages of pulsed-Doppler ultrasonography, PVFR30 was measured using two-dimensional (2D) phase-shift (PS) magnetic resonance imaging (MRI). PVFR30 was measured in 17 patients and 7 volunteers: 13 with liver cirrhosis (LC) and 11 without LC (non-LC). Portal venous flow could be measured in all patients without any disturbance of intestinal gas or patient fat, or the high degree of technical skill that Doppler US requires. PVFR30 was significantly lower in the LC group than in the non-LC group. In addition, it correlated significantly with other indicators of liver function, including the indocyanine green clearance test, prothrombin time, hepaplastin test, and cholinesterase activity. These results suggest that PVFR30 measured by 2D PS MRI can be used to estimate liver function, and that this MRI method can be performed more easily than pulsed-Doppler US.     

Guillain-barré syndrome associated Campylobacter jejuni serotype Penner 2: a case report]

We described a 15-year-old male who had Guillain-Barré syndrome(GBS). Nine days after watery diarrhea, the patient developed pain and weakness of foot muscles. On admission, the nerve conduction studies revealed peripheral neuropathy with axonal degeneration and demyelination. Campylobacter jejuni(C. jejuni) with serotype of Lior 4, Penner 2 was isolated from his stool culture. IgM anti-GM 1 antibody and other various anti-ganglioside antibodies were detected in his serum. After receiving plasma exchange and intravenous immunoglobulin therapy, he was able to walk without assistance. In general, C. jejuni with the serotype Penner 19 has been isolated from many GBS patients. In this patient, C. jejuni with the serotype of Penner 2 was isolated. The serotype is detected commonly in Miller Fisher syndrome.     

Masks for influenza patients: measurement of airflow from the mouth

In case of a new influenza pandemic, provision of masks to coughing patients could be used to slow expansion of the epidemic. To quantitatively assess the efficacy of different masks, we used an ultrasonic anemometer to measure the velocity of airflow from the mouth in coughing. We found that even the cheapest paper masks reduced the air speed to less than 1/10, implicating their effectiveness in decreasing viral spread. We therefore propose that governments provide free masks to coughing patients in the general population upon the emergence of a new human influenza virus.     

Improved generation of HLA class I/peptide tetramers

As a tetrameric major histocompatibility complex (MHC) class I-peptide complex (tetramer) is capable of detecting antigen-specific cytotoxic T lymphocyte (CTL) by flow cytometry, significant information about the generation of in vivo immunity can be obtained. It is, however, difficult to make a soluble wild type of MHC class I heavy chain by the prokaryotic expression system. Therefore, we developed a new method for making soluble mutant HLA-A*2402 heavy chain. In this method, signal sequences were deleted, and the codon was changed to silent mutated nucleotide sequences that bacteria could use as preferable codon. When purified mutant HLA-A*2402 molecules were examined for the protein generation by SDS-polyacrylamide gel electrophoresis (PAGE) and western blotting using anti-HLA class I monoclonal antibody (mAb) as compared with wild type, a large amount of mutant heavy chain could be detected. In contrast, the expression of wild-type stable HLA-A*2402 heavy chain molecule was not detected in this system. Consequently, by using mutant HLA-A*2402/peptide tetramers, CTL precursors (CTLp) that specifically recognize antigenic peptide derived from the X;18 chromosomal translocations of synovial sarcoma were detected in patients' PBL.     

High-throughput screening of low molecular weight NS3-NS4A protease inhibitors using a fluorescence resonance energy transfer substrate

Hepatitis C virus (HCV) NS3-NS4A protease is an attractive target for anti-HCV agents because of its important role in replication. An optimized fluorescence resonance energy transfer (FRET) substrate for NS3-NS4A protease, based on the sequence of the NS5A-5B cleavage site, was designed and synthesized. High-throughput screening of in-house compound libraries was performed using a FRET substrate FS10 (MOCAcDKIVPC-SMSYK-Dnp) and MBP-NS3-NS4A fusion protein. Several hit compounds were found, including YZ-9577 (2-oxido-1,2,5-oxadiazole-3,4-diyl) bis (phenylmethanone) with potent inhibitory activity (IC50=1.6 microM) and good selectivity against other human serine proteases.     

Fluid resuscitation of trauma patients: how fast is the optimal rate?

The Advanced Trauma Life Support guidelines recommend an initial rapid infusion of fluid (1-2 L) in trauma and hemorrhage victims as a diagnostic procedure to aid treatment decisions. Although patient response to initial fluid resuscitation is the key to determining therapeutic strategies, the appropriate rate of infusion is not clearly defined. Ninety-nine adult (age >16 years) blunt trauma victims with hypotension were enrolled. Patients were classified into 3 groups according to hemodynamic state after initial fluid resuscitation and requirement of surgical intervention. Total volume and rate of infusion differed significantly between the groups (P < .05). Patients requiring fluid administration at higher rate were all hemodynamically unstable and required immediate surgical intervention. Moreover, rate of infusion was the best predictor of the patients who required immediate surgical intervention. Moderate fluid infusion rate should be considered to allow identification of the patient's response to initial fluid resuscitation.     

Investigation of the relationship between atherosclerosis and paraoxonase or homocysteine thiolactonase activity in patients with type 2 diabetes mellitus using a commercially available assay

BACKGROUND: It is known that serum paraoxonase activity decreases in patients with atherosclerotic diseases such as type 2 diabetes mellitus. Paraoxonase hydrolyzes homocysteine thiolactone, which is thought to be an endogenous substrate. The relationship between the status of atherosclerosis and homocysteine thiolactonase activity in patients with type 2 diabetes mellitus was investigated. METHOD: Serum homocysteine thiolactonase activities in 207 patients with type 2 diabetes mellitus were measured with a recently developed commercially available assay. RESULTS: The coefficient of variation for the assay was <0.7%. Serum paraoxonase and arylesterase activities were dependent on the PON1 192 polymorphism expressed by the subjects but homocysteine thiolactonase activity was not affected. A tendency for a negative association between homocysteine thiolactonase activity and the thickness of the carotid intima media was observed in patients with type 2 diabetes mellitus. CONCLUSIONS: The serum homocysteine thiolactonase activity may represent the status of atherosclerosis in patients with type 2 diabetes mellitus. Since the assay can be used with a routine analyzer, the activities of thousands of samples can be measured with excellent precision in a few hours.