RNA-Seq analysis in giant pandas reveals the differential expression of multiple genes involved in cataract formation.
The giant panda (Ailuropoda melanoleuca) is an endangered mammalian species native to China. Fewer than 2500 giant pandas are known to exist, many of which are bred in captivity as a means to preserve and repopulate the species. Like other captive mammals, giant pandas acquire age-related cataracts, reducing their quality of life. Recent comparative genome-wide methylation analysis revealed 110 differentially methylated genes associated with cataract formation including six also associated with the formation of age-related cataracts in humans. To investigate the pathological pathway in greater detail, here we used RNA-Seq analysis to investigate the differential expression profiles of genes in three giant pandas with cataracts and three healthy controls. We identified more than 700 differentially expressed genes, 29 of which were selected for further analysis based on their low q-value. We found that many of the genes encoded regulatory and signaling proteins associated with the control of cell growth, migration, differentiation and apoptosis, supporting previous research indicating a key role for apoptosis in cataract formation. The identification of genes involved in the formation of age-related cataracts could facilitate the development of predictive markers, preventative measures and even new therapies to improve the life of captive animals.


The development of the cricoid cartilage and its implications for the use of endotracheal tubes in the pediatric population.
The rigid cricoid cartilage is functionally the narrowest portion of the larynx. There is some controversy over the shape of the pediatric cricoid cartilage in the transverse plane. It is important to understand the development of the cricoid cartilage so that endo-traceheal tubes can be used more safely. To determine changes in the internal diameter and shape of the cricoid cartilage during development and explore the implications of those changes for the selection of ETT type and size for children. The cervical computed tomography scans were reviewed in patients aged 1-20 years. After performing the multiplanar reconstruction and correcting the slant, the transverse and anteroposterior internal diameters of the inlet and outlet of the cricoid cartilage were measured, respectively. The angle between the arch and the lamina of the cricoid cartilage in the middle sagittal plane was measured. The ratios of transverse to anteroposterior diameter for the inlet and outlet of the cricoid cartilage were calculated, respectively. In females, the internal diameters of the cricoid cartilage increased linearly with age. In males, the internal diameters of the cricoid cartilage exhibited a growth spurt during adolescence. The transverse diameter of the inlet was the smallest diameter of the cricoid cartilage, and the predicting formula of the transverse diameter of the inlet for children aged 1-12 was 0.4 × age (year) + 5.1, R2 = .758. The angle between the arch and lamina of the cricoid cartilage and the ratios of transverse to anteroposterior diameter correlated weakly with age. The transverse inner diameter of the inlet is the smallest diameter of the cricoid cartilage. The "funnel shape" of the cricoid cartilage remains unchanged during development. The outer diameter should be considered when selecting an endotracheal tube.


The octopus vertical lobe modulates short-term learning rate and uses LTP to acquire long-term memory.
Analyzing the processes and neuronal circuitry involved in complex behaviors in phylogenetically remote species can help us understand the evolution and function of these systems. Cephalopods, with their vertebrate-like behaviors but much simpler brains, are ideal for such an analysis. The vertical lobe (VL) of Octopus vulgaris is a pivotal brain station in its learning and memory system. To examine the organization of the learning and memory circuitry and to test whether the LTP that we discovered in the VL is involved in behavioral learning, we tetanized the VL to induce a global synaptic enhancement of the VL pathway. The effects of tetanization on learning and memory of a passive avoidance task were compared to those of transecting the same pathway. Tetanization accelerated and transection slowed short-term learning to avoid attacking a negatively reinforced object. However, both treatments impaired long-term recall the next day. Our results suggest that the learning and memory system in the octopus, as in mammals [9], is separated into short- and long-term memory sites. In the octopus, the two memory sites are not independent; the VL, which mediates long-term memory acquisition through LTP, also modulates the circuitry controlling behavior and short-term learning.


Prehospital Whole Blood Resuscitation Reduces Fluid Requirement While Maintaining Critical Physiology in a Model of Penetrating Traumatic Brain Injury and Hemorrhage: Implications on Resource-Limited Combat Casualty Care.
Prehospital resuscitation using whole blood (WB) is the standard of care for hemorrhagic shock (HS) but there is no consensus recommendation for resuscitation in the presence of traumatic brain injury (TBI) due to a lack of sufficient evidence. In order to evaluate the optimal resuscitation strategies for TBI+HS, Sprague-Dawley rats were randomized into four groups based on resuscitation fluid and prehospital mean arterial pressure (MAP) threshold (n = 9-10/group): Lactated Ringer's (LR)-60 mm Hg (LR60), LR-70 mm Hg (LR70), WB-60 mm Hg (WB60), WB-70 mm Hg (WB70). All groups received a frontal penetrating ballistic-like brain injury followed by a 35-min period of HS. During the prehospital phase, rats received an initial bolus of resuscitation fluid (WB or LR) followed by LR as needed to maintain MAP above the designated threshold for 90 min. During the in-hospital phase, rats received definitive resuscitation with shed WB. Physiological parameters were recorded continuously and cerebral edema was measured at 3 and 24 h postinjury. The WB60 group demonstrated a significantly lower prehospital fluid requirement compared WB70, LR60, and LR70 (P < 0.05). Compared to the respective LR groups, both the WB60 and WB70 groups also demonstrated improved MAP, cerebral perfusion pressure, brain tissue oxygen tension, and cerebral edema. The edema benefits were observed at 3 h, but not 24 h postinjury, and were localized to the injury site. Together, these results provide evidence that prehospital WB resuscitation and lower MAP resuscitation thresholds can reduce the prehospital fluid requirement while still maintaining critical cerebral physiology in a model of HS and concomitant TBI.


Low-Cost and Simple Frontal Sinus Surgical Cutting Guide Modeling for Anterior Cranioplasty in Facial Feminization Surgery: How To Do It.
This study demonstrates a novel and simple design to create a low cost frontal sinus surgical cutting guide, using patient's frontal sinus cavities as references, to perform an anterior cranioplasty in facial feminization surgery. A clinical series demonstrates its clinical use and safety. Authors used a 4 open-source software protocol for the virtual surgical modeling (VSP). Retrospectively, pre- and post-operative complications were reviewed, and a FACE-Q questionnaires were used to evaluate patient's postoperative "Satisfaction with forehead and eyebrows." Recordings of the VSP and surgical technic are presented. Sixteen patients were operated between November 2018 and November 2020 using in-house surgical guides. All were performed by authors in 5 to 10 minutes using Blender open-source Software. No complications were reported and authors always found an optimal retention of the surgical guide on the convex bony surface of the forehead and frontonasal area. FACE-Q questionnaire reported a very high satisfaction for all patients (mean score: 22/24). This simplify organic design can be performed efficiently by any surgeon even without previous training in home staging VSP. Patient satisfaction was very high, along with an absence of postoperative complications.


DTP vaccine reactions: effect of prior reactions on rate of subsequent reactions.
It is generally presumed that children who have had reactions to diphtheria-tetanus-pertussis (DTP) immunization will be more likely to have similar reactions or more severe reactions upon subsequent immunization. To evaluate this contention, we studied the rates of selected reactions occurring within 48 hours of primary DTP immunization in 1,241 infants less than one year of age. Both local and systemic reactions were significantly more frequent following subsequent DTP immunization if present following a prior immunization. The rates of local reactions following subsequent DTP immunization as a function of previous reactions were as follows (no prior reactions/prior reaction): local redness greater than or equal to 2.5 cm, 12.5%/25.5% (p less than 0.0001); local swelling greater than or equal to 2.5 cm, 16.8%/29.0% (p less than 0.0001); local pain, 37.4%/56.4%; (p less than 0.0001). The rates of systemic reactions as a function of previous reactions were as follows (no prior reactions/prior reaction): drowsiness, 24.9%/42.8% (p less than 0.0001); fretfulness, 47.5%/64.7% (p less than 0.0001); vomiting, 4.8%/11.2% (p = 0.0084); anorexia 16.0%/26.3% (p = 0.0001); fever greater than or equal to 38 degrees C, 37.6%/58.5% (p less than) 0.0001); persistent crying, 2.6%/4.5% (p = 0.3557). In addition, infants who experienced a fever greater than or equal to 38 degrees C on the first of two immunizations were more likely to have a temperature greater than or equal to 39 degrees C following the second immunization, 4.2% vs. 12.4% (p = 0.0017). These data strongly support the presumption that children who have had previous reactions following DTP immunization are more likely to have similar reactions upon subsequent immunization.


Hyaluronan production in human rheumatoid fibroblastic synovial lining cells is increased by interleukin 1 beta but inhibited by transforming growth factor beta 1.
To investigate the regulatory roles of interleukin 1 beta (IL1 beta), tumour necrosis factor alpha (TNF alpha), interferon gamma (IFN gamma) or transforming growth factor beta 1 (TGF beta 1) on hyaluronan (HA) synthesis by human fibroblastic synovial lining cells. Concentrations of HA in culture supernatants of fibroblastic synovial lining cell line (RAMAK-1 cell line) with or without stimulation by IL1 beta, TNF alpha, IFN gamma or TGF beta 1 were measured by sandwich binding protein assay. Levels of HA synthase mRNA of the cells with or without stimulation were detected by reverse transcribed polymerase chain reaction. Molecular weights of HA in the culture supernatants of the cells with or without stimulation were measured using high performance gel permeation liquid chromatography. HA synthesis by the cells was not significantly augmented by TNF alpha or by IFN gamma. It was significantly stimulated by IL1 beta but inhibited by TGF beta 1. Molecular weights of HA in the culture supernatants of the cells were unchanged by stimulation with TNF alpha. They were remarkably increased by stimulation with IL1 beta and IFN gamma, but reduced with TGF beta 1. IL 1 beta is an up regulator of HA synthesis, while TGF beta 1 is a down regulator. HA production in the synovial lining cells of inflamed joints (for example, rheumatoid arthritis) might be regulated by the balance of these cytokines.


Oxidative phosphorylation in human muscle in patients with ocular myopathy and after general anaesthesia.
The fuel preference of human muscle mitochondria has been given. Substrates which are oxidized with low velocity cannot be used to detect defects in oxidative phosphorylation. After general anaesthesia, the oxygen uptake with the different substrates is much lower than after local analgesia. The latter was therefore used in the subsequent study. In 15 out of 18 patients with ocular myopathy, defects in oxidative phosphorylation could be detected in isolated muscle mitochondria prepared from freshly biopsied tissue. Measurement of the activity of segments of the respiratory chain in homogenate from frozen muscle showed no, or minor defects. In two of these patients showing exercise intolerance, decreased oxidation of NAD(+)-linked substrates and apparently normal mitochondrial DNA, further study revealed deficiency of pyruvate dehydrogenase in a girl with ptosis and a high Km of complex I for NADH in a man. Both patients responded to vitamin therapy.


Collagen synthesis promoting pullulan-PEI-ascorbic acid conjugate as an efficient anti-cancer gene delivery vector.
Cationized pullulan (pullulan-PEI; PP) was synthesized and further modified with an anti-oxidant molecule, ascorbic acid (PPAA) at various ratios. The nanoplexes formed at an optimum ratio of 4:1 was within a size of 150nm and had a zeta potential of 9-14mV. The nanoplexes at this ratio was used for further investigations. The cell internalization and transfection efficiency of these nanoplexes were determined in presence of serum. The internalization and transfection efficiency were found to be unaffected by the presence of fetal bovine serum. Another interesting observation was that this polymer was found to have collagen synthesis promoting property. The collagen synthesis effect of these polymers was quantified and observed that PPAA3 promoted the highest. Transfection efficiency was evaluated by assessing the p53 gene expression in C6 rat glioma cells and cell death was quantified to be 96% by flow cytometry, thus establishing the high efficacy of this polymer.


[The effect of fluoride on dental alloys in different concentrations of fluoride artificial saliva].
To study the changes in surface morphology of cobalt-chromium alloy, pure titanium and high-cobalt chromium molybdenum alloy immersed in artificial saliva with different concentrations of fluoride, and to analyze the corrosion resistance of these metals. The three kinds of metal were polished and then were immersed in artificial saliva with different concentrations of fluoride (0%, 0.05%, 0.2%). All specimens were immersed for 3 weeks at (37.0 +/- 1.0) degrees C. Metal surface roughness before and after immersion were measured and analyzed by scanning electron microscope (SEM). Metal surface roughness was increased with the concentration of fluoride. The surface roughness of the Co-Cr group and Ti group had a statistically significant difference between before and after immersion at the fluoride concentration of 0.05% (P<0.01), and the difference between Co-Cr group and vitallium2000 group, Ti group and vitallium2000 group were statistically significant difference (P<0.01). The differences of three groups before and after immersion were statistically significant at the fluoride concentration of 0.2% (P<0.01), and the difference among three groups was statistically significant (P<0.01). Patients who frequently use fluoride products should not use pure titanium or cobalt-chromium alloy prosthesis. Patients with these three metal prostheses are not advised to use high concentration fluoride products.


Early spring leaf out enhances growth and survival of saplings in a temperate deciduous forest.
Saplings of many canopy tree species in winter deciduous forests receive the major portion of their light budget for their growing season prior to canopy closure in the spring. This period of high light may be critical for achieving a positive carbon (C) gain, thus contributing strongly to their growth and survival. This study of saplings of Aesculus glabra and Acer saccharum in Trelease Woods, Illinois, USA, tested this hypothesis experimentally by placing tents of shade cloth over saplings during their spring period of high light prior to canopy closure in three consecutive years. Leaf senescence began 16 days (year 0) and 60 days (year 1) earlier for shaded A. glabra saplings than control saplings. No change in senescence occurred for A. saccharum. The annual absolute growth in stem diameter of both species was negligible or negative for shaded saplings, but positive for control saplings. Only 7% of the shaded A. glabra saplings were alive after 2 years, while all control saplings survived for 3 years; only 20% of the shaded A. saccharum saplings survived for 3 years, while 73% of control saplings were alive after the same period. Early spring leaf out is a critical mechanism that allows the long-term persistence of saplings of these species in this winter deciduous forest. Studies and models of C gain, growth, and survival of saplings in deciduous forests may need to take into account their spring phenology because saplings of many species are actually "sun" individuals in the spring prior to their longer period in the summer shade.


Nucleotide sequence and deduced amino acid sequence of a Plasmodium falciparum actin gene.
The nucleotide sequence of a Plasmodium falciparum actin gene has been established. The gene codes for a protein of 376 amino acids and is not interrupted by introns. The nucleotide sequence reveals an extreme bias in codon usage. Not less than 85% of the codons possess an A or T at the third position. As has been found for the actins in other unicellular eukaryotes, P. falciparum actin is related both to vertebrate cytoplasmic and vertebrate muscle specific actins. However, the malarial actin is one of the most alpha-like actins hitherto found in lower eukaryotes.


High-performance biochar-loaded MgAl-layered double oxide adsorbents derived from sewage sludge towards nanoplastics removal: Mechanism elucidation and QSAR modeling.
Utilization of sewage sludge for the fabrication of environmental functional materials is highly desirable to achieve pollution mitigation and resource recovery. In the present work, we introduced a novel MgAl-layered double oxide (LDO)@biochar composite adsorbent in-situ fabricated from Al-rich sewage sludge, and its excellent application in nanoplastics adsorption. Initially, fifteen model contaminants with varied conjugate structures, hydrogen bonding and ionic properties were selected for an investigation of adsorption behavior and adsorption selectivity on LDO@biochar. Structural variation of LDO@biochar suggested reconstruction of the layered double hydroxide (LDH) during the adsorption process due to the "memory effect". Under the synergy of LDH and biochar, the contaminants were adsorbed via multiple adsorbent-adsorbate interactions, including anion exchange, electrostatic interaction, hydrogen bonding and π-π conjugation. Then, a quantitative structure-activity relationship (QSAR) model was constructed by integrating the number of hydrogen bond acceptors, polarity surface area, number of aromatic rings, and Fukui index f(-)x together to reflect the affinity of each contaminant to the adsorbent. Guided by the QSAR model, the negatively charged polystyrene nanoplastics with continuously conjugated aromatic rings were predicted to be effectively adsorbed on LDO@biochar. Experimental tests confirmed a great capacity of LDO@biochar towards the polystyrene nanoplastics, given the equilibrium adsorption capacity as high as 360 mg g-1 at 30-50 °C. This work not only opened up a new avenue for sustainable utilization of sewage sludge towards high-performance environmental functional materials, but also demonstrated the potential of the QSAR analysis as a rapid and accurate approach for guiding the application of an adsorbent to new emerging containments.


Translational regulation of the novel haploid-specific transcripts for the c-abl proto-oncogene and a member of the 70 kDa heat-shock protein gene family in the male germ line.
Expression of the c-abl proto-oncogene in the mouse testis is characterized by the production of a unique 4.7-kb transcript present in germ cells that have entered the haploid phase of spermatogenesis. A similar developmental stage specificity of expression is observed for a member of the 70-kDa heat-shock protein (hsp 70) gene family. A unique-sized hsp 70 transcript (T-hsp 70) is produced in haploid spermatids and is stable throughout spermatogenesis. In the present study, we examined the regulation of expression of these genes by examining their association with polyribosomes. The germ cell-specific c-abl and T-hsp 70 mRNA variants were both associated with the polysomal fractions of mouse testis cells, suggesting that they are functional mRNAs. However, both c-abl and T-hsp 70 mRNAs were also found in the ribonucleoprotein particle fractions. The distribution of these mRNAs in both the polysomal and nonpolysomal fractions is comparable to that seen for the mRNA of protamine-1, a gene whose expression in the testis is known to be regulated at the level of translation. In contrast, transcripts from the beta-tubulin gene were seen predominantly in the polyribosomal fractions. These findings suggest that translation of the novel c-abl and T-hsp 70 transcripts is confined to subpopulations of testicular cells.


Prevalence of asymptomatic incidental aneurysms: a review of 2,685 computed tomographic angiograms.
Intracranial aneurysms are a public health issue with a potential for rupturing, causing significant morbidity/mortality. The prevalence of unruptured intracranial aneurysms, including those that are asymptomatic, varies widely, as it has been determined through autopsy studies as well as conventional angiography. However, computed tomography angiography, a less invasive procedure, has replaced the use of conventional angiography in the recent decades. The objective of the present study is to determine the prevalence of incidental aneurysms. The present study reviews all computed tomography angiography cases within the past decade at a single institution. Of 2195 cases included in the study, 39 (1.8%) were found to have asymptomatic unruptured aneurysms. The numbers of asymptomatic incidental aneurysms have increased.


Changes in prefrontal cortical behaviour depend upon familiarity on a bimanual co-ordination task: an fNIRS study.
To investigate neurocognitive mechanisms associated with task-related expertise development, this paper investigates serial changes in prefrontal activation patterns using functional near infrared spectroscopy (fNIRS). We evaluate cortical function in 62 healthy subjects with varying experience during serial evaluations of a knot-tying task. All tasks were performed bimanually and self paced, with fixed episodes of motor rest for five repetitions. Improvements in technical skill were evaluated using dexterity indices to quantify time, total movements and pathlength required to complete trials. Significant improvements in technical skills were observed in novices between the 2nd and 3rd trials, associated with increasing task familiarity. In trained subjects, minimal fluctuation in task-related oxyhaemoglobin (HbO(2)) and deoxyhaemoglobin (HHb) changes were observed in association with more stable task performance. In contrast, two significant transitions in prefrontal haemodynamic change were observed in novices. Greater task-related increases in HbO(2) and decreases in HHb were identified on the second trial compared to the first. Relative decreases in HbO(2) and increases in HHb change were observed between the third and fourth, and fourth and fifth trials respectively. These data suggest that prefrontal processing across five knot-tying trials is influenced by the level of experience on a task. Modifications in prefrontal activation appear to confer technical performance adaptation in novices.


AVP V1 receptor-mediated decrease in Cl- efflux and increase in dark cell number in choroid plexus epithelium.
The cerebrospinal fluid (CSF)-generating choroid plexus (CP) has many V1 binding sites for arginine vasopressin (AVP). AVP decreases CSF formation rate and choroidal blood flow, but little is known about how AVP alters ion transport across the blood-CSF barrier. Adult rat lateral ventricle CP was loaded with 36Cl-, exposed to AVP for 20 min, and then placed in isotope-free artificial CSF to measure release of 36Cl-. Effect of AVP at 10(-12) to 10(-7) M on the Cl- efflux rate coefficient (in s-1) was quantified. Maximal inhibition (by 20%) of Cl- extrusion at 10(-9) M AVP was prevented by the V1 receptor antagonist [beta-mercapto-beta, beta-cyclopentamethyleneproprionyl1,O-Me-Tyr2,Arg8]vasopressin. AVP also increased by more than twofold the number of dark and possibly dehydrated but otherwise morphologically normal choroid epithelial cells in adult CP. The V1 receptor antagonist prevented this AVP-induced increment in dark cell frequency. In infant rats (1 wk) with incomplete CSF secretory ability, 10(-9) M AVP altered neither Cl- efflux nor dark cell frequency. The ability of AVP to elicit functional and structural changes in adult, but not infant, CP epithelium is discussed in regard to ion transport, CSF secretion, intracranial pressure, and hydrocephalus.


Fluorescence Methods for Unraveling Oligomeric Amyloid Intermediates.
Amyloid oligomers are considered to be the relevant toxic species in many amyloid diseases and much research effort has been devoted to fully characterize these oligomers. Despite their importance, oligomers have proven to be difficult to characterize structurally. Information on their aggregation number is scarce, largely because standard techniques struggle to provide reliable results. In this chapter, we present two different methods that reproducibly yield fluorescently labeled α-Synuclein oligomers. We then discuss a new approach, combining single-molecule photobleaching and sub-stoichiometric fluorescent labeling, that we have developed to determine the aggregation number of supramolecular protein assemblies.


Using the angiogenic factors sFlt-1 and PlGF with Doppler ultrasound of the uterine artery for confirming preeclampsia.
The aim of this study is to assess the value of the angiogenic factors for diagnosing preeclampsia and predicting the severity of manifestation. A secondary aim is assessing the combination of the uterine artery Doppler with the angiogenic factors for improving the diagnostic power. This is a prospective single center study in a tertiary referral hospital. This study includes 728 individual patients. Inclusion criteria were singleton pregnancies, a referral to the hospital with suspicion of preeclampsia and any one or combination of the following symptoms: headache, upper abdominal pain, edema, and hypertension. Patients with complications that would affect the course of the pregnancy, such as placenta praevia, premature preterm rupture of membranes, breech presentation, and fetal chromosomal or structural anomalies, were excluded from the study. Blood samples collection and uterine artery Doppler ultrasound were performed at time of recruitment. The differences in sFlt-1, PlGF, and their quotient among normal collective and patients with preeclampsia were analyzed. Doppler ultrasound was performed by one of four highly qualified sonographers. Wilcoxon-Mann-Whitney U test, Spearman's rank correlation, receiver operating characteristic curves, Chi-square test, and logistic regression were used in the analysis. A total of 1003 individual samples for the angiogenic factors were included in the analysis. 584 out of the recruited 728 patients had follow-up data with delivery information at the study hospital. Patients with preeclampsia show a significant increase in sFlt-1, which directly correlate with the increased severity of manifestation (Spearman's ρ 0.49). The sFlt-1 cut-off value of 5424 pg/ml confirms preeclampsia with 83.7 % sensitivity, 68.1 % specificity, and 24 % misclassification rate. Preeclampsia patients also show a significant decrease in PlGF, which negatively correlates with the increased severity of manifestation (Spearman's ρ -0.39). A PlGF cut-off value of 118 pg/ml confirms preeclampsia with 47.6 % sensitivity, 71.4 % specificity, and 27 % misclassification rate. Logistic regression shows that a combination of the quotient from sFlt-1/PlGF with notching and uterine artery PI provides a valid model for diagnosing preeclampsia with a diagnostic power of 74.4 %. The study confirms the use of the sFlt-1 and PlGF for diagnosing preeclampsia. It also shows their significance in differentiating between different categories of preeclampsia according to severity. This study shows that the use of angiogenic factors in combination with ultrasound findings provides valid models for confirming preeclampsia.


Evaluation of a new chromogenic medium for the detection of methicillin-resistant Staphylococcus aureus carriage on nasal and perianal specimens.
A new chromogenic medium, denim blue (DB), was compared with methicillin-resistant Staphylococcus aureus (MRSA) Select (MRSAS) for detection of MRSA from surveillance specimens. On DB, MRSA colonies are larger (0.57-0.8 versus 0.45-0.6 mm at 18 and 24 h, respectively). Despite this, sensitivities of DB were 77% and 96% at 18 and 24 h, respectively, and those of MRSAS were 63% and 97.5%. Specificities were significantly higher for MRSAS than DB. The study demonstrates that DB and MRSA are equal for the detection of MRSA from surveillance specimens provided the plates are read after 24 h of incubation. Positive predictive values of both media were less than 95% requiring confirmation of MRSA, by another method, from all new patients.


CircMMP1 promotes the progression of glioma through miR-433/HMGB3 axis in vitro and in vivo.
Circular RNAs (circRNAs) are non-coding RNAs that have shown to regulate the progression of human diseases, including a variety of cancers. We aimed to investigate the function and the underlying working mechanism of circRNA matrix metallopeptidase 1 (circMMP1; hsa_circ_0024108) in glioma progression. Quantitative real-time polymerase chain reaction (qRT-PCR) was performed to examine the expression of circMMP1, microRNA-433-3p (written as "miR-433") and high mobility group box 3 (HMGB3). Nanoparticle tracking analysis (NTA) was used to analyze the relative concentration and size distribution of serum exosomes. Cell proliferation was analyzed via cell counting kit-8 (CCK8) assay and colony formation assay. Transwell migration and invasion assays were used to examine the metastasis ability of glioma cells. Cell apoptosis was assessed by flow cytometry. Dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay were used to verify the interactions of circMMP1/miR-433 and miR-433/HMGB3 messenger RNA (mRNA). Xenograft mouse model was built to clarify the role of circMMP1 in vivo. The results showed that high serum exosomal circMMP1 level might predict poor prognosis of glioma patients. CircMMP1 promoted the proliferation and motility and impeded the apoptosis of glioma cells. MiR-433 was a direct target of circMMP1, and circMMP1 silencing-mediated influences in glioma cells were partly alleviated by the knockdown of miR-433. MiR-433 directly interacted with HMGB3 mRNA, and HMGB3 overexpression partly counteracted the effects of miR-433 up-regulation in glioma cells. CircMMP1 enhanced HMGB3 level through sponging miR-433 in glioma cells. CircMMP1 silencing suppressed the progression of glioma in vivo. CircMMP1 promoted the progression of glioma through acting as a competitive endogenous RNA (ceRNA) of miR-433 to up-regulate HMGB3. CircMMP1 level in serum exosomes might be a potential marker for the diagnosis of glioma patients. Targeting circMMP1/miR-433/HMGB3 signaling might be a novel insight for glioma treatment.


Comparision of Hem-o-lok polymeric clip and tri-staple in laparoscopic splenectomy.
This study aimed to compare the hem-o-lok polymeric clip (HC) and tri-staple (TS) methods used in dividing the splenic hilum in terms of results, and to reveal their superiority to each other. Medical records of patients undergoing elective laparoscopic splenectomy at the Ondokuz Mayis University Faculty of Medicine General Surgery Clinic between March 2011 and March 2020 were retrospectively analyzed. Forty-two laparoscopic splenectomy cases performed using hem-o-lok polymeric clip (HC) or tri-staple (TS) were included in this study. Demographic features, primary diagnoses, splenic size, intraoperative data and postoperative complications, as well as the clip and stapler prices used in the surgery were analyzed. The mean operative time was significantly longer for HC group than TS group (116.7 min vs. 87.6 min, p<0.05). The mean cost of surgical instruments used to divide the splenic hilum was significantly lower for HC group than TS group (34.1 usd vs. 165.4 usd, p<0.05). There was no postoperative mortality, with a morbidity rate 6 (26.1%) for TS group and 4 (21.1%) for HC group (p>0.05). No significant difference existed in the complication rates. In the HC group, the operation time was longer, but the surgical cost was significantly lower. There was no significant difference when comparing other perioperative results. Although both techniques can be applied safely, we would like to emphasize that hemostasis is the most important factor for good results. Hem-o-lok polymeric clip, Laparoscopic splenectomy, Tri-staple.


Evidence of the receptor nature of the binding sites induced in Tetrahymena by insulin treatment. A quantitative cytofluorimetric technique for the study of binding kinetics.
Tetrahymena pyriformis GL cells pretreated (imprinted) and not pretreated with insulin showed dissimilar quantitative relations of FITC-insulin binding. Displacement of FITC-insulin by unlabelled insulin was considerably less in the control than in the imprinted series. The curve for saturation of the binding sites with FITC-insulin resembled a true saturation curve. The imprinted cells bound considerably more hormone in a shorter time than the control cells at identical levels of exposure. The dissociation of bound hormone from the imprinted cells increased over the control at 23 degrees C, and to a still greater degree at 4 degrees C. The effect of the pH of the medium on the dissociation of bound FITC-insulin also differed between the imprinted and not imprinted cells. Thus the proposed cytofluorimetric assay of binding kinetics demonstrated the actual conditions of receptor activity, and indicated that the induced insulin binding sites of Tetrahymena behaved similarly to 'classical' receptors.


Efficient and durable uranium extraction from uranium mine tailings seepage water via a photoelectrochemical method.
Current photocatalytic uranium (U) extraction methods have intrinsic obstacles, such as the recombination of charge carriers, and the deactivation of catalysts by extracted U. Here we show that, by applying a bias potential on the photocatalyst, the photoelectrochemical (PEC) method can address these limitations. We demonstrate that, owing to efficient spatial charge-carriers separation driven by the applied bias, the PEC method enables efficient and durable U extraction. The effects of multiple operation conditions are investigated. The U extraction proceeds via single-step one-electron reduction, resulting in the formation of pentavalent U, which can facilitate future studies on this often-overlooked U species. In real seepage water the PEC method achieves an extraction capacity of 0.67 gU m-3·h-1 without deactivation for 156 h continuous operation, which is 17 times faster than the photocatalytic method. This work provides an alternative tool for U resource recovery and facilitates future studies on U(V) chemistry.


Simple, rapid and effective preservation and reactivation of anaerobic ammonium oxidizing bacterium "Candidatus Brocadia sinica".
It is still the biggest challenge to secure enough seeding biomass for rapid start-up of full-scale (anaerobic ammonium oxidation) anammox processes due to slow growth. Preservation of active anammox biomass could be one of the solutions. In this study, biomass of anammox bacterium, "Candidatus Brocadia sinica", immersed in various nutrient media were preserved at -80 °C, 4 °C and room temperature. After 45, 90 and 150 days of preservation, specific anammox activity (SAA) of the preserved anammox biomass was determined by measuring (29)N2 production rate and transcription levels of hzsA gene encoding hydrazine synthase alpha subunit. Storage in nutrient medium containing 3 mM of molybdate at room temperature with periodical (every 45 days) supply of NH4(+) and NO2(-) was proved to be the most effective storage technique for "Ca. Brocadia sinica" biomass. Using this preservation condition, 96, 92 and 65% of the initial SAA was sustained after 45, 90 and 150 days of storage, respectively. Transcription levels of hzsA gene in biomass correlated with the SAA (R(2) = 0.83), indicating it can be used as a genetic marker to evaluate the anammox activity of preserved biomass. Furthermore, the 90-day-stored biomass was successfully reactivated by immobilizing in polyvinyl alcohol (6%, w/v) and sodium alginate (2%, w/v) gel and then inoculated to up-flow column reactors. Total nitrogen removal rates rapidly increased to 7 kg-N m(-3) d(-1) within 35 days of operation. Based on these results, the room temperature preservation with molybdate addition is simple, cost-effective and feasible at a practical scale, which will accelerate the practical use of anammox process for wastewater treatment.


Incidence of new cardiovascular events in patients with and without peripheral arterial disease seen in a vascular surgery clinic.
To investigate the incidence of death and of new cardiovascular events at long-term follow-up of patients with and without PAD seen in a vascular surgery clinic. We investigated the incidence of death, new stroke/transient ischemic attack, new myocardial infarction, new coronary revascularization, new carotid endarterectomy, new peripheral arterial disease (PAD) revascularization, or at least one of the above outcomes at long-term follow-up of patients with and without PAD followed in a vascular surgery clinic. At least one of the above outcomes occurred in 259 of 414 patients (63%) with PAD at 33-month follow-up and in 21 of 89 patients (24%) without PAD at 48-month follow-up (p<0.0001). Death occurred in 112 of 414 patients (27%) with PAD and in 10 of 89 patients (11%) without PAD (p=0.002). Stepwise Cox regression analysis for the time to at least one of the 6 outcomes showed that significant independent risk factors were men (hazard ratio =1.394; 95% CI, 1.072-1.813; p=0.013), estimated glomerular filtration rate (hazard ratio =0.992; 95% CI, 0.987-0.997; p=0.003), and PAD (hazard ratio =3.520; 95% CI, 2.196-5.641; p<0.0001). Stepwise Cox regression analysis for the time to death showed that significant independent risk factors were age (hazard ratio =1.024; 95% CI, 1.000-1.049; p=0.048), estimated glomerular filtration rate (hazard ratio =0.985; 95% CI, 0.974-0.996; p=0.007), and PAD (hazard ratio =2.157; 95% CI, 1.118-4.160; p=0.022). Patients with PAD have a significantly higher incidence of cardiovascular outcomes, especially death, new PAD revascularization, and new carotid endarterectomy, than patients without PAD followed in a vascular surgery clinic.


Biohydrogen and carboxylic acids production from wheat straw hydrolysate.
Hydrolyzed wheat straw was converted into carboxylic acids and biohydrogen using digesting bacteria. The fermentations were carried out using both free and membrane-encased thermophilic bacteria (55°C) at various OLRs (4.42-17.95g COD/L.d), in semi-continuous conditions using one or two bioreactors in a series. The highest production of biohydrogen and acetic acid was achieved at an OLR of 4.42g COD/L.d, whilst the highest lactic acid production occurred at an OLR of 9.33g COD/L.d. Furthermore, the bioreactor with both free and membrane-encased cells produced 60% more lactic acid compared to the conventional, free-cell bioreactor. In addition, an increase of 121% and 100% in the production of acetic and isobutyric acid, respectively, was achieved in the 2nd-stage bioreactor compared to the 1st-stage bioreactor.


Infantile spasms and neuronal heterotopias. A report on six cases.
Six infants with infantile spasms are presented, three primarily considered idiopathic and three symptomatic. Autopsy unexpectedly revealed heterotopias, cortical dyslamination, and other brain malformations in all six cases. No history of any etiologic moment during the neuronal migratory period of early gestation was found. It is concluded that wide-spread disturbances of CNS morphology should always be considered in cases of infantile spasms.


Improving CoQ10 productivity by strengthening glucose transmembrane of Rhodobacter sphaeroides.
Several Rhodobacter sphaeroides have been widely applied in commercial CoQ10 production, but they have poor glucose use. Strategies for enhancing glucose use have been widely exploited in R. sphaeroides. Nevertheless, little research has focused on the role of glucose transmembrane in the improvement of production. There are two potential glucose transmembrane pathways in R. sphaeroides ATCC 17023: the fructose specific-phosphotransferase system (PTSFru, fruAB) and non-PTS that relied on glucokinase (glk). fruAB mutation revealed two effects on bacterial growth: inhibition at the early cultivation phase (12-24 h) and promotion since 36 h. Glucose metabolism showed a corresponding change in characteristic vs. the growth. For ΔfruAΔfruB, maximum biomass (Biomax) was increased by 44.39% and the CoQ10 content was 27.08% more than that of the WT. glk mutation caused a significant decrease in growth and glucose metabolism. Over-expressing a galactose:H+ symporter (galP) in the ΔfruAΔfruB relieved the inhibition and enhanced the growth further. Finally, a mutant with rapid growth and high CoQ10 titer was constructed (ΔfruAΔfruB/tac::galPOP) using several glucose metabolism modifications and was verified by fermentation in 1 L fermenters. The PTSFru mutation revealed two effects on bacterial growth: inhibition at the early cultivation phase and promotion later. Additionally, biomass yield to glucose (Yb/glc) and CoQ10 synthesis can be promoted using fruAB mutation, and glk plays a key role in glucose metabolism. Strengthening glucose transmembrane via non-PTS improves the productivity of CoQ10 fermentation.


A magnetic resonance imaging-compatible, large-scale array for trans-skull ultrasound surgery and therapy.
Advances in ultrasound transducer array and amplifier technologies have prompted many intriguing scientific proposals for ultrasound therapy. These include both mildly invasive and noninvasive techniques to be used in ultrasound brain surgery through the skull. In previous work, it was shown how a 500-element hemisphere-shaped transducer could correct the wave distortion caused by the skull with a transducer that operates at a frequency near 0.8 MHz. Because the objective for trans-skull focusing is its ultimate use in a clinical context, a new hemispheric phased-array system has now been developed with acoustic parameters that are optimized to match the values determined in preliminary studies. The transducer was tested by focusing ultrasound through ex vivo human skulls and into a brain phantom by means of a phase-adaptive focusing technique. Simultaneously, the procedure was monitored by the use of magnetic resonance guidance and thermometry. The ultrasound focus of a 500-element 30-cm-diameter, 0.81-MHz array could be steered electronically through the skull over a volume of approximately 30 x 30 x 26 mm. Furthermore, temperature monitoring of the inner and outer surfaces of the skull showed that the array could coagulate targeted brain tissue without causing excessive skull heating. The successful outcome of these experiments indicates that intensities high enough to destroy brain tissue can be produced without excessive heating of the surrounding areas and without producing large magnetic resonance noise and artifacts.


Knowledge toward ovarian cancer symptoms among women in Syria: Cross-sectional study.
Ovarian cancer is the second most prevalent malignancy in women over 40, especially in low-income nations. For every 100,000 women in Syria, 473 new cases of ovarian cancer are diagnosed. This study aims to investigate the knowledge of ovarian cancer symptoms among Syrian women and determine the factors associated with good knowledge. An online cross-sectional was performed between July 29 and August 17, 2022. The inquired participants in the study were Syrian females above 18 years. The questionnaire consists of 41 questions organized into three sections: sociodemographic information, Confidence in recognizing ovarian cancer symptoms, and women's Awareness of the symptoms of ovarian cancer. This research included 557 Syrian women, and the average age was 23. Only 20.5% of involved women demonstrated a good knowledge of the symptoms of ovarian cancer. The participants who agreed that abdominal pain and pelvic pain are ovarian cancer symptoms formed (36.8%), and (63.9%), respectively. Regarding the additional presenting symptoms of ovarian cancer, "extreme generalized fatigue" was the most often reported symptom (66.1%). Divorced women showed greater knowledge scores than other marital status groups (7.13 ± 3.31, P-value<0.05), while public sector participants scored higher than other occupational groups (6.38 ± 2.5, P-value<0.05). Our findings indicate that Syrian females have inadequate knowledge regarding ovarian cancer symptoms. More ovarian cancer awareness programs for Syrian women of all ages are needed to increase the early identification of this illness.


Substance P in the uterine cervix, dorsal root ganglia and spinal cord during pregnancy and the effect of estrogen on SP synthesis.
Prior to parturition the non-pliable uterine cervix undergoes a ripening process ("softens" and dilates) to allow a timely passage of the fetus at term. The exact mechanism(s) triggering and involved in cervical ripening are unknown, though evidence for a role for sensory neurons and their contained neuropeptides is emerging. Moreover, an apparent increase in neuropeptide immunoreactive nerves occurs in the cervix during pregnancy, maternal serum estrogen levels rise at term and uterine cervix-related L6-S1 dorsal root ganglia (DRG) sensory neurons express estrogen receptor (ER) and neuropeptides. Thus, we sought to test the hypothesis that the neuropeptide substance P (SP) changes biosynthesis and release over pregnancy, that estrogen, acting via the ER pathway, increases synthesis of SP in DRG, and that SP is utilized in cervical ripening at late pregnancy. Using immunohistochemistry, in situ hybridization, reverse transcriptase-polymerase chain reaction (RT-PCR) and radioimmunoassay (RIA), we investigated coexpression of ER-alpha/beta and SP; differential expression of ER-alpha and -beta mRNA in DRG neurons; SP synthesis in DRG; and changes in SP concentration in the cervix, DRG and spinal cord over pregnancy. In addition, the effect of exogenous estrogen on SP synthesis in L6-S1 DRG of ovariectomized rats was examined. SP-immunoreactive neurons expressed ER-alpha and ER-beta. SP synthesis (expressed as beta-PPT mRNA label) was prominent in small DRG neurons. SP concentration increased in the L6-S1 DRG and spinal cord segments, with a peak at Day 20 of gestation, but decreased in the cervix during the first two trimesters, with a rise over the last trimester to Day 10 levels. SP and ER-alpha mRNA synthesis increased in DRG over pregnancy but ER-beta mRNA levels were largely unchanged. When ovariectomized rats were treated with exogenous estrogen, SP mRNA synthesis in the DRG increased in a dose-related manner, an effect blocked by ER blocker ICI 182 780. From these results, we postulate that synthesis of SP in L6-S1 DRG and utilization in the cervix increase over pregnancy and this synthesis is under influence of the estrogen-ER system, most likely ER-alpha. We postulate that SP may play a role in cervical ripening and, consequently in the birth process.


Carotid artery diameter in men and women and the relation to body and neck size.
To explore relationships among gender, body size, neck size, and the diameters of the common carotid artery (CCA) and internal carotid artery (ICA). Using multivariate regression, the best predictors of sonographic diameters of CCA and ICA were determined based on age, height, weight, body mass index, body surface area, neck circumference, neck length, and blood pressure. Measurements were obtained in 500 consecutive patients (age 52+/-15 years; 61% women). Mean diameters of ICA (4.66+/-0.78 mm) and CCA (6.10+/-0.80 mm) in women were significantly smaller than in men: 5.11+/-0.87 mm and 6.52+/-0.98 mm, respectively. Sex significantly influenced the diameters after controlling for body size, neck size, age, and blood pressure. Carotid arteries are smaller in women even after adjusting for body and neck size, age, and blood pressure.


Similar behavioral and biochemical effects of long-term haloperidol and caerulein treatment in albino mice.
Behavioral and biochemical experiments on male albino mice have revealed similar effects after the cessation of repeated (15 days) haloperidol (0.5 mg/kg daily IP) and caerulein (0.1 mg/kg daily SC) treatment. Tolerance developed to the action of muscimol (a GABA-A agonist, 1 mg/kg IP), caerulein (a CCK-8 agonist, 15 micrograms/kg SC) and flumazenil (a benzodiazepine antagonist, 10 mg/kg IP). Muscimol and caerulein were not able to suppress the motor activity of mice after 15 days treatment with haloperidol and caerulein. Flumazenil, which increased motor activity in saline-treated animals, also failed to affect activity after extended haloperidol or caerulein treatment. In contrast, the motor excitation induced by amphetamine (an indirect dopamine agonist, 3 mg/kg IP) was increased after haloperidol or caerulein administration. In radioligand binding studies the density of dopamine-2-receptors in striatum, opioid receptors in mesolimbic structures, and benzodiazepine and GABA-A receptors in brainstem was significantly elevated after long-term haloperidol or caerulein treatment. Simultaneously, the number of CCK-8, benzodiazepine and GABA-A receptors in cerebral cortex was decreased. It is probable that CCK-8-ergic mechanisms are involved closely in the action of repeated haloperidol treatment. CCK-8 seems to modulate the action of haloperidol through altering the sensitivity of dopamine, opioid, GABA-A and benzodiazepine receptors.


Trends in emergency department visits for mental health disorder diagnoses before and during the COVID-19 pandemic: a retrospective cohort study 2018-2021.
As the pandemic started, Saudi Arabia applied extreme measures in responses to the pandemic crises, which have adversely affected individuals' mental health. Therefore, the aim of this study is to describe trends in mental health disorder ED visit before and after the pandemic in two medical centers in Saudi Arabia. A retrospective cohort study using data from electronic health records in the Ministry of National Guard Health Affairs' (MNG-HA) in Saudi Arabia. Multiple logistic regression model was used to examine the age and sex differences in mental health disorder ED visits before and during the COVID19 pandemic. A total of 1117 ED psychiatric visits, a sharp increase in the number of psychiatric ED visits during the pandemic with an average increase of 25.66% was observed. During the pandemic, psychiatric ED visits were significantly higher in females (adjusted AOR 1.41, 95% CI 1.08, 1.83) than males. During the COVID19 pandemic, generalized anxiety disorder was the most common psychiatric ED disorder with reported increase of visits by 53.34% (p-value 0.005). ED psychiatric visits have consistently increased before and during the COVID19 pandemic. Interventions for mental health related disorders are urgently needed for preventing mental health problems among females.


Chronic granulocytic leukemia: correlation of blastic transformation type with karyotypic evolution.
Over a 3-year period, 26 patients with Philadelphia chromosome-positive chronic granulocytic leukemia were studied cytogenetically in both the chronic and blastic transformation phases of the disease; a further three patients were studied only after blastic transformation. Sixteen were considered to have adequate evidence of the type of transformation and form the basis of the report, where chromosome changes have been correlated with the morphological type of blastic transformation. Seven patients developed a myeloblastic transformation, seven a lymphoblastic transformation, and two an erythroblastic transformation. All patients in the myeloid group acquired one or more of the nonrandom changes associated with CGL blastic transformation, viz. +8,i(17q), +19, +22q-. Patients in the lymphoblastic group acquired structural abnormalities, apparently random in nature and usually in a small percentage of cells. The two patients with erythroblastic transformation developed markedly hyperdiploid cells (greater than 50 chromosomes) with both numerical and structural abnormalities. Patients in the lymphoblastic group appeared to have a slightly better prognosis than the myeloid group, whilst the patients with erythroblastic transformation had a very poor prognosis.


Paired associate learning in early infantile autism and receptive developmental aphasia.
A comparative study of nonverbal MA matched groups of five autistic, five aphasic, five normal and five retarded children was made on an auditory-visual and a visual-visual paired-associate learning task. The results showed that, although the autistic and receptive aphasic children were both unable to associate sounds with their visual counterparts at the same rate as normal children, the aphasic children overcame this deficit at a significantly faster rate than autistic children. This study pinpoints one dysfunction that could subsume the severe verbal comprehension defect in autistic and aphasic children.


The prediction of fetal oxygenation by an on-line computer analysis of fetal monitor output.
The deceleration delay index (DDI) is a statistical parameter computed by an on-line system which analyzes fetal monitor output for the temporal relationship between each uterine contraction and any associated deceleration of the fetal heart rate. A similar parameter describing this relationship is known to be quantitatively predictive of fetal oxygenation in the rhesus monkey, a species for which the level of fetal oxygenation is also known to be directly and progressively related to aberrations in the vital signs, to brain damage, and to fetal death. Thus, the DDI should be of significant, practical value in predicting fetal hypoxia at a time early enough to avert severe neonatal depression and/or hypoxic brain damage. A blind clinical trial of 106 parturients is presented which supports this concept by showing the DDI to be highly correlated with another oxygenation-associated parameter, the one-minute Apgar score.


The National Weight Control Registry: is it useful in helping deal with our obesity epidemic?
The National Weight Control Registry (NWCR) consists of over 4800 individuals who have been successful in long-term weight loss maintenance. The purpose of establishing the NWCR was to identify the common characteristics of those who succeed in long-term weight loss maintenance. We found very little similarity in how these individuals lost weight but some common behaviors in how they are keeping their weight off. To maintain their weight loss NWCR participants report eating a relatively low-fat diet, eating breakfast almost every day, weighing themselves regularly, and engaging in high levels (about 1 hour/day) of physical activity. Because this is not a random sample of those who attempt weight loss, the results have limited generalizability to the entire population of overweight and obese individuals. The value of this project lies in identifying potential strategies that may help others be more successful in keeping weight off.


Mitral Valve Regurgitation with a Rotary Left Ventricular Assist Device: The Haemodynamic Effect of Inlet Cannulation Site and Speed Modulation.
Mitral valve regurgitation (MVR) is common in patients receiving left ventricular assist device (LVAD) support, however the haemodynamic effect of MVR is not entirely clear. This study evaluated the haemodynamic effect of MVR with LVAD support and the influence of inflow cannulation site and LVAD speed modulation. Left atrial (LAC) and ventricular (LVC) cannulation was evaluated in a mock circulation loop with no, mild, moderate and severe MVR with constant speed and speed modulation (±600 RPM) modes. The use of an LVAD relieved pulmonary congestion during severe MVR, by reducing left atrial pressure from 20.5 to 10.8 (LAC) and 11.5 (LVC) mmHg. However, LAC resulted in decreased left ventricular stroke work (-0.08 J), ejection fraction (-7.9%) and higher MVR volume (+12.7 mL) and pump speed (+100 RPM) compared to LVC. This suggests that LVC, in addition to reducing MVR severity, also improves ventricular washout over LAC. LVAD speed modulation in synchrony with ventricular systole reduced MVR volume and increased ejection fraction with LAC and LVC, thus demonstrating the potential benefits of this mode, despite a reduction in cardiac output.


XX ovarian dysgenesis is caused by a PSMC3IP/HOP2 mutation that abolishes coactivation of estrogen-driven transcription.
XX female gonadal dysgenesis (XX-GD) is a rare, genetically heterogeneous disorder characterized by lack of spontaneous pubertal development, primary amenorrhea, uterine hypoplasia, and hypergonadotropic hypogonadism as a result of streak gonads. Most cases are unexplained but thought to be autosomal recessive. We elucidated the genetic basis of XX-GD in a highly consanguineous Palestinian family by using homozygosity mapping and candidate-gene and whole-exome sequencing. Affected females were homozygous for a 3 bp deletion (NM_016556.2, c.600_602del) in the PSMC3IP gene, leading to deletion of a glutamic acid residue (p.Glu201del) in the highly conserved C-terminal acidic domain. Proteasome 26S subunit, ATPase, 3-Interacting Protein (PSMC3IP)/Tat Binding Protein Interacting Protein (TBPIP) is a nuclear, tissue-specific protein with multiple functions. It is critical for meiotic recombination as indicated by the known role of its yeast ortholog, Hop2. Through the C terminus (not present in yeast), PSMC3IP also coactivates ligand-driven transcription mediated by estrogen, androgen, glucocorticoid, progesterone, and thyroid nuclear receptors. In cell lines, the p.Glu201del mutation abolished PSMC3IP activation of estrogen-driven transcription. Impaired estrogenic signaling can lead to ovarian dysgenesis both by affecting the size of the follicular pool created during fetal development and by failing to counteract follicular atresia during puberty. PSMC3IP joins previous genes known to be mutated in XX-GD, the FSH receptor, and BMP15, highlighting the importance of hormonal signaling in ovarian development and maintenance and suggesting a common pathway perturbed in isolated XX-GD. By analogy to other XX-GD genes, PSMC3IP is also a candidate gene for premature ovarian failure, and its role in folliculogenesis should be further investigated.


Superior mesenteric artery syndrome following initiation of cisplatin-containing chemotherapy: a case report.
Superior mesenteric artery syndrome is a rare cause of upper intestinal obstruction resulting from compression of the duodenum by the superior mesenteric artery and abdominal aorta. We describe a case of superior mesenteric artery syndrome in a 61-year-old Japanese man with non-small cell lung cancer who had been treated with cisplatin-containing chemotherapy and had lost 7 kg in weight. The diagnosis was confirmed by the typical findings of abdominal computed tomography showing distended stomach resulting from compression of the third portion of the duodenum and reduction of an aortomesenteric distance and aortomesenteric angle. This case highlights the importance of considering the possibility of superior mesenteric artery syndrome in patients treated with chemotherapy, especially those presenting with a low body mass index and showing weight loss during chemotherapy.


Calcium and vitamin D enriched diets increase and preserve vertebral mineral content in aging laboratory rats.
To assess the long-term effect of vitamin D or calcium supplementation on the skeletal metabolism of aging laboratory rodents, 1.5-month-old female Wistar rats were fed with diets containing twice the concentration of vitamin D (group 2) and of calcium (group 3) as in the usual rat chow. Follow-up to 24 months of age did not show significant differences between the enriched-diet groups and the controls (group 1) in terms of the vertebral body weight and protein content. Significantly higher bone mineral contents were found in groups 2 and 3 than were found in controls, as revealed by an increased bone mineral density (BMD: +62%, group 2; +48%, group 3) and vertebral calcium content (+73%, group 2; +84%, group 3). The vertebral alkaline phosphatase enzymatic activity was significantly lower in the enriched diet groups than in controls (-47%, group 2; -45%, group 3). The ratio alkaline phosphatase/acid phosphatase activity was markedly reduced in groups 2 and 3 (-57% and -59%, respectively), which might indicate a diminished rate of bone turnover. The trabecular bone volume (BV/TV) decreased in all groups during senescence, being significantly elevated in group 3 as compared to controls. Vitamin D and calcium dietary supplementations increase the axial mineral bone content in laboratory rats and might reduce the bone turnover. Their influence on the trabecular bone volume has yet to be examined.


A clinical coagulopathy score concurrent with viscoelastic testing defines opportunities to improve hemostatic resuscitation and enhance blood product utilization during liver transplantation.
An NIH clinical coagulopathy score has been devised for trauma patients, but no such clinical score exists in transplantation surgery. We hypothesize that that this coagulopathy score can effectively identify laboratory defined coagulopathy during liver transplantation and correlates to blood product utilization. TEGs were performed and coagulopathy scores (1, normal bleeding - 5, diffuse coagulopathic bleeding) were assigned by the surgeons at 5 intra-operative time points. Blood products used during the case were recorded between time points. Statistical analyses were performed to identify correlations between coagulopathy scores, TEG-detected abnormalities, and blood product utilization. Transfusions rarely correlated with the appropriate TEG measurements of coagulation dysfunction. Coagulopathy score had significant correlation to various transfusions and TEG-detected coagulopathies at multiple points during the case. High aggregate coagulopathy scores identified patients receiving more transfusions, re-operations, and longer hospital stays CONCLUSION: The combination of viscoelastic testing and a standardized clinical coagulopathy score has the potential to optimize transfusions if used in tandem as well as standardize communication between surgery and anesthesia teams about clinically evident coagulopathy.


Oral administration of all-trans retinoic acid suppresses experimental periodontitis by modulating the Th17/Treg imbalance.
A T-helper 17 (Th17)/regulatory T (Treg) imbalance has been suggested recently to play a role in the development of periodontitis. All-trans retinoic acid (ATRA) has been reported to modulate Th17/Treg imbalances in some diseases. However, the effect of ATRA on periodontitis remains unknown. This study observes the effect of ATRA on Th17/Treg imbalance modulation in experimental periodontitis. Experimental periodontitis was induced in mice by oral infection with Porphyromonas gingivalis (P. gingivalis). ATRA was orally administered every other day. Alveolar bone resorption (ABR) was estimated by measuring the distance from the cemento-enamel junction to the alveolar bone crest. CD4(+) T-cell subsets in the cervical lymph nodes (CLNs) and spleen were analyzed by flow cytometry. Th17/Treg cell-related cytokine messenger ribonucleic acid expression was quantified by real-time reverse transcription-polymerase chain reaction. The present data shows that ATRA suppressed ABR and inhibited inflammatory cell infiltration into periodontal tissues. These effects were closely associated with reduced CD4(+) retinoid-related orphan receptor γτ(+) cells and increased CD4(+) forkhead box P3(+) cells in the CLNs. Furthermore, ATRA downregulated interleukin (IL)-17A expression and upregulated IL-10 and transforming growth factor-β1 expression in both the CLNs and P. gingivalis-infected gingival tissues. These results suggest that ATRA modulation of the Th17/Treg imbalance provides protection against periodontitis by enhancing Treg cell activation and inhibiting Th17 cell activation. These results indicate the potential for clinical prevention of periodontitis.


Varicose bleeding after liver transplantation in a patient with severe portosystemic shunts.
Recipients for liver transplantation often have portosystemic shunts due to portal hypertension. It is an important problem whether such shunts should be ligated during operations. Ligating the shunts seems of benefit for increasing portal blood flow to the liver, but it is sometimes difficult technically, and it is invasive to the patient. We experienced a recipient with huge portosystemic shunts and no esophageal varices before living-related liver transplantation. Some shunts were ligated during operation to increase portal blood flow to the graft. Unfortunately, the patient suffered severe bleeding from esophagogastric varices after he underwent retransplantation owing to accidental liver failure. Based on our experience, extreme care should be exercised to avoid varicose bleeding after ligating the portosystemic shunts of liver transplantation patients.


Mindfulness to Manage Moral Injury: Rationale and development of a live online 7-week group intervention for veterans with moral injury.
Military service puts service members at risk for moral injury. Moral injury is an array of symptoms (e.g., guilt, shame, anger) that develop from events that violate or transgress one's moral code. We describe adaption of in-person mindfulness training program, Mindfulness to Manage Chronic Pain (MMCP), to address symptoms of moral injury to be delivered live via the web. We discuss how we will assess benchmarks (i.e., recruitment, credibility and acceptability, completion rates, and adherence) of the Mindfulness to Manage Moral Injury (MMMI) program. Aim 1: To develop and then adapt the MMCP program based on feedback from experts and veterans who took part in Study 1. Aim 2: To develop an equally intensive facilitator-led online Educational Support (ES) program to serve as a comparison intervention and conduct a run-through of each program with 20 veterans (10 MMMI; 10 ES). Aim 3: To conduct a small-scale randomized controlled trial (N = 42 veterans; 21 MMMI; 21 ES) in which we will collect pre-post-test and weekly benchmark data for both refined intervention arms. Study 1 and 2 are completed. Data collection for Study 3 will be completed in 2022. MMMI is designed to provide a live facilitated mindfulness program to address symptoms of moral injury. If Study 3 demonstrates good benchmarks, with additional large-scale testing, MMMI may be a promising treatment that can reach veterans who may not seek traditional VAMC care and/or who prefer a web-based program.


Cytomegalovirus anterior uveitis and occlusive retinal vasculitis without retinitis in a patient on immunomodulatory therapy.
To describe unusual clinical features and therapeutic management of a case of cytomegalovirus (CMV) ocular disease in a patient on immunomodulatory therapy. Moorfields Eye Hospital NHS Foundation Trust, London, UK. Medical history, clinical findings, investigation results, and multimodal imaging were retrospectively collected. A 61-year-old, South-East Asian man, developed CMV-related endotheliitis and occlusive retinal vasculitis, diagnosed by wide-angle fluorescein angiography. No retinitis was present on the fundus examination. Suspicion of CMV etiology was based on anterior segment findings, especially the presence of coin-shaped endothelial lesions. The diagnosis was confirmed by aqueous polymerase chain reaction (PCR) analysis which was positive for CMV DNA. The combined use of topical and systemic valganciclovir resulted in significant improvement of the picture. CMV can manifest in the eye as occlusive retinal vasculitis without the presence of typical retinitis.


A phase I trial of trimetrexate glucuronate (NSC 352122) given every 3 weeks: clinical pharmacology and pharmacodynamics.
Trimetrexate glucuronate (TMTX), a non-classic folate antagonist, has been evaluated clinically on several schedules. We studied TMTX given as an i.v. bolus over 5-30 min every 3 weeks in 44 patients with advanced solid tumors; it was given at doses ranging from 20 to 275 mg/m2. The maximal tolerated dose (MTD) on this schedule is 220 mg/m2, which we also recommend as a starting dose for phase II studies in patients without extensive prior therapy. Because of wide individual differences in drug tolerance, dose escalation in 25% increments is recommended for non-toxic patients. The principal dose-limiting toxicity was myelosuppression, although in some patients a flu-like syndrome precluded dose escalation. Significant rash and mucositis also frequently occurred in toxic patients. TMTX plasma concentrations were measured after the first dose and the data were fit by two- or three-compartment mammillary pharmacokinetic models. The TMTX clearance rate was 36.5 +/- 21 ml/min per m2 and did not change with dose; non-linearities with increasing dose were apparent in the steady-state volume of distribution (Vss) and in the terminal disposition half-life (t1/2). The difference between pre-treatment and nadir leucocyte counts was correlated with TMTX dose (r = 0.58; P = 0.0006) and with the area under the concentration vs time curve (AUC) (r = 0.41; P = 0.02). Pre-treatment plasma albumin concentrations correlated weakly with the nadir white blood count (r = -0.36; P = 0.047). Optimal schedules for the administration of TMTX have not been established and phase II trials using both bolus and daily X 5 schedules are under way.


Mid-pregnancy, perinatal, and neonatal reproductive endocrinology: a prospective cohort study in twins and singleton control subjects.
To answer the questions: Are perinatal reproductive hormone profiles different in case of a twin compared with a singleton pregnancy? Are reproductive endocrine profiles of twin girls influenced by their male co-twin and vice versa? Prospective cohort study from January 2004 to October 2009. Not applicable. A total of 204 mothers of twins and 248 singleton control subjects, aged >18 years, pregnant with a twin or singleton and no endocrine disease or malignancy. Blood samples were collected at mid-gestation from the mother and at delivery from the mothers and the umbilical cords. Estrogens, androgens, sex hormone-binding globulin, progesterone, and gonadotropins were measured. Hormonal profiles were compared between singletons and twins, different types of twins, and opposite-sex and same-sex twins. Estrogen and progesterone concentrations were higher in mothers of twins compared with singletons, but twin babies had lower estrogen and progesterone concentrations at birth. Opposite-sex twin girls did not have higher androgens in cord blood compared with same-sex twin girls. Boys of an opposite-sex twin had lower luteinizing hormone concentrations compared with dizygotic twin boys with a brother as a co-twin. Children from a twin are not overexposed to sex steroids at the time of birth, despite higher concentrations in their mothers, and girls from opposite sex twins do not show androgenic influences from their male co-twin. The female co-twin may influence the hypothalamic-pituitary-testicular axis of her brother via central inhibition.


Porphyrin test as an alternative to benzidine test for detecting cytochromes in catalase-negative gram-positive cocci.
A total of 66 strains of gram-positive cocci, including 21 catalase-negative members of the family Streptococcaceae and strains of Stomatococcus mucilaginosus, were investigated for the ability to produce porphobilinogen and porphyrin from delta-aminolevulinic acid as an alternative to the benzidine test for detecting the presence of cytochromes. Production of porphobilinogen correlated 100% with membership in the family Micrococcaceae.


Comparing the Use of a Mobile App and a Web-Based Notification Platform for Surveillance of Adverse Events Following Influenza Immunization: Randomized Controlled Trial.
Vaccine safety surveillance is a core component of vaccine pharmacovigilance. In Canada, active, participant-centered vaccine surveillance is available for influenza vaccines and has been used for COVID-19 vaccines. The objective of this study is to evaluate the effectiveness and feasibility of using a mobile app for reporting participant-centered seasonal influenza adverse events following immunization (AEFIs) compared to a web-based notification system. Participants were randomized to influenza vaccine safety reporting via a mobile app or a web-based notification platform. All participants were invited to complete a user experience survey. Among the 2408 randomized participants, 1319 (54%) completed their safety survey 1 week after vaccination, with a higher completion rate among the web-based notification platform users (767/1196, 64%) than among mobile app users (552/1212, 45%; P<.001). Ease-of-use ratings were high for the web-based notification platform users (99% strongly agree or agree) and 88.8% of them strongly agreed or agreed that the system made reporting AEFIs easier. Web-based notification platform users supported the statement that a web-based notification-only approach would make it easier for public health professionals to detect vaccine safety signals (91.4%, agreed or strongly agreed). Participants in this study were significantly more likely to respond to a web-based safety survey rather than within a mobile app. These results suggest that mobile apps present an additional barrier for use compared to the web-based notification-only approach. ClinicalTrials.gov NCT05794113; https://clinicaltrials.gov/show/NCT05794113.


SU6668 in idiopathic myelofibrosis--a rational therapeutic approach targeting several tyrosine kinases of importance for the myeloproliferation and the development of bone marrow fibrosis and angiogenesis.
Idiopathic myelofibrosis is a chronic myeloproliferative disorder being featured by progressive accumulation of connective tissue in concert with marked neovascularization (angiogenesis) of the bone marrow. Both fibrogenesis and angiogenesis are considered to develop consequent to the intramedullary release of various growth-promoting factors from rapidly proliferating and dysplastic megakaryocytes. Among these growth factors are platelet-derived growth factor (PDGF), basic fibroblast growth factor (bFGF), transforming growth factor beta (TGF-beta) and vascular endothelial growth factor (VEGF). The protein kinase inhibitor SU6668 is a potent antiangiogenic inhibitor of receptor tyrosine kinases, including those of VEGFR, PDGFR, bFGFR, and c-kit. The hypothesis is that SU6668 may be an effective agent in the treatment of idiopathic myelofibrosis. This compound has an inhibitory target profile on several tyrosine kinases involved in the myeloproliferation, the development of myeloid metaplasia (bFGFR, PDGFR, VEGFR, and c-kit) and the development of the major stromal changes in the bone marrow - fibrosis and angiogenesis (bFGFR, PDGFR, and VEGFR).


Favourable involvement of α2A-adrenoreceptor antagonism in the I₂-imidazoline binding sites-mediated morphine analgesia enhancement.
Aim of the present study was to obtain novel α(2)-adrenoreceptor (α(2)-AR) antagonists, possibly endowed with subtype-selectivity. Therefore, inspired by the non subtype-selective α(2)-AR antagonist idazoxan, we designed 1,4-dioxane derivatives bearing an aromatic area in position 5 or 6 and the imidazoline nucleus in position 2. Among the novel molecules 1-6, compound 2, with a trans stereochemical relationship between 5-phenyl and 2-imidazoline groups, was able to antagonize the sole α(2A)-subtype. Moreover, 2 showed an affinity at I(2)-imidazoline binding sites (I(2)-IBS) comparable to that at α(2A)-AR. In in vivo studies 2 strongly increased morphine analgesia. This interesting behaviour appeared to be induced by the favourable involvement of α(2A)-AR antagonism in the I(2)-IBS-mediated morphine analgesia enhancement.


Investigation of the understanding on tropical infectious diseases and oral health among dental professionals in China.
With the increasing exchange of domestic and international personnel, local infections of tropical infectious diseases are continuing in tropics and imported infections are emerging in non-tropics, some of which are accompanied by oral manifestations. Therefore, it is essential for dental professionals to identify the related oral manifestations, who are working for domestic medical service, international medical assistance, peace-keeping medical service or medical support of international joint military exercises. This study aims to investigate the attitude and knowledge of Chinese dental professionals on tropical infectious diseases and oral health, and to explore the difference between different genders, education backgrounds, professional identities, professional titles and tropics working experience. Network questionnaire was used to evaluate the knowledge and attitude of 236 Chinese dental professionals towards tropical infectious diseases and oral health. The scores of the participants on tropical infectious diseases and oral health were quite low. Although working experience in the tropics partially affected the understanding, there was no difference between different genders, education backgrounds, professional identities and professional titles. The understanding of dental professionals on tropical infectious diseases and oral health was insufficient. It is necessary to improve the clinical education and management specified with tropical infectious diseases and oral health.


Integration of responses within and across Arabidopsis natural accessions uncovers loci controlling root systems architecture.
Phenotypic plasticity is presumed to be involved in adaptive change toward species diversification. We thus examined how candidate genes underlying natural variation across populations might also mediate plasticity within an individual. Our implementation of an integrative "plasticity space" approach revealed that the root plasticity of a single Arabidopsis accession exposed to distinct environments broadly recapitulates the natural variation "space." Genome-wide association mapping identified the known gene PHOSPHATE 1 (PHO1) and other genes such as Root System Architecture 1 (RSA1) associated with differences in root allometry, a highly plastic trait capturing the distribution of lateral roots along the primary axis. The response of mutants in the Columbia-0 background suggests their involvement in signaling key modulators of root development including auxin, abscisic acid, and nitrate. Moreover, genotype-by-environment interactions for the PHO1 and RSA1 genes in Columbia-0 phenocopy the root allometry of other natural variants. This finding supports a role for plasticity responses in phenotypic evolution in natural environments.


Incubation of liver nuclei from hepatectomised rats with radioactive amino acids shows that the rates of synthesis of the globular and residual protein fractions increase linearly for up to 24 hr after hepatectomy, whereas the histones show maximal incorporation at 15 hr.


Terlipressin bolus induces systemic vasoconstriction in septic shock.
To report the use of a synthetic, long-acting, vasopressin analog, terlipressin, as an effective vasoconstrictor in septic shock. Case report. A 22-bed pediatric intensive care unit in a tertiary referral center. An 11-yr-old male with multiple-organism Gram-negative septic shock with high normal cardiac output as assessed by pulse contour analysis and low systemic vascular resistance despite norepinephrine infusion. Two peripherally administered doses of terlipressin (0.5 mg). Each dose of terlipressin was associated with a rapid increase in systemic vascular resistance, despite weaning and discontinuation of norepinephrine infusion from 0.15 microg.kg(-1).min(-1) lasting approximately 6 hrs. Terlipressin may be useful for sepsis-induced vasodilation.


The Effect of Hand Dominance on Patient-Reported Outcomes of Carpal Tunnel Release in Patients with Bilateral Carpal Tunnel Syndrome.
There is a paucity of studies in published literature that examines the effect of hand dominance on the resolution of symptoms following a carpal tunnel release. The objective of this study is to examine the effect of hand dominance on the resolution of symptoms following surgical decompression in patients with severe and moderate carpal tunnel syndrome. Bilateral carpal tunnel release (total 90 open and 84 endoscopic) was performed on 87 patients (11 males, 76 females) presenting with bilateral severe or moderate carpal tunnel syndrome of equal severity. Patient-reported outcome of resolution of symptoms were recorded, with patients followed up until complete resolution of symptoms or last recorded consultation (mean follow-up duration 11.4 months, range 3.1 to 32.4 months). In patients with bilateral severe carpal tunnel syndrome, a larger proportion of non-dominant hand (75.4%) achieved complete resolution compared to dominant hand (72.1%), and did so at a statistically shorter time (mean: 52.3 days) than the dominant hand (mean: 81.0 days). However, there was no statistically significant difference between proportion of patients and time taken before complete resolution of symptoms between dominant and non-dominant hand in patients with bilateral moderate carpal tunnel syndrome. Symptoms in the non-dominant hand resolved faster after carpal tunnel release in patients with severe carpal tunnel syndrome. We postulate that greater daily activity by the dominant hand compared to the non-dominant hand may be a contributing factor to its slower rate of symptoms resolution post-surgically in patients with bilateral severe carpal tunnel syndrome. This effect of hand dominance is not evident in post-surgical patients with moderate carpal tunnel syndrome.


Methionine-restricted diet inhibits growth of MCF10AT1-derived mammary tumors by increasing cell cycle inhibitors in athymic nude mice.
Dietary methionine restriction (MR) improves healthspan in part by reducing adiposity and by increasing insulin sensitivity in rodent models. The purpose of this study was to determine whether MR inhibits tumor progression in breast cancer xenograft model and breast cancer cell lines. Athymic nude mice were injected with MCF10AT1 cells in Matrigel® and fed a diet containing either 0.86 % methionine (control fed, CF), or 0.12 % methionine (MR) for 12 weeks. Plasma amino acid concentrations were measured by UPLC, and proliferation and apoptosis were examined using RT-PCR, immunohistochemistry, and Cell Titer 96® Aqueous One Solution Cell Proliferation assay. Mice on the MR diet had reduced body weight and decreased adiposity. They also had smaller tumors when compared to the mice bearing tumors on the CF diet. Plasma concentrations of the sulfur amino acids (methionine, cysteine, and taurine) were reduced, whereas ornithine, serine, and glutamate acid were increased in mice on the MR diet. MR mice exhibited decreased proliferation and increased apoptosis in cells that comprise the mammary glands and tumors of mice. Elevated expression of P21 occurred in both MCF10AT1-derived tumor tissue and endogenously in mammary gland tissue of MR mice. Breast cancer cell lines MCF10A and MDA-MB-231 grown in methionine-restricted cysteine-depleted media for 24 h also up-regulated P21 and P27 gene expression, and MDA-MB-231 cells had decreased proliferation. MR hinders cancer progression by increasing cell cycle inhibitors that halt cell cycle progression. The application of MR in a clinical setting may provide a delay in the progression of cancer, which would provide more time for conventional cancer therapies to be effective.


A soluble form of the interleukin 4 receptor in biological fluids.
Murine biological fluids and murine cell culture supernatants were analyzed for the presence of soluble murine interleukin 4 receptor (sIL4R) with the use of two monoclonal antibodies directed against the receptor. Mouse urine, serum, ascitic fluid, and cell culture supernatants contained varying levels of immunoreactive protein. All of the immunoreactive protein possessed interleukin 4 (IL 4) binding activity. Following partial purification of ascitic fluid a protein was isolated that binds IL 4 with high affinity. This data is consistent with the fact that murine biological fluids contain a soluble version of the murine IL 4 receptor that arises via secretion of the soluble receptor and/or via shedding of the extracellular portion of the full-length receptor from the cell surface.


Addressing racial and phenotypic bias in human neuroscience methods.
Despite their premise of objectivity, neuroscience tools for physiological data collection, such as electroencephalography and functional near-infrared spectroscopy, introduce racial bias into studies by excluding individuals on the basis of phenotypic differences in hair type and skin pigmentation. Furthermore, at least one methodology-electrodermal activity recording (skin conductance responses)-may be influenced not only by potential phenotypic differences but also by negative psychological effects stemming from the lived experience of racism. Here we situate these issues within structural injustice, urge researchers to challenge racism in their scientific work and propose procedures and changes that may lead to more equitable science.


Role of DNA topology in uptake of polyplex molecules by dendritic cells.
Dendritic cells (DCs) are an attractive target for DNA vaccines as they are potent antigen presenting cells. This study demonstrated how non-viral gene delivery to DCs involving complexes of poly-l-lysine (PLL) and plasmid DNA (pDNA) (polyplexes) showed dependence on DNA vector topology. DNA topology is of importance from both production and regulatory viewpoints. In our previous study with CHO cells we demonstrated that polyplex uptake was dependent on DNA topology whereby complexes containing supercoiled (SC) pDNA were smaller, more resistant to nucleases and more effectively condensed by PLL than open circular (OC) and linear-pDNA complexes. In this study polyplex uptake in DCs was measured qualitatively and quantitatively by confocal microscopy along with gene expression studies and measurement of DC phenotype. PLL is known for its ability to condense DNA and serve as an effective gene delivery vehicle. Quantification studies revealed that by 1h following uptake 15% (±2.59% relative standard error [RSE]) of SC-pDNA polyplexes were identified to be associated (fluorescent co-localisation) with the nucleus, in comparison to no nuclear association identified for OC- and linear-pDNA complexes. By 48 h following uptake, 30% (±1.82% RSE) of SC-pDNA complexes associated with the nucleus in comparison to 16% (±4.40% RSE) and 12% (±6.97% RSE) of OC- and linear-pDNA polyplexes respectively. Confocal microscopy images showed how DNA and PLL remained associated following uptake by dual labelling. Polyplex (containing 20 μg pDNA) gene expression (plasmid encoded lacZ [β-galactosidase] reporter gene) in DCs was greatest for SC-pDNA polyplexes at 14.12% unlike that of OC- (9.59%) and linear-pDNA (7.43%). DCs express cell surface markers which contribute towards antigen presentation. Polyplex gene expression did not alter DC phenotype through surface marker expression. This may be due to the pDNA dose employed (20μg) as other studies have used doses as high as 200 μg pDNA to induce DC phenotypic changes. Although no change in DC phenotype occurred, this could be advantageous in terms of biocompatibility. Collectively these results indicate that DNA topology is an important parameter for DC vector design, particularly pDNA in the SC conformation in regards to DNA vaccination studies.


A new colorimetric method for the determination of NADH/NADPH dependent glutathione reductase in erythrocytes and in plasma.
A simple and rapid colorimetric method for the assay of erythrocyte and plasma glutathione reductase (GR) activity is described. The method is based on the colorimetric measurement of reduced glutathione (GSH) (1) produced when the enzyme is incubated with oxidised glutathione (GSSG) in the presence of either reduced nicotinamide adenine dinucleotide (NADH) or reduced nicotinamide adenine dinucleotide phosphate (NADPH). Results of investigations on the effects of substrate and coenzyme concentrations, pH, EDTA, sodium/potassium chloride, and time, on enzyme activity are presented. Erythrocyte and plasma NADH-GR and NADPH-GR activities in 100 healthy blood donors, and 85 cord blood samples and plasma NADH-GR and NADPH-GR levels in patients with various disease conditions are given.


The positive relationship of serum paraoxonase-1 activity with apolipoprotein E is abrogated in metabolic syndrome.
High density lipoproteins (HDL) contain paraoxonase-1 (PON-1), which has strong anti-oxidative properties. Apolipoprotein E (apoE) may enhance PON-1 activity in vitro, but the extent to which PON-1 activity is determined by circulating apoE levels is unknown. Here we determined relationships of serum PON-1 activity with apoE in subjects without and with metabolic syndrome (MetS). We measured PON-1 activity (arylesterase activity), plasma apoE and serum amyloid A (SAA) in 93 subjects without and in 75 subjects with MetS (25 and 54 subjects with Type 2 diabetes mellitus (T2DM), respectively; p < 0.001). PON-1 activity was lower in MetS (p < 0.005) coinciding lower HDL cholesterol, apoA-I (p < 0.001)) and SAA levels (p < 0.01), whereas apoE was increased in relation to higher triglycerides (p < 0.01). In subjects without MetS, PON-1 activity was correlated positively with apoE (r = 0.376, p < 0.001), but this relationship was absent in MetS subjects (r = 0.085, p = 0.47). Multiple linear regression analysis showed that the relationship of PON-1 activity with apoE was different in subjects with MetS compared to subjects without MetS (β = -0.270, p = 0.014 for the interaction between apoE and MetS), independently from age, sex, T2DM, use of glucose lowering drugs, anti-hypertensives and the inverse relation with SAA levels (p = 0.008). Of the individual MetS components, apoE only interacted with low HDL-C on PON-1 activity (β = -0.175, p = 0.074). The relationship of apoE with PON-1 activity was neither modified by T2DM (p = 0.49), nor by SAA (p = 0.79). Higher apoE levels may confer higher PON-1 activity. The relationship of PON-I activity with total plasma apoE is apparently abrogated in MetS.


Glycosylation does not affect in-vitro biological activity of interleukin-3.
Murine interleukin-3 is secreted by activated T cells in three major molecular mass classes, which differ from one another in the extent of their N-linked glycosylation. Experiments were performed to determine whether carbohydrate content of different IL-3 glycoforms will affect their biological activity. IL-3 produced by activated T cells was biosynthetically labeled with 35S-methionine and the three major IL-3 glycoforms forms, with M(r) values of 22,000, 28,000 and 36,000, were purified using antibody affinity chromatography and preparative SDS-PAGE. Portions of these IL-3 glycoforms were enzymatically deglycosylated with N-glycanase and the bioactivity of each IL-3 glycoform and the corresponding deglycosylated fraction was compared in cell proliferation assays. The amount of 35S-label present in the samples was used as an index of protein amount so that equivalent concentrations of the various IL-3 forms could be compared. Our results indicate that the three major glycoforms have identical specific activity and that removal of N-linked carbohydrate does not change the specific in-vitro activity of IL-3. In addition to the three major glycoforms, small amounts of non-glycosylated IL-3 were also recovered from the affinity purified T-cell derived material. Using again the level of incorporated 35S as reference point, no difference in bioactivity compared with glycosylated IL-3 was detected. There is potential heterogeneity in IL-3 receptor complexes present on the many different cell types responsive to IL-3. We therefore tested whether the three IL-3 glycoforms differed in their interaction with various IL-3 responsive cell lines.(ABSTRACT TRUNCATED AT 250 WORDS)


Genetic analysis of developmental mechanisms in hydra. XVIII. Mechanism for elimination of the interstitial cell lineage in the mutant strain Sf-1.
The interstitial cell lineage in mutant strain sf-1 of hydra is temperature sensitive and is lost rapidly from tissue when the animal is cultured at a restrictive temperature of 23 degrees C or higher. The mechanism responsible for this cell elimination process was investigated. Sf-1 polyps were treated at a restrictive temperature of 27 degrees C for varying lengths of time, their tissues were macerated, and the resultant dissociated cells were examined for evidence of phagocytosis after Feulgen staining. It was found that large phagocytic vacuoles were present in the cytoplasm of some epithelial cells. These vacuoles contained partially degraded cells, whose nuclei had highly-condensed and intensely Feulgen-positive chromatin granules. This indicated that, as in colchicine-treated (Campbell, 1976) or starved (Bosch and David, 1984) wild-type hydra, the epithelial cells in strain sf-1 engulfed and disintegrated other cells in the phagocytic vacuoles. The incidence of phagocytosis was higher in sf-1 tissue maintained at elevated temperature than in sf-1 tissue maintained at normal temperature. However, the observed incidence was relatively low (maximally 0.14 phagocytosed cells per epithelial cell) and appeared to be too low to account for the very rapid interstitial cell loss occurring in this strain. We concluded that elimination of the interstitial cell lineage at a restrictive temperature in strain sf-1 takes place in part by phagocytosis and in part by other yet-unidentified mechanisms (cf., Marcum et al., 1980).


Identification of vaccinia promoters by heterologous expression of hepatitis B surface antigen in mouse cells infected by recombinant vaccinia viruses.
DNA fragments preceding open reading frames in a conserved segment of the vaccinia virus genome (Plucienniczak A., et al. (1985) Nucleic Acids Res. 13, 985-998) were cloned into plasmids upstream of the S gene of the hepatitis B virus encoding the surface antigen (HBsAg). Recombinant vaccinia virus obtained after insertion of these constructs into the thymidine kinase gene were used to infect mouse 1D cells. HBsAg was assayed in cellular supernatants. A strong promoter was thus identified in a 295 bp fragment preceding the coding region of the 147 kDa subunit of the vaccinia RNA polymerase.


Circadian rhythms of locomotor activity in zebrafish.
As part of an effort to characterize the circadian system of the zebrafish, we examined the circadian regulation of locomotor activity in adult males and females. Gross locomotor activity was measured using infrared movement detectors. The effects of light, dark, and temperature on the amplitude, phase, and free-running periods of locomotor rhythms were determined. When zebrafish were maintained in a 12-h light:12 h dark cycle at 25 degrees C, 86% of the fish were most active during the light phase of the cycle. The phases of free-running rhythms measured after transfer of fish from light cycles to constant conditions indicate that this diurnal activity profile reflects entrained circadian rhythmicity. When animals were maintained in constant conditions, the proportion that expressed significant circadian rhythmicity depended on ambient temperature. At 21 degrees C, 73% of the animals were rhythmic in constant darkness, and 65% were rhythmic in constant light. Fewer (28-59%) were rhythmic at 18 degrees, 25 degrees, and 28.5 degrees C. The free-running period of rhythmic animals was not affected by temperature within this range. The average period was shorter in constant light (LL; 12 lx) than in constant darkness (DD) in all but one experiment, and the difference was statistically significant for animals held at 21 degrees C. These data indicate that zebrafish locomotor activity is regulated by a circadian clock that is temperature compensated. Because rhythmicity is most robust at 21 degrees C, this would be the optimal temperature for future studies of the physiological basis of zebrafish behavioral rhythms.


Competition in Bembicium auratum (Gastropoda) and its effect on microalgal standing stock in mangrove muds.
Field experiments were undertaken on the intertidal gastropod Bembicium auratum, which is one of the dominant organisms in sheltered bays and mangroves in New South Wales. Animals were caged at various densities ranging from natural levels to about four times normal density. Increased density resulted in increased mortality (particularly of juveniles), reduced body weight, and (less conclusively) a decline in growth rate. Chlorophyll levels of the mangrove mud within the cages were monitored as an index of food availability, and increased rapidly in the control cages in the absence of Bembicium, remained steady at normal Bembicium densities, and declined at higher densities. Parallel experiments were conducted on the effects of substratum on Bembicium. One series of animals was allowed access to a hard substratum and the other only to mud. Body weights and survivorship were proportionally lower in animals deprived of a hard substratum. Juveniles were particularly susceptible to a shortage of food and more dependent on a hard substratum than adults. The population structure also suggests a high juvenile mortality but high adult survival and longevity. Bembicium is very abundant low on the shore but numbers decline and body weights increase higher up. Chlorophyll levels and amounts of hard substratum also decline up the shore. Possibly juveniles settle selectively (or survive better) low on the shore where oysters provide a dense hard substratum. Adults are less dependent on the hard substratum and may disperse up the shore, explaining the patterns of abundance and size.As Bembicium limits its own food supply and seemingly is limited by it, predation is unlikely to play an important role in restricting population density.