An extravascular fluid transport system based on structural framework of fibrous connective tissues in human body.

OBJECTIVE: Interstitial fluid in extracellular matrices may not be totally fixed but partially flow through long-distance oriented fibrous connective tissues via physical mechanisms. We hypothesized there is a long-distance interstitial fluid transport network beyond vascular circulations. MATERIALS AND METHODS: We first used 20 volunteers to determine hypodermic entrant points to visualize long-distance extravascular pathway by MRI. We then investigated the extravascular pathways initiating from the point of thumb in cadavers by chest compressor. The distributions and structures of long-distance pathways from extremity ending to associated visceral structures were identified. RESULTS: Using fluorescent tracer, the pathways from right thumb to right atrium wall near chest were visualized in seven of 10 subjects. The cutaneous pathways were found in dermic, hypodermic and fascial tissues of hand and forearm. The perivascular pathways were along the veins of arm, axillary sheath, superior vena cava and into the superficial tissues on right atrium. Histological and micro-CT data showed these pathways were neither blood nor lymphatic vessels but long-distance oriented fibrous matrices, which contained the longitudinally assembled micro-scale fibres consistently from thumb to superficial tissues on right atrium. CONCLUSIONS: These data revealed the structural framework of the fibrous extracellular matrices in oriented fibrous connective tissues was of the long-distance assembled fibres throughout human body. Along fibres, interstitial fluid can systemically transport by certain driving-transfer mechanisms beyond vascular circulations.

Human ultraweak photon emission and the yin yang concept of Chinese medicine.

The relationship between connective tissue and meridian function is discussed in terms of energy transmission. The network of hydrogen-bonded water molecules interspersed within the collagen fibrillar matrix is especially significant for both the sensitivity of connective tissue to weak signals of mechanical pressure, heat, or electricity and the electrical intercommunication that may correlate with the meridian acupuncture system. Special electromagnetic properties of connective tissue have similar collective properties of ultraweak photon emission. A relationship between ultraweak photon emission and yin yang dynamics is based on three types of ultraweak photon emission studies, focusing on diurnal and annual dynamics, diseased states, and acupuncture points. A novel concept explains the functional (health) integrity of physiologic systems in relation to the left-right balance in ultraweak photon emission by pointing to, (1) balanced corticoneuromusculoskeletal activities and triboluminescent aspects of ultraweak photon emission by skeletal structures, and (2) local fine-tuning in oxygen supply and the formation of radical oxygen species. This approach offers testable hypotheses for further validation utilizing a combination of human photon recording techniques and specialized metabolomics for the estimation of organ-specific oxidative states.

Changes in the composition of the thoracic aortic wall in spontaneously hypertensive rats treated with losartan or spironolactone.

1. In the present study, we compared the elastin and collagen content of thoracic aortic medial and adventitial layers from Wistar-kyoto (WKY) and spontaneously hypertensive rats (SHR). In addition, the effects of losartan, an angiotensin II receptor antagonist, and spironolactone, a mineralocorticoid receptor antagonist, on collagen and elastin content were determined. 2. Prehypertensive (4-week-old) and hypertensive (16-week-old) SHR were randomly divided into three groups treated with either 0.9% NaCl, losartan (20 mg/kg per day) or spironolactone (200 mg/kg per day). Prehypertensive and hypertensive SHR were treated for 12 and 16 weeks, respectively. Age-matched WKY rats were not treated with NaCl, losartan or spironolactone and served as the control group. 3. The medial and adventitial layers of the thoracic aorta were composed mainly of elastin and collagen, respectively, in both SHR and WKY rats. Compared with WKY rats, SHR exhibited greater collagen and elastin content in the media, but decreased collagen and elastin content in the adventitial layer. Both medial and adventitial collagen and elastin content increased significantly with age in both strains and was greater in 32-week-old rats compared with 16-week-old rats. Spironolactone treatment decreased collagen content in the media of thoracic aortas from prehypertensive SHR, whereas losartan decreased collagen content in the media of aortas from hypertensive SHR. In contrast, neither spironolactone nor losartan had any effect on adventitial collagen content in prehypertensive and hypertensive SHR. Medial collagen and elastin were positively related to pulse pressure (PP), but there was no correlation between adventitial mass or collagen content and PP or mean arterial pressure in untreated and treated SHR and WKY rats. 4. In conclusion, the composition of the medial and adventitial layers of the thoracic aorta differs and treatment of SHR with losartan and spironolactone decreases collagen content when delivered at the hypertensive or prehypertensive stage, respectively. However, neither drug has any effect on adventitial collagen content in SHR.

An in vitro assay of collagen fiber alignment by acupuncture needle rotation.

BACKGROUND: During traditional acupuncture therapy, soft tissues attach to and wind around the acupuncture needle. To study this phenomenon in a controlled and quantitative setting, we performed acupuncture needling in vitro. METHODS: Acupuncture was simulated in vitro in three-dimensional, type I collagen gels prepared at 1.5 mg/ml, 2.0 mg/ml, and 2.5 mg/ml collagen, and either crosslinked with formalin or left untreated. Acupuncture needles were inserted into the gels and rotated via a computer-controlled motor at 0.3 rev/sec for up to 10 revolutions while capturing the evolution of birefringence under cross-polarization. RESULTS: Simulated acupuncture produced circumferential alignment of collagen fibers close to the needle that evolved into radial alignment as the distance from the needle increased, which generally matched observations from published tissue explant studies. All gels failed prior to 10 revolutions, and the location of failure was near the transition between circumferential and radial alignment. Crosslinked collagen failed at a significantly lower number of revolutions than untreated collagen, whereas collagen concentration had no effect on gel failure. The strength of the alignment field increased with increasing collagen concentration and decreased with crosslinking. Separate studies were performed in which the gel thickness and depth of needle insertion were varied. As gel thickness increased, gels failed at fewer needle revolutions. For the same depth of insertion, alignment was greater in thinner gels. Alignment increased as the depth of insertion increased. CONCLUSION: These results indicate that the mechanostructural properties of soft connective tissues may affect their response to acupuncture therapy. The in vitro model provides a platform to study mechanotransduction during acupuncture in a highly controlled and quantitative setting.

Mapping chemical gradients within and along a fibrous structural tissue, mussel byssal threads.

The byssal thread of a mussel is an extraorganismic connective tissue that exhibits a striking end-to-end gradient in mechanical properties and thus provides a unique opportunity for studying how gradients are made. Mfp-1 (Mytilus foot protein-1) is a conspicuous component of the protective outer cuticle of byssal threads given its high 3,4-dihydroxyphenylalanine (Dopa) content at 10-15 mol %. Amino acid analysis of mfp-1 extracted from successive foot sections of Mytilus galloprovincialis reveals a post-translationally mediated gradient with highest Dopa levels present in mfp-1 from the accessory gland near the tip of the foot decreasing gradually toward the base. The Dopa content of successive segments of byssal threads decreases from the distal to the proximal end and thus reflects the trend of mfp-1 in the foot. Inductively coupled plasma analysis indicates that certain metal ions including iron follow the trend in Dopa along the thread. Energy-dispersive x-ray spectrometry showed that iron, when present, was concentrated in the cuticle of the threads but sparse in the core. The axial iron gradient appears most closely correlated with the Dopa gradient. The direct incubation of mussels and byssal threads in Fe(3+) supplemented seawater showed that byssal threads are unable to sequester iron from the seawater. Instead, particulate/soluble iron is actively taken up by mussels during filter feeding and incorporated into byssal threads during their secretion. Our results suggest that mussels may exploit the interplay between Dopa and metals to tailor the different parts of threads for specific mechanical properties.

Acupoints [corrected] and meridians: a histochemical study.

The aim of this study was to elucidate the relationship between the structural specificities of acupoints and meridians as well as their clinical effects. We processed 356 specimens, 287 of which from 48 adult and 2 newborn cadavers and the remaining 69 from living patients; samples were taken at three different levels: (1) beneath acupoints; (2) between meridians; (3) at a distance from meridians. We performed seven different staining to show the distribution of collagen fibers, reticular fibers, mucopolysaccharides (MPS), connective tissue, nerve threads, and blood vessels in specimens obtained from different areas. We found that some structural and biochemical discrepancies associated with acupoints and meridians including: (1) mucopolysaccharides (MPS), in particular acid MPS; (2) collagen fibers; (3) nerve endings. We discussed these findings from an anatomo-clinical point of view.

Visualization and detectability of elements rarely contained in soft tissue by X-ray scanning analytical microscopy and electron-probe micro analysis.

The detectability of elements rarely contained in soft tissue was compared using X-ray scanning analytical microscope (XSAM) and electron-probe micro analysis (EPMA). Mapping images of Ca, S and P in normal soft tissue of the rat and dissolved Ni in Ni implanted soft tissue could be obtained by XSAM and EPMA. EPMA was more sensitive in detection of P, while XSAM was superior for Ca, S and Ni mapping. The high detectability for heavier elements by XSAM was explained by the large volume of characteristic X-ray generation in XSAM and low attenuation of the characteristic X-rays from heavier elements. XSAM could provide clearer mapping images for heavier elements whose concentration was low without radiation damage to specimens.

Raised tissue copper levels in oral submucous fibrosis.

Oral submucous fibrosis (OSF) is a well-recognised-potentially malignant condition of the oral cavity associated with areca nut chewing. Areca nut has been shown to have a high copper content compared to other commonly eaten nuts, and chewing areca nut for 5-30 min significantly increases soluble copper in whole mouth fluids. Our aims were to determine if tissue and serum concentrations of copper were raised in patients with OSF as a result of chewing areca nut. A panel of buccal mucosal biopsies from patients with OSF from Nagpur, India, was used to measure the tissue concentrations of copper by mass absorption spectrometry (MAS). By MAS, the mean tissue copper level was 5.5+/-2.9 microg/g in the OSF specimens (n=11) compared with 4+/-1.9 microg/g in the non-areca chewing controls (n=7) (P=0.2). Energy dispersive x-ray microanalysis (EDX) was used to identify the presence and distribution of the metal element. EDX showed distinct peaks corresponding to copper (Kalpha 8.04 keV; Kbeta, 8.91 keV) in the epithelium (21/23) and in the connective tissue (17/23) of the OSF specimens compared to spectra obtained from control oral biopsies from non-areca chewing subjects (n=7). These findings were confirmed by secondary ion mass spectrometry (SIMS) analysis in a small number of samples. Serum copper (17.23+/-1.80 pmol/l), caeruloplasmin (0.32+/-0.04 g/l) levels and urinary copper (0.52+/-0.26 micromol/l) in OSF patients (n=14) were within the laboratory reference ranges. The finding of copper in OSF tissue supports the hypothesis of copper as an initiating factor in OSF, playing a role in stimulating fibrogenesis by the upregulation of lysyl oxidase activity.

From cadavers to implants: silicon tissue assays of medical devices.

A plethora of data has been used to condemn and defend the role of silicone and its association with "adjuvant disease." In the ongoing attempt to enhance our knowledge, we have chosen to identify tissue silicon levels in patients with saline implants or tissue expanders. We have compared these levels with tissue samples from a variety of patients with and without medicinal silicone devices from both the northeast and southwest United States over a 4-year period. All specimens were harvested by a "no touch" technique, non-formalin fixed, frozen, and shipped to an independent toxicology laboratory for analysis. Inductively coupled plasma atomic emission spectroscopy was used to obtain the tissue silicon measurements. Silicon tissue values in cadaveric tissue (n = 20 cadavers; n = 120 specimens) averaged 2.2 mcg/gm of tissue with undetectable silicon levels in over 50 percent of the specimens (range 0 to 45 mcg/gm; median = 0). Silicon levels surrounding port-a-catheter devices (n = 15 patients; n = 15 specimens) averaged 8.04 mcg/gm of tissue (range 0 to 41 mcg/gm; median = 0). Tissue levels in the capsules surrounding saline (n = 10 patients; n = 22 specimens) and silicone implants (n = 31 patients; n = 58 specimens) averaged 292 mcg/gm (range 0 to 1380 mcg/gm; median = 110) and 1439 mcg/gm (range 0 to 9800 mcg/gm, median = 490), respectively. Tissue levels, however, from distant sites (n = 22 specimens) in these same patients were equivalent to the cadaveric nonaugmented values (average = 3.2 mcg/gm; range 0 to 5.8 mcg/gm; median = 2.7). The results imply that there is a continuum of exposure to silicone medical devices based on the mechanical properties of silicone. The data seem to suggest that there may be a progression of measurable tissue silicon levels based on the amount of environmental or device-related silicone exposure a person has over his or her lifetime. It is our hope that these levels will serve as a baseline for our continuing knowledge of implantable medical devices.

Silicon tissue assay: a measurement of capsular levels from chemotherapeutic port-a-catheter devices.

A plethora of data has been used to condemn and defend the role of silicone and its association with "adjuvant disease." In the ongoing attempt to enhance our knowledge, we have chosen to identify tissue silicon levels (n = 15) in capsules that form around chemotherapeutic port-a-catheter devices, which consist of a metal dome encapsuled by silicone. We have compared these levels with previously established silicon levels in augmented breast capsules, distant tissue sites in these same augmented women, and nonaugmented cadaveric tissues from various geographic locations in the United States. All specimens were harvested by a "no touch" technique, not formalin fixed, frozen, and shipped to an independent toxicology laboratory for analysis. Inductively coupled plasma atomic emission spectroscopy was employed to obtain the tissue silicon measurements. Results demonstrated silicon values ranging from nondetectable in 9 patients to as high as 41 micrograms/gm. These values fell in between our cadaveric (0.5 to 6.8 micrograms/gm) and augmented tissue silicon levels (18 to 8700 micrograms/gm). Although the sample size is small and the power of statistical analysis is low, there was no correlation between the patient's silicon level and age, type of cancer, type of chemotherapeutic agent, radiation therapy, or length of time the port-a-catheters were in place. Although detectable levels of silicon identified around port-a-catheter devices were higher than expected, it is impossible to make any conclusions about these levels and the role of a potential collagen-vascular disease. What we have shown, however, is that silicone breast implants may not be the only medical device that can elevate tissue silicon levels. Our data seem to suggest that there may be a progression of measurable tissue silicon levels based on the amount of environmental or device-related silicon exposure a person has had at a particular time in his or her life. It is our belief that as we identify these tissue silicon levels, they will serve as a baseline and reference for further scientific studies.

Influence of incremental zeranol implant doses on the chemical and physical characteristics of third metacarpal bone and chemical composition of liver and soft tissue from feedlot steers.

Chemical and physical characteristics of third metacarpal bones and liver and rib soft tissue composition from feedlot steers were determined. Steers were selected (32 from each experimental location) to represent the range in slaughter weight and composition for each treatment group in three (total n = 1,088) feedlot experiments. Steers were implanted with 0, 24, 36, 48, 60, 72, 84, or 96 mg of zeranol at approximately 140 d before slaughter. Cattle at each location were fed for the same number of days and slaughtered as a group. Zeranol dose had no effect on the chemical composition of bone, liver, or rib soft tissue with the following exceptions: zeranol decreased (P < .01) bone Ca concentration and increased (P < .07) liver P concentration. Zeranol implantation decreased medullary cavity anterioposterior (AP) diameters and AP cortical width (P < .08). Loads withstood by the bones up to flexure (P < .08) and the strain at flexure (P < .09) were inversely related to the quadratic of zeranol dose. However, modulus of elasticity at flexure and breaking increased numerically with zeranol dose. Stress withstood by bones at flexure was greater (P < .09) for implanted steers. Strain data indicate that metacarpals from steers receiving zeranol would exhibit less deformation upon loading to flexure (P < .09) than controls. These data indicate that administration of intermediate doses of zeranol altered bone deposition of Ca, which resulted in modified third metacarpal physical and mechanical characteristics.

Olive oil decreases liver damage in rats caused by carbon tetrachloride (CCl4).

Oral carbon tetrachloride (CCl4) poisoning of the liver of male F-344 rats was modified by dissolving CCl4 in various oils (sunflower, corn, fish and olive). After 8 weeks of CCl4 treatment (3 x 0.2 ml/kg body weight every other day, dissolved in aliquots of 0.5 ml of each types of oil) the rats were sacrificed and the ratio of connective tissue in the liver was determined by morphometry after picrosirius staining. The percentage of collagen fibres increased in all CCl4-treated groups compared to the controls. This increase was almost the same (6-8%) in the case of CCl4 dissolved in sunflower, corn or fish oil, but when olive oil was applied as a solvent, the collagen ratio was only 2-4 percent. On the bases of this finding olive oil is considered as less harmful to the liver in acute CCl4 poisoning than other oils.

Molecular genetic analyses of the Tyrolean Ice Man.

An approximately 5000-year-old mummified human body was recently found in the Tyrolean Alps. The DNA from tissue samples of this Late Neolithic individual, the so-called "Ice Man," has been extracted and analyzed. The number of DNA molecules surviving in the tissue was on the order of 10 genome equivalents per gram of tissue, which meant the only multi-copy sequences could be analyzed. The degradation of the DNA made the enzymatic amplification of mitochondrial DNA fragments of more than 100 to 200 base pairs difficult. One DNA sequence of a hypervariable segment of the mitochondrial control region was determined independently in two different laboratories from internal samples of the body. This sequence showed that the mitochondrial type of the Ice Man fits into the genetic variation of contemporary Europeans and that it was most closely related to mitochondrial types determined from central and northern European populations.

Scanning electron microscopy, chemical and histologic analysis in human periarthritic shoulder biopsy.

In the present study biopsies were analyzed, taken from the left shoulder of a patient who, according to the radiological diagnosis, was suffering from calcific periarthritis. In both optical and electronic microscopy the mineralogical observations showed crystalline aggregations, while the chemical analysis, carried out with an Edax EDS spectrophotometer, confirmed the presence of Ca and P in them, in the ratio typical of biological apatites. The histological observations clearly indicate a change in the metabolism of the tissues present. In the samples observed the damage to the muscular tissues is easily seen while the connective tissue, though apparently less compromised, shows the presence of numerous calcifications whose damage cannot be revealed solely through histologic observations. The authors, linking the structural histologic alterations observed to the presence of hydroxyapatite granules as well as to the patient's painful symptoms, believe all these observations are the result of a chronic process.

Morphometric and microanalytical analyses of alumina wear particles in hip prostheses.

We analysed tissues harvested during 24 retrievals of hip joint prostheses, with one or both articular components made of alumina. We describe the morphology of wear particles, measure their size and analyse their chemical composition. We relate histopathological aspects to the parameters that characterize ceramic wear particles, and notice that tissue reaction relates to the physical aspect and amount of wear debris and does not necessarily depend on their chemical composition.