Pretreatment prostate-specific antigen velocity is associated with development of distant metastases and prostate cancer mortality in men treated with radiotherapy and androgen-deprivation therapy.
In men with prostate cancer, pretreatment prostate-specific antigen (PSA) velocity (PSAV) has been demonstrated as a predictor of biochemical and survival outcomes in patients undergoing radical prostatectomy (RP). The utility of pretreatment PSAV in predicting outcomes after radiotherapy (RT), with or without androgen-deprivation therapy (ADT), is less certain. This study was undertaken to determine whether pretreatment PSAV is associated with biochemical disease-free survival, patterns of recurrence, and survival outcomes in men treated with radiation therapy and ADT. Two hundred seventy-seven patients with intermediate- and high-risk prostate cancer treated with RT and ADT formed the study cohort. Kaplan-Meier survival estimates and Cox regression analyses were used to evaluate whether PSAV was associated with disease outcomes. The median age of diagnosis was 70 years, and the median follow-up was 6.8 years. Men with a PSAV in the highest quartile tended to have higher risk disease at presentation (P = .028). After adjustment for known prognostic factors and duration of ADT, men who had a PSAV in the highest quartile had an increased risk of distant metastasis (hazard ratio [HR], 4.0; 95% confidence interval [95% CI], 1.61-9.9 [P = .003]) and prostate cancer-specific mortality (HR, 2.75; 95% CI, 1.27-5.95 [P = .01]) compared with men who had a lower PSAV, but had no increase in the risk of local recurrence (P = .76). A high pretreatment PSAV was associated with distant metastasis and prostate cancer-specific mortality but not with local recurrence. A high pretreatment PSAV may signify the presence of occult metastatic disease. Randomized trials are needed to determine whether more aggressive intervention is required in men who present with high pretreatment PSAV.


Effectiveness of bio-dispersant in homogenizing hydroxyapatite for proliferation and differentiation of osteoblast.
Hydroxyapatite (HA), an inorganic compound, plays an essential role in the proliferation and differentiation of bone cells. Using cellulose nanocrystals (CNCs) as green dispersants to improve homogenization of HA is promising in the fabrication of nanocomposite scaffolds with biocompatibility for bone tissue engineering. The HA/CNC (HC) nanoparticle suspension was incorporated in polyvinyl alcohol (PVA)-based scaffold to investigate the physical and chemical properties. The PVA/HC composites demonstrated high porous structure and swelling ability for cell attachment and a 3-fold improvement in compressive modulus compared with free HC scaffold. Moreover, the presence of HC nanoparticles has promoted the proliferation and mineralization of pre-osteoblast. Our findings could provide an effective strategy by using bio-dispersants to incorporate mineral elements into synthetic polymers for the fabrication of functional tissue engineering scaffolds.


The low-frequency response of inner hair cells in the guinea pig cochlea: implications for fluid coupling and resonance of the stereocilia.
AC receptor potentials within the inner hair cells of the basal turn of the guinea pig cochlea have been recorded for stimuli in the frequency range 20 Hz to 3200 Hz. Comparison of these potentials with potentials recorded in scala media suggests that the stereocilia of many inner hair cells are stimulated by the transverse velocity of the cochlear partition for very low frequency, but above a transition frequency in the range 400 Hz to 1000 Hz they become entrained with partition displacement. It is suggested that such a transition is probably a simple consequence of the fluid coupling that drives these cells, and that mechanical resonance of the free-standing stereocilia of the inner hair cells does not occur in the basal turn of the guinea pig. These results do not, however, preclude the possibility of mechanical resonance involving the stereocilia of the outer hair cells. The results also indicate that the bodies of these cells low-pass filter the intracellular receptor potential, with a cutoff frequency of approximately 1000 Hz.


[The middle fossa approaches].
Describe the surgical systematization of the different variants of the middle fossa approach. The search for the blue line of the superior canal at the arcuate eminence is the most reliable method to identify the internal auditory canal. When this is demonstrated, each surgical variant can then be undertaken with the surgical landmarks described below. The classical suprapetrosal approach allows one to directly control the entire internal acoustic meatus from the fundus of the canal to the porus. Enlarged, it provides access to the cerebello pontine angle nearby to the porus. Finally, the petrous apex can be approached while preserving the bony labyrinth via the transpetrosal middle fossa approach. Difficult to learn, the middle fossa approach depends on recognizing the surgical landmarks of the anterosuperior surface of the petrous bone. Extradural, preserving the bony labyrinth, and controlling the entire internal acoustic meatus, it is an important approach in skull base surgery.


Higher activity by opaque endometriotic lesions than nonopaque lesions.
Higher activity by early endometriosis than advanced endometriosis has been reported. However, the pattern of activity in individual colored endometriotic lesions in pelvic cavity is unknown. We investigated the variation in activity of the different colored morphologic lesions as proposed by the current revised American Society of Reproductive Medicine (ASRM) classification in women with endometriosis. Peritoneal fluid (PF) and biopsies were collected during laparoscopy from 45 infertile women with pelvic endometriosis and 15 women without endometriosis. PF concentrations of hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), interleukin-6 (IL-6) and monocyte chemotactic protein-1 (MCP-1) were measured by enzyme-linked immunosorbent assay (ELISA). The immunohistochemical staining of HGF and proliferating cell nuclear antigen (PCNA) in opaque and nonopaque endometriotic lesions was examined using respective antibodies, and a computer-analyzed modified quantitative-histogram score (Q-H score) was used to quantify the immunoreaction. Opaque red lesions were defined as nontransparent lesions, including polypoid excrescence, blood bleb or ecchymosis, and nonopaque lesions were defined as translucent or transparent lesions, including vesicular bleb or clear papule. The women with endometriosis containing dominant opaque red lesions showed significantly higher PF concentrations of HGF, VEGF, IL-6 or MCP-1 than those with either nonopaque lesions or black lesions or control women. There was no difference in any of these cytokine or chemokine levels between women harboring nonopaque lesions and black lesions. Interestingly, PF concentrations of all these markers were significantly reduced in women containing true black lesions and true white lesions after excluding the presence of coexisting red spot from dominant black or white lesions. In a parallel fashion, the immunoexpressions of HGF and PCNA in intact tissue as shown by the Q-H score were significantly stronger in the opaque red lesions than in the nonopaque lesions or black lesions. Opaque red lesions display a higher activity than with nonopaque red lesions in both PF and intact tissue. This indicates different histogenesis in their generation in the pelvic cavity.


In vitro activation, purification, and characterization of Escherichia coli expressed aryl-alcohol oxidase, a unique H2O2-producing enzyme.
Aryl-alcohol oxidase (AAO), a flavoenzyme with unique spectral and catalytic properties that provides H2O2 for fungal degradation of lignin, has been successfully activated in vitro after Escherichia coli expression. The recombinant AAO (AAO*) protein was recovered from inclusion bodies of E. coli W3110 transformed with pFLAG1 containing the aao cDNA from Pleurotus eryngii. Optimization of in vitro refolding yielded 75% active enzyme after incubation of AAO* protein (10 microg/ml) for 80 h (at 16 degrees C and pH 9) in the presence of glycerol (35%), urea (0.6 M), glutathione (GSSG/GSH molar ratio of 2), and FAD (0.08 mM). For large-scale production, the refolding volume was 15-fold reduced and over 45 mg of pure active AAO* was obtained per liter of E. coli culture after a single anion-exchange chromatographic step. Correct FAD binding and enzyme conformation were verified by UV-visible spectroscopy and circular dichroism. Although the three enzymes oxidized the same aromatic and aliphatic polyunsaturated primary alcohols, some differences in physicochemical properties, including lower pH and thermal stability, were observed when the activated enzyme was compared with fungal AAO from P. eryngii (wild enzyme) and Emericella nidulans (recombinant enzyme), which are probably related to the absence of glycosylation in the E. coli expressed AAO.


Water reuse: >90% water yield in MBR/RO through concentrate recycling and CO2 addition as scaling control.
Over 1.5 years continuous piloting of a municipal wastewater plant upgraded with a double membrane system (ca. 0.6 m(3) d(-1) of product water produced) have demonstrated the feasibility of achieving high water quality with a water yield of 90% by combining a membrane bioreactor (MBR) with a submerged ultrafiltration membrane followed by a reverse osmosis membrane (RO). The novelty of the proposed treatment scheme consists of the appropriate conditioning of MBR effluent prior to the RO and in recycling the RO concentrates back to the biological unit. All the 15 pharmaceuticals measured in the influent municipal sewage were retained below 100 ng L(-1), a proposed quality parameter, and mostly below detection limits of 10 ng L(-1). The mass balance of the micropollutants shows that these are either degraded or discharged with the excess concentrate, while only minor quantities were found in the excess sludge. The micropollutant load in the concentrate can be significantly reduced by ozonation. A low treated water salinity (<10 mM inorganic salts; 280 ± 70 μS cm(-1)) also confirms that the resulting product has a high water quality. Solids precipitation and inorganic scaling are effectively mitigated by lowering the pH in the RO feed water with CO(2) conditioning, while the concentrate from the RO is recycled to the biological unit where CO(2) is stripped by aeration. This causes precipitation to occur in the bioreactor bulk, where it is much less of a process issue. SiO(2) is the sole exception. Equilibrium modeling of precipitation reactions confirms the effectiveness of this scaling-mitigation approach for CaCO(3) precipitation, calcium phosphate and sulfate minerals.


Respiratory Center Function and Its Impact in Obesity Hypoventilation Syndrome Treatment.
Patients with obesity hypoventilation syndrome (OHS) need treatment with positive pressure either with continuous (CPAP) or double pressure (NIV). The apnea-hypopnea index (AHI) is considered a key data for making therapeutic decisions. We hypothesized that HR may be an useful tool to establish different phenotypes and individualize treatment in patients with OHS. Our objective was to analyze the role of the respiratory center response to hypercapnia (HR) in the adequacy of positive airway pressure therapy. We included subjects with OHS treated with CPAP or NIV according to AHI and baseline pCO2. We analyzed therapeutic effectiveness and treatment changes prioritizing CPAP if AHI>30/h. Therapy was considered adequate if it was effective after two years. HR was measured with the p0.1/pEtCO2 ratio and its capability to select therapy was analyzed. The statistical study was performed by means comparison (Student's t) and multivariate analysis (logistic regression). 67 subjects were included of 68(11) years old, 37 (55%) males, initially 45 (67%) treated with NIV and 22 (33%) with CPAP, one case was excluded and in 25 (38%) the treatment was changed. Finally, CPAP was adequate for 29 subjects (44%) and NIV for 37 (56%). The CPAP group showed AHI 57/h (24) and p0.1/pEtCO2 0.37cmH2O/mmHg (0.23), NIV group AHI 43/h (35) and p0.1/pEtCO2 0.24 (0.15) with p=0.049 and 0.006. In multivariate analysis, p0.1/pEtCO2 (p=0.033) and AHI>30 (p=0.001) were predictors of adequate therapy. Measuring the RH of the respiratory center helps to select the most appropriate treatment for patients with OHS.


Circadian differences in emergence from volatile anaesthesia in mice: involvement of the locus coeruleus noradrenergic system.
Circadian differences in the induction, maintenance, or emergence from volatile anaesthesia have not been well studied. The minimal alveolar concentration (MAC) for preventing movement in response to a painful stimulus, MAC for loss of righting reflex (MACLORR), and MAC for recovery of righting reflex (MACRORR) in C57BL/6J male mice with isoflurane or sevoflurane exposure were measured during either the light or dark phase. Time to onset of loss of righting reflex (TimeLORR) and recovery of righting reflex (TimeRORR) upon exposure to 1 MAC of isoflurane or sevoflurane were determined. EEG was also monitored in the light and dark phase under isoflurane or sevoflurane exposure. The noradrenergic toxin N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine (DSP-4) was used to deplete noradrenergic neurones in the locus coeruleus to explore the impact of norepinephrine on these measurements. MACLORR, TimeLORR, and MAC did not show light- or dark-phase-dependent variations for either isoflurane or sevoflurane exposure. However, MACRORR was higher and TimeRORR was shorter in the dark phase than in the light phase for both isoflurane and sevoflurane exposure. The EEG delta wave power was higher but theta wave power was lower in the light phase than that in the dark phase during the rest state and emergence of anaesthesia. These light- and dark-phase-dependent changes in emergence were abolished in DSP-4-treated mice. Our data show that circadian differences exist during emergence but not during induction or maintenance of sevoflurane or isoflurane anaesthesia. The locus coeruleus noradrenergic system may contribute to these differences.


Immune consequences of protracted host-tumor interactions in a transgenic mouse model of mammary carcinoma.
A transgenic mouse model of autochthonous mammary carcinoma was chosen to study the impact of tumor progression on the immune system over an extended period. We found: i) that splenocyte numbers, particularly myeloid cells, increased concurrently with tumor burden; ii) the percentage of tumor-infiltrating Treg cells was similar to that in human breast cancer; iii) suppressed T cell proliferation and cytokine production and; iv) significantly elevated MCP-1 and TNF-alpha in the sera of tumor-bearing mice. The modified immune status in these tumor-bearing hosts is consistent with a "syndrome" that likely impacts the efficacy of cancer immunosurveillance and response to therapy.


Site-specific oligonucleotide binding represses transcription of the human c-myc gene in vitro.
A 27-base-long DNA oligonucleotide was designed that binds to duplex DNA at a single site within the 5' end of the human c-myc gene, 115 base pairs upstream from the transcription origin P1. On the basis of the physical properties of its bound complex, it was concluded that the oligonucleotide forms a colinear triplex with the duplex binding site. By means of an in vitro assay system, it was possible to show a correlation between triplex formation at -115 base pairs and repression of c-myc transcription. The possibility is discussed that triplex formation (site-specific RNA binding to a DNA duplex) could serve as the basis for an alternative program of gene control in vivo.


Variation in plasma leptin levels in response to fasting in Antarctic fur seals (Arctocephalus gazella).
Plasma leptin levels were determined in 8 lactating female and 20 pup Antarctic fur seals (Arctocephalus gazella) during fasting periods of normal duration. Plasma leptin levels ranged from 1.35 3.19 ng x ml(-1) in lactating females and 1.79-4.80 ng x ml(-1) in pups and were not positively correlated with body mass or condition. A negative trend, however, was observed between plasma leptin levels and body condition in lactating females upon their arrival at the colony following a foraging trip (beginning of fast). In accordance with findings in other species, plasma leptin levels dropped significantly (P < 0.02) in response to the 17-19% drop in body mass experienced by pups during fasting. In contrast, plasma leptin levels in lactating females increased during the first 24 h of fasting before decreasing throughout the remaining 48 h of the fast. This unexpected result could be due to the high level of energy expenditure by seals as they swim back to the colony (i.e. post-exercise response) or may be influenced by the intense suckling activity experienced by females during the onshore fasting periods. The results of this study support recent findings in other carnivore species which suggest the primary physiological role of leptin in these species may not necessarily be as a signal of the magnitude of body energy reserves.


Eye movements of teenage children of schizophrenics: a possible inherited marker of susceptibility to the disease.
Teenage children each having one schizophrenic parent showed deficits in oculomotor control which are also found in schizophrenic subjects compared with controls. They made significantly more saccadic eye movements interrupting smooth pursuit ocular tracking. They also made significantly more double-jump saccadic movements in a looking task. Since this latter difference was true only when the visual target was illuminated during the saccade (p less than 0.001), the cause was likely visual processing and not a fixation stability problem. The two measures correlated significantly for all subjects (r = 0.63), and half the experimentals had scores more than two standard deviations above the mean for controls. This suggests that a pattern of oculomotor dysfunction found in schizophrenics which comes from a single source may act as a marker for susceptibility of their offspring to the disease.


Small-bowel tumors in the elderly 65+ years: 10 years of experience.
Primary, secondary, benign or malignant tumorous diseases of the small intestine are rare. They are very often diagnosed by accident or as a cause of acute abdomen. Less serious symptoms can be overseen mainly in older patients. This work represents a prospective evaluation of a diagnostic and therapeutic algorithm in the management of the small-intestine tumor below the ligamentum Treitzi in patients over 65 years of age. A population of 96 patients who underwent surgery for a small-intestinal tumor in our hospital from 1996 to 2006 is presented. Thirty-four of these patients were older than 65 years. Ultrasound of the small intestine was sufficient to make the diagnosis in 21 of them. Elective surgery (laparoscopy was highly preferred) could be offered to 31 of these older patients. Perioperative mortality was zero. The small-bowel ultrasound can be recommended as the first choice method. All complaints regarding the gastrointestinal tract should be verified. Older patients can be safely offered an elective laparoscopic surgery with acceptable morbidity and mortality.


Emotion-related hemisphere asymmetry: subjective emotional responses to laterally presented films.
To investigate whether the cerebral hemispheres differ in their subjective emotional responses 54 adult subjects were presented two films of different emotion-related qualities (positive and negative film) either to their left or right hemisphere. The films were exposed by means of a technique for the lateralization of visual input that allows prolonged viewing while permitting free ocular scanning. Subjective emotional responses were assessed by means of a continuous rating of emotional arousal experienced during the movie as well as by retrospective ratings of ten different emotional qualities. Presenting both films to the right hemisphere resulted in stronger subjective responses in the continuous emotion rating as well as in some retrospectively assessed ratings compared to left-hemispheric presentation. The effects were more pronounced for the negative film. Taken together, the findings suggest a higher responsiveness of the right hemisphere in subjective emotional experience.


Survival improvements for advanced stage nonbronchioloalveolar carcinoma-type nonsmall cell lung cancer cases with ipsilateral intrapulmonary nodules.
Survival improvements have been demonstrated for patients with bronchioloalveolar (BAC) nonsmall cell lung cancer (NSCLC) with intrapulmonary satellite T4 nodules compared with other patients with stage IIIB disease, and for ipsilateral intrapulmonary M1 tumors versus contralateral or distant metastasis. However, it is not known whether these differences are observed in patients with non-BAC NSCLC. A case-only analysis of the U.S. Surveillance, Epidemiology, and End Results (SEER) data (1999-2003) was conducted. Overall survival (OS) and lung cancer-specific survival (LCSS) univariate analyses were conducted using the Kaplan-Meier method. Multivariate survival analyses were performed using Cox proportional hazards ratios. A total of 27,435 incident cases of histologically confirmed, advanced stage NSCLC were identified. Cases with stage IIIB NSCLC due to multiple lesions in the same lobe (n = 633) had a significantly improved median OS (21 months) and LCSS (31 months) compared with other cases with stage IIIB NSCLC (n = 7695), with an OS of 7 months and an LCSS of 9 months (P < .0001 for both comparisons). Among cases with stage IV NSCLC, those with intrapulmonary nodules (n = 3010) had a significantly improved median OS (9 months) and LCSS (10 months) compared with those with distant metastasis (n = 16,097), with an OS of 4 months and an LCSS of 5 months (P < .0001 for both comparisons). These survival differences persisted after adjustment for age, sex, ethnicity, surgery, and radiotherapy. Among cases with stage IV NSCLC, those with ipsilateral intrapulmonary nodules (n = 1120) had improved OS (12 months) compared with those with bilateral intrapulmonary nodules (n = 1890), with an OS of 7 months (P < .0001). Cases with stage IIIB and IV NSCLC with ipsilateral intrapulmonary nodules were found to have improved survival outcomes compared with other cases with stage IIIB and IV disease. The results of the current study add additional support for modifications to the current NSCLC staging system.


Child safety seat use for infants with Pierre Robin sequence.
To determine what child restraints would accommodate infants with Pierre Robin sequence who often require special attention in motor vehicle travel since microagnathia usually requires a prone position to keep the infant's airway open. Dynamic testing and clinical trial. An Indiana children's hospital providing primary and tertiary care. Four patients with Pierre Robin sequence are described to illustrate use of the modified infant car seat and the appropriateness of the car bed restraints for meeting requirements for prone positioning during travel. Convenience sample. Selected restraints were loaned to families through a clinical setting until the patient was able to use a conventional child restraint. Three child restraint systems were determined to accommodate the prone position necessary to keep the airway open for children with Pierre Robin sequence. Dynamic crash testing demonstrated the crashworthiness of an infant car seat modified to allow for prone positioning. Through a clinical trial, two car bed restraints were also found to provide safe prone positioning of infants. To enable safe transportation for infants with Pierre Robin sequence, health care providers can direct parents to appropriate resources for travel and can monitor the airway and oxygenation of the infant with Pierre Robin sequence before hospital discharge.


Impaired catabolism of very low-density lipoprotein-triglyceride in a family with primary hypertriglyceridemia.
In this report, kinetic studies of plasma very low-density lipoprotein-triglyceride (VLDL-TG) were examined in five brothers (three affected and two unaffected) from a family with primary hypertriglyceridemia. Synthesis and catabolism of VLDL-TG were studied by in vivo labelling of plasma TG with 3H-glycerol, and multicompartmental analysis of the plasma die-away curves. Results of the kinetic studies revealed the following information: (1) one brother, who had the highest plasma TG level and was obese, had both overproduction and a reduced fractional catabolic rate (FCR) of VLDL-TG; (2) second brother, who had moderate hypertriglyceridemia, had a low FCR and high-normal synthesis of VLDL-TG; (3) a third, who had only mildly elevated TG, had a low FCR and normal synthesis of VLDL-TG; and (4) the two normolipidemic brothers had neither overproduction nor decreased FCR of VLDL-TG. The composition of the soluble apoproteins of VLDL was normal. The apoprotein E phenotypes were E4/3 in four brothers, and E3/2 in the fifth. We have reached the following conclusions regarding this family: (1) the common kinetic abnormality of VLDL-TG metabolism in the hypertriglyceridemic brothers was a low clearance of VLDL-TG; (2) impaired catabolism of VLDL could not be explained by the apoprotein C or E patterns; and (3) the most severe hypertriglyceridemia occurred when the decreased FCR was present in conjunction with VLDL-TG overproduction due to obesity. Thus, a moderate defect in catabolism of plasma TG appears to be responsible for one familial form of primary hypertriglyceridemia.


Fluorescence quenching and time-resolved fluorescence studies on Trichosanthes dioica seed lectin.
Fluorescence quenching and time-resolved fluorescence studies have been carried out on the Trichosanthes dioica seed lectin (TDSL). The emission lambdamax of native TDSL, seen at 328nm, shifts to 343nm upon denaturation with 6M guanidinium chloride. Quenching titrations were performed with neutral (acrylamide and succinimide) and ionic (I(-) and Cs(+)) quenchers in order to probe the exposure and accessibility of tryptophan residues of the protein. Maximum quenching was observed with acrylamide, followed by succinimide, iodide and Cs(+). Dramatic increase in the extent of quenching and other quenching parameters by all the quenchers were observed upon denaturation of TDSL, suggesting that all the tryptophan residues in native TDSL are buried in the hydrophobic core of the protein. Increase in the extent of quenching upon denaturation of TDSL was maximum with I(-) and minimum with Cs(+), suggesting the presence of positively charged residue(s), near at least one tryptophan residue. Addition of saccharide ligands such as methyl-beta-d-galactopyranoside and lactose led to a small, but reproducible decrease in the fluorescence intensity of the lectin. The presence of lactose provided a partial protection against quenching by I(-), Cs(+) and succinimide, but not acrylamide. In time-resolved fluorescence measurements the fluorescence decay curves could be best fitted to biexponential patterns with lifetimes of 4.09 and 1.53ns for native lectin, 3.40 and 1.65ns for the lectin in presence of 0.1M lactose and 3.50 and 1.40ns for denatured lectin.


The Changing Epidemiology of Central Nervous System Infection: Can Radiologists Keep Up?
Diagnostic radiologists can increase their clinical value by supplementing image pattern recognition with knowledge of epidemiology and geographic distribution of central nervous system (CNS) infections and their causative organisms. This article reviews the changing global disease patterns, as well as zoonotic outbreaks of henipaviruses, coronaviruses, and other emerging, reemerging, and vector-borne organisms; case examples highlight typical imaging features of CNS infections and their mimics. Technical advances in neuroimaging help to enhance the value of radiologists to the multidisciplinary team and the responses to future pandemic preparation.


Effects of Alpinae Oxyphyllae Fructus on microglial polarization in a LPS-induced BV2 cells model of neuroinflammation via TREM2.
As one of the important traditional Chinese medicines, Alpinia oxyphylla could warm and tonify the kidney and spleen. It has been used as anti-salivation, anti-diarrhea in various diseases. In recent years, many studies have reported the significant effect of Alpinia oxyphylla on improving cognitive ability, anti oxidative stress and protecting neurons. In this paper, we studied whether AE and its main active components could improve M1 and M2 polarization, inhibit neuroinflammation through triggering receptor expressed on myeloid cells 2 (TREM2), and exert anti-inflammatory effects. In this paper, the concentrations of inflammatory cytokines such as NO, TNF-α, IL-10 were assessed using detection kits respectively. Arg-1 and Iba-1, as polarized markers of M1 and M2, were detected by Immunofluorescence staining. CD86 and CD206 were tested by flow cytometry as surface markers of M1 and M2. Furthermore, RT-PCR was performed to determinate TNF-α, IL-10, Arg-1, and Iba-1. Western blot was used to test the activation of PI3K/AKT/GSK3β and BDNF/TrkB/TLR4 signaling pathways. TREM2 siRNA treatment further verified the action target of Chrysin, the main active ingredient of Alpinia oxyphylla. Molecular docking study was performed to investigate the binding mode between Chrysin and the human TREM2. We found that AE could promote the phenotypic transformation of microglia from M1 to M2, and similar effects of Chrysin were observed. Furthermore, downregulation of TREM2 blocked the anti-neuroinflammation of Chrysin, and inhibited the shift of M1 phenotype to M2 phenotype. Additionally, TREM2-siRNA suppressed the effects of Chrysin on PI3K/AKT/GSK3β and BDNF/TrkB/TLR4 signaling pathways. Our findings indicated that AE could improve the polarization response of microglia. TREM2 plays a vital role in the microglial repolarization effects of Chrysin through PI3K/AKT/GSK3β and BDNF/TrkB/TLR4 signaling pathways regulated by neuroinflammation.


Fasting and postprandial serum bile acid concentrations in 10 healthy female red-eared terrapins (Trachemys scripta elegans).
The fasting and postprandial serum concentrations of bile acids and other blood constituents were measured in a group of 10 clinically healthy, female, six-year-old captive red-eared terrapins (Trachemys scripta elegans). The terrapins were housed in a temperate room and maintained in four aquaria in which the water temperature ranged from 24 to 27 degrees C and the temperature above the basking site ranged from 27 to 30 degrees C. The serum concentrations of bile acids were measured four times in a period of five months, and at the second sampling the fasting and two postprandial (after 24 and 48 hours) serum concentrations of total protein, albumin, glucose, uric acid, cholesterol, triglycerides, alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase, and bile acids were determined. Coelioscopy revealed vitellogenic and previtellogenic follicles on the ovaries of all the terrapins, and eggs with calcified shells were detected in two of them. The livers were mostly pink to deep yellow in colour, with sharp edges, a smooth serosal surface, distinct large superficial vessels, and multifocal melanin deposits. Liver biopsies revealed fine, more or less oil red O-positive lipid droplets in all the hepatocytes, but in none of the cases was it considered to be pathological lipidosis. The mean (sd) bile acid concentrations ranged from 7.35 (4.52) to 10.04 (7.40) micromol/l. The fasting and postprandial concentrations were 3.1 (2.3), 4.5 (5.4) (24 hours) and 2.2 (1.5) (48 hours) micromol/l. High concentrations between 27.6 and 66.6 micromol/l were associated with lipaemia. There were no significant differences between the biochemical profiles of the fasting and postprandial serum samples.


Glide insertion technique for donor cornea lenticule during Descemet's stripping automated endothelial keratoplasty.
We describe a simple technique for inserting the donor corneal lenticule in Descemet's stripping automated endothelial keratoplasty. Unlike the standard insertion method, which requires folding the donor lenticule, this technique uses a modified intraocular lens (IOL) glide to guide the unfolded donor lenticule through the small incision with an intraocular forceps. This prevents significant endothelial trauma and surgical difficulties when the lenticule is unfolded. Additional endothelial protection is provided by an ophthalmic visco-surgical device on the IOL glide, which protects the donor endothelium from the corneal wound edge and prevents iris prolapse during insertion.


Differential adsorption of variants of the Thermomyces lanuginosus lipase on a hydrophobic surface suggests a role for local flexibility.
Lipases are activated at interfaces between aqueous and hydrophobic phases, where they typically undergo conformational changes leading to significant activity increase. Here I use a quartz crystal microbalance with dissipation (QCM-D) to study changes in layer thickness and viscosity during the adsorption of variants of the Thermomyces lanuginosus lipase (TlL) onto a methyl-terminated hydrophobic surface. Unlike wildtype TlL, the variant Mut1, which shows improved performance under certain test conditions, shows a large dissipation increase during the binding process, leading to a significantly thicker layer. This altered adsorption behaviour may be linked to Mut1's changes in secondary structure. This is corroborated by the fact that four other TlL mutants with unaltered secondary structure showed wildtype-like absorption behaviour. Unlike wildtype TlL and the other variants, Mut1 contains several consecutive basic residues introduced into the C-terminal region which is close in space to the N-terminal part of the protein, which also contains several basic residues. Electrostatic repulsion between these two regions leading to local structural flexibility may facilitate altered adsorption behaviour and ultimately to improved enzymatic performance on a solid surface. QCM-D thus provides a good approach to screen protein variants for their adsorption properties on hydrophobic surfaces.


What are the correlates of substance abuse treatment counselor salaries?
This article identifies and measures the key correlates of substance abuse treatment (SAT) counselor salaries. The data come from the 2002-2003 wave of the National Treatment Center Study that consists of approximately 1,500 full-time counselors from nationally representative samples of public and private SAT facilities. We used interval regression models to estimate the relationship between annual salary and numerous counselor and organizational characteristics. We found that counselor characteristics associated with annual salary include education, tenure in the SAT field, licensure, race, recovery status, and administrative responsibilities above and beyond duties as a full-time counselor. Organizational characteristics associated with counselor salary include accreditation status, hospital ownership, and the population density of the county where the facility is located. Taken together, these factors explain approximately 50% of the total variation in SAT counselor salaries.


Implementing machine learning in bipolar diagnosis in China.
Bipolar disorder (BPD) is often confused with major depression, and current diagnostic questionnaires are subjective and time intensive. The aim of this study was to develop a new Bipolar Diagnosis Checklist in Chinese (BDCC) by using machine learning to shorten the Affective Disorder Evaluation scale (ADE) based on an analysis of registered Chinese multisite cohort data. In order to evaluate the importance of each item of the ADE, a case-control study of 360 bipolar disorder (BPD) patients, 255 major depressive disorder (MDD) patients and 228 healthy (no psychiatric diagnosis) controls (HCs) was conducted, spanning 9 Chinese health facilities participating in the Comprehensive Assessment and Follow-up Descriptive Study on Bipolar Disorder (CAFÉ-BD). The BDCC was formed by selected items from the ADE according to their importance as calculated by a random forest machine learning algorithm. Five classical machine learning algorithms, namely, a random forest algorithm, support vector regression (SVR), the least absolute shrinkage and selection operator (LASSO), linear discriminant analysis (LDA) and logistic regression, were used to retrospectively analyze the aforementioned cohort data to shorten the ADE. Regarding the area under the receiver operating characteristic (ROC) curve (AUC), the BDCC had high AUCs of 0.948, 0.921, and 0.923 for the diagnosis of MDD, BPD, and HC, respectively, despite containing only 15% (17/113) of the items from the ADE. Traditional scales can be shortened using machine learning analysis. By shortening the ADE using a random forest algorithm, we generated the BDCC, which can be more easily applied in clinical practice to effectively enhance both BPD and MDD diagnosis.


Hormonal modulation of phonotaxis and advertisement-call preferences in the gray treefrog (Hyla versicolor).
Hormonal levels fluctuate during the breeding season in many anurans, but the identity of the hormones that modulate breeding behavior and their effects remain unclear. We tested the influence of a combined treatment of progesterone and prostaglandin on phonotaxis, the key proceptive reproductive behavior of female anurans. First, we found that female gray treefrogs (Hyla versicolor) treated with progesterone and prostaglandin exhibited phonotaxis to synthetic male advertisement signals significantly more often than animals treated with ringers vehicle or uninjected controls. Responsive females had greater levels of plasma progesterone and estradiol compared to both control groups, suggesting that these steroids may be promoting phonotaxis. Second, we found that the selectivity of hormonally-induced phonotaxis in H.versicolor was similar to that observed in freshly captured breeding animals. Females made the same choices between acoustic signals after hormone treatments in tests of frequency, call rate and pulse rate, compared to their responses without treatment immediately after collection from the breeding chorus. The preference for a longer call was, however, significantly weaker after hormonal induction of phonotaxis. Hormonally primed females were also less likely to respond in any test and took longer to respond than did freshly collected females. Consequently, our study shows how progesterone-prostaglandin induced phonotaxis in female treefrogs influences both the quality and quantity of phonotaxis, relative to that exhibited by naturally breeding females.


Plasma levels of platelet factor 4 measured by radioimmunoassay.
A radioimmunoassay has been developed to measure platelet factor 4 (PF4) in biological fluids both in vitro and in vivo. The assay has been shown to be highly specific for PF4 and has a sensitivity of 0.08 ng/assay tube and 1.6 ng/ml of plasma. The preparation of plasma for the measurement of in vivo levels of PF4 requires the use of an anticoagulant containing EDTA, theophylline and prostaglandin E1, the immediate cooling of the blood and high speed or prolonged centrifugation to reduce platelet contamination. Plasma levels of PF4 are normally between 4 and 24 ng/ml with a median of 7.4 ng/ml. Plasma PF4 levels are markedly increased during cardiopulmonary bypass surgery with shortened 51Cr-labelled platelet survival times and during arterial thrombosis. However, despite similarly shortened platelet survival times, the level of PF4 is normal in immune thrombocytopenia. Elevations of plasma PF4 levels are found following surgery, acute myocardial infarction and frequently during acute infections and in inflammatory states. On the contrary, normal levels are usual in disseminated malignancy, in severe hepatic and renal disease and in chronic arterial disease. The measurement of PF4 is a useful addition to the study of platelet pathophysiology. It is apparent, however, that raised levels are by no means specific for thromboembolic disease and similarly platelet destruction is not invariably associated with abnormally increased plasma PF4 levels.


Evaluation of circulating microRNA profiles in blood as potential candidate biomarkers in a subacute ruminal acidosis cow model - a pilot study.
Subacute ruminal acidosis (SARA) is a metabolic disorder often observed in high-yielding dairy cows, that are fed diets high in concentrates. We hypothesized that circulating miRNAs in blood of cows could serve as potential candidate biomarkers to detect animals with metabolic dysbalances such as SARA. MicroRNAs (miRNAs) are a class of small non-coding RNAs, serving as regulators of a plethora of molecular processes. To test our hypothesis, we performed a pilot study with non-lactating Holstein-Friesian cows fed a forage diet (FD; 0% concentrate, n = 4) or a high-grain diet (HG; 65% concentrate, n = 4) to induce SARA. Comprehensive profiling of miRNA expression in plasma and leucocytes were performed by next generation sequencing (NGS). The success of our model to induce SARA was evaluated based on ruminal pH and was evidenced by increased time spent with a pH threshold of 5.8 for an average period of 320 min/d. A total of 520 and 730 miRNAs were found in plasma and leucocytes, respectively. From these, 498 miRNAs were shared by both plasma and leucocytes, with 22 miRNAs expressed exclusively in plasma and 232 miRNAs expressed exclusively in leucocytes. Differential expression analysis revealed 10 miRNAs that were up-regulated and 2 that were down-regulated in plasma of cows when fed the HG diet. A total of 63 circulating miRNAs were detected exclusively in the plasma of cows with SARA, indicating that these animals exhibited a higher number and diversity of circulating miRNAs. Considering the total read counts of miRNAs expressed when fed the HG diet, differentially expressed miRNAs ( log2 fold change) and known function, we have identified bta-miR-11982, bta-miR-1388-5p, bta-miR-12034, bta-miR-2285u, and bta-miR-30b-3p as potential candidates for SARA-biomarker in cows by NGS. These were further subjected to validation using small RNA RT-qPCR, confirming the promising role of bta-miR-30b-3p and bta-miR-2285. Our data demonstrate that dietary change impacts the release and expression of miRNAs in systemic circulation, which may modulate post-transcriptional gene expression in cows undergoing SARA. Particularly, bta-miR-30b-3p and bta-miR-2285 might serve as promising candidate biomarker predictive for SARA and should be further validated in larger cohorts.


Separation of isolectins by high-performance hydrophobic interaction chromatography.
High-performance hydrophobic interaction chromatography (HP-HIC) was found to be an effective method for the separation of lectins into isolectin fractions. All of the purified lectins used in this study, Phaseolus vulgaris haemagglutinin (PHA), wheat germ agglutinin (WGA), Ricinus communis agglutinin (RCA), and Arachis hypogaea agglutinin (AHA), were prepared by affinity chromatography. HP-HIC was performed on a column (15 X 2.1 cm) of TSK gel Phenyl-5PW at room temperature. The lectin sample, dissolved in 1.0 or 0.5 M ammonium sulphate in phosphate buffered saline (pH 7.4) (PBS), was applied to the column and eluted with a linear gradient from 1.0 or 0.5 M ammonium sulphate in PBS to 0 M ammonium sulphate in PBS at a flow-rate of 4 ml/min. In the case of RCA, addition of glycerol to the elution buffer resulted in sharper isolectin peaks. PHA, WGA, RCA, and AHA were rapidly separated into 5, 5, 4, and 6 isolectins, respectively.


Evolution of morphology in electrodeposited nanocrystalline Co-Ni films by in-situ high magnetic field application.
The effect of high magnetic fields up to 12 T, applied during electrodeposition process, on the morphology of nanocrystalline CoNi films has been investigated. The magneto-induced dramatic modifications in the morphology were observed by using field-emission scanning electronic microscopy and atomic force microscopy. Along with the increase of magnetic flux density (B), the grain size and the surface roughness of the films increased to reach a maximum value at a field of 9 T. Meanwhile, higher magnetic flux density could improve cobalt atomic percentage in the film due to the impacts of magnetohydrodynamic effect. However, at a high field of 12 T, the paramagnetic force played a predominant role in a decrease of mass transport, resulting in minimum grain size and roughness, even smaller than that of the ordinarily (B=0 T) sample.


Delineating an epigenetic continuum in head and neck cancer.
A tissue field of somatic genetic alterations precedes the histopathological phenotypic changes of carcinoma. Genomic changes could be of potential use in the diagnosis and prognosis of pre-invasive squamous head and neck carcinoma (HNSCC) lesions and as markers for cancer risk assessment. Studies of sequential molecular alterations and genetic progression of pre-invasive HNSCC have not been clearly defined. Studies have shown recurring alterations at chromosome 9p21 (location of the CDKN2A) and TP53 mutations in the early stages of HNSCC. However, gene silencing via hypermethylation is still a relatively new idea in the development of HNSCC and little is known about the contribution of epigenetics to the development of neoplasia, its transformation, progression, and recurrence in HNSCC. This review examines the role of promoter hypermethylation of tumor suppressor genes in the progression continuum from benign papillomas to malignancy in HNSCC.


Extracellular vesicles and chronic obstructive pulmonary disease (COPD): a systematic review.
Chronic Obstructive Pulmonary Disease (COPD) is a common inflammatory disease of the airways characterized by irreversible airflow limitation, ranking the third highest cause of death worldwide. Extracellular vesicles (EVs) are important intercellular communication mediators released by cells into their extracellular environment with the capacity to transfer biological signals. EVs involved in COPD hold great potential to understand disease pathogenesis and identify important biomarkers. This systematic review aims to examine all available research on EVs in the pathogenesis and diagnosis of COPD to identify existing knowledge and support further research within the field. Publications were searched using PubMed and EMBASE with the search terms (Exosomes or extracellular vesicles or microvesicles or microparticles or ectosomes) AND (chronic obstructive pulmonary disease or COPD or emphysema or bronchitis). Initial search yielded 512 papers of which 142 were manually selected for review and 43 were eligible for analyses. The studies were divided into groups according to the role of EVs in pathogenesis, EV origin and cargo, their role in COPD exacerbations and their diagnostic utility. EVs were found to be involved in the mechanism of pathogenesis of COPD, derived from various cell types, as well as containing modified levels of miRNAs. EVs also varied according to the pathophysiological status of disease, therefore presenting a possible method for COPD diagnosis and progress monitoring. The current findings show the limited but good quality research looking at the role of EVs in COPD, demonstrating the need for more studies to better define and provide further insight into the functional characteristics of EV in COPD pathogenesis.


Amino sugar contents and phage inactivating properties of lipopolysaccharide from cholera and El Tor vibrios.
Cholera and El Tor lipopolysaccharide (LPS) were identical in respect of chemical composition except that the hexosamine content was significantly lower and galactosamine was absent in El Tor LPS. Mukerjee's group IV cholera phage phi149 was inactivated by cholera LPS but was resistant to El Tor LPS.


An empirical analysis of the dimensions of health status measures.
The objective of this study is to verify empirically the existence of separate dimensions in the overall concept of health status by analyzing 10 variables included in a questionnaire that was applied to all adults in a simple random sample of households in St John's, Newfoundland. The response rate was 85% for a total of 3300 subjects. These data were analyzed by frequencies and by associations with sex, age and education. Nonparametric correlation, factor and cluster analyses on variables were used to verify if health status had identifiable dimensions. All these methods produced similar results showing five distinct factors. The first factor is composed of variables related to disease (disability/chronic conditions/worry about health); the second, to happiness (happiness/emotional); the third, to subjective appraisal of health (physical condition/comparative level of energy/self-rated health status). Finally, the fourth and fifth factors were single variables; restriction of normal activities and social contacts. An interesting finding was that self-rated health status was distributed with almost equal weight in both the first and third factors. A validation of the 10 variables and the 5 factors was undertaken by studying their association with health care utilization. Two measures of utilization were used; number of physicians' visits in a year and number of hospital days in a 4-year period. Number of chronic conditions, disability and self-rated health status were associated with both measures of utilization; factor 1 was the only summary construct showing association with utilization. This paper demonstrates that self-rated health status is valid as a single measure of overall health status in this sample, being associated with both disease and subjective assessment components.


Gamma-secretase inhibitor enhances the cytotoxic effect of bortezomib in multiple myeloma.
Targeting of Notch signaling with γ-secretase inhibitors (GSIs) has been considered a promising strategy for the treatment of hematological malignancies including multiple myeloma (MM). Here we investigated whether the cytotoxic effect of bortezomib, an agent commonly used in MM, could be enhanced by the addition of a GSI. MM cells were treated with GSI, bortezomib or the combination thereof. Apoptosis of MM cells, proteasome activity and Notch signaling activation were determined. The effect of the drug combination was also evaluated in MM cells transfected with the active domain of Notch-1. Using MM cell lines and primary MM cells isolated from the bone marrow of patients with MM we found a strong synergistic effect of bortezomib in combination with one of the GSIs studied. We next investigated the mechanism underlying this synergistic effect and determined that the effect of the drug combination was mainly dependent on the ability of the selected GSI to inhibit proteasome activity in MM cells. Our study demonstrates that selected GSIs that inhibit proteasome activity may be successfully used in combination with bortezomib enhancing its anti-MM effect.


Galactofuranosyl groups are immunodominant in Aspergillus fumigatus galactomannan.
Aspergillus fumigatus galactomannan was prepared and its structure partially characterized. Galactofuranosyl groups were immunodominant when this polysaccharide antigen was reacted with antibody raised in rabbits.


Progabide in the treatment of hyperkinetic extrapyramidal movement disorders.
The ability of the selective GABA-receptor agonist, progabide, to suppress abnormal involuntary movements was evaluated in a preliminary open pilot study. 17 patients, 10 males and 7 females, aged 10-78 years, with hyperkinetic movement disorders were included in the study. Daily doses of progabide ranged from 900 to 3600 mg (median 2400 mg) corresponding to 14-45 mg/kg (median 45 mg/kg), while the duration of treatment varied from 2 to 52 weeks. Improvement, with a reduction of involuntary movements exceeding 25%, occurred in two of four patients with Gilles de la Tourette's syndrome, and in two of three patients with postanoxic intention myoclonus, while no consistent beneficial effects were registered in ten patients with Huntington's chorea, postanoxic choreoathetosis, torsion dystonia, tardive dyskinesia, action tremor, essential myoclonus, or oro-branchio-respiratory myoclonus.


The Effect of Lanthanum (III) Nitrate on the Osteogenic Differentiation of Mice Bone Marrow Stromal Cells.
To study the species of lanthanum (III) nitrate (La[NO3]3) dispersed in cell media and the effect on the osteoblast differentiation of bone marrow stroma cells (BMSCs). Different La-containing precipitations were obtained by adding various concentrations of La(NO3)3 solutions to Dulbecco's modified Eagle medium (DMEM) or DMEM with fetal bovine serum (FBS). A series of characterisation methods, including dynamic light scattering, scanning electron microscopy, transmission electron microscopy, energy-dispersive X-ray spectroscopy, and protein quantification were employed to clarify the species of the different La-containing precipitations. The primary BMSCs were isolated, and the cell viability, alkaline phosphatase activity, and the formation of a mineralised nodule of BMSCs were tested when treated with different La-containing precipitations. The La(NO3)3 solutions in DMEM could form LaPO4, which exits in the particle formation, while the La(NO3)3 solutions in DMEM with FBS could form a La-PO4-protein compound. When treated with La(NO3)3 solutions in DMEM, the cell viability of the BMSCs was inhibited at the concentrations of 1, 10, and 100 μM at 1 day and 3 days. Meanwhile, the supernatant derived from the La(NO3)3 solutions in DMEM did not affect the cell viability of the BMSCs. In addition, the precipitate derived from the La(NO3)3 solutions in DMEM added to the complete medium inhibited the cell viability of the BMSCs at concentrations of 10 μM and 100 μM. When treated with La(NO3)3 solutions in DMEM with FBS, the derived precipitate and supernatant did not affect the cell viability of the BMSCs, except for the concentration of 100 μM La(NO3)3. The La-PO4-protein formed from the La(NO3)3 solutions in DMEM with FBS inhibited the osteoblast differentiation of BMSCs at the concentration of 1 μM La(NO3)3 (P < 0.05) but had no effect on either the osteoblast differentiation at the concentrations of 0.001 and 0.1 μM or on the formation of a mineralised nodule at all tested concentrations of La(NO3)3. Overall, La(NO3)3 solutions in different cell culture media could form different La-containing compounds: La-PO4 particles (in DMEM) and a La-PO4-protein compound (in DMEM with FBS). The different La-containing compounds caused different effects on the cell viability, osteoblast differentiation, and the formation of a mineralised nodule of the BMSCs. The La-containing precipitation inhibited the osteoblast differentiation by inhibiting the expression of osteoblast-related genes and proteins, providing a theoretical basis for clinical doctors to apply phosphorus-lowering drugs such as lanthanum carbon.


Methicillin-resistant Staphylococcus schleiferi subspecies coagulans associated with otitis externa and pyoderma in dogs.
Dermatological infections are the most common cases in the daily pet clinic. Since its discovery in 1990, Staphylococcus schleiferi subspecies coagulans have been reported more frequently in canine otitis externa and pyoderma and even in cases of zoonoses. Detect the presence of S. schleiferi subsp. coagulans of canine otitis externa and pyoderma, its antimicrobial resistance, and the presence of mecAgen. Three-hundred-thirty-one swabs from dogs with otitis externa and pyoderma were cultured on bacteriological agar for bacterial isolation and subsequent biochemical and molecular identification. The identified S. schleiferi subsp. coagulans were evaluated for their antimicrobial susceptibility using the Kirby-Bauer technique, including an oxacillin disk, and subsequently, a PCR was run to identify which ones had the mecA gene. Thirty-four (22.97%) and twelve (6.56%) isolates were identified as S. schleiferi subspecies coagulans from otitis externa and pyoderma, respectively. Fluoroquinolones, the most widely used group of antibiotics in Peru, showed a susceptibility of 58.82% (20/34) in cases of otitis externa and 50% (6/12) in cases of canine pyoderma. Meanwhile, nitrofurantoin was the antibiotic with the best efficacy in both cases, with 97% (33/34) in otitis externa and 83% (10/12) in pyoderma. Furthermore, 40% (13/34) of S. schleiferi subsp. coagulans isolated from otitis externa were resistant to methicillin, and 85.29% (29/34) had the mecA gene. On the other hand, the only methicillin-resistant isolate from pyoderma was also the only one with a mecA gene. This study is the first report of S. schleiferi subsp. coagulans in Peru, finding a higher percentage than reported in other South American countries.


Characterization of lipoproteins containing a truncated form of apolipoprotein B, apolipoprotein B32.
We have characterized the lipoproteins isolated from the plasma of a human subject who was heterozygous for a mutation yielding a truncated apolipoprotein B (apo-B32). The apo-B32 lipoproteins were isolated from the plasma high density lipoprotein (HDL) fraction by heparin-Sepharose chromatography. Although the chemical composition of the apo-B32-containing lipoproteins was similar to that of normal HDL, the mean diameter of the apo-B32 lipoproteins was larger than typical apo-AI-containing HDL particles. On agarose gels, the apo-B32 lipoproteins had pre-beta mobility similar to that of normal very low density lipoproteins. Analysis of the purified apo-B32 lipoproteins by SDS-polyacrylamide gel electrophoresis revealed the presence of both apo-AI and apo-E. In order to further analyze the properties of apo-B32, we developed an apo-B32 expression vector and generated stable rat hepatoma cell lines expressing apo-B32. In these cell lines, the apo-B32 protein was secreted in a d > 1.21 g/ml lipoprotein. Oleic acid supplementation of the cell-culture media had no measurable affect on the density distribution of the apo-B32 lipoproteins that were secreted by the cells.


Monoamines, amino acids and acetylcholine in the preoptic area and anterior hypothalamus of rats: measurements of tissue extracts and in vivo microdialysates.
A microbore column high-performance liquid chromatography (HPLC) system was used to measure neurotransmitters in tissue extracts and in vivo microdialysates obtained from the preoptic area (PO) and anterior hypothalamus (AH) of rats. The extracts contained norepinephrine, epinephrine, 3,4-dihydroxyphenylacetic acid (DOPAC), dopamine, 5-hydroxyindoleacetic acid (5-HIAA), homovanillic acid (HVA), 5-hydroxytryptamine (5-HT), aspartate, glutamate, GABA, acetylcholine (ACh) and choline. The microdialysates obtained from the PO and AH of freely moving rats contained all of these substances except for norepinephrine, epinephrine, dopamine, and 5-HT. During collection of microdialysate from the PO and AH, core body temperature and locomotor activity were simultaneously measured by means of telemetry. The locomotor activity and body temperature increased during the night. This was accompanied by increased levels of 5-HIAA. The results suggest that serotonergic neuronal mechanisms in the PO and AH may be involved in hypothalamic regulation of spontaneous behaviors and body temperature.


Early influx of horseradish peroxidase into axons of the hypoglossal nerve during Wallerian degeneration.
Horseradish peroxidase (HRP) was applied around mouse hypoglossal nerves which were damaged by a crush or a ligature. HRP was then visualized distal to the lesions by light- and electron microscopic histochemistry. At the injury the enzyme entered axons and could also be detected several millimetres down in the distal segment. By 24 h reaction product (r.p.) was either diffusely distributed in the axoplasm or present in various vesicular organelles. Our results indicate that there is a rapid influx of macromolecules into axons after a lesion to a nerve. A similar uptake of 'wound substances' into axons distal to an injury might well have some relation to the process by which axonal breakdown is initiated during Wallerian degeneration.


Molecular docking-based virtual screening, molecular dynamic simulation, and 3-D QSAR modeling of some pyrazolopyrimidine analogs as potent anti-filarial agents.
Lymphatic filariasis and onchocerciasis are common filarial diseases caused by filarial worms, which co-habit symbiotically with the Wolbachia organism. One good treatment method seeks Wolbachia as a drug target. Here, a computer-aided molecular docking screening and 3-D QSAR modeling were conducted on a series of Fifty-two (52) pyrazolopyrimidine derivatives against four Wolbachia receptors, including a pharmacokinetics study and Molecular Dynamic (MD) investigation, to find a more potent anti-filarial drug. The DFT approach (B3LYP with 6-31G** option) was used for the structural optimization. Five ligand-protein interaction pairs with the highest binding affinities were identified in the order; 23_7ESX (-10.2 kcal/mol) > 14_6EEZ (- 9.0) > 29_3F4R (- 8.0) > 26_6W9O (- 7.7) ≈ doxycycline_7ESX (- 7.7), with good pharmacological interaction profiles. The built 3-D QSAR model satisfied the requirement of a good model with R2 = 0.9425, Q2 LOO = 0.5019, SDEC = 0.1446, and F test = 98.282. The selected molecules (14, 23, 26, and 29) perfectly obeyed Lipinski's RO5 for oral bio-availability, and showed excellent ADMET properties, except 14 with positive AMES toxicity. The result of the MD simulation showed the great stability associated with the binding of 23 onto 7ESX's binding pocket with an estimated binding free energy (MM/GBSA) of - 60.6552 kcal/mol. Therefore, 23 could be recommended as a potential anti-filarial drug molecule, and/or template for the design of more prominent inhibitors. The online version contains supplementary material available at 10.1007/s40203-022-00136-y.


Stem cell therapy combined with core decompression versus core decompression alone in the treatment of avascular necrosis of the femoral head: a systematic review and meta-analysis.
Accumulated clinical trials had been focused on stem cell therapy in combination of core decompression (CD) in the treatment of avascular necrosis of the femoral head (ANFH). Nonetheless, the results were inconclusive. Here, we performed a systematic review and meta-analysis of previous randomized controlled trials (RCTs) and retrospective studies to assess whether combined stem cell augmentation with CD improved the outcomes of ANFH compared with CD alone. The current study included 11 RCTs and 7 retrospective studies reporting the clinical outcomes of a total of 916 patients and 1257 hips. 557 and 700 hips received CD and CD plus stem cell therapy, respectively. To compare CD with CD plus stem cell therapy, we examined the clinical evaluating scores, the occurrence of the femoral head, radiologic progression and conversion to total hip arthroplasty (THA). Only 10 studies reported significantly greater improvement in hip functions while combining stem cell procedure with CD. The pooled results in subgroup analysis indicated that stem cell group had a lower collapse rate on a mid-term basis (P = 0.001), when combined with mechanical support (P < 0.00001), and with extracted stem cells (P = 0.0002). Likewise, stem cell group had a lower radiographic progression rate at 2- to 5-year follow-up [P = 0.003], when combined with structural grafting (P < 0.00001), and with extracted stem cells (P = 0.004). Stem cell therapy resulted in an overall lower THA conversion rate (P < 0.0001) except that at a follow-up longer than 5 years. Stem cell therapy combined with core decompression was more effective in preventing collapse, radiographic progression and conversion to THA. Trial Registration The current protocol has been registered in PROSPERO with the registration number: CRD42023417248.


Respiratory Disorders Among Workers in Slaughterhouses.
Workers in slaughterhouses are exposed to a wide range of biological contaminants, such as bacteria and fungi, due to their working environment. This study aimed to assess the prevalence of respiratory disorders among workers in slaughterhouses. This study was conducted on 81 workers in slaughterhouses and 81 healthy office workers as a reference group. The American Thoracic Society standard respiratory symptoms questionnaire was used to determine the prevalence of respiratory disorders. Besides, lung function tests were conducted using a calibrated spirometer at the beginning (preshift) and at the end (postshift) of the 1st working day. Single-stage Anderson sampler was used to measure the concentration of bioaerosols in different parts of slaughterhouses. The prevalence of respiratory disorders, such as cough, productive cough, breathlessness, phlegm, and wheezing, was 3.17, 4.02, 3.07, 4.66, and 3.94 times, respectively, higher among workers in slaughterhouses compared with the reference group. The prevalence of respiratory disorders was significantly higher among workers in slaughterhouses. Thus, the significant reduction in the percentage predicted lung function among workers in slaughterhouses might be associated with exposure to bioaerosols in their work environment.


Small Molecule Dysregulation of TEAD Lipidation Induces a Dominant-Negative Inhibition of Hippo Pathway Signaling.
The transcriptional enhanced associate domain (TEAD) family of transcription factors serves as the receptors for the downstream effectors of the Hippo pathway, YAP and TAZ, to upregulate the expression of multiple genes involved in cellular proliferation and survival. Recent work identified TEAD S-palmitoylation as critical for protein stability and activity as the lipid tail extends into a hydrophobic core of the protein. Here, we report the identification and characterization of a potent small molecule that binds the TEAD lipid pocket (LP) and disrupts TEAD S-palmitoylation. Using a variety of biochemical, structural, and cellular methods, we uncover that TEAD S-palmitoylation functions as a TEAD homeostatic protein level checkpoint and that dysregulation of this lipidation affects TEAD transcriptional activity in a dominant-negative manner. Furthermore, we demonstrate that targeting the TEAD LP is a promising therapeutic strategy for modulating the Hippo pathway, showing tumor stasis in a mouse xenograft model.


Plasma equilin concentrations in an oophorectomized woman following ingestion of conjugated equine oestrogens (Premarin).
The plasma equilin concentration was 25.2 nmol/l (6.7 ng/ml) 4 h after an oral dose of Premarin (1.25 mg), and a maximum concentration of 20.5 nmol/l (5.5 ng/ml) was recorded 3 h after a smaller dose (0.625 mg) was given. 24 h later, the plasma equilin was still considerable, being 6.1 nmol/l (1.6 ng/ml) and 3.7 nmol/l (1.2 ng/ml) for the larger and smaller dosages respectively. The findings for equilin are considerably higher than physiological oestrogen concentrations and in addition show that daily therapy with Premarin could lead to cumulation of the steroid.


Old Fragments of Forest Inside an Urban Area Are Able to Keep Orchid Bee (Hymenoptera: Apidae: Euglossini) Assemblages? The Case of a Brazilian Historical City.
Retention of habitat fragments within the urban matrix can provide critical resources for the maintenance of regional biodiversity while still providing socio-economic value. Euglossini bees are important components in a community as they are important pollinators for economically valuable plants as well as hundreds of orchid species. However, some species are very sensitive to environmental impacts like urbanization. This study presents the role of antique urban fragments in a historical city in Brazil and compares it with a conservation area on the aspects of orchid bee assemblage, such as richness, composition, and abundance. Four fragments inside the city of Ouro Preto and three inside Parque Estadual do Itacolomi (PEIT) were sampled for Euglossini bees. Sorensen similarity index was used to compare community composition. The Mantel test was applied to verify the hypothesis that an urban center is a barrier for the mobility of the individuals. Fourteen Euglossini species from the region were registered. Close to 75% of the sampled bees were collected from the PEIT sampling areas. The fragments presented differences in Euglossini richness and abundance. A majority of the sampled fragments were dominated by the Eulaema cingulata Fabricius, Eulaema nigrita Lepeletier, and Euglossa securigera Dressler species. We found differences on community composition between the fragments localized in PEIT and those located in the urban center. The data suggest that there is a possible flux of individuals between the sampled fragments. The various small forest fragments in Ouro Preto, primarily in backyards, may also serve as stepping stones between sampled fragments.


Switching to ziprasidone in the clinical practice setting: an open-label study.
This observational study evaluates the long-term outcome of switching to ziprasidone in patients with schizophrenia in the clinical practice setting. Patients (208) with schizophrenia who had been switched to ziprasidone monotherapy due to partial response or tolerability problems were followed for 1 year. Efficacy was assessed at baseline and months 1, 3, and 12 with Brief Psychiatric Rating Scale (BPRS), Clinical Global Impression-severity (CGI-S), and CGI-improvement. Quality of life, functionality, and safety measures, including metabolic parameters, were also assessed; 195 subjects comprised the per protocol analysis population. A reduction > or = 30% in BPRS total score was observed in 42.5% of the subjects. Mean scores of the BPRS (global and positive and negative clusters), CGI-S and CGI-I significantly decreased at endpoint (p < 0.001). Ziprasidone treatment was also associated with statistically significant improvements in the GAF, WHO-DAS-II, and SF-12. After 1-year follow-up, a mean weight decrease of -1.6 kg (p < 0.05) was observed. Mean levels of LDL cholesterol and triglycerides also decreased (p < 0.01) while HDL cholesterol levels increased (p < 0.05) at endpoint. No significant changes in mean glucose levels at study end were detected. These findings suggest that switching to ziprasidone is effective and well tolerated in patients with schizophrenia requiring a change in antipsychotic medication.


Intestinal brush border calmodulin: key role in the regulation of NaCl transport in Giardia lamblia infected mice.
Activity of calmodulin and uptake of Ca2+, Na+ and Cl- was studied in control and Giardia lamblia infected mice. The activity of calmodulin was found to be significantly increased (p less than 0.001) in the experimental group as compared to control group. The uptake of Ca2+ increased significantly (p less than 0.001) while that of Na+ and Cl- decreased (p less than 0.001) in brush border membrane (BBM) vesicles from experimental group as compared to control group. In the presence of calmodulin inhibitor, trifluoperazine (TFP), the transport of Na+ and Cl- increased significantly (p less than 0.05) as compared to in the absence of inhibitor while transport of Ca2+ remained unaltered.


Diagnostic-driven antifungal therapy in neutropenic patients using the D-index and serial serum galactomannan testing.
Invasive mold disease is an important complication of patients with hematologic malignancies, and is associated with high mortality. A diagnostic-driven approach has been an alternative to the classical empiric antifungal therapy. In the present study we tested an algorithm that incorporated risk stratification using the D-index, serial serum galactomannan and computed tomographic-scan to guide the decision to start antifungal therapy in neutropenic patients. Between May 2010 and August 2012, patients with acute leukemia in induction remission were prospectively monitored from day 1 of chemotherapy until discharge or death with the D-index and galactomannan. Patients were stratified in low, intermediate and high risk according to the D-index and an extensive workup for invasive mold disease was performed in case of positive galactomannan (≥0.5), persistent fever, or the appearance of clinical manifestations suggestive of invasive mold disease. Among 29 patients, 6 (21%), 11 (38%), and 12 (41%) were classified as high, intermediate, and low risk, respectively. Workup for invasive mold disease was undertaken in 67%, 73% and 58% (p=0.77) of patients in each risk category, respectively, and antifungal therapy was given to 67%, 54.5%, and 17% (p=0.07). Proven or probable invasive mold disease was diagnosed in 67%, 45.5%, and in none (p=0.007) of high, intermediate, and low risk patients, respectively. All patients survived. A risk stratification using D-index was a useful instrument to be incorporated in invasive mold disease diagnostic approach, resulting in a more comprehensive antifungal treatment strategy, and to guide an earlier start of treatment in afebrile patients under very high risk.


Cold Stress Induced a Higher Level of Fat Oxidation in Women.
Niu, X, Han, P, Tang, Z, Huang, J, Li, L, He, H, Zhang, W, Zhao, L, and Zhao, L. Cold stress induced a higher level of fat oxidation in women. J Strength Cond Res 37(1): 114-122, 2023-To investigate women's fat oxidation under cold stress during incremental exercise testing and compare the effect of cold stress on fat oxidation between the sexes. Twenty-six healthy subjects performed 2 incremental exercise tests to determine maximal oxygen uptake (V̇ o2 max) on a treadmill in different ambient temperatures. Cardiopulmonary variables were continuously recorded during incremental exercise tests. Maximal fat oxidation (MFO) and the corresponding exercise intensity (Fat max ) were determined from the fat oxidation curve constructed by indirect calorimetry. Both men and women relied more on fat oxidation in cold environment ( p < 0.05). Compared with men, fat oxidation was significantly greater in women in the cold environment from 50 to 70% V̇ o2 max ( p < 0.05). There were no significant differences in the change of fat oxidation as exercise intensity increased between the sexes ( p > 0.05). Women had a greater MFO ( p < 0.05) and Fat max ( p < 0.05) than men in the cold environment. When MFO was the dependent variable, sex, fat-free mass, fat mass, V̇ o2 max, and temperature accounted for 48% of its variability. We conclude that cold stress enhances fat oxidation in women. Compared with men, women have significantly higher value and rely more on fat oxidation to supply energy in the cold environment, although the increasing level of fat oxidation was similar between the sexes. These factors may have important implications in the individualization of exercise prescription in cold conditions for both men and women.


Spontaneous epidural hematoma of thoracic spine presenting as Brown-Séquard syndrome: report of a case with review of the literature.
Spontaneous spinal epidural hematoma (SSEH) is an uncommon clinical entity. It produces a severe neurological deficit and prompt decompression is usually the first choice of treatment. Brown-Séquard syndrome is commonly seen in the setting of spinal trauma or an extramedullary spinal neoplasm, but rarely caused by SSEH. Case report and literature review. A previously healthy man presented with Brown-Séquard syndrome below T5-T6 cord segment secondary to spontaneous epidural hematoma. He opted for conservative treatment, which was followed by rapid resolution. Although Brown-Séquard syndrome as a presenting feature of SSEH is rare, it does exist in exceptional case, which should be taken into consideration for differential diagnosis. Prompt surgical decompression is an absolute surgical indication widely accepted for patient with progressive neurological deficit. However, SSEH presenting with incomplete neurological insult such as Brown-Séquard syndrome might have a benign course. Successful non-operative management of this problem does not make it a standard of care, and surgical decompression remains the standard treatment for SSEH.


A novel prenatal index predicting the probability of neonatal intensive care in pregnants: amnion progesterone receptor to alfa fetoprotein rate.
Amniocentesis (AC) is the most used interventional procedure for prenatal diagnosis. The study aims to evaluate the pregnancy outcomes undergoing AC and the potential of amnion progesterone receptor (aPR) to alfa fetoprotein (AFP) rate for predicting the probability of neonatal intensive care unit (NICU). This prospective cross-sectional study population consisted of 85 pregnant women who underwent mid-trimester AC. All cases were screened by ultrasound before AC. Maternal venous and amniotic samples were obtained simultaneously to evaluate the serum progesterone (sPRG), aPR, and aAFP and analyzed with patient results. Unlike sPRG and aAFP, aPR showed a positive correlation with NICU and a negative correlation with parity. In linear regression, the aPR-AFP rate showed strong linearity with NICU and parity. In an aPR-AFP rate analysis, we saw a strong predictivity for NICU compared to the other three parameters. It presented 73.4% specificity and 79% sensitivity at 0.0075 cut-off (AUC: 0.78; p = 0.003; 95% CI: 0.608-0.914). Evaluating the PR either alone or in a rational combination with AFP will provide physicians with valuable information about the advanced process of pregnancy and postpartum complications. The physicians might use the aPR-AFP rate to predict NICU potential for pregnancy and need further studies to make more vital predictions on postpartum complications.


Measurement and Management of Cognitive Load in Surgical Education: A Narrative Review.
Cognitive load should be considered in designing optimal educational programs in health care. Despite the highly demanding nature of surgery and surgical education, a consensus on how to manage cognitive load has not been established. The objective of this review is to map out how recent surgical education research incorporates cognitive load. A literature search was performed using keywords related to cognitive load and digital education up to December 2021. Studies published in English relevant to assessment and management of cognitive load in surgical education were included. Terminology, assessment tools, association with different surgical procedures and training modalities, and programs considering cognitive load were reported. We identified several terms to describe cognitive load. Cognitive load was measured by subjective, self-reported questionnaires and by objective measurements, such as physiological parameters or estimated by reaction time to secondary tasks. Subjective measurements reported cognitive load in one or multiple dimensions. Correlations between subjective and objective measurements were shown in multiple studies. Overall, higher cognitive load was observed in training for more complex tasks and high-fidelity modalities, and among less experienced trainees. Cognitive load theory has been lately incorporated into designing teaching programs. A broad range of terms and assessment tools were identified for cognitive load. To maximize the learning outcome, management of cognitive load is necessary in surgical education. This review summarizes the current knowledge in assessment and management of cognitive load in surgical education and provides suggestions for future studies.


Raman spectroscopic peculiarities of Icelandic poorly crystalline minerals and their implications for Mars exploration.
In this work, we have analyzed natural samples collected at three hydrothermal areas of Iceland by Raman spectroscopy. The studied high-latitude regions are considered environmentally and mineralogically appropriate Martian analogues since they are rich in weathered basalts that have been altered by hydrothermalism to mineral phases such as silica, clay minerals, sulfates, oxides, and sulfur. The main objective of this work was to assess the relation of the spectroscopic signatures of alteration to hydrothermal processes and biomediation, considering previous studies focused on the detection of lipid biomarkers in the same samples. The recorded Raman spectra, taken with optical parameters similar to the ExoMars 2022 Raman spectrometer, showed structural modifications in all secondary minerals in the form of peak shifts (in the case of sulfur and clay minerals), changes in the relative ratio intensity (in anatase) and/or shape broadening (in sulfates and hematite). These results reveal the suitability of Raman spectroscopy to examine areas rich in water-altered minerals, where a mixture of crystalline and amorphous phases can co-exist. The detection of silica is singularly interesting since, on the one hand, it can imply the past existence of hydrothermal hot springs rich in nutrient and redox gradients and, on the other hand, provides excellent matrix for biosignature preservation. The data can be helpful as an astrobiological database for the forthcoming missions to Mars, where potential upwelling groundwater systems could have altered the mineral phases in a similar way to that observed in this work.


Insulin-like growth factor-1 protects peroxynitrite-induced cell death by preventing cytochrome c-induced caspase-3 activation.
We investigated the effect of IGF-1 on cell death induced by peroxynitrite in human neuroblastoma SH-SY5Y cells. Exposure of the cells to 3-morpholinosydnonimine (SIN-1), a peroxynitrite donor, caused cytochrome c release from the mitochondria, caspase-3-like activation, and cell death. Pre-incubation of the cells with the caspase-3 inhibitor partially prevented SIN-1-induced cell death. Simultaneous addition of IGF-1 reduced SIN-1-induced caspase-3-like activation and cell death, whereas IGF-1 failed to reduce the release of cytochrome c. IGF-1 increased Akt phosphorylation, and Akt phosphorylation was inhibited by wortmannin, an inhibitor of phosphatidylinositol 3-kinase. In addition, wortmannin prevented IGF-1-evoked inhibition of cell death and caspase-3-like activation. In a cell-free system, addition of cytochrome c to cytosolic fraction resulted in caspase-3-like activation. The activation was reduced when the cytosolic fraction prepared from IGF-1-treated cells was used. These results suggest that IGF-1 protects peroxynitrite-induced cell death downstream of cytochrome c release through the inhibition of caspase-3-like activation.


Modest cortex and promiscuous medulla for thymic repertoire formation.
Recent advances in the understanding of medullary thymic epithelial cell biology, and especially of their promiscuous gene expression, have highlighted the indispensable role of thymic medulla in shaping a self-tolerant T-cell repertoire. Additionally, our recent results have shown that cortical thymic epithelial cells possess a unique proteasome (the so-called thymoproteasome), which seems to possess limited protein degradation capability for generating peptides that are loaded onto class I major histocompatibility complex molecules. We discuss here the unique role of thymoproteasomes in the development and repertoire formation of CD8+ T cells, focusing on the stepwise and contrasting roles of cortical epithelial cells and medullary epithelial cells. These results could offer fundamental new insights into the molecular mechanisms of T-cell repertoire selection.


Anti-Diabetic Potential of Chlorella Pyrenoidosa-Based Mixture and its Regulation of Gut Microbiota.
The aim of the present study was to investigate the anti-diabetic effect of CGSGCG and its beneficial effects on gut microbiota in type 2 diabetes (T2D) mice induced by streptozotocin and high sucrose and high fat diet. The results showed that treatment with CGSGCG reduced fasting blood glucose, improved oral glucose tolerance test, protected the liver from injury, and reduced inflammation in T2D mice. The contents of acetic acid, propionic acid, butyric acid, isobutyric acid, valeric acid and isovaleric acid in CGSGCG group were 2.49-, 1.74-, 3.31-, 1.93-, 1.36- and 1.30-fold than that of the model group. Moreover, administration of CGSGCG up-regulated the expression of INSR/IRS-1/PI3K/AKT/GLUT4 and mTOR but down-regulated the P38MAPK expression. Furthermore, the abundance of beneficial bacteria such as Verrucomicrobia, Proteobacteria, Osillibacter, Dubosiella and Lactococcus in intestinal tract increased, indicating that CGSCGG regulated and improved the bacterial community structure of T2D mice, which were closely related to glycometabolism.


Antibiotic therapy and outcome from immune-checkpoint inhibitors.
Sensitivity to immune checkpoint inhibitor (ICPI) therapy is governed by a complex interplay of tumor and host-related determinants. Epidemiological studies have highlighted that exposure to antibiotic therapy influences the probability of response to ICPI and predict for shorter patient survival across malignancies. Whilst a number of studies have reproducibly documented the detrimental effect of broad-spectrum antibiotics, the immune-biologic mechanisms underlying the association with outcome are poorly understood. Perturbation of the gut microbiota, an increasingly well-characterized factor capable of influencing ICPI-mediated immune reconstitution, has been indicated as a putative mechanism to explain the adverse effects attributed to antibiotic exposure in the context of ICPI therapy. Prospective studies are required to validate antibiotic-mediated gut perturbations as a mechanism of ICPI refractoriness and guide the development of strategies to overcome this barrier to an effective delivery of anti-cancer immunotherapy.


Central action of CART induces neuronal activation in the paraventricular and dorsomedial hypothalamus of diet-induced obese and lean mice.
The cocaine- and amphetamine-regulated transcript (CART) is a peptide commonly studied in the feeding behavior, but it exerts an important role in the autonomic and cardiovascular control as well. It is known that exogenous administration of CART in the central nervous system can elicit increase in blood pressure of both conscious and anesthetized rodents, but little is known whether these central effects might differ between lean and obese animals. We have recently shown that diet-induced obese mice that developed hypertension presented an upregulation of CART levels in the dorsomedial nucleus of hypothalamus, while obese normotensive ones had not. Herein we investigate whether the central action of CART could activate differently the hypothalamic nuclei of diet-induced obese mice compared to the lean counterparts by using Fos protein expression, C57BL/6 mice were randomly assigned to two cohorts, one fed with a high-fat diet for 8 weeks (obese), and the other fed with regular rodent chow (lean). Both groups received an ICV injection of CART at the dose of 400μM, 1 mM or vehicle. Subsequently, the brains were processed for Fos protein immunohistochemical in order to identify hypothalamic neuronal activation. Significantly greater numbers of Fos-positive neurons were observed in the PVN and DMH of obese mice that received CART 1 mM, when compared to the lean control. These results indicate that the central action of CART induces neuronal activation in the hypothalamic nuclei of obese and lean mice, and this could be relevant to the different autonomic and cardiovascular adjustments that an organism exposed to different diet and metabolic condition.


Cadherin-6B stimulates an epithelial mesenchymal transition and the delamination of cells from the neural ectoderm via LIMK/cofilin mediated non-canonical BMP receptor signaling.
We previously provided evidence that cadherin-6B induces de-epithelialization of the neural crest prior to delamination and is required for the overall epithelial mesenchymal transition (EMT). Furthermore, de-epithelialization induced by cadherin-6B was found to be mediated by BMP receptor signaling independent of BMP. We now find that de-epithelialization is mediated by non-canonical BMP signaling through the BMP type II receptor (BMPRII) and not by canonical Smad dependent signaling through BMP Type I receptor. The LIM kinase/cofilin pathway mediates non-canonical BMPRII induced de-epithelialization, in response to either cadherin-6B or BMP. LIMK1 induces de-epithelialization in the neural tube and dominant negative LIMK1 decreases de-epithelialization induced by either cadherin-6B or BMP. Cofilin is the major known LIMK1 target and a S3A phosphorylation deficient mutated cofilin inhibits de-epithelialization induced by cadherin-6B as well as LIMK1. Importantly, LIMK1 as well as cadherin-6B can trigger ectopic delamination when co-expressed with the competence factor SOX9, showing that this cadherin-6B stimulated signaling pathway can mediate the full EMT in the appropriate context. These findings suggest that the de-epithelialization step of the neural crest EMT by cadherin-6B/BMPRII involves regulation of actin dynamics via LIMK/cofilin.


Modulation of protein synthesis by nerve growth factor.
The effects of nerve growth factor (NGF) and two analogs of cAMP have been studied in the PC12 line of rat pheochromocytoma cells. We have used two-dimensional gel electrophoresis to detect more than 800 proteins from control and NGF-treated cells. Of these proteins, none were qualitatively repressed in response to NGF, and no new proteins appeared after NGF treatment. Visual inspection of the gels showed that approximately 5% of the proteins were detectably increased or decreased in rate of synthesis by NGF, and each of these changes was mimicked by both cAMP analogs. The two-dimensional gel data were further analyzed by a computerized scanning system. This analysis has revealed many significant changes that are smaller than those detected by eye. Approximately 25 to 30% of the proteins analyzed were found to be altered in rate of synthesis by 30% or more. Statistical analysis has shown that the response to NGF and the response to dibutyryl cAMP are highly correlated, even down to changes as small as 30%. No proteins were found to be significantly altered by both dibutyryl cAMP and 8-bromo cAMP, but not by NGF. These results show that NGF causes only quantitative modulations of protein synthesis in PC12 cells, and these data strongly suggest that the response of PC12 cells to NGF is mediated by cAMP.


Dominant and recessive polycystic kidney disease in children: classification by intravenous pyelography, ultrasound, and computed tomography.
Both dominant and recessive polycystic kidney disease appear in childhood. We have analyzed findings of intravenous pyelography, ultrasound and computed tomography in genetically classified cases of dominant (13 children) and recessive polycystic kidney disease (5 children) and thus defined criteria by which sporadic cases of childhood polycystic kidney disease can be classified to dominant or recessive polycystic kidney disease.


Evidence for polymorphism of MB3 antigens among three HLA-D clusters associated with HLA-DR4.
In a previous study, we showed that the three hitherto serologically indistinguishable HLA-D specificities associated with HLA-DR4, HLA- DYT , HLA- DKT2 , and HLA-Dw4 can be distinguished on the basis of their reactivity with two distinct Ia-like-specific monoclonal antibodies, HU-18 and HU-23. In this study, we attempted to identify and characterize Ia-like molecules recognized by HU-18 and HU-23 on a molecular level because Ia subsets (HLA-DR, MB, MT, or SB) identified by them remained unknown. The results of sequential coprecipitation assays and two-dimensional gel analyses showed that both HU-18 and HU-23 recognize antigenic determinants borne on MB3 but not on HLA-DRw6.2 molecules. Because the two monoclonal antibodies, specific for determinants carried on MB3 molecules, show distinct reactivity against homozygous typing cells defining HLA- DYT , HLA- DKT2 , and HLA-Dw4, all of which share DR4- MB3 , the data indicate that these three HLA-D clusters associated with HLA-DR4 possess distinct MB3 molecules, suggesting the existence of polymorphism in MB3 antigens.


Evolutionary Dynamics of Homophily and Heterophily.
Most social interactions do not take place at random. In many situations, individuals choose their interaction partners on the basis of phenotypic cues. When this happens, individuals are often homophilic, that is, they tend to interact with individuals that are similar to them. Here we investigate the joint evolution of phenotypic cues and cue-dependent interaction strategies. By a combination of individual-based simulations and analytical arguments, we show that homophily evolves less easily than earlier studies suggest. The evolutionary interplay of cues and cue-based behaviour is intricate and has many interesting facets. For example, an interaction strategy like heterophily may stably persist in the population even if it is selected against in association with any particular cue. Homophily persisted for extensive periods of time just in those simulations where homophilic interactions provide a lower (rather than a higher) payoff than heterophilic interactions. Our results indicate that even the simplest cue-based social interactions can have rich dynamics and a surprising diversity of evolutionary outcomes.


Negatively charged polysulfone membranes with hydrophilicity and antifouling properties based on in situ cross-linked polymerization.
Polysulfone (PSf) membrane has been widely used in water separation and purification, although, membrane fouling is still a serious problem limiting its potential. We aim to improve the antifouling of PSf membranes via a very simple and efficient method. In this work, antifouling PSf membranes were fabricated via in situ cross-linked polymerization coupled with non-solvent induced phase separation. In brief, acrylic acid (AA) and vinyltriethoxysilane (VTEOS) were copolymerized in PSf solution, then directly casted into membranes without purification. With the increase of monomers concentration, the morphology of the as-cast membranes changed from a finger-like morphology to a fully sponge-like structure due to the increased viscosity and decreased precipitation rate of the polymer solutions. Meanwhile, the hydrophilicity and electronegativity of modified membranes were highly improved leading to inhibited protein adsorption and improved antifouling property. Furthermore, in order to further find out the different roles player by AA and VTESO, the modified membrane without VTEOS was prepared and characterized. The results indicated that AA is more effective in the membrane hydrophilicity improvement, VTEOS is more crucial to improve membrane stability. This work provides valuable guidance for fabricating PSf membranes with hydrophilicity and antifouling property via in situ cross-linked polymerization.


In vitro fertilization versus conversion to intrauterine insemination in the setting of three or fewer follicles: how should patients proceed when follicular response falls short of expectation?
To determine whether in vitro fertilization (IVF) cycles with suboptimal response should be converted to intrauterine insemination (IUI) or proceed to oocyte retrieval (OR). Retrospective cohort. Academic medical center. All patients initiating IVF from January 2004 through December 2011. OR versus conversion to IUI. A total of 1,098 patients were identified whose IVF cycles were characterized by recruitment of three or fewer follicles, excluding patients with bilateral tubal disease or severe male factor. Cycles with three follicles were defined as those with three follicles ≥ 14 mm with no fourth follicle ≥ 10 mm. Cycles with two or fewer follicles were similarly defined. Outcomes were compared for patients proceeding with OR (n = 624) versus converting to IUI (n = 474). Age-adjusted relative risks for pregnancy were calculated, stratifying for number of follicles. The likelihood of retrieving at least one mature oocyte (82.9% vs. 94.8% vs. 96.2%), having at least one zygote (61.9% vs. 76.8% vs. 84.2%), and undergoing transfer (57.1% vs. 73.0% vs. 83.3%) increased significantly with increasing follicle number. Patients with three or fewer follicles were 2.6 times more likely to achieve a live birth with IVF versus IUI (9.3% vs. 3.4%). This benefit was only apparent when at least two follicles were present. No benefit was gained by performing OR in the setting of one follicle. IVF compared with IUI presents superior pregnancy rates in the setting of two or more follicles. Assisted reproduction programs may benefit their patients by pursuing IVF in this scenario.


Dopamine receptor 1 expressing B cells exert a proinflammatory role in female patients with rheumatoid arthritis.
Rheumatoid arthritis (RA) is a chronic rheumatic disease with a clear sex-bias. Recent data indicated a role for dopamine in RA pathogenesis, while dopaminergic pathways can be modulated by estrogens. As defined mechanism of action of dopamine on B cell function in RA are unclear, we aimed to elucidate this, with special focus on sex-differences. Healthy controls (HC, n = 64) and RA patients (n = 61) were recruited. Expression of D1-D5 dopamine receptors (DRs) was investigated by flow cytometry on peripheral blood mononuclear cells (PBMCs). D1-like DRs were stimulated in vitro to assess effects on B cell activation and proliferation. Secretion of cytokines and dopamine content were measured by ELISA. All DRs were expressed on PBMCs of HC and RA patients. Dopamine content in PBMCs, and frequency of D1DR expressing B cells were significantly higher in RA females (p < 0.001). Expression of D1DR on RA B cells correlated positively with disease duration and severity only in women. Combined B cell and D1-like DR stimulation induced higher IL-8 and CCL-3 secretion from PBMCs of female RA patients compared to HC. These results indicate sex-specific differences in dopaminergic pathway in RA, with a proinflammatory feature of the D1DR pathway in women.