Does the endometrial gene expression of fertile women vary within and between cycles?

STUDY QUESTION: Does gene expression of putative endometrial implantation markers vary in expression between menstrual cycles? SUMMARY ANSWER: In fertile women the expression of certain genes exhibits a pattern of stable regulation.which is not affected even when sampled twice in one cycle. WHAT IS KNOWN ALREADY: Successful implantation occurs in a minority of IVF embryo transfers. In contrast to knowledge regarding the ovulatory process, there is a sparse understanding of endometrial genes critical to implantation. This lack of knowledge hinders progress in this field. STUDY DESIGN, SIZE, DURATION: Endometrial pipelle samples were collected based on blood endocrinological markers at 2 and 7 days post initial LH surge. Five samples were collected over four cycles where the interval between collections ranged from sequential months to three years. PARTICIPANTS/MATERIALS, SETTING, METHODS: Six fertile women attending an IVF clinic for male factor infertility, had samples collected. Global gene expression profiles were obtained from laser-microdissected, endometrial glands and stroma. Nineteen potential proliferation, cytokine and adhesion markers based on previous validated reports were studied. MAIN RESULTS AND THE ROLE OF CHANCE: There was a significant modification between LH+2 and LH+7 of expression for 23 genes-11 in 8 in glands and stroma, 4 in stroma only and 3 in glands only suggesting stable, controlled regulation. Nevertheless, genes exhibited individual characteristics, e.g MKI67 exhibited lower expression at LH+7 than LH+2 and CCL4 higher, whereas TRO expressed limited difference in both cell types. Stability between cycles was demonstrated for gene expression at both LH+2-more than 60% of genes had <25% variation and at LH+7-60% had <30% variation. Further, effects of prior collection of an LH+2 sample on gene expression at LH+7 were not detected. The range of mRNA expression suggested that a clinical/diagnostic sample at LH+2 and LH+7 is likely to be a better index of endometrial function than a single sample. The possibility of redundancy suggests a panel would be more informative than a single marker. LARGE SCALE DATA: Raw and normalized microarray data have been deposited with the EMBL's European Genome-Phenome Archive for collaborative analysis, reference ega-box-815 (Lappalainen I, Almeida-King J, Kumanduri V, Senf A, Spalding JD, Ur-Rehman S, Saunders G, Kandasamy J, Caccamo M, Leinonen R et al. The European Genome-phenome Archive of human data consented for biomedical research. Nat Genet 2015;47:692-695.) [https://www.ebi.ac.uk/ega/home]. LIMITATIONS, REASONS FOR CAUTION: This type of research has difficulties of recruitment of fertile women for multiple blood testing and repeat endometrial biopsies. Therefore, these data had decreased statistical power due to the overall participant numbers. However, the inclusion of four cycles for each participant permitted the aim of obtaining information on intercycle and intracycle variability to be achieved. WIDER IMPLICATIONS OF THE FINDINGS: Our results support the feasibility of a clinical means of identification of a functional receptive endometrium. The robustness of data from individual women suggests that samples from one cycle can generally be applied to subsequent cycles. STUDY FUNDING/COMPETING INTEREST(S): Funding was granted from the Tertiary Education Commission of New Zealand, Contract I.D.:UOOX06007. There are no competing interests.

Short, Medium and Long Term Complications After Total Anatomical Shoulder Arthroplasty.

Total shoulder arthroplasty (TSA) is an effective approach for the treatment of a variety of clinical conditions affecting the shoulder, including osteoarthritis, inflammatory arthritis and osteonecrosis, and the number of TSA implanted has grown exponentially over the past decade. This review gives an update of the major complications, mainly infections, instability and loosening, encountered after TSA, based on a corpus of recent publications and a dynamic approach: The review focuses on the causes of glenoid loosening, which account for 80% of the complication, and underlines the importance of glenoid positioning in the recovery of early shouder function and in the long term survival rate of TSA.

Shoulder Arthroplasty Imaging: What's New.

BACKGROUND: Shoulder arthroplasty, in its different forms (hemiarthroplasty, total shoulder arthroplasty and reverse total shoulder arthroplasty) has transformed the clinical outcomes of shoulder disorders. Improvement of general clinical outcome is the result of stronger adequacy of the treatment to the diagnosis, enhanced surgical techniques, specific implanted materials, and more accurate follow up. Imaging is an important tool in each step of these processes. METHOD: This article is a review article declining recent imaging processes for shoulder arthroplasty. RESULTS: Shoulder imaging is important for shoulder arthroplasty pre-operative planning but also for post-operative monitoring of the prosthesis and this article has a focus on the validity of plain radiographs for detecting radiolucent line and on new Computed Tomography scan method established to eliminate the prosthesis metallic artefacts that obscure the component fixation visualisation. CONCLUSION: Number of shoulder arthroplasties implanted have grown up rapidly for the past decade, leading to an increase in the number of complications. In parallel, new imaging system have been established to monitor these complications, especially component loosening.

In the secretory endometria of women, luminal epithelia exhibit gene and protein expressions that differ from those of glandular epithelia.

OBJECTIVE: To characterize the transcriptome of luminal epithelia (LE) of fertile secretory endometria and compare the results with those from glandular epithelia (GE). DESIGN: Endometrial samples were collected at 2 and 7 days after initial blood LH surge in separate menstrual cycles. LE were obtained with the use of laser microdissection. mRNA was amplified with the use of linear polymerase chain reaction and hybridized to Agilent 4×44 microarrays. Gene analysis was used to identify differentially expressed mRNAs. Immunohistochemistry was used to assess nine proteins. SETTING: One IVF clinic. PATIENT(S): Seven Caucasian fertile cycling women. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Cycle dating with the use of blood endocrinologic markers, microarrays of laser-microdissected LE, immunohistochemical analysis. RESULT(S): One hundred sixty-one (of 401) differentially expressed mRNAs in LE were identified from the metabolism pathway. Increased selective protein expression in LE at 7 days after initial LH surge was observed. LE mRNA expression was the converse of that in GE. The two cell types each had a different significant biologic pathway identified. CONCLUSION(S): Our results introduce a new concept that LE differentially expressed mRNAs are in the converse direction to that of GE, indicating different biologic processes despite the GE being continuous with the luminal monolayer. This probable distinction of biologic roles has not been noted previously. Further investigations must take cognizance of this observation.

Surgical treatment of three and four-part proximal humeral fractures.

Three- and four-part fractures of the proximal humerus are usually treated surgically. Open reduction with internal fixation (ORIF) is the method of choice in younger patients. Anatomic reduction of the tuberosities is crucial to ensure that, in the event of poorly tolerated avascular necrosis of the humeral head, hemiarthroplasty can be performed under optimal conditions. Suboptimal outcomes may occur after ORIF, as less-than-perfect reduction and fixation is poorly tolerated at the shoulder. Preoperative computed tomography must be performed routinely to analyse fragment displacement and comminution, classify the fracture, assess humeral head vitality, and evaluate the mechanical properties of the underlying bone. Fracture reduction relies on principles that are shared by the various available techniques. Reduction of each fragment should be assessed separately. Reduction of the humeral head to the shaft should be performed before reduction of the tuberosities. The fixation technique should ensure stability of the anatomic reduction, with secure fixation of the tuberosities and a minimal risk of material migration into the joint. Here, we provide a detailed discussion of the various techniques, with their advantages and drawbacks, to help surgeons select the method that is most appropriate to each individual patient.

Gene and protein expression signature of endometrial glandular and stromal compartments during the window of implantation.

OBJECTIVE: To map the changes in messenger RNA (mRNA) and protein abundance during the window of implantation in specifically endometrial stromal and glandular epithelial cells obtained using laser microdissection microscopy (LDM). DESIGN: Endometrial samples were collected from two menstrual cycles at 2 and 7 days after first significant rise in blood LH, and separate cell populations were obtained using LDM. A new generation linear polymerase chain reaction (PCR) amplified the mRNA, which were hybridized to both Affymetrix U133 Plus2 and Agilent 4x44K microarrays followed by gene set analysis. Immunohistochemistry assessed protein expression between the two collection times. SETTING: In vitro fertilization clinic. PATIENT(S): Nine Caucasian, fertile, cycling women. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Cycle dating using blood markers; microarrays on laser microdissected glands and stroma; dual platform microarray confirmation; immunohistochemical analysis of cell cycle proteins. RESULT(S): The two microarray platforms showed concordance. During the window of implantation, a statistically significant network of 22 mRNA associated with the cell cycle was down-regulated. Immunohistochemistry identified altered localization in stroma. CONCLUSION(S): Microarrays demonstrated glands and stroma have distinct mRNA signatures, each dependent on the day of the cycle. We characterized two compartments of the receptive endometrium with a transcriptomic signature identifying regulation of only the cell cycle. Immunohistochemical analysis of cell cycle proteins identified a signature staining pattern of nuclear relocalization of a group of cyclins of stromal cells, which may be clinically applicable.

The effect of exercise on the presence of leukocytes, erythrocytes and collagen fibers in skeletal muscle after contusion.

OBJECTIVE: To determine the comparative rate of resolution of a contusion resulting from mechanical trauma to skeletal muscle, as a function of one of four exercise regimens. DESIGN: Randomized control trial. The four exercise regimens were: running with its onset immediately after injury, running with a 72 hr delay after injury, swimming with immediate onset, or swimming with a 72 hr delay. Control did not exercise. SETTING: Small-animal laboratory. PARTICIPANTS: Male Sprague-Dawley rats. INTERVENTION: A small animal traumatizing machine applied to the biceps femoris. MAIN OUTCOME MEASURES: The rate of contusion resolution was determined by a manual count of erythrocytes, leukocytes and collagen fibers in the contusion, and the data were analyzed using analysis of variance. RESULTS: Exercise of any type produced a greater decline in erythrocyte count (28.2% after 32 days vs. control) than no exercise. Immediate onset of any of the exercise regimens after injury resulted in a greater decline in erythrocyte count (32.7% after 32 days vs. control) and in leukocyte count (17.3% after 32 days vs. control) than delayed onset. Running with either immediate or delayed onset of exercise after injury produced a greater decline in erythrocyte count (36.2% after 32 days vs. control) than swimming. Running with its onset immediately after injury produced the greatest overall rate of decrease in erythrocyte count (44.8% after 32 days vs. control), and the second greatest overall rate of decrease in leukocyte count (15.0% after 32 days vs. control). CONCLUSIONS: Running with immediate onset is the regimen of choice. Any of the given exercises is preferable to no exercise, immediate onset of exercise is preferable to delayed onset, and running is preferable to swimming.

Temporomandibular disorder associated with sacroiliac sprain.

A case of the external derangement-type temporomandibular disorder (TMD), temporarily relieved following chiropractic sacro-occipital technique (SOT) treatment, including SOT category II blocking to reduce sacroiliac sprain, is presented. Symptom exacerbation midway through the course of treatment followed additional dental work; symptom remission followed additional SOT treatment. Freedom from symptoms is maintained with a 3-wk treatment interval. There appears to be a cause-effect relationship between external derangement-type TMD and sacroiliac sprain. Concurrent, coordinated chiropractic and dental treatments may improve the success rate of TMD resolution.

Changes in open-field behavior of rats following spinal nerve root sectioning.

The open-field behavior of of 30 laboratory albino rats was measured after selective unilateral sectioning of either the L2, L3, L4 or L5 spinal nerve root using a novel abdominal entry. Although treated subjects displayed observable motor dysfunction of the operated limb, no differences were found in open-field ambulation, rearing, defecation or urination based either on the level of the sectioned nerve root or on the day the behavior was measured, suggesting that there was an effect of the treatment not measurable by standard statistical procedures, quite possibly due to the small sample size.

A Mathematical Model for Dental Caries: A Coupled Dissolution-Diffusion Process.

Demineralization of tooth mineral in the caries process was studied using a computer model that simulates a diffusion controlled dissolution process. The model consists of a two-compartment system. An acidic solution in the outer ("plaque") compartment was assumed to be large in volume so that its composition remained constant during the process. The solution in the inner ("lesion") compartment was in equilibrium with the tooth mineral, but its composition changed in response to diffusion of ions between the two solutions through an infinitely thin barrier. The permselectivity of the diffusion barrier to cations and anions can be modified as desired thus allowing the effects of membrane on the diffusion-dissolution process to be examined. Because the losses of calcium (Ca) and phosphate (P) from the "lesion" to the "plaque" generally does not occur at a molar ratio of 5/3, the Ca to P ratio of the dissolving mineral, the composition of the "lesion" fluid can change significantly from the starting composition, and this in turn modifies the Ca and P fluxes. A steady state condition is eventually reached under which the ratio of flux of Ca to that of P becomes 5/3. The results of the simulation show that for a given "plaque" pH, the rate of demineralization at steady state was the highest for cation and the lowest for anion permselective membranes. These results were in good agreement with those from an experimental study under comparable conditions.

Micro-analysis of mineral saturation within enamel during lactic acid demineralization.

In this study, the physicochemical factors responsible for caries-like lesion propagation were investigated by means of a micro-analytical system used to study the fluid within a lesion during a simulation of the decay process. Four 500-microns-thick serial sections prepared from a single human molar were mounted between glass plates with only the natural surface of the tooth exposed. Microwells were then drilled into sound and pre-existing carious regions of the section through one of the plates. These microwells were then filled with fluid under mineral oil, and after a week of equilibration, the natural surface of the section was exposed to a lesion-producing fluid. The concentrations of calcium, phosphate, and hydrogen ions of the fluid in the wells were then followed as a function of time as the lesion advanced. The results of this study, in which lactic acid was used to demineralize enamel, were consistent with those previously reported (Vogel et al., 1987a): The solution within the lesion remained saturated during the acid attack. Differences in initial mobilities of the calcium and phosphate and other ions, a result of the permselectivity of the enamel, increased the concentrations within the lesion and permanently changed the ratio of these ions in the lesion solution. Based on these results, we suggest that the ionic permselectivity of tooth enamel can have a profound effect on the transport of mineral from a caries lesion.

The buffer capacity of single-site, resting, human dental-plaque fluid.

A carbonate equilibration method was used to measure the buffer capacity of resting plaque fluid collected from single buccal or interproximal sites of upper and lower first molars or anterior teeth. The maximum buffer capacity was 26 m-equiv./l at pH 7.1. The buffer contribution from the measured concentrations of phosphate and carbonate was calculated for each sample. These values were compared with the buffering actually measured and with that expected from organic acids, proteins, and amino acids at average values, as taken from reports in the literature. Relative contributions of buffer species at the average pH of the samples (6.86) were: 35 per cent phosphate, 10 per cent carbonate, 10 per cent protein, 10 per cent organic acids, 2 per cent amino acids, 30 per cent unidentified. There were no significant differences in the buffer capacities of samples originating from sites that differ in their accessibility to saliva. Buffering in resting plaque fluid is more than twice that in saliva and did not show differences correlated with the intra-oral location of the samples.

Solubility of CaHPO4 · 2H2O in the Quaternary System Ca(OH)2 - H3PO4 - NaCl - H2O at 25 °C.

Solubility of CaHPO4 · 2H2O was determined in the quaternary system Ca(OH)2 - H3PO4 - NaCl - H2O at 25 °C in the pH range 4.39-6.38; ionic strengths of the saturated solutions varied from 0.00485 to 0.545, Satisfactory constancy in the solubility product, ( K sp = [ Ca 2 + ] ⋅ [ HPO 4 2 - ] ⋅ γ Ca 2 + ⋅ γ HPO 4 2 - = 2.49 ± 0.05 × 10 - 7   mol 2 × 1 - 2 ) was obtained when (i) the ion activity coefficients, γ i , were calculated with the Debye-Hückel equation, log   γ i = - A Z i 2 I / ( 1 + B α i I ) + 0.0626   I ; the value 0.0626 for the coefficient in the linear term was derived from the solubility data by utilizing a statistical procedure, and (ii) formation of an ion pair NaHPO 4 - was taken into account; a statistically derived value for the stability constant of this ion pair is 7.0 ± 2.4 1 × mol-1. The ion pair NaHPO 4 - appears to have significant concentrations in physiological fluids.

Solubility of ß-Ca3(PO4)2 in the System Ca(OH)2-H3PO4-H2O at 5, 15, 25, and 37 °C.

Solubility isotherms of beta-tricalcium phosphate, ß-Ca(PO4)2, prepared by heating mixtures of CaCO3 and CaHPO4 above 800 °C, were determined in the ternary system Ca(OH)2-H3PO4-H2O at 5, 15, 25, and 37 °C in the pH range 6.0-7.5 by equilibration with dilute H3PO4 solutions. The results indicate that ß-Ca3(PO4)2 has a negative thermal coefficient of solubility. The solubility product, K s , was determined as a function of temperature by a generalized least-squares procedure; the resulting equation is log K s = - 45723.26 / T + 287.4536 - 0.546763 T ; the values of K s and its dispersion at 25 and 37 °C are 1.20(0.056), and 0.283(0.011) × 10-29. Thermodynamic functions for the dissolution of the salt at the four experimental temperatures are reported. When treated as an adjustable constant, the Ca/P ratio in these ß-Ca3(PO4)2 solutions was found to have the value 1.514(0.010), confirming that the stoichiometry of the high temperature form of this salt is correctly indicated by the above formula. The computed isotherms at 25 °C for several calcium phosphates are compared; it is shown that ß-Ca3(PO4)2 is, next to Ca5(PO4)3OH, the most stable at pH's above 6.36. The singular points of the ß-Ca3(PO4)2 isotherm with those of CaHPO4 · 2H2O and CaHPO4 are at pH's 5.88 and 6.36, respectively.

Calculation of Diffusion Coefficients in Ternary Systems from Diaphragm Cell Experiments.

Using a generalized least square procedure, a method is developed to estimate diffusion coefficients, D ij , from concentration measurements made in a set of experiments of varying duration but with the same initial conditions. The method requires neither approximations nor restrictions in the flow equations. The D ij and their errors are calculated simultaneously with the adjustment of the weighted observables - concentrations of the two solute components and time. A procedure is described to insure convergence in a given experimental set. The effect of the duration of the longest experiment, and errors in the observables on the estimated values of D ij and their errors, are investigated. Application of the method to available data yielded values for D ij in agreement with those previously calculated by more involved procedures.

Preparation and Solubility of Hydroxyapatite.

Two portions of a synthetic hydroxyapatite (HA), Ca5OH(PO4)3, fully characterized by x-ray, infrared, petrographic, and chemical analyses, were heated at 1,000 °C in air and steam atmospheres, respectively. Solubility isotherms for these two samples in the system Ca(OH)2-H3PO4-H2O were determined in the pH range 5 to 7 by equilibrating the solids with dilute H3PO4 solutions. Both samples of HA dissolved stoichiometrically. The activity products ( Ca + + ) 5 ( OH - ) ( PO 4 ≡ ) 3 and their standard errors-obtained by a least squares adjustment of the measurements (Ca and P concentrations and pH of the saturated solutions) subject to the conditions of electroneutrality, constancy of the activity product, and stoichiometric dissolution - were 3.73 ± 0.5 × 10-58 for the steam-heated HA and 2.51 ± 0.4 × 10-55 for the air-heated HA. Allowance was made in the calculations for the presence of the ion pairs [CaHPO4]0 and [CaH2PO4]+. The higher solubility product for the air-heated HA is ascribed either to a change in the heat of formation brought about by partial dehydration or to a state of fine subdivision resulting from a disproportionation reaction. The solubility product constants were used to calculate the points of intersection (i.e., singular points) of the two HA solubility isotherms with the isotherms of CaHPO4 · 2H2O and CaHPO4; it was found that the pH's of the singular points for the air-heated HA were a full unit higher than those of the steam-heated preparation. Conditions are described for the precipitation of HA crystals suitable for solubility measurements.