Effects of freezing rates and cryoprotectant on thermal expansion of articular cartilage during freezing process.

The intact articular cartilage has not yet been successfully preserved at low temperature most likely due to the volume expansion from water to ice during freezing. The objective of this current study focuses on examining thermal expansion behavior of articular cartilage (AC) during freezing from 0 degree C to -100 degree C. Thermo Mechanical Analysis (TMA) was used to investigate the effects of different concentrations of dimethyl sulphoxide (DMSO) (0%, 10%, 30% and 60% v/v) and different freezing rates (1 C/min, 3 C/min and 5 C/min). The results showed that: (1) the inhomogeneous thermal expansion (or contraction) presents due to inhomogeneous water distributions in articular cartilage during freezing, which also may be the most likely reason that the matrix has been damaged in cryopreserved intact articular cartilage; (2) at the phase transition temperature range, the maximum thermal strain change value for 5C/min is approximately 1.45 times than that for 1 C/min, but the maximum thermal expansion coefficient of the later is about six times than that of the former; (3) the thermal expansion coefficient decreases with increasing cooling rate at the unfrozen temperature region, but some opposite results are obtained at the frozen temperature region; (4) the higher the DMSO concentration is, at the phase change temperature region, the smaller the thermal strain change as well as the maximum thermal expansion coefficient are, but DMSO concentration exhibits little effect on the thermal expansion coefficient at both unfrozen and frozen region. Once the DMSO concentration increasing enough, e.g. 60% v/v, the thermal strain decreases linearly and smoothly without any abrupt change due to little or no ice crystal forms (i.e. vitrification) in frozen articular cartilage. This study may improve our understanding of the thermal expansion (or contraction) behavior of cryopreserved articular cartilage and it may be useful for the future study on cryopreservation of intact articular cartilage.

Improvement of osseointegration of titanium dental implants by a modified sandblasting surface treatment: an in vivo interfacial biomechanics study.

To study the effects of a modified sandblasting surface treatment on the osseointegration of dental implants at the level of interfacial biomechanics, an in vivo pullout test was conducted using bone-interfacial shear strength as a criterion. Titanium implants were inserted into the medialis condyli of dogs and harvested 2, 4, and 12 weeks after insertion. Shear strength was determined with an Instron pullout tester. Observation and analysis of the surface of modified sandblasted implants after pullout at 12 weeks were performed with scanning electron microscopy and x-ray spectroscopy. Results showed that the shear strength of implants with a modified sandblasted surface was about five times as high as that of implants with a smooth surface. We concluded that the rough surface of titanium dental implants created by the modified sandblasting treatment can greatly enhance the shear strength at the dental implant-bone interface and that, with this enhancement, the secondary micropores play a much more important role in implant-bone bonding.

Molecular cloning and disease association of hepatitis G virus: a transfusion-transmissible agent.

An RNA virus, designated hepatitis G virus (HGV), was identified from the plasma of a patient with chronic hepatitis. Extension from an immunoreactive complementary DNA clone yielded the entire genome (9392 nucleotides) encoding a polyprotein of 2873 amino acids. The virus is closely related to GB virus C (GBV-C) and distantly related to hepatitis C virus, GBV-A, and GBV-B. HGV was associated with acute and chronic hepatitis. Persistent viremia was detected for up to 9 years in patients with hepatitis. The virus is transfusion-transmissible. It has a global distribution and is present within the volunteer blood donor population in the United States.

Anti-IL-6 monoclonal antibodies protect against lethal Escherichia coli infection and lethal tumor necrosis factor-alpha challenge in mice.

Potentially fatal physiologic and metabolic derangements can occur in response to bacterial infection in animals and man. Recently it has been shown that alterations in the levels of circulating cytokines such as IL-6 and TNF-alpha occur shortly after bacterial challenge. To understand better the role of IL-6 in inflammation, we investigated the effects of in vivo anti-mouse IL-6 antibody treatment in a mouse model of septic shock. Rat anti-mouse IL-6 neutralizing mAb was produced from splenocytes of an animal immunized with mouse rIL-6. This mAb, MP5-20F3, was a very potent and specific antagonist of mouse IL-6 in vitro bioactivity, demonstrated using the NFS60 myelomonocytic and KD83 plasmacytoma target cell lines, and also immunoprecipitated radiolabeled IL-6. Anti-IL-6 mAb pretreatment of mice subsequently challenged with lethal doses of i.p. Escherichia coli or i.v. TNF-alpha protected mice from death caused by these treatments. Pretreatment of E. coli-challenged mice with anti-IL-6 led to an increase in serum TNF bioactivity, in comparison to isotype control antibody, implicating IL-6 as a negative modulator of TNF in vivo. Anti-TNF-alpha treatment of mice challenged i.p. with live E. coli resulted in a 70% decrease in serum IL-6 levels, determined by immunoenzymetric assay, compared to control antibody, thereby supporting a role for TNF-alpha as a positive regulator of IL-6 levels. We conclude that IL-6 is a mediator in lethal E. coli infection, and suggest that antagonists of IL-6 may be beneficial therapeutically in life-threatening bacterial infection.

Antineoplastic agents, 103. The isolation and structure of hypoestestatins 1 and 2 from the East African Hypoëstes verticillaris.

The East African shrub Hypoëstes verticillaris representing a heretofore chemically unexplored. Acanthaceae genus has been found to contain two new cell growth inhibitory (murine P-388 lymphocytic leukemia) phenanthroindolizidine alkaloids termed hypoestestatin 1 (1a) and hypoestestatin 2 (1b). Both substances were found to markedly inhibit growth of the murine P-388 cell line (ED50 = 10(-5) micrograms/ml). The structures of hypoestestatins 1 and 2 were assigned primarily on the basis of results from extensive spectral studies.

Antineoplastic agents, 88. Pimelea prostrata.

The aerial portion of Pimelea prostrata (Thymelaeaceae) collected in New Zealand was evaluated as a source of substances that inhibit growth of the murine P-388 lymphocytic leukemia (PS). Simplexin (1) and Pimelea factor P2 (2) were found to strongly inhibit growth (ED50 5 X 10(-3) and 8 X 10(-4) micrograms/ml, respectively) of the PS in vitro cell line. The cyclic orthoester (2) was also found to inhibit growth (T/C 132 at 20 micrograms/kg) of the PS in vivo system. Detailed 1H-nmr (at 400 MHz) and 13C-nmr studies combined with fast atom bombardment mass spectral evidence were employed to confirm the structural assignments.

A non-surgical technique for the transcervical administration of physiological and pharmacological agents into rat uteri.

A technique for the instillation of solutions into the rat uterine lumen is described. The method has been tested by following the receptor depletion/replenishment cycle after oestradiol injection and by checking on the stoichiometry of hormone/receptor translocation from the cytoplasmic compartment into the nucleus. Both crude sediments from frozen uteri and nuclei purified by a novel procedure were analyzed and gave identical results. The limitations and the advantages of the technique are discussed.