[The S2k guideline: Indications for knee endoprosthesis : Evidence and consent-based indications for total knee arthroplasty]. / S2k-Leitlinie: Indikation Knieendoprothese : Evidenz- und konsensbasierte Indikationsstellung in der Knie-Endoprothetik (EKIT-Knie).

Total knee arthroplasty (TKA) is one of the most frequent surgical procedures in orthopaedic surgery. Until now there have not been any standardized indication criteria, which might contribute to the large geographical differences in the frequency of TKA. This guideline aims to consent minimal requirements (main criteria), additional important aspects (minor criteria), as well as relative and absolute contraindications for TKA. The following main criteria have been consented: knee pain, radiological confirmation of osteoarthritis or osteonecrosis, inadequate response to conservative treatment, adverse impact of knee disease on the patient's quality of life and the burden of suffering due to the knee disease. Relative contraindications have been consented as severe general disease with reduced life expectancy and a BMI ≥40; absolute contraindications are an active infection and if the patient is not able to undergo major surgery.

The impact of ovarian stimulation on the expression of candidate reprogramming genes in mouse preimplantation embryos.

Ovarian stimulation with gonadotrophins is an integral part of assisted reproductive technologies in human subfertility/infertility treatment. Recent findings have associated ovarian stimulation with the increased incidence of imprinting disorders in humans as well as defects in genome-wide methylation reprogramming and, in particular, imprinting in mice. Here, we present the first study that determined the impact of ovarian stimulation on the expression of developmentally important reprogramming genes (Apex1, Lig1, Lig3, Mbd2, Mbd3, Mbd4, and Polb) in single early mouse morula embryos (16-cell stage). Using absolute quantification of mRNA by quantitative real-time PCR, we observed an association of ovarian stimulation with a downregulation of mRNAs encoding the base excision repair proteins APEX1 and POLB as well as the 5-methyl-CpG-binding domain protein MBD3 in individual morula embryos. Whole mount immunofluorescence staining of early and late morula embryos with an antibody against APEX1 revealed individual embryos with lower protein expression levels after ovarian stimulation and a correlation of mRNA expression with protein abundance. Our data argue for a negative impact of ovarian stimulation during female gametogenesis and/or early embryo development affecting the expression of candidate reprogramming factors.

United States (US) multi-center study to assess the validity and reliability of the Spinal Cord Independence Measure (SCIM III).

STUDY DESIGN: Multi-center, prospective, cohort study. OBJECTIVES: To assess the validity and reliability of the Spinal Cord Independence Measure (SCIM III) in measuring functional ability in persons with spinal cord injury (SCI). SETTING: Inpatient rehabilitation hospitals in the United States (US). METHODS: Functional ability was measured with the SCIM III during the first week of admittance into inpatient acute rehabilitation and within one week of discharge from the same rehabilitation program. Motor and sensory neurologic impairment was measured with the American Spinal Injury Association Impairment Scale. The Functional Independence Measure (FIM), the default functional measure currently used in most US hospitals, was used as a comparison standard for the SCIM III. Statistical analyses were used to test the validity and reliability of the SCIM III. RESULTS: Total agreement between raters was above 70% on most SCIM III tasks and all κ-coefficients were statistically significant (P<0.001). The coefficients of Pearson correlation between the paired raters were above 0.81 and intraclass correlation coefficients were above 0.81. Cronbach's-α was above 0.7, with the exception of the respiration task. The coefficient of Pearson correlation between the FIM and SCIM III was 0.8 (P<0.001). For the respiration and sphincter management subscale, the SCIM III was more responsive to change, than the FIM (P<0.0001). CONCLUSION: Overall, the SCIM III is a reliable and valid measure of functional change in SCI. However, improved scoring instructions and a few modifications to the scoring categories may reduce variability between raters and enhance clinical utility.

[Exulcerated, nodular tumor on the occiput of a 29-year-old patient]. / Exulzerierter, nodulärer Tumor am Hinterkopf eines 29-jährigen Patienten.

Cranial fasciitis is a rare variation of nodular fasciitis that occurs in the region of the capillitium. We report on a 29-year-old patient who presented with a 2-month history of a tumor progressively increasing in size located on the occiput. Histological examination revealed a tumor, consisting of tightly packed spindle-shaped cells with underlying myxoid stroma, which extended from the dermis to the subcutis. Actin and vimentin were detected by immunohistochemistry. We established a diagnosis of cranial fasciitis and excised the tumor. Especially when a child or young adult presents with a tumor in the skull area, consideration should be given to cranial fasciitis in the differential diagnosis to avoid unnecessary and possibly very invasive treatment approaches.

Regulation of surfactant phosphatidylcholine secretion from alveolar type II cells during Pneumocystis carinii pneumonia in the rat.

We used an immunosuppressed rat model to test the hypothesis that normal mechanisms regulating surfactant phosphatidylcholine synthesis and secretion in alveolar type II cells are aberrant in Pneumocystis carinii pneumonia. Animal groups included: group 1, healthy controls; group 2, immunosuppressed, without pneumocystosis; group 3, immunosuppressed with pneumocystosis; group 4, immunosuppressed with well-established pneumocystosis treated with trimethoprim-sulfamethoxazole (TMP-SMX). Type II cells were isolated from rats in each group and compared for [3H]choline incorporation into phospholipid and response of the type II cells to secretagogues. Incorporation of [3H]choline into phospholipid subclasses exhibited significant differences. Incorporation into phosphatidylcholine fell from 89.3 +/- 2.2% of total incorporation in group 1 control rats to 79.6 +/- 3.1% in group 3 rats with P. carinii pneumonia, while incorporation into sphingomyelin rose from 5.6 +/- 1.2% in group 1 animals to 15.2 +/- 2.7% in group 3 rats. Incorporation of [3H]choline into phospholipid subclasses in cells from group 2 and group 4 animals was not different from incorporation for group 1 animals. Type II cells from group 1 and group 2 (immunosuppressed control) rats responded appropriately to the secretagogues ATP, TPA, and terbutaline with a marked increase in surfactant phosphatidylcholine secretion; the effect of ATP was also blocked by the lectin, concanavalin A. In contrast, type II cells from group 3 rats failed to respond to the secretagogues with a significant increase in phospholipid secretion. Although treatment of group 4 rats with TMP-SMX markedly reduced the P. carinii organism burden, type II cells from these animals also responded poorly to the secretagogues. The depressed type II cell function described here provides a mechanism for the observed decrease in surfactant phospholipids from bronchoalveolar lavage fluid of experimental animals and patients with P. carinii pneumonia. The data also suggest this defect may become irreversible with advanced disease.

Delayed profound local brain hypothermia markedly reduces interleukin-1beta gene expression and vasogenic edema development in a porcine model of intracerebral hemorrhage.

White matter (lobar) intracerebral hemorrhage (ICH) can cause edema-related deaths and life-long morbidity. In our porcine model, ICH induces oxidative stress, acute interstitial and delayed vasogenic edema, and up-regulates interleukin-1beta (IL-1beta), a proinflammatory cytokine-linked to blood-brain barrier (BBB) opening. In brain injury models, hypothermia reduces inflammatory cytokine production and protects the BBB. Clinically, however, hypothermia for stroke treatment using surface and systemic approaches can be challenging. We tested the hypothesis that an alternative approach, i.e., local brain cooling using the ChillerPad System, would reduce IL-1beta gene expression and vasogenic edema development even if initiated several hours after ICH. We infused autologous whole blood (3.0 mL) into the frontal hemispheric white matter of 20 kg pentobarbital-anesthetized pigs. At 3 hours post-ICH, we performed a craniotomy for epidural placement of the ChillerPad. Chilled saline was then circulated through the pad for 12 hours to induce profound local hypothermia (14 degrees C brain surface temperature). We froze brains in situ at 16 hours after ICH induction, sampled perihematomal white matter, extracted RNA, and performed real-time RT-PCR. Local brain cooling markedly reduced both IL-1beta RNA levels and vasogenic edema. These robust results support the potential for local brain cooling to protect the BBB and reduce injury after ICH.

Identity of a cytosolic neutral cholesterol esterase in rat liver with the bile salt stimulated cholesterol esterase in pancreas.

A neutral cholesterol esterase has been purified to homogeneity from the cytosolic fraction of rat liver. The 105,000 x g supernatant fraction of rat liver was applied to a DEAE-cellulose column to isolate a partially purified fraction of hepatic cholesterol esterase. Immunoblot analysis of the partially purified liver fraction with the anti-porcine pancreatic cholesterol esterase IgG demonstrated a single band with a molecular weight of 67,000. The hepatic protein was then isolated by immunoaffinity chromatography technique using a column constructed with antibodies prepared against the pancreatic cholesterol esterase. Characterization of the hepatic cholesterol esterase revealed that the hepatic enzyme shared antigenic epitopes with the pancreatic cholesterol esterase and was similarly activated by addition of bile salt such as taurocholate. Moreover, amino-terminal sequencing analysis of the hepatic cholesterol esterase showed an identical sequence with the pancreatic enzyme. Taken together, these results showed that the cholesterol esterases in the liver and the pancreas are very similar and possibly identical proteins.

Analysis of Pneumocystis carinii organism burden, viability and antigens in bronchoalveolar lavage fluid in AIDS patients with pneumocystosis: correlation with disease severity.

OBJECTIVES: We examined 96 bronchoalveolar lavage fluid (BALF) specimens from AIDS patients with proven Pneumocystis carinii pneumonia (PCP) in order to compare the relationship of organism burden, viability and antigen expression with disease severity at the time of clinical presentation. METHODS: Tinctorial analysis of BALF specimens with proven PCP using Diff-Quik, cresyl echt violet and erythrosin B stains to evaluate organism burden and viability. P. carinii antigen examination was performed by Western blot analysis. RESULTS: P. carinii cluster ratios were more sensitive than cyst counts as an indicator of organism burden, and correlated well with the alveolar-arterial oxygen gradient as a measure of disease severity. Erythrosin B, the vital stain used to measure P. carinii viability, displayed a wide range of values and provided little useful information. Antigens of 35-45 and 95kD, which were specific for P. carinii, were found by immunoblot analysis in BALF cellular fraction of most patients with pneumocystosis. By contrast, antigens of 52 and 66 kD, which were found in both BALF supernatant and cellular fractions of P. carinii patients and controls, most likely represented albumin and immunoglobulin G heavy chain, respectively, of host origin. The 35-45 kD antigen was found in 88% of the BALF specimens and appeared to represent an important marker of P. carinii infection. The 95 kD antigen was detected in 49% of the specimens. CONCLUSIONS: We conclude that analysis of P. carinii characteristics in BALF specimens of patients with pneumocystis may provide additional information. These data will also be helpful in developing more sensitive assays and in targeting specific P. carinii factors for future investigation.

Retinoic acid decreases attachment of JAR choriocarcinoma spheroids to a human endometrial cell monolayer in vitro.

The ability of retinoic acid (RA) to modulate attachment of JAR choriocarcinoma multicellular spheroids to monolayers of a human uterine epithelial cell line (RL95-2) was examined using a centrifugal force-based adhesion assay. Exposure of choriocarcinoma spheroids to RA (10(-7) to 10(-5) M) over a 3-day culture period resulted in a dose-dependent decrease of attachment. Significant decreases in attachment were detected after 30 min (75 per cent versus 25 per cent) and 1 h (92 per cent versus 26 per cent) of confrontation-culture between choriocarcinoma and uterine cells for control versus 10(-5) M RA; by 5 h 100 per cent spheroid attachment was detected in all treatment groups. RA had no effect on cell proliferation in JAR spheroids, but 10(-5) M RA treatment induced a fivefold increase in secretion of human chorionic gonadotrophin (hCG), a known marker of conversion of cytotrophoblast to syncytiotrophoblast-like cells. These findings demonstrate that RA modulates cellular attachment and differentiation in choriocarcinoma spheroids in vitro.

Stereoselectivity of lipases in supercritical carbon dioxide. I. Dependence of the regio- and enantioselectivity of porcine pancreas lipase on the water content during the hydrolysis of triolein and its partial glycerides.

The stereoselectivity of porcine pancreas lipase (PPL) was investigated during the enzymatic hydrolysis of triolein and its partial glycerides in the presence of supercritical carbon dioxide (SCCO2) as reaction medium. The water content of the immobilized lipases was varied. The partial glycerides were separated into mono- and diglycerides by TLC, converted to their 3,5-dinitrophenylurethane derivatives and subsequently resolved into sn-1,2 and sn-2,3 enantiomers (estimation of dioleins) or into sn-1 and sn-3 enantiomers (estimation of monooleins) by HPLC on a chiral stationary phase (Sumichiral OA-4100). In all reactions under the conditions employed, PPL revealed a distinct preference for the sn-3 position of the glycerol. However, the stereoselectivity depends on the reaction time, the substances initially used and the enzyme water content. It seems that the effect of the enzyme water content on the activity and selectivity of porcine pancreas lipase in SCCO2 is based on a modification of the "micro-environment' of the enzyme by the solution of CO2 in water, causing a decrease of the pH value.

Quantitation of human choriocarcinoma spheroid attachment to uterine epithelial cell monolayers.

Adhesive interactions of trophoblast cells with endometrium are essential for embryo implantation in the uterus. Choriocarcinoma cells, the malignant counterpart of trophoblast, show pronounced invasiveness and are of interest for model studies. We describe here an in vitro model system for the study of adhesion of human JAR choriocarcinoma multicellular spheroids to different human endometrial epithelial cell lines (RL95-2, HEC-1A, KLE, AN3-CA) grown as monolayers. Cell characterization showed JAR spheroids to s++ecrete the placental hormones human chorionic gonadotropin and progesterone into the culture medium; distinct patterns of keratin, vimentin, and uvomorulin expression were seen in the endometrial cell lines. Spheroid attachment to endometrial monolayers was quantified using a centrifugal force-based adhesion assay, and morphology was examined by light and electron microscopy. Results showed the JAR spheroids to attach to three of the endometrial monolayers (RL95-2, HEC-1A, KLE) progressively over a 24-h period (by which time > or = 80% of the spheroids attached). Significant differences in spheroid attachment were most pronounced at 5 h (RL95-2 > HEC-1A > KLE and poly-D-lysine control, i.e. 90:45:17:17% attached). JAR spheroids did not attach to the endometrial cell line AN3-CA. Morphology revealed choriocarcinoma cells to begin to intrude between the uterine RL95-2 epithelial cells at 5 h. At 24 h, this intrusive type of penetration continued to be seen only with the RL95-2 monolayer. The assay system thus identifies differences in attachment properties between choriocarcinoma cells and various endometrial cell lines and forms the basis for further studies on the molecular interactions involved.

Quantitation of human choriocarcinoma spheroid attachment to uterine epithelial cell monolayers.

Adhesive interactions of trophoblast cells with the endometrium are essential for embryo implantation in the uterus. Choriocarcinoma cells, the malignant counterpart of trophoblast, show pronounced invasiveness and are of interest for model studies. We describe here an in vitro model system for the study of adhesion of human JAR choriocarcinoma multicellular spheroids to different human endometrial epithelial cell lines (RL95-2, HEC-1A, KLE, AN3-CA) grown as monolayers. Cell characterization showed JAR spheroids to secrete the placental hormones human chorionic gonadotropin and progesterone into the culture medium; distinct patterns of keratin, vimentin, and uvomorulin expression were seen in the endometrial cell lines. Spheroid attachment to endometrial monolayers was quantified using a centrifugal force-based adhesion assay, and morphology was examined by light and electron microscopy. Results showed the JAR spheroids to attach to three of the endometrial monolayers (RL95-2, HEC-1A, KLE) progressively over a 24-h period (by which time ≥80% of the spheroids attached). Significant differences in spheroid attachment were most pronounced at 5 h (RL95-2 > HEC-1A > KLE and poly-D-lysine control, i.e. 90:45:17:17% attached). JAR spheroids did not attach to the endometrial cell line AN3-CA. Morphology revealed choriocarcinoma cells to begin to intrude between the uterine RL95-2 epithelial cells at 5 h. At 24 h, this intrusive type of penetration continued to be seen only with the RL95-2 monolayer. The assay system thus identifies differences in attachment properties between choriocarcinoma cells and various endometrial cell lines and forms the basis for further studies on the molecular interactions involved.

A survey of birds in Denmark for the presence of Pneumocystis carinii.

One hundred eighty-three toluidine blue O-stained necropsy lung imprint smears from different avian species were examined microscopically for Pneumocystis carinii. No cyst forms of the organism could be identified. Seventy-eight serum samples from a total of 155 chickens were examined by a competition enzyme-linked immunosorbent assay (ELISA) for antibodies to P. carinii; 53 serum samples were from individual chickens, and 25 samples were pools of sera from two to five chickens. Diluted 1:50, the 78 serum samples showed a specific ELISA-inhibition of 4% to 56% (the 95% confidence limit being 25% to 30% inhibition). Diluted 1:50, nine serum pools representing 34 chickens and 17 of the 53 individual serum samples (32.1%) showed an inhibition greater than 30%. No specific pneumocyst DNA could be detected in serum from 13 of the 53 chickens using polymerase chain reaction and dihydrofolate reductase gene as a specific probe. Specific antibodies to a 116,000-molecular-weight antigen of rat pneumocysts were shown in two (13.3%) of 15 individual chicken serum samples. The results indicate that P. carinii organisms do not commonly reside in the lungs of birds, although some birds may be exposed to external sources of organisms.

[Echocardiography and right heart catheterization in pulmonal hypertension]. / Echokardiographie und Rechtsherzkatheterisierung bei pulmonaler Hypertonie.

BACKGROUND AND OBJECTIVE: Since pulmonary hypertension is defined haemodynamically with a mean pulmonary artery pressure (mPAP) of at least 25 mmHg, right heart catheterization is mandatory for diagnosis of PH. However, it remains unclear if echocardiography can always detect a PH and to what extent echocardiographic and invasive parameters correlate. We aimed to determine the frequency of right heart valve insufficiencies, the correlation of mPAP measured invasively and estimated by echocardiography and the correlation of other echocardiographic parameters with invasively measured cardiac output (CO) and pulmonary vascular resistance (PVR) in patients who presented at our center for pulmonary hypertension. METHODS: Retrospective analysis of echocardiographic and invasively measured parameters in consecutive patients who presented with dyspnea and suspected PH. RESULTS: 10% of the patients with invasively confirmed PH had no tricuspid valve insufficiency, 61.3% had no pulmonary valve insufficiency. The highest correlation of invasively measured mPAP was found for mPAP estimated non-invasively via RVSP (R = 0.80; < 0.0001). Correlation of non-invasively estimated mPAP by the pulmonary valve regurgitation (R = 0,72, p < 0,004) and mPAP measured by the right ventricular outflow tract flow acceleration time (R = 0,54, p < 0.0001) with invasively measured mPAP were inferior. Left ventricular eccentricity index and systolic tissue Doppler velocity of tricuspid annulus correlated highly significant with PVR, TAPSE correlated with PVR and CO. Right ventricular myocardial performance index correlated with CO. CONCLUSION: In 10% of invasively proven PH, the diagnosis can be missed by estimation of RVSP by echocardiography alone, due to a lack of tricuspid valve insufficiency. Echocardiographic assessment of mPAP remains difficult. Echocardiography can deliver qualitative information about CO and PVR. Right heart catheterization remains mandatory to confirm or to rule out PH.

Social isolation-induced epigenetic changes in midbrain of adult mice.

Social isolation and loneliness increase the risk of death as much as well-established risk factors for mortality such as cigarette smoking and alcohol consumption. The underlying molecular mechanisms are poorly understood. In the present study, 3 months old male C57BL/6 mice were socially isolated by individual housing for another 3 months. At the age of 6 months, epigenetic changes were analyzed in midbrain. Social isolation of male adult mice led to an increased global DNA methylation, which was associated with enhanced activity of DNA methyltransferase. Di- and trimethylation of global histone H3 lysine 4 (H3K4) were increased in midbrain of socially isolated mice, accompanied by enhanced H3K4 histone methyltransferase activity. In addition, social isolation of adult mice led to activation of histone acetyltransferases as well as of histone deacetylases (HDAC) resulting in a net enhancement of histone H3 lysine 9 (H3K9) acetylation. Gene-specific effects were observed for Hdac1, Hdac3 and the serotonin transporter Slc6a4. Social isolation led to an up-regulation of Hdac1 and Hdac3, associated with decreased DNA methylation in the CpG island of the respective genes. On the contrary, the Slc6a4 gene was down-regulated, which was associated with enhanced DNA methylation. Collectively, the results from the present study demonstrate for the first time that social isolation of adult mice leads to a wide range of global epigenetic changes and these effects may have profound impact on gene expression pattern and phenotype of the socially isolated animals.

Leaf surface structures enable the endemic Namib desert grass Stipagrostis sabulicola to irrigate itself with fog water.

The Namib grass Stipagrostis sabulicola relies, to a large degree, upon fog for its water supply and is able to guide collected water towards the plant base. This directed irrigation of the plant base allows an efficient and rapid uptake of the fog water by the shallow roots. In this contribution, the mechanisms for this directed water flow are analysed. Stipagrostis sabulicola has a highly irregular surface. Advancing contact angle is 98° ± 5° and the receding angle is 56° ± 9°, with a mean of both values of approximately 77°. The surface is thus not hydrophobic, shows a substantial contact angle hysteresis and therefore, allows the development of pinned drops of a substantial size. The key factor for the water conduction is the presence of grooves within the leaf surface that run parallel to the long axis of the plant. These grooves provide a guided downslide of drops that have exceeded the maximum size for attachment. It also leads to a minimum of inefficient drop scattering around the plant. The combination of these surface traits together with the tall and upright stature of S. sabulicola contributes to a highly efficient natural fog-collecting system that enables this species to thrive in a hyperarid environment.

Evaluation of portal venous gas detected by ultrasound examination for diagnosis of necrotising enterocolitis.

BACKGROUND: Early diagnosis of necrotising enterocolitis (NEC) is difficult but essential for timely therapy. The diagnostic hallmarks and specific radiological signs for NEC are pneumatosis intestinalis (PI) and portal venous gas (PVG), but PVG in abdominal ultrasound (PVG-US) has been proposed as an effective tool in the diagnosis of NEC as well. OBJECTIVE: To prospectively assess the value of PVG-US for the diagnosis of NEC. METHODS: The study screened 352 neonates for PVG-US (n = 796 routine examinations) and performed 48 additional screenings in 34/352 neonates with suspected (stage I, n = 28) or definite NEC (stage > or =II, n = 20). Sensitivity and specificity of PVG-US for detection of NEC were computed by using NEC stage > or =II as the reference standard. RESULTS: PVG-US was only present in cases of suspected or definite NEC. The study observed PVG-US in 4/28 NEC stage I and in 9/20 NEC stage > or =II episodes corresponding to a 86% specificity and a 45% sensitivity for diagnosis of NEC stage > or =II. However, 7/20 patients with NEC stage > or =II showed intraoperative findings other than NEC and another 3/20 infants presented with radiologically unspecific intestinal dilatation. None of these 10 infants had detectable PVG-US. Thus, with application of specific radiological signs the sensitivity of PVG-US for diagnosis of NEC stage > or =II increased to 90%. CONCLUSION: Screening for PVG-US is a useful, easy and quick bedside test with a high specificity for NEC. Moreover, these results question the value of the Walsh criteria in the diagnosis of NEC.