The Role of Muscle Loading on Bone (Re)modeling at the Developing Enthesis

Muscle forces are necessary for the development and maintenance of a mineralized skeleton. Removal of loads leads to malformed bones and impaired musculoskeletal function due to changes in bone (re)modeling. In the current study, the development of a mineralized junction at the interface between muscle and bone was examined under normal and impaired loading conditions. Unilateral mouse rotator cuff muscles were paralyzed using botulinum toxin A at birth. Control groups consisted of contralateral shoulders injected with saline and a separate group of normal mice. It was hypothesized that muscle unloading would suppress bone formation and enhance bone resorption at the enthesis, and that the unloading-induced bony defects could be rescued by suppressing osteoclast activity. In order to modulate osteoclast activity, mice were injected with the bisphosphonate alendronate. Bone formation was measured at the tendon enthesis using alizarin and calcein fluorescent labeling of bone surfaces followed by quantitative histomorphometry of histologic sections. Bone volume and architecture was measured using micro computed tomography. Osteoclast surface was determined via quantitative histomorphometry of tartrate resistant acid phosphatase stained histologic sections. Muscle unloading resulted in delayed initiation of endochondral ossification at the enthesis, but did not impair bone formation rate. Unloading led to severe defects in bone volume and trabecular bone architecture. These defects were partially rescued by suppression of osteoclast activity through alendronate treatment, and the effect of alendronate was dose dependent. Similarly, bone formation rate was increased with increasing alendronate dose across loading groups. The bony defects caused by unloading were therefore likely due to maintained high osteoclast activity, which normally decreases from neonatal through mature timepoints. These results have important implications for the treatment of muscle unloading conditions such as neonatal brachial plexus palsy, which results in shoulder paralysis at birth and subsequent defects in the rotator cuff enthesis and humeral head.     

Aspects of Mineral Structure in Normally Calcifying Avian Tendon

Structural characteristics of normally calcifying leg tendons of the domestic turkey Meleagris gallopavo have been observed for the first time by tapping mode atomic force microscopy (TMAFM), and phase as well as corresponding topographic images were acquired to gain insight into the features of mineralizing collagen fibrils and fibers. Analysis of different regions of the tendon has yielded new information concerning the structural interrelationships in vivo between collagen fibrils and fibers and mineral crystals appearing in the form of plates and plate aggregates. TMAFM images show numerous mineralized collagen structures exhibiting characteristic periodicity (54-70 nm), organized with their respective long axes parallel to each other. In some instances, mineral plates (30-40 nm thick) are found interspersed between and in intimate contact with the mineralized collagen. The edges of such plates lie parallel to the neighboring collagen. Many of these plates appear to be aligned to form larger aggregates (475-600 nm long x 75-90 nm thick) that also retain collagen periodicity along their exposed edges. Intrinsic structural properties of the mineralizing avian tendon have not previously been described on the scale reported in this study. These data provide the first visual evidence supporting the concept that larger plates form from parallel association of smaller ones, and the data fill a gap in knowledge between macromolecular- and anatomic-scale studies of the mineralization of avian tendon and connective tissues in general. The observed organization of mineralized collagen, plates, and plate aggregates maintaining a consistently parallel nature demonstrates the means by which increasing structural complexity may be achieved in a calcified tissue over greater levels of hierarchical order.     

Functional Grading of Mineral and Collagen in the Attachment of Tendon to Bone

Attachment of dissimilar materials is a major challenge because high levels of localized stress may develop at their interfaces. An effective biologic solution to this problem exists at one of nature's most extreme interfaces: the attachment of tendon (a compliant, structural “soft tissue”) to bone (a stiff, structural “hard tissue”). The goal of our study was to develop biomechanical models to describe how the tendon-to-bone insertion derives its mechanical properties. We examined the tendon-to-bone insertion and found two factors that give the tendon-to-bone transition a unique grading in mechanical properties: 1), a gradation in mineral concentration, measured by Raman spectroscopy; and 2), a gradation in collagen fiber orientation, measured by polarized light microscopy. Our measurements motivate a new physiological picture of the tissue that achieves this transition, the tendon-to-bone insertion, as a continuous, functionally graded material. Our biomechanical model suggests that the experimentally observed increase in mineral accumulation within collagen fibers can provide significant stiffening of the partially mineralized fibers, but only for concentrations of mineral above a “percolation threshold” corresponding to formation of a mechanically continuous mineral network within each collagen fiber (e.g., the case of mineral connectivity extending from one end of the fiber to the other). Increasing dispersion in the orientation distribution of collagen fibers from tendon to bone is a second major determinant of tissue stiffness. The combination of these two factors may explain the nonmonotonic variation of stiffness over the length of the tendon-to-bone insertion reported previously. Our models explain how tendon-to-bone attachment is achieved through a functionally graded material composition, and provide targets for tissue engineered surgical interventions and biomimetic material interfaces.     

Developing Univariate Distributions from Data for Risk Analysis

The importance of fitting distributions to data for risk analysis continues to grow as regulatory agencies, like the Environmental Protection Agency (EPA), continue to shift from deterministic to probabilistic risk assessment techniques. The use of Monte Carlo simulation as a tool for propagating variability and uncertainty in risk requires specification of the risk model's inputs in the form of distributions or tables of data. Several software tools exist to support risk assessors in their efforts to develop distributions. However, users must keep in mind that these tools do not replace clear thought about judgments that must be made in characterizing the information from data. This overview introduces risk assessors to the statistical concepts and physical reasons that support important judgments about appropriate types of parametric distributions and goodness-of-fit. In the context of using data to improve risk assessment and ultimately risk management, this paper discusses issues related to the nature of the data (representativeness, quantity, and quality, correlation with space and time, and distinguishing between variability and uncertainty for a set of data), and matching data and distributions appropriately. All data analysis (whether “Frequentist” or “Bayesian” or oblivious to the distinction) requires the use of subjective judgment. The paper offers an iterative process for developing distributions using data to characterize variability and uncertainty for inputs to risk models that provides incentives for collecting better information when the value of information exceeds its cost. Risk analysts need to focus attention on characterizing the information appropriately for purposes of the risk assessment (and risk management questions at hand), not on characterization for its own sake.     

Comparison of Approaches for Developing Distributions for Carcinogenic Slope Factors

In recent years, risk assessors have increasingly been moving away from deterministic risk assessment approaches and are applying probabilistic approaches that incorporate distributions of possible values for each input parameter. This paper reviews several approaches that are being used or that could potentially be used to develop distributions for carcinogenic slope factors (CSFs). Based on the primary tool or framework that is applied, these approaches have been divided into the following three categories: the statistical framework, the decision analysis framework, and the biological framework. Work that has been done on each approach is summarized, and the aspects of variability and uncertainty that are incorporated into each approach are examined. The implications of the resulting distributional information for calculating risk estimates or risk-based concentrations is explored. The approaches differ in their stage of development, the degree to which they diverge from the U.S. Environmental Protection Agency's (EPA) current practice in establishing CSF values, their flexibility to accommodate varying data sets or theories of carcinogenicity, and their complexity of application. In some cases, wide ranges of potential potency estimates are indicated by these approaches. Such findings suggest widely divergent risk assessment implications and the need for additional evaluation of the goals of developing CSF distributions for use in risk assessment applications and the types of information that should be reflected in such distributions. Some combination of the features offered by these approaches may best support risk assessment and risk management decisions.     

Distributions of Actin, Vinculin and Fibronectin in the Duodenum of Developing Chick Embryos: Immunohistochemical Studies at the Light Microscope Level

Microscopic studies were made on the localizations of three different cytoskeletal proteins, actin, vinculin and fibronectin, in the duodenum of developing chick embryos and chicks by an indirect immuno-fluorescent staining method with specific antibodies.
Topographical changes in the distributions of these three proteins seemed to be related to stages in morphogenesis of the duodenum. In early stages of embryonic development, findings suggested interaction between actin and vinculin in the apical region of epithelial cells and between actin and fibronectin in the basal region of these cells. From this stage, vinculin and fibronectin seemed to be of importance in determination and continuity of the polarity of the duodenal epithelium, and in control of the intracellular arrangement of actin. This relation between actin and vinculin seemed to continue throughout embryogenesis.
The main role of actin in epithelial cells seemed to change on day 12 from that of forming constricting bundles for morphogenesis of previllous ridges to that of microfilaments in the microvilli and the terminal web.     

Virio- and Bacterioplankton Microscale Distributions at the Sediment-Water Interface

The marine sediment-water interface is an important location for microbially controlled nutrient and gas exchange processes. While microbial distributions on the sediment side of the interface are well established in many locations, the distributions of microbes on the water side of the interface are less well known. Here, we measured that distribution for marine virio- and bacterioplankton with a new two-dimensional technique. Our results revealed higher heterogeneity in sediment-water interface biomass distributions than previously reported with a greater than 45– and 2500-fold change cm⁻¹ found within bacterial and viral subpopulations compared to previous maxima of 1.5- and 1.4-fold cm⁻¹ in bacteria and viruses in the same environments. The 45-fold and 2500-fold changes were due to patches of elevated and patches of reduced viral and bacterial abundance. The bacterial and viral hotspots were found over single and multiple sample points and the two groups often coincided whilst the coldspots only occurred over single sample points and the bacterial and viral abundances showed no correlation. The total mean abundances of viruses strongly correlated with bacteria (r = 0.90, p<0.0001, n = 12) for all three microplates (n = 1350). Spatial autocorrelation analysis via Moran’s I and Geary’s C revealed non-random distributions in bacterial subpopulations and random distributions in viral subpopulations. The variable distributions of viral and bacterial abundance over centimetre-scale distances suggest that competition and the likelihood of viral infection are higher in the small volumes important for individual cell encounters than bulk measurements indicate. We conclude that large scale measurements are not an accurate measurement of the conditions under which microbial dynamics exist. The high variability we report indicates that few microbes experience the ‘average’ concentrations that are frequently measured.     

不同类型水稻不育系药隔发育过程中Ca2+的分布变化

采用焦锑酸钾沉淀法研究了不同类型水稻不育系与其相应保持系及其可育花药药隔发育过程中Ca^2＋的分布变化。结果显示在正常花药的发育过程中,药隔中的Ca^2＋呈以下变化规律：（1）花药发育早期药隔中的Ca^2＋沉淀很少;（2）随着花药的发育,药隔中的Ca^2＋沉淀增加,木质部细胞的次生加厚壁上有较多的Ca^2＋沉淀,连接组织中的Ca^2＋沉淀也大大增加;（3）到了花粉内容物充实后期及成熟花粉时期,药隔中的Ca^2＋又略有减少。而不育花药药隔中的Ca^2＋在花药发育的各时期均比可育花药要多。败育类型不同,药隔中出现大量Ca^2＋沉淀的时期也不同,无花粉型不育系在花粉母细胞时期药隔中就有大量Ca^2＋沉淀,而农垦58S从单核花粉时期约隔中才出现大量Ca^2＋沉淀。实验结果表明：Ca^2＋在不同类型的不育水稻中呈现出一定的规律性变化,药隔中Ca^2＋的异常分布可能与水稻花粉的败育有关。     

Life-Cycle Assessment and Temporal Distributions of Emissions: Developing a Fleet-Based Analysis

Although the product-centered focus of life-cycle assessment has been one of its strengths, this analytical perspective embeds assumptions that may conflict with the realities of environmental problems. This article demonstrates, through a series of mathematical derivations, that all the products in use, rather than a single product, frequently should be the appropriate unit of analysis. Such a "fleet-centered" approach supplies a richer perspective on the comparative emissions burdens generated by alternative products, and it eliminates certain simplifying assumptions imposed upon the analysis by a product-centered approach.
A sample numerical case, examining the comparative emissions of steel-intensive and aluminum-intensive automobiles, is presented to contrast the results of the two approaches. The fleet-centered analysis shows that the "crossover time" (i.e., the time required before the fuel economy benefits of the lighter aluminum vehicle offset the energy intensity of the processes used to manufacture the aluminum in the first place) can be dramatically longer than that predicted by a product-centered life-cycle assessment.
The fleet-centered perspective explicitly introduces the notion of time as a critical element of comparative life-cycle assessments and raises important questions about the role of the analyst in selecting the appropriate time horizon for analysis. Moreover, with the introduction of time as an appropriate dimension to life-cycle assessment, the influences of effects distributed over time can be more naturally and consistently treated.     

Tissue Mineral Distributions are Differentially Modified by Dietary Protein Deficiency and a Murine Nematode Infection

This study was designed to investigate whether mineral concentrations in the spleen, serum, and liver were modified by challenge infection with a gastrointestinal nematode, by infection dose, or by protein deficiency despite adequate dietary intakes of minerals. BALB/c mice fed protein-sufficient (PS, 24%) or protein-deficient (PD, 3%) diets were infected with 100 L3 of Heligmosomoides bakeri, drug-treated, and then re-infected with either 0, 100, or 200 L3. Protein deficiency and infection, but not dose, independently modified tissue mineral distributions. H. bakeri infection lowered serum iron concentrations in both diet groups. Despite this, PD mice had elevated iron and calcium concentrations and Ca/Zn ratio in the spleen as well as Fe/Zn ratio in liver, but they had reduced calcium, zinc, copper, and sulfur concentrations, and Cu/Zn ratio in the liver. Infection reduced calcium and iron concentrations and the Ca/Zn ratio in the spleen. We suggest that tissue mineral distribution is a consequence of Th2 immune and inflammatory responses induced by infection in PS mice and the switch to predominant Th1 inflammation in PD, nematode-infected mice.     

Stochastic Interdigitation as a Toughening Mechanism at the Interface between Tendon and Bone

Reattachment and healing of tendon to bone poses a persistent clinical challenge and often results in poor outcomes, in part because the mechanisms that imbue the uninjured tendon-to-bone attachment with toughness are not known. One feature of typical tendon-to-bone surgical repairs is direct attachment of tendon to smooth bone. The native tendon-to-bone attachment, however, presents a rough mineralized interface that might serve an important role in stress transfer between tendon and bone. In this study, we examined the effects of interfacial roughness and interdigital stochasticity on the strength and toughness of a bimaterial interface. Closed form linear approximations of the amplification of stresses at the rough interface were derived and applied in a two-dimensional unit-cell model. Results demonstrated that roughness may serve to increase the toughness of the tendon-to-bone insertion site at the expense of its strength. Results further suggested that the natural tendon-to-bone attachment presents roughness for which the gain in toughness outweighs the loss in strength. More generally, our results suggest a pathway for stochasticity to improve surgical reattachment strategies and structural engineering attachments.     

Tendon collagen synthesis at rest and after exercise in women.

In general, there is a higher incidence of musculoskeletal injuries during physical activity in women than in men. We hypothesized that in women rates of tendon collagen synthesis would be lower than in men at rest and after exercise, especially in the later luteal phase when estrogen and progesterone concentrations are higher than the early follicular phase. We studied tendon collagen fractional synthesis rate (FSR) in 15 young, healthy female subjects in either the early follicular (n = 8) or the late luteal phase (n = 7) 72 h after an acute bout of one-legged exercise (60 min kicking at 67% workload maximum) (72 h) and compared the results with those previously obtained for men. Samples were taken from the patellar tendon in both the exercised and rested legs to determine collagen FSR by the incorporation of [15N]proline into tendon collagen hydroxyproline. There was no effect of menstrual phase on tendon collagen synthesis either at rest or after exercise. However, there was a significant difference between women and men at rest (women = 0.025 +/- 0.002%/h, men = 0.045 +/- 0.008%/h; P < 0.05) and 72 h after exercise (women = 0.027 +/- 0.005%/h; men = 0.058 +/- 0.008%/h). Furthermore, rest and 72-h tendon collagen synthesis were not different in women, whereas in men tendon collagen synthesis remained significantly elevated 72 h after exercise. It is concluded that both in the resting state and after exercise, tendon collagen FSR is lower in women than in men, which may contribute to a lower rate of tissue repair after exercise.     

Tendon disorders of the hand

LEARNING OBJECTIVES: After reading this article, the participant should be able to: 1. Make decisions on flexor tendon repair based on current evidence. 2. Perform some important tendon transfers after viewing Dr. Kozin's videos. 3. Inject local anesthesia for wide-awake flexor tendon repair after viewing the appropriate videos in the article. 4. Use relative motion extension splints for the postoperative management of extensor tendon injuries. SUMMARY: This article provides a practical, clinically useful overview of some of the current best techniques and evidence available to the plastic surgeon in the treatment of flexor and extensor tendon injuries, tendon transfers, trigger fingers, mallet fingers, boutonniere deformities, and De Quervain tenosynovitis. Twelve short movies and drawings emphasize important points of diagnosis and treatment of tendon disorders.     

Temporal and Vertical Distributions of Bacterioplankton at the Gray's Reef National Marine Sanctuary

Large spatial scales and long-term shifts of bacterial community composition (BCC) in the open ocean can often be reliably predicted based on the dynamics of physical-chemical variables. The power of abiotic factors in shaping BCC on shorter time scales in shallow estuarine mixing zones is less clear. We examined the diurnal variation in BCC at different water depths in the spring and fall of 2011 at a station in the Gray''s Reef National Marine Sanctuary (GRNMS). This site is located in the transition zone between the estuarine plume and continental shelf waters of the South Atlantic Bight. A total of 234,516 pyrotag sequences of bacterial 16S rRNA genes were recovered; they were taxonomically affiliated with >200 families of 23 bacterial phyla. Nonmetric multidimensional scaling analysis revealed significant differences in BCC between spring and fall samples, likely due to seasonality in the concentrations of dissolved organic carbon and nitrate plus nitrite. Within each diurnal sampling, BCC differed significantly by depth only in the spring and differed significantly between day and night only in the fall. The former variation largely tracked changes in light availability, while the latter was most correlated with concentrations of polyamines and chlorophyll a. Our results suggest that at the GRNMS, a coastal mixing zone, diurnal variation in BCC is attributable to the mixing of local and imported bacterioplankton rather than to bacterial growth in response to environmental changes. Our results also indicate that, like members of the Roseobacter clade, SAR11 bacteria may play an important role in processing dissolved organic material in coastal oceans.     

Tendon homeostasis: the right pull

Mechanotransduction, the conversion of a biophysical force into a cellular response, allows cells and tissues to respond to their mechanical milieu. How muscle force is translated through TGF-β signaling to regulate tendon homeostasis offers an interesting in vivo example of mechanotransduction.     

Causes of Ecosystem Transformation at the End of the Pleistocene: Evidence from Mammal Body-Mass Distributions

Animal body sizes reflect the discontinuous architecture of the landscapes in which they live, and consequently their body-mass distributions are distinctly clumped rather than continuous. This architectural discontinuity is generated by ecological processes that discretely operate over micro-, meso-, and macroscales. Therefore, changes in these important scale-specific processes for a given geographical region over time should be reflected by corresponding changes in faunal body-mass clump patterns. In this study, we utilized this hypothesis to investigate the terminal Pleistocene mammal extinction event. Specifically, we analyzed the body-mass distributions of latest Pleistocene and modern mammal faunas from northern Florida and southern California to determine the nature of any changes in the clump structures of these regions during the Pleistocene–Holocene transition. In both regions, despite their wide geographical separation and faunal distinctiveness, body-mass clumps below the 40-kg level were remarkably stable across the Pleistocene–Holocene transition despite suffering extinctions. Larger clumps, in contrast, were either orderly truncated or completely eliminated rather than chaotically fragmented. Based on these findings, we argue that the terminal Pleistocene mammal extinctions were caused, at least in part, by changes in key mesoscale aspects of the landscape crucial to supporting a diversity of large mammals. Received 23 April 1997; accepted 14 October 1997.