Manual and manipulation techniques for rheumatic disease.

Manipulation is practiced primarily by chiropractors and osteopaths and is one of the most commonly utilized alternative treatments for rheumatic diseases. Low back pain and neck pain are the most frequently treated disorders, but manipulation is also used to treat a broad range of rheumatic diseases. Manipulation has been shown to decrease joint pain and normalize function. The mechanisms of action, however, are not well understood. Current theories propose an imbalance of muscle activity is a source of pain that manipulation can relieve through reflexive actions. Such muscle imbalances would exacerbate rheumatic and arthritic conditions, suggesting that manipulation may be an important therapy that is appropriate for early conservative care as part of a comprehensive treatment program.

INK4a/ARF mutations accelerate lymphomagenesis and promote chemoresistance by disabling p53.

The INK4a/ARF locus encodes upstream regulators of the retinoblastoma and p53 tumor suppressor gene products. To compare the impact of these loci on tumor development and treatment response, the Emu-myc transgenic lymphoma model was used to generate genetically defined tumors with mutations in the INK4a/ARF, Rb, or p53 genes. Like p53 null lymphomas, INK4a/ARF null lymphomas formed rapidly, were highly invasive, displayed apoptotic defects, and were markedly resistant to chemotherapy in vitro and in vivo. Furthermore, INK4a/ARF(-/-) lymphomas displayed reduced p53 activity despite the presence of wild-type p53 genes. Consequently, INK4a/ARF and p53 mutations lead to aggressive tumors by disrupting overlapping tumor suppressor functions. These data have important implications for understanding the clinical behavior of human tumors.

Clinical implications of p53 mutations.

The ultimate goal of basic cancer research is to provide a theoretical foundation for rational approaches to improve cancer therapy. Our extensive insight into the biology of the p53 tumour suppressor and the clinical behaviour of tumours harbouring p53 mutations indicates that information concerning p53 will be useful in diagnosis and prognosis, and may ultimately produce new therapeutic strategies. At the same time, efforts to understand the clinical implications of p53 mutations have revealed conceptual and technical limitations in translating basic biology to the clinic. The lessons learned from p53 may lay the groundwork for future efforts to synthesize cancer gene function, cancer genetics and cancer therapy.

Chronic neck pain, standing balance, and suboccipital muscle atrophy--a pilot study.

OBJECTIVE: To study the relationship between chronic neck pain, standing balance and suboccipital muscle atrophy. We hypothesize that patients with chronic neck pain have more somatic dysfunction in the cervical spine than control subjects without neck pain. We also hypothesize that patients with chronic neck pain and somatic dysfunction exhibit more atrophy of suboccipital muscles. Lastly, because suboccipital muscles have a high density of proprioceptors, we hypothesize that chronic pain patients exhibit a loss in standing balance. DESIGN: Randomized, controlled, partially blind study examining chronic neck pain patients and control subjects for differences in degree of upper cervical somatic dysfunction, standing balance and suboccipital muscle atrophy. SETTING: Subjects were recruited from a clinical practice at Michigan State University; controls were recruited from the faculty, staff and students. PARTICIPANTS: Seven chronic neck pain patients and seven asymptomatic control subjects. MAIN OUTCOME MEASURES: Palpation was used to diagnose somatic dysfunction in the upper cervical spine. Balance parameters were calculated using a force platform; muscle atrophy was judged with magnetic resonance images. RESULTS: Chronic neck pain patients had almost twice as many somatic dysfunctions as controls (p = .028). Force platform results showed a decrease in standing balance in patients compared with control subjects (p = .004). MRI showed that chronic neck pain subjects had marked atrophy of the rectus capitis posterior major and minor muscles, including fatty infiltration. CONCLUSIONS: This study suggests that there is a relationship between chronic pain, somatic dysfunction, muscle atrophy and standing balance. We hypothesize a cycle initiated by chronic somatic dysfunction, which may result in muscle atrophy, which can be further expected to reduce proprioceptive output from atrophied muscles. The lack of proprioceptive inhibition of nociceptors at the dorsal horn of the spinal cord would result in chronic pain and a loss of standing balance.

Enzymatic and metabolic studies on retrograde regulation mutants of yeast.

Two nuclear genes, RTG1 and RTG2, which sense the functional state of yeast mitochondria, have been described recently. Yeast strains with null alleles of either of these two genes (delta rtg1, delta rtg2) cannot grow on acetate as the sole carbon source and are auxotrophic for glutamate and aspartate. We report here a series of metabolic experiments and enzyme activity measurements that were made in an attempt to determine the reason for the acetate- phenotype and the glutamate/aspartate auxotrophy. Decreases in the activities (approximately 50%) in mitochondrial citrate synthase (CS1), acetyl-CoA synthetase, NAD isocitrate dehydrogenase, and pyruvate carboxylase were noted. When CS1 was overexpressed in the delta rtg1 and delta rtg2 mutants, these strains could grow on acetate but were still auxotrophic for glutamate/aspartate. We propose that, in the mutant strain, CS1 activity becomes limiting for efficient acetate utilization, but that other complex metabolic interactions are affected, limiting production of intermediates that would allow synthesis of glutamic and aspartic acids.

The audible release associated with joint manipulation.

OBJECTIVE: The objective of this paper is to review the literature on the audible release associated with manipulation. DATA SOURCES: Bibliographic information in pertinent articles and papers located in the MEDLINE database containing the keywords joint, joints, cartilage, crack, cracking, cavitation, crepitus and noise. STUDY SELECTION: All articles relevant to the objectives were selected. DATA EXTRACTION: All available data was used. DATA SYNTHESIS: The audible release is caused by a cavitation process whereby a sudden decrease in intracapsular pressure causes dissolved gasses in the synovial fluid to be released into the joint cavity. Once a joint undergoes cavitation, the force-displacement curve changes and the range of motion of the joint increases. The gasses released from the synovial fluid make up about 15% of the joint volume and consist of approximately 80% carbon dioxide. Habitual joint cracking does not correlate with arthritic changes, but does correlate with loss of grip strength and soft-tissue swelling. During the "crack" associated with a joint manipulation, there is a sudden joint distraction that occurs in less time than that required to complete the stretch reflexes of periarticular muscles. Theories on the cavitation mechanism were reviewed and new information on the cavitation process is introduced. In this paper, it is proposed that the cavitation process is generated by an elastic recoil of the synovial capsule as it "snaps back" from the capsule/synovial fluid interface. CONCLUSIONS: Because the sudden joint distraction during a manipulation occurs in a shorter time period than that required to complete the stretch reflexes of the periarticular muscles, there is likely to be a high impulse acting on the ligaments and muscles associated with the joint. This is an important conclusion, because others have proposed that reflex actions from high threshold periarticular receptors are associated with the many beneficial results of manipulation. This suggests that the cavitation process provides a simple means for initiating the reflex actions and that without the cavitation process, it would be difficult to generate the forces in the appropriate tissue without causing muscular damage.

Metabolic studies on Saccharomyces cerevisiae containing fused citrate synthase/malate dehydrogenase.

We have constructed two different fusion proteins consisting of the C-terminal end of CS1 fused in-frame to the N-terminal end of MDH1 and HSA, respectively. The fusion proteins were expressed in mutants of Saccharomyces cerevisiae in which CS1 and MDH1 had been deleted and the phenotypes of the transformants characterized. The results show that the fusion proteins are transported into the mitochondria and that they restore the ability for the yeast mutants CS1-, MDH1-, and CS1-/MDH1- to grow on acetate. Determination of CS1 activity in isolated mitochondria showed a 10-fold increase for the strain that expressed native CS1, relative to the parental. In the transformant with CS1/MDH1 fusion protein, parental levels of CS1 were observed, while one-fifth this amount was observed for the strain expressing the CS1/HSA conjugate. Oxygen consumption studies on isolated mitochondria did not show any significant differences between parental-type yeast and the strains expressing the different fusion proteins or native CS1. [3(-13)C]Propionate was used to study the Krebs TCA cycle metabolism of yeast cells containing CS1/MDH1 fusion constructs. The 13C NMR study was performed in respiratory-competent parental yeast cells and using the genetically engineered yeast cells consisting of CS1- mutants expressing native CS1 and the fusion proteins CS1/MDH1 and CS1/HSA, respectively. [3(-13)C]Propionate is believed to be metabolized to [2(-13)C]succinyl-CoA before it enters the TCA cycle in the mitochondria. This metabolite is then oxidized through two symmetrical intermediates, succinate and fumarate, followed by conversion to malate, oxalacetate, and other metabolites such as alanine.(ABSTRACT TRUNCATED AT 250 WORDS)

Inter- and intra-examiner reliability of motion palpation for the thoracolumbar spine.

An inter- and intra-examiner reliability study was conducted to determine the reliability of a standardized method of motion palpation. Eight senior students in the Palmer College Public Clinic palpated the thoracolumbar spine of 32 student volunteers. Each volunteer was palpated twice by each examiner to allow inter- and intra-examiner analysis. Each examiner had at least 1 yr of experience using the procedure. Only the most hypomobile motor unit of the spine was recorded. Analysis of the data revealed statistically significant agreement for intra-examiner reliability. Inter-examiner reliability was not statistically significant. T9-T10 was chosen significantly more often by the palpators as the most hypomobile motor unit.

Translational control and the cytoskeleton in Physarum polycephalum.

Translationally active plasmodia of the syncytial slime mold Physarum polycephalum develop into translationally dormant sclerotia during starvation. Although functional mRNA and ribosomes exist in sclerotia, protein synthesis is suppressed at the level of initiation. To test the possibility that alterations in the cytoskeleton may limit protein synthesis, we have examined the distribution of polysomes and actin mRNA in the cytoskeletal (CSK) and soluble (SOL) fractions of Triton X-100-extracted plasmodia and sclerotia. Most of the polysomes and actin mRNA were located in the CSK of plasmodia, while most of the ribosomes and actin mRNA were located in the SOL of sclerotia. The results suggest that ribosomes and mRNA shift from the CSK to the SOL as protein synthesis is suppressed during starvation. Plasmodia and sclerotia can be induced to accumulate excess polysomes by treatment with low levels of the elongation inhibitor cycloheximide. Treatment of plasmodia with cycloheximide caused excess polysomes to accumulate in the SOL, suggesting that the CSK contains a limited capacity for binding translational components and that the association of polysomes with the cytoskeleton is not required for protein synthesis. Treatment of sclerotia with cycloheximide, however, caused polysomes and actin mRNA to accumulate in the CSK, suggesting that the sclerotial cytoskeleton, although depleted in ribosomes and mRNA, is capable of binding translational components. It is concluded that alterations in the sclerotial cytoskeleton are not involved in translational control.

Stability of an injectable disulfiram formulation sterilized by gamma irradiation.

Stability of an injectable disulfiram suspension sterilized by gamma(gamma) irradiation was tested. Single doses of disulfiram powder in plastic syringes were subjected to 50,000 rads of gamma radiation. Culture media were inoculated with the irradiated drug to test for growth of bacteria, fungi, and mycobacteria. The irradiated drug and nonirradiated controls were analyzed by high-performance liquid chromatography (HPLC) for disulfiram and its major degradation product, diethyldithiocarbamate (DDC). Ultraviolet absorption spectra of irradiated and nonirradiated disulfiram were obtained. No organisms grew in any of the culture media. HPLC analysis indicated that disulfiram content of the irradiated specimens was not reduced, and DDC was not detected. There were no important differences between the ultraviolet spectra of the irradiated and nonirradiated samples. Disulfiram can be sterilized by gamma irradiation without chemical degradation.

Localization of actin messenger RNA during early ascidian development.

The spatial distribution of RNA sequences during early development of the ascidian, Styela plicata, was determined by in situ hybridization with poly(U) and cloned DNA probes. Styela eggs and embryos contain three colored cytoplasmic regions of specific morphogenetic fates, the ectoplasm, endoplasm, and myoplasm. These cytoplasmic regions participate in ooplasmic segregation after fertilization and are distributed to different cell lineages during early embryogenesis. n situ hybridization with poly(U) suggests that poly(A)+RNA is unevenly distributed in eggs and embryos, with about 45% in the ectoplasm, 50% in the endoplasm, and only 5% in the myoplasm. In situ hybridization with a histone DNA probe showed that histone RNA sequences were not localized in eggs or embryos and distributed between the three cytoplasmic regions according to their volumes. In situ hybridization with an actin DNA probe showed actin RNA was localized in the myoplasm and ectoplasm of eggs and embryos with about 45% present in the myoplasm, 40% in the ectoplasm, and only 15% in the endoplasm. These results suggest that a large proportion of the egg actin mRNA is localized in the myoplasm, participates in ooplasmic segregation after fertilization, and is differentially distributed to the mesodermal cell lineages during embryogenesis. Analysis of the translation products of egg mRNA suggests that the localized mRNA codes for a cytoplasmic actin isoform.

Relative increase in polysomal mRNA for R1 cAMP-binding protein in neuroblastoma cells treated with 1,N6-dibutyryl-adenosine 3',-5'-phosphate.

Polysomal RNAs were isolated from control neuroblastoma cells and those treated with 1,N6-dibutyrl-adenosine 3',5'-phosphate (Bt2cAMP) and translated in wheat germ lysates. Comparison of proteins synthesized in vitro on two-dimensional gel electrophoretograms showed that there was a specific induction in the synthesis of a protein, Mr 48000, by the polysomal RNAs from Bt2cAMP-treated cells. This protein was identified as the R1 cAMP-binding protein by its coelectrophoresis with unlabelled binding protein and by its specific retention on 8-(6-aminohexylamino)-adenosine 3',5'-phosphate linked to Sepharose. Quantification of the proteins synthesized in vitro with subsaturating inputs of polysomal RNAs showed that there was a 1.4--1.7-fold increase in the synthesis of the R1 cAMP-binding protein by polysomal RNAs isolated from Bt2cAMP-treated cells. There was a similar increase when purified polyadenylated mRNA populations were compared. showing there was no change in the ratio of adenylated to nonadenylated mRNAs in the induced mRNA population. There was no corresponding increase in the synthesis of the R2 cAMP-binding protein although the relative synthesis of several other proteins was also increased and the synthesis of actin and the alpha and beta-tubulin subunits was decreased. The increased levels of the R1 cAMP-binding protein found in Bt2cAMP-treated neuroblastoma cells are therefore partly caused by a specific accumulation of its mRNA on polysomes. The mRNA content of the cytoplasmic messenger ribonucleoprotein (mRNP) population of control cells was insufficient to account for this increase by a translocation of R1 mRNA from the mRNP to the polysome fraction in Bt2cAMP-treated cells. The increase in polysomal R1 mRNA is therefore caused by its increased transcription of post-transcriptional processing or its decreased rate of degradation in Bt2cAMP-treated cells. Although the R1 and R2 binding proteins have identical molecular weights and similar pI values, the specific induction of the mRNA for R1 cAMP-binding protein and the differential distribution of the R1 and R2 mRNAs between the polysomal and messenger ribonucleoprotein compartments show that these two cAMP-binding proteins are encoded by different mRNA populations.

Soluble antigens of virulent and attenuated biotypes of Brucella abortus.

Several methods were used to characterize three Brucella abortus biotypes (1, 5, and 7), including the attenuated vaccine strain S-19. Chemical analysis did not reveal remarkable differences among these strains, and only minor differences were noted in elution patterns of soluble extracts subjected to column chromatography. Qualitative and quantitative differences in extract components were demonstrated, however, by polyacrylamide gel isoelectric focusing. A distinctive difference was the presence of components in extracts from one or more of the virulent biotypes that were absent in similar preparations from the attenuated strain. In addition, one component common to all virulent strains was absent in strain S-19. Results of immunodiffusion experiments employing adsorbed and unadsorbed antisera also suggested that the quantity, quality, and surface distribution of various cellular antigens differed among the biotypes studied.