Computational studies of Family A and Family B GPCRs.
A full picture of the similarities between Family A and Family B GPCRs (G-protein coupled receptors) has been frustrated by the lack of clear homology between the respective sequences. Here, we review previous computational studies on GPCR dimerization in which the putative dimerization interfaces have been analysed using entropy, the ET (evolutionary trace) method and related methods. The results derived from multiple sequence alignments of Family A subfamilies have been mapped on to the rhodopsin crystal structure using standard alignments. Similarly, the results for the Family B alignments have been mapped on to the rhodopsin crystal structure using the 'cold-spot' alignment. For both Family A and Family B GPCRs, the sequence analysis indicates that there are functional sites on essentially all transmembrane helices, consistent with the parallel daisy chain model of GPCR oligomerization in which each GPCR makes interactions with a number of neighbouring GPCRs. The results are not too sensitive to the quality of the alignment. Molecular Dynamics simulations of the activation process within a single transmembrane bundle of the rhodopsin and the beta(2)-adrenergic receptor have been reviewed; the key observation, which is consistent with other computational studies, is that there is a translation and bending of helix 6, which contributes to a significant opening out of the intracellular face of the receptor, as shown in the accompanying movies. The simulations required the application of specific experiment-derived harmonic and half-harmonic distance restraints and so the application of such simulations to Family B GPCRs requires considerable care because of the alignment problem. Thus, in order to address the alignment problem, we have exploited the observation that GCR1, a plant GPCR, has homology with Family A, Family B and Family E GPCRs. The resulting alignment for transmembrane helix 3 is presented.


The role of sperm oxidative stress in male infertility and the significance of oral antioxidant therapy.
Oxidative stress in the male germ line is thought to affect male fertility and impact upon normal embryonic development. Accordingly, fertility specialists are actively exploring the diagnosis of such stress in spermatozoa and evaluating the possible use of antioxidants to ameliorate this condition. In this review, evidence for the presence of oxidative stress in human spermatozoa, the origins of this phenomenon, its clinical significance in the aetiology of male infertility and recent advances in methods for its diagnosis and treatment are re-examined. Moreover, an extensive review of the results presented in published clinical studies has been conducted to evaluate the overall impact of oral antioxidants on measures of sperm oxidative stress and DNA damage. Administration of antioxidants to infertile men has been assessed in numerous clinical studies with at least 20 reports highlighting its effect on measures of oxidative stress in human spermatozoa. A qualitative but detailed review of the results revealed that 19 of the 20 studies conclusively showed a significant reduction relating to some measure of oxidative stress in these cells. Strong evidence also supports improved motility, particularly in asthenospermic patients. However, of these studies, only 10 reported pregnancy-related outcomes, with 6 reporting positive associations. Adequately powered, placebo-controlled comprehensive clinical trials are now required to establish a clear role for antioxidants in the prevention of oxidative stress in the male germ line, such that the clinical utility of this form of therapy becomes established once and for all.


Quantifying transduction efficiencies of unmodified and tyrosine capsid mutant AAV vectors in vitro using two ocular cell lines.
With the increasing number of retinal gene-based therapies and therapeutic constructs, in vitro bioassays characterizing vector transduction efficiency and quality are becoming increasingly important. Currently, in vitro assays quantifying vector transduction efficiency are performed predominantly for non-ocular tissues. A human retinal pigment epithelial cell line (ARPE19) and a mouse cone photoreceptor cell line, 661W, have been well characterized and are used for many retinal metabolism and biologic pathway studies. The purpose of this study is to quantify transduction efficiencies of a variety of self-complementary (sc) adeno-associated virus (AAV) vectors in these biologically relevant ocular cell lines using high-throughput fluorescence-activated cell sorting (FACS) analysis. ARPE19 and 661W cells were infected with sc-smCBA-mCherry packaged in unmodified AAV capsids or capsids containing single/multiple tyrosine-phenylalanine (Y-F) mutations at multiplicity of infections (MOIs) ranging from 100 to 10,000. Three days post infection fluorescent images verified mCherry expression. Following microscopy, FACS analysis was performed to quantify the number of positive cells and the mean intensity of mCherry fluorescence, the product of which is reported as transduction efficiency for each vector. The scAAV vectors containing cone-specific (sc-mCARpro-green fluorescent protein [GFP]), rod-specific (sc-MOPS500-eGFP), retinal pigment epithelium (RPE)-specific (sc-VMD2-GFP), or ubiquitous (sc-smCBA-GFP) promoters were used to infect both cell lines at an MOI of 10,000. Three days post infection, cells were immunostained with an antibody raised against GFP and imaged. Finally, based on our in vitro results, we tested a prediction of transduction efficiency in vivo. Expression from unmodified scAAV1, scAAV2, scAAV5, and scAAV8 vectors was detectable by FACS in both ARPE19 and 661W cells, with scAAV1 and scAAV2 being the most efficient in both cell lines. scAAV5 showed moderate efficiency in both ARPE19 and 661W cells. scAAV8 was moderately efficient in 661W cells and was by comparison less so in ARPE19 cells; however, transduction was still apparent. scAAV9 performed poorly in both cell types. With some exceptions, the Y-F capsid mutations generally increased the efficiency of scAAV vector transduction, with the increasing number of mutated residues improving efficiency. Results for single scAAV1 and scAAV8 capsid mutants were mixed. In some cases, efficiency improved; in others, it was unchanged or marginally reduced. Retinal-specific promoters were also active in both cell lines, with the 661W cells showing a pattern consistent with the in vivo activity of the respective promoters tested. The prediction based on in vitro data that AAV2 sextuple Y-F mutants would show higher transduction efficiency in RPE relative to AAV2 triple Y-F capsid mutants was validated by evaluating the transduction characteristics of the two mutant vectors in mouse retina. Our results suggest that this rapid and quantifiable cell-based assay using two biologically relevant ocular cell lines will prove useful in screening and optimizing AAV vectors for application in retina-targeted gene therapies.


The Coup-TFII orphan nuclear receptor is an activator of the γ-globin gene.
The human fetal γ-globin gene is repressed in the adult stage through complex regulatory mechanisms involving transcription factors and epigenetic modifiers. Reversing γ-globin repression, or maintaining its expression by manipulating regulatory mechanisms, has become a major clinical goal in the treatment of β-hemoglobinopathies. Here, we identify the orphan nuclear receptor Coup-TFII (NR2F2/ARP-1) as an embryonic/fetal stage activator of γ-globin expression. We show that Coup-TFII is expressed in early erythropoiesis of yolk sac origin, together with embryonic/fetal globins. When overexpressed in adult cells (including peripheral blood cells from human healthy donors and β039 thalassemic patients) Coup-TFII activates the embryonic/fetal globins genes, overcoming the repression imposed by the adult erythroid environment. Conversely, the knock-out of Coup-TFII increases the β/γ+β globin ratio. Molecular analysis indicates that Coup-TFII binds in vivo to the β-locus and contributes to its conformation. Overall, our data identify Coup-TFII as a specific activator of the γ-globin gene.


Novel method to establish molecular identity using inter-simple sequence repeat markers in cotton (Gossypium hirsutum) cultivars.
Plant molecular identity (ID) is used to describe molecular characteristics of plants, which should contain all of the necessary information. Using inter-simple sequence repeat (ISSR) primers, molecular ID can be described in a way that reflects the polymerase chain reaction (PCR) conditions, annealing temperature, and the bands obtained in PCR amplification. A new complete molecular ID system is described in this study, which can be easily used and expanded to include more information. Using three cotton cultivars, we analyzed the products of PCR with ISSR primers and discussed the strategy for establishing their molecular ID. Using the segmented naming method, we designate the simple names and the full name systems of these three cultivars.


[Postoperative necrotizing sclerokeratitis].
Two cases of necrotizing sclerokeratitis following uncomplicated extracapsular cataract extraction are reported. Enucleation became necessary in the first case despite initially successful immunosuppressive treatment. In the second case, a stable condition was achieved by covering the affected area with a patch of lyophilized dura. Two years later, however, phthisis bulbi developed.


The buccal bifurcation cyst: in non-surgical treatment an option?
The mandibular buccal bifurcation cyst (BBC) was first described by Stoneman and Worth in 1983. It is a lesion with a specific location, classically the buccal surface of the mandibular first molar and less frequently the mandibular second molar. There have been numerous reports in the literature describing the BBC and its treatment. Treatment advocated thus far includes extraction of the involved first molar, marsupialization and enucleation of the cyst. In their most recent article, Pompura, Sándor and Stoneman reported on the successful treatment of 44 cysts with enucleation without tooth extraction. This article will describe the diagnostic features of the BBC and present three cases with a total of five cysts, which were treated non-surgically and ultimately resolved. The authors, therefore, propose that a more conservative non-surgical approach to these lesions may be considered.


Zone 1 and 2 flexor tendon repair via a longitudinal incision crossing the volar distal interphalangeal joint: functional and cosmetic outcomes.
The primary aim of this study was to present patient-reported functional outcomes after zone 1 and 2 flexor tendon repair using a novel longitudinal volar approach crossing the distal interphalangeal joint (DIPJ) perpendicularly. The secondary aims were to investigate the cosmetic outcomes, satisfaction and complication rates associated with this technique. Over a 6-year period, 75 patients underwent flexor tendon repair using this technique. The quick version of the Disabilities of the Arm, Shoulder and Hand (QuickDASH), five-level EQ-5D (EQ-5D-5L), Patient and Observer Scar Assessment Scale (POSAS) and satisfaction questionnaires were collected. QuickDASH scores were available for 59 patients (79%), and POSAS scores were available for 45 patients (60%) at a mean follow-up of 48 months. Concomitant nerve repairs were undertaken in 24 cases. The median QuickDASH score was 4.5 (interquartile range [IQR] 1.1-12.5) and median POSAS score was 12 (IQR 8-16). No significant difference was observed in median QuickDASH score between patients with and without a concomitant nerve repair (4.5 vs 6.8; p = 0.36). Fifty-six patients (94%) were satisfied. Two patients required open tenolysis, and a single case of early re-rupture occurred that required revision open repair. Utilising validated patient-reported functional outcomes, this study demonstrated that a direct longitudinal incision over the DIPJ for zone 1 and 2 flexor tendon repairs can result in excellent functional and cosmetic outcomes, with high rates of patient satisfaction and low rates of complications. This challenges previously held dogma that longitudinal incisions crossing flexion creases should be avoided.


Drug-resistant Staphylococcus aureus bacteria detection by combining surface-enhanced Raman spectroscopy (SERS) and deep learning techniques.
Over the past year, the world's attention has focused on combating COVID-19 disease, but the other threat waiting at the door-antimicrobial resistance should not be forgotten. Although making the diagnosis rapidly and accurately is crucial in preventing antibiotic resistance development, bacterial identification techniques include some challenging processes. To address this challenge, we proposed a deep neural network (DNN) that can discriminate antibiotic-resistant bacteria using surface-enhanced Raman spectroscopy (SERS). Stacked autoencoder (SAE)-based DNN was used for the rapid identification of methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-sensitive S. aureus (MSSA) bacteria using a label-free SERS technique. The performance of the DNN was compared with traditional classifiers. Since the SERS technique provides high signal-to-noise ratio (SNR) data, some subtle differences were found between MRSA and MSSA in relative band intensities. SAE-based DNN can learn features from raw data and classify them with an accuracy of 97.66%. Moreover, the model discriminates bacteria with an area under curve (AUC) of 0.99. Compared to traditional classifiers, SAE-based DNN was found superior in accuracy and AUC values. The obtained results are also supported by statistical analysis. These results demonstrate that deep learning has great potential to characterize and detect antibiotic-resistant bacteria by using SERS spectral data.


Mechanisms of bone loss.
We have seen rapid and important advances in the understanding of bone resorption processes at the cellular and molecular levels. Indeed, the field of molecular and cellular biology of bone is proliferating at a virtually exponential rate. Osteoclastic bone resorption, the major process involved in Type 1 osteoporosis, is the target of several pharmacological agents for the treatment of osteoporosis, and is the potential target of several agents that are proposed for the treatment of osteoporosis. This review will focus on (1) the factors known or postulated to affect bone loss; (2) the function of the differentiated osteoclast and the pharmacological effects on osteoclast function; (3) the processes involved in the generation and differentiation of mature osteoclasts and, again, the pharmacological implications of these processes.


Paleoclimatic Variation and Brain Expansion during Human Evolution.
One of the major adaptations during the evolution of Homo sapiens was an increase in brain size. Here we present evidence that a significant and substantial proportion of variation in brain size may be related to changes in temperature. Based on a sample of 109 fossilized hominid skulls, we found that cranial capacities were highly correlated with paleoclimatic changes in temperature, as indexed by oxygen isotope data and sea-surface temperature. Indeed, as much as 52% of the variance in the cranial capacity of these skulls could be accounted for by temperature variation at 100 ka intervals. As an index of more short-term seasonal fluctuations in temperature, we examined the latitude of the sites from which the crania originated. More than 22% of the variance in cranial capacity of these skulls could be accounted for by variation in equatorial distance.


Neuroendocrine Tumor of the Fallopian Tube and Serous Adenocarcinoma of the Ovary: Multicentric Primary Tumors.
Neuroendocrine tumors (NETs) comprise a large group of tumors that are most often localized in the gastrointestinal tract and lungs. They are rarely found in the organs of the female reproductive tract; such NETs are primarily localized in the ovaries. We present a case of multicentric primary low-grade NET of the fallopian tube and high-grade ovarian serous adenocarcinoma. In both tumor regions, the histotypes of neoplasms were determined by morphological and immunohistochemical investigations. The NET of the fallopian tube was diffusely positive for chromogranin A and CD56, but wild type for p53 and negative for CK7, CK20, and ER; Ki-67 expression was observed in 3% of the neoplastic cells. The ovarian serous adenocarcinoma was positive for CK7 and ER, mutant for p53, but negative for chromogranin A, CK20, and CD56; Ki-67 expression was observed in 45% of the tumor cells. These results support the possibility that NET can occur in the female reproductive tract and coexist with other malignant tumors.


Before your very eyes: illness, agency, and the management of Tourette Syndrome.
In this article, I examine the ways that people with Tourette Syndrome (TS) manage the motor and vocal tics characteristic of this neurological disorder. To mitigate the powerful stigmas associated with TS, individuals must either remove tics from public view or strive to recast the way that they are perceived. Drawing on ethnographic research with TS sufferers in Indiana, I elaborate three strategies by which this is done, strategies referred to here as displacement, misattribution, and contextualization. These processes strongly affect both the symptoms themselves and the subjective experience of the illness. They also affect the perception of TS in the larger culture, associating the disease with florid symptoms like cursing--symptoms that, although not at all typical of TS, are the ones most resistant to these kinds of management. These patterns highlight how individual agency may actively shape the cultural construction of illness.


A descriptive study of individuals with membranoproliferative glomerulonephritis.
Membranoproliferative Glomerulonephritis (MPGN) is one of a group of glomerulonephritides that often begins in childhood and progresses to renal failure. The purpose of this paper is to describe the initial results of a patient-driven database on individuals with MPGN. Patients with MPGN Types I, II, and III and their family members were recruited to this survey study. A population survey design was used. A survey was developed for this study that obtained information from the individual with MPGN or a guardian on: patient information, family/patient health history, history of PGN, medications, course of MPGN, history of dialysis, and history of transplant. Surveys were completed either on-line or in paper format. Fifty-nine individuals who are primarily white (80%) and female (61%) with MPGN Type II (52%) participated in this study. The average age of onset of MPGN in this sample was 12 years. Ten (71%) of the total 14 patients with onset of less than 10y ears of age were female. Among these 10 female, 8 (80%) were diagnosed with MPGN II and developed end stage renal disease. Health screenings for blood pressure, urinary dipstick for both proteinuria and hematuria play important roles in early detection for MPGN. Nurses must provide emotional and information support to this population.


[Group separation of caseins from cow milk on hydroxyapatite].
Ion exchange-chromatography method with the use of hydroxy apatite is considered one of the methods used for the casein fractionation particularly those containing phosphate in its composition. Simplicity of hydroxyapatite-preparation and its easily use in casein fractionation in batches are advantages of this method. Sephadex G-25 or Agarose is used to easement the sample passage through the column. By using of hydroxy-apatite in batches, alpha s-casein can easily separated with the use of buffer solution 0.006 M phosphate at pH 6.6 and the elution is carried out by buffer solution containing 0.25 M KH2PO4, 0.2 M KCl, 4.5 M urea and 0.002 M di-thiotreitol. Before the elution of alpha s-casein, beta- and kappa-casein are separated using buffer solutions containing 0.005 M and 0.1 M KH2PO4 in addition to the previously mentioned components.


Meiosis-specific stable binding of augmin to acentrosomal spindle poles promotes biased microtubule assembly in oocytes.
In the oocytes of many animals including humans, the meiotic spindle assembles without centrosomes. It is still unclear how multiple pathways contribute to spindle microtubule assembly, and whether they are regulated differently in mitosis and meiosis. Augmin is a γ-tubulin recruiting complex which "amplifies" spindle microtubules by generating new microtubules along existing ones in mitosis. Here we show that in Drosophila melanogaster oocytes Augmin is dispensable for chromatin-driven assembly of bulk spindle microtubules, but is required for full microtubule assembly near the poles. The level of Augmin accumulated at spindle poles is well correlated with the degree of chromosome congression. Fluorescence recovery after photobleaching shows that Augmin stably associates with the polar regions of the spindle in oocytes, unlike in mitotic cells where it transiently and uniformly associates with the metaphase spindle. This stable association is enhanced by γ-tubulin and the kinesin-14 Ncd. Therefore, we suggest that meiosis-specific regulation of Augmin compensates for the lack of centrosomes in oocytes by actively biasing sites of microtubule generation within the spindle.


Treatment of endometrial carcinoma.
The optimal treatment for endometrial carcinoma is radical hysterectomy and hemivaginectomy with pre-operative caesium sources in the uterus and vaginal vault. Block pelvic lymphadenectomy is unsatisfactory, and gland extension is best treated by radiotherapy to the lateral pelvic and para-aortic fields. Surgery has a definite place in the treatment of advanced cases.


Laparoscopic pancreaticoduodenectomy for tumors of the head of pancreas; 10 cases for a single center experience.
The tumors of the head of the pancreas are one of the leading causes of cancer-related death in Western countries. The current gold standard for these tumors is a Whipple procedure. This procedure did not change in its surgical steps since when it was initially introduced in 1935. More recently, a laparoscopic approach with similar outcomes has been described. The aim of this paper is to describe the laparoscopic surgical technique performed in our unit, reporting single center postoperative outcomes. From the 1st January 2013 to the 31st December 2015 a database was created. Data about patients who underwent a laparoscopic pancreaticoduodenectomy (LPD) were collected prospectively. All patients were preoperatively assessed with blood samples, tumor markers, CT chest abdomen and pelvis and/or MRI pancreas. Only patients with specific characteristics were considered eligible for an LPD: performance status 0, body mass index (BMI) less than 30 kg/m2, a small neoplastic lesion (< 3.5 cm) confined to the pancreas, the absence of infiltrated organs and/or blood vessels (T1 or T2). Postoperative data and complications were recorded and described according to the Clavien-Dindo classification and the international study group of pancreatic surgery definitions. In a time interval of 36 months, 31 patients with an initially considered resectable pancreatic cancer were referred. 11 patients were found to have metastasis during the preoperative workout. Only 10 patients were considered eligible for a LPD. Six of them were men (60%). The mean BMI was 25.01 kg/m2 (19.6-29.8). 5 patients, who underwent to LPD did not have any comorbidities. An overall 50% of all patients were jaundice at the time of diagnosis with a mean bilirubin level of 181.3 µmol/L (119.7-307.8). All patients with a direct bilirubin greater than 250 µmol/L underwent a preoperative percutaneous biliary drainage. In the majority of the LPD performed (50%), the histology reported a pancreatic adenocarcinoma. Other postoperative histology described were: IPMN (20%), ampullar neoplasia (20%) and neuroendocrine tumor (10%). Neo-adjuvant chemotherapy was never considered indicated. The reported postoperative complications were: 1 anastomotic bleeding, 2 pancreatic fistula, 1 infected intra-abdominal collection and 1 delay gastric emptying. The pancreatic fistulas were considered grade A and grade B. One fatality after LPD occurred because of an uncontrollable, diffuse severe hemorrhagic gastritis associated with a GJ anastomosis bleeding in the POD 25. The mean hospital stay was 12.3 days (8-25). The mean operative time was 224 min (170-310). There were no intraoperative complications. The main intraoperative blood loss was 220 ml (180-400) and intraoperative blood transfusions were not required. The resection margins were negative (R0) in 100% of cases and the mean lymph nodes harvested were 24 (18-40). The LPD is still a not common practice. Our results are comparable with those reported in literature about the open technique. These remarkable surgical outcomes are probably related to the extremely careful preoperative patient selection performed. The indication for a laparoscopic vs. an open pancreaticoduodenectomy was based on a CT scan pancreas performed less than 30 days before the planned date of surgery and a careful preoperative assessment. A low complication rate and a relative short stay in hospital were associated to a good quality of life in the early postoperative period and an early referral for postoperative chemotherapy. Good clinical outcomes were associated with outstanding oncological results. Laparoscopic pancreaticoduodenectomy is a feasible surgical procedure. Remarkable oncological and surgical outcomes can be achieved with a morbidity and mortality rate in line with the data reported by the large series of open procedures.


Evolving global risk assessment of ocular hypertension to glaucoma.
To discuss current knowledge of global risk assessment in ocular hypertension. The ophthalmologist treating patients with ocular hypertension is frequently faced with the clinical dilemma of which patients to treat and how vigorous treatment should be. The goal of risk assessment for glaucoma is to identify patients at greatest risk for symptomatic vision loss. Risk factors can be identified by history such as age, race, and family history or can be clinically observed by examination such as elevated intraocular pressure, optic nerve head appearance, central corneal thickness, and visual field abnormalities. Risk assessment is a well accepted tool in other fields of medicine. Parallels can be drawn between the evolution of risk assessment for coronary artery disease and glaucoma. Validated risk calculators for ocular hypertension are currently available mostly derived from the Ocular Hypertension Treatment Study. The aim of assessing global risk for conversion from ocular hypertension to glaucoma is to identify patients who are most likely to benefit from early treatment. Calculation of risk should be accompanied by thorough analysis of risks, benefits, and alternatives for the individual patient.


Delineation of a conserved arrestin-biased signaling repertoire in vivo.
Biased G protein-coupled receptor agonists engender a restricted repertoire of downstream events from their cognate receptors, permitting them to produce mixed agonist-antagonist effects in vivo. While this opens the possibility of novel therapeutics, it complicates rational drug design, since the in vivo response to a biased agonist cannot be reliably predicted from its in cellula efficacy. We have employed novel informatic approaches to characterize the in vivo transcriptomic signature of the arrestin pathway-selective parathyroid hormone analog [d-Trp(12), Tyr(34)]bovine PTH(7-34) in six different murine tissues after chronic drug exposure. We find that [d-Trp(12), Tyr(34)]bovine PTH(7-34) elicits a distinctive arrestin-signaling focused transcriptomic response that is more coherently regulated across tissues than that of the pluripotent agonist, human PTH(1-34). This arrestin-focused network is closely associated with transcriptional control of cell growth and development. Our demonstration of a conserved arrestin-dependent transcriptomic signature suggests a framework within which the in vivo outcomes of arrestin-biased signaling may be generalized.


A cryptic activity in the Nudix hydrolase superfamily.
The Nudix hydrolase superfamily is identified by a conserved cassette of 23 amino acids, and it is characterized by its pyrophosphorylytic activity on a wide variety of nucleoside diphosphate derivatives. Of the 13 members of the family in Escherichia coli, only one, Orf180, has not been identified with a substrate, although a host of nucleoside diphosphate compounds has been tested. Several reports have noted a strong similarity in the three-dimensional structure of the unrelated enzyme, isopentenyl diphosphate isomerase (IDI) to the Nudix structure, and the report that a Nudix enzyme was involved in the synthesis of geraniol, a product of the two substrates of IDI, prompted an investigation of whether the IDI substrates, isopentenyl diphosphate (IPP), and dimethylallyl diphosphate (DAPP) could be substrates of Orf180. This article demonstrates that Orf180 does have a very low activity on IPP, DAPP, and geranyl pyrophosphate (GPP). However, several of the other Nudix enzymes with established nucleoside diphosphate substrates hydrolyze these compounds at substantial rates. In fact, some Nudix hydrolases have higher activities on IPP, DAPP, and GPP than on their signature nucleoside diphosphate derivatives.


Activation reduction in anterior temporal cortices during repeated recognition of faces of personal acquaintances.
Repeated recognition of the face of a familiar individual is known to show semantic repetition priming effect. In this study, normal subjects were repeatedly presented faces of their colleagues, and the effect of repetition on the regional cerebral blood flow change was measured using positron emission tomography. They repeated a set of three tasks: the familiar-face detection (F) task, the facial direction discrimination (D) task, and the perceptual control (C) task. During five repetitions of the F task, familiar faces were presented six times from different views in a pseudorandom order. Activation reduction through the repetition of the F tasks was observed in the bilateral anterior (anterolateral to the polar region) temporal cortices which are suggested to be involved in the access to the long-term memory concerning people. The bilateral amygdala, the hypothalamus, and the medial frontal cortices, were constantly activated during the F tasks, and considered to be associated with the behavioral significance of the presented familiar faces. Constant activation was also observed in the bilateral occipitotemporal regions and fusiform gyri and the right medial temporal regions during perception of the faces, and in the left medial temporal regions during the facial familiarity detection task, which are consistent with the results of previous functional brain imaging studies. The results have provided further information about the functional segregation of the anterior temporal regions in face recognition and long-term memory.


Treatment gap in common mental disorders: the Singapore perspective.
The problem of wide treatment gaps in mental disorders is endemic world wide. The study aims to establish the treatment gap of common mental disorders in Singapore. A national sample of 6616 persons aged 18 years and above was surveyed with the World Mental Health Composite International Diagnostic Interview in which for each diagnostic module, respondents were asked a series of questions regarding treatment contact. Treatment gap varied considerably between disorders; alcohol abuse had the largest treatment gap (96.2%), followed by obsessive compulsive disorder (89.8%) and alcohol dependence (88.3%). The disorder for which people were most likely to seek help was major depressive disorder. Women with dysthmia were more likely than men to seek help but this help seeking behavior was reversed among those with alcohol abuse and dependence. Age of onset was significantly associated with treatment contact with those who had an earlier age of onset less likely to have treatment contact than those with late age of onset for all disorders except obsessive compulsive disorder. Our findings suggest that treatment gaps are wide even in an economically developed country like Singapore and other than sociodemographic factors, cultural influences might play an important role in help seeking behavior.


Comparion of stability in titanium implants with different surface topographies in dogs.
A few of studies which compared and continuously measured the stability of various surface treated implants in the same individual had been performed. We aim to find the clinical significance of surface treatments by observing the differences in the stabilization stages of implant stability. Eight different surface topographies of dental implants were especially designed for the present study. Machined surface implants were used as a control group. 4 nano-treated surface implants (20 nm TiO(2) coating surface, heat-treated 80 nm TiO(2) coating surface, CaP coating surface, heat treated CaP coating surface) and 3 micro-treated surface implants [resorbable blast media (RBM) surface, sandblast and acid-etched (SAE) surface, anodized RBM surface] were used as experiment groups. All 24 implants were placed in 3 adult dogs. Periotest® & ISQ values measured for 8 weeks and all animals were sacrificed at 8 weeks after surgery. Then the histological analyses were done. In PTV, all implants were stabilized except 1 failed implants. In ISQ values, The lowest stability was observed at different times for each individual. The ISQ values were showed increased tendency after 5 weeks in every groups. After 4 to 5 weeks, the values were stabilized. There was no statistical correlation between the ISQ values and PTV. In the histological findings, the bone formation was observed to be adequate in general and no differences among the 8 surface treated implants. In this study, the difference in the stability of the implants was determined not by the differences in the surface treatment but by the individual specificity.


Surgical Continuum Manipulator Control Using Multiagent Team Deep Q Learning.
Continuum manipulator has shown great potential in surgical applications. The flexibility of the continuum manipulator helps it achieve many complicated surgeries, such as neurosurgery, vascular surgery, abdominal surgery, etc. In this paper, we propose a Team Deep Q learning framework (TDQN) to control a 2-DoF surgical continuum manipulator with four cables, where two cables in a pair form one agent. During the learning process, each agent shares state and reward information with the other one, which namely is centralized learning. Using the shared information, TDQN shows better targeting accuracy than multiagent deep Q learning (MADQN) by verifying on a 2-DoF cable-driven surgical continuum manipulator. The root mean square error during tracking with and without disturbance are 0.82mm and 0.16mm respectively using TDQN, whereas 1.52mm and 0.98mm using MADQN respectively.Clinical Relevance-The proposed TDQN shows a promising future in improving control accuracy under disturbance and maneuverability in robotic-assisted endoscopic surgery.


Non-invasive assessment of fluid volume status in the interstitium after haemodialysis.
During dialysis excess fluid is removed from uraemic patients. The excess fluid is mainly located in the skin and subcutaneous tissues. In this study we wished, with two noninvasive techniques, the IM (impression method) and BIA (bioimpedance analysis), to study what mechanical (IM) and electrical cellular membrane (BIA) effects the fluid withdrawal has on these tissues. The IM measures the resistive force of the tissues when mechanically compressed. From the force curve two parameters are calculated, the F(0), indicative of interstitial tissue pressure and the FT corresponding to the translocation of tissue fluid (interstitial movable water). The BIA phase angle shift (phi), i.e. geometrical angular transformation of the ratio between reactance and resistance, which has been associated with cellular membrane function, was used as a measurement of electrical cellular membrane effects. Twenty patients were studied before and after haemodialysis measuring the F(0), FT and phi. Theresults showed that the patients lost a median of 3.7 kg during the haemodialysis. F(0) increased until after dialysis, but did not reach significant values, whereas FT increased significantly after dialysis, p < 0.001, as compared with before. After a peak at one hour postdialysis the FT value returned to predialysis values at four hours after termination of dialysis. Also phi increased from before to after dialysis, p < 0.001, but already after one hour it returned to predialysis values. It is common knowledge that dialysis alters the dynamics of fluid in the interstitium of the skin and subcutis. We conclude that the impression method is sensitive enough to detect and chronicle these changes. Furthermore, with the BIA, (phase angle) signs of changes in the electrical properties of the tissues, possibly reflecting cellular membrane function, could be detected.


Solving the Structures of Polycrystalline Materials: from the Debye-Scherrer Camera to SwissFEL.
The development of powder diffraction (PD) techniques for structure analysis is traced from its inception almost 100 years ago to the present day, with a brief glimpse of what SwissFEL can contribute in the near future. Although PD data were used in the early days to deduce some simple high-symmetry structures, it was not until computers, instrumentation and synchrotrons arrived on the scene that the true potential of PD data could be realized. In the last 25 years, PD has blossomed into a viable method, not only for structure refinement, but also for structure solution. This means that scientists with polycrystalline materials that cannot be grown as single crystals can still obtain the structural information they need. Historically, structure solution from PD data began with model building, progressed through the application of single-crystal methods to simpler structures and the adaptation of those methods to the special problems posed by PD data, the development of automated model-building algorithms, and most recently to the application of charge flipping. As X-ray sources and detectors continue to develop, the boundary between a powder and a single crystal is narrowing. Laue microdiffraction techniques and the prospects offered by SwissFEL will allow single-crystal data to be collected on some polycrystalline materials.


A prospective clinical evaluation of the longevity of resorbable sutures in oral surgical procedures.
The objectives of this prospective randomized study were to clinically evaluate the longevity of resorbable sutures (chromic catgut and vicryl) and determine the effect of chlorhexidine mouth wash on their absorption time in oral surgical procedures. Both sutures were of size 3/0 with round body needle and were placed using the standard technique. One hundred patients selected for the study were divided into two groups of 50 patients each (vicryl and chromic catgut groups). Each group was subsequently subdivided into chlorhexidine (17 cases); warm saline mouth wash (17 cases) and warm water mouth wash (16 cases (control). The sutures were placed during various minor surgical procedures e.g. third molar surgery (65 cases), incisional biopsy (7 cases), excisional biopsy (8 cases), sutured lacerations (10 cases) and malar elevations through intraoral upper buccal sulci approach (8 cases) and 2 cases of cystic enucleations. The mean longevity of chromic catgut for chlorhexidine was 11.4 days with a range of 5-16 days. The patients, who used warm saline mouth wash, had a mean longevity of 11.7 days with a range of 7-24 days. The mean longevity of vicryl in patients that used chlorhexidine was 22.7 days, with a range of 14-36 days and that of warm saline mouth wash was 24.5 days with a range of 14-47 days. The resorbable sutures investigated have a mean longevity, which was slightly shorter than the figure stated by the manufacturer. Chlorhexidine was found to have no appreciable effect on absorption time of the sutures.


Hippocampal cell death following ischemia: effects of brain temperature and anesthesia.
The effect of brain temperature and anesthesia on ischemic neuronal damage was studied in the hippocampal formation using the four vessel occlusion model in awake and anesthetized rats. Neuronal damage was assessed by immunocytochemistry and silver impregnation of tissue sections. The degree of ischemia was monitored by recording spontaneous and evoked electrical activity from the hippocampus and dentate gyrus in all animals. In addition, the hippocampal temperature and oxygen tension were also recorded using a chamber-type thin-film microelectrode in the anesthetized animals. Fifteen minutes ischemia in the awake animals caused greater neuronal damage and mortality of animals than 30 min ischemia in anesthetized rats. The temperature of the brain was found to drop by 4-6 degrees C during complete forebrain ischemia in the latter group. Neuronal damage was observed infrequently in the hippocampus of these animals. When the brain temperature was kept constant at the preischemic level during 30 min occlusion, all animals died within a day, while after 15 min occlusion the majority showed an almost complete degeneration of CA1 pyramidal cells and hilar somatostatin immunoreactive neurons. Following 15 min ischemia, the awake animals showed a similar cell loss in the CA1 region and the hilus. It is concluded that, in the anesthetized animals prepared for acute recording, the decreased temperature of the brain during ischemia is a major factor in protecting neurons from damage, but that Equithesin anesthesia also has a significant protective effect. Consistent ischemic degeneration occurs in awake animals by four vessel occlusion, if the brain temperature is controlled and the completeness of ischemia is monitored by recording spontaneous and evoked electrical activity with chronic electrodes.


Fibrinogen induces adhesion, spreading, and microfilament organization of human endothelial cells in vitro.
Human umbilical vein endothelial cells (ECs) have been shown to attach to a substratum of fibrinogen (fg). Later, ECs undergo spreading, organization of thick microfilament bundles of the stress fiber type, and formation of focal contacts (adhesion plaques) that correspond to accumulation of vinculin at the cytoplasmic aspect of the ventral membrane. The rate of attachment to fg and the type of spreading is virtually identical to that obtained on substrata coated with fibronectin (FN). Antibodies to fg, but not to FN, prevent EC adhesion to fg; conversely, antibodies to FN, but not to fg, prevent adhesion of ECs to a FN-coated substratum. The removal of residual FN contamination from fg preparations by means of DEAE-cellulose chromatography does not result in any difference in EC adhesion on fg. Moreover, pretreatment of cells with inhibitors of synthesis and release of proteins does not impair their adhesion capacity on an fg-coated substratum. In contrast, human arterial smooth muscle cells do not adhere and spread on fg substrata but do so on FN. The synthetic peptides (Gly-Arg-Gly-Asp[GRGD] and Gly-Arg-Gly-Asp-Ser-Pro[GRGDSP]) containing the tripeptide Arg-Gly-Asp (RGD), originally found to be responsible for the cell binding activity of FN, have been found to inhibit EC spreading and the redistribution of their cytoskeleton, including the formation of stress fibers and the localization of vinculin either on fg or on FN. Conversely, the synthetic peptide Arg-Gly-Gly (RGG) was completely uneffective in inhibiting the adhesion and the sequence of events leading to spreading and cytoskeletal organization. These results indicate that ECs, but not smooth muscle cells, specifically adhere and spread on an fg substratum and this occurs by recognition mechanisms similar to those reported for FN.


The potential of genes and other markers to inform about risk.
Advances in biotechnology have raised expectations that biomarkers, including genetic profiles, will yield information to accurately predict outcomes for individuals. However, results to date have been disappointing. In addition, statistical methods to quantify the predictive information in markers have not been standardized. We discuss statistical techniques to summarize predictive information, including risk distribution curves and measures derived from them, that relate to decision making. Attributes of these measures are contrasted with alternatives such as receiver operating characteristic curves, R(2), percent reclassification, and net reclassification index. Data are generated from simple models of risk conferred by genetic profiles for individuals in a population. Statistical techniques are illustrated, and the risk prediction capacities of different risk models are quantified. Risk distribution curves are most informative and relevant to clinical practice. They show proportions of subjects classified into clinically relevant risk categories. In a population in which 10% have the outcome event and subjects are categorized as high risk if their risk exceeds 20%, we identified some settings where more than half of those destined to have an event were classified as high risk by the risk model. Either 150 genes each with odds ratio of 1.5 or 250 genes each with odds ratio of 1.25 were required when the minor allele frequencies are 10%. We show that conclusions based on receiver operating characteristic curves may not be the same as conclusions based on risk distribution curves. Many highly predictive genes will be required to identify substantial numbers of subjects at high risk.


Nanocellulose: A comprehensive review investigating its potential as an innovative material for water remediation.
Rapid growth in industrialization sectors, the wastewater treatment plants become exhausted and potentially not able to give desirable discharge standards. Many industries discharge the untreated effluent into the water bodies which affects the aquatic diversity and human health. The effective disposal of industrial effluents thus has been an imperative requirement. For decades nanocellulose based materials gained immense attraction towards application in wastewater remediation and emerged out as a new biobased nanomaterial. It is light weighted, cost effective, mechanically strong and easily available. Large surface area, versatile surface functionality, biodegradability, high aspect ratio etc., make them suitable candidate in this field. Majorly cellulose based nanomaterials are used in the form of cellulose nanocrystals (CNCs), cellulose nanofibers (CNFs), or bacterial nanocellulose (BNC). This review specifically describes about a variety of extraction methods to produced nanocellulose and also discusses the modification of nanocellulose by adding functionalities in its surface chemistry. We majorly focus on the utilization of nanocellulose based materials in water remediation for the removal of different contaminants such as dyes, heavy metals, oil, microbial colony etc. This review mainly emphasizes in ray of hope towards nanocellulose materials to achieve more advancement in the water remediation fields.


Preferences in choosing between breast reconstruction options: a survey of female plastic surgeons.
Female plastic surgeons are well suited to make a personal choice regarding breast reconstruction options, based on their knowledge of the actual procedures and first-hand experience with results. The authors surveyed this group to elicit their personal views on various modalities of breast reconstruction and to ascertain which types of reconstruction they would choose if faced with such a decision. All board-certified female plastic surgeons in the United States and Canada were surveyed by means of e-mail. This survey included questions regarding basic demographic and practice data. Respondents were requested to rank desired methods of reconstruction for themselves and to cite reasons for these choices. A total of 435 surveys were sent: 350 were delivered (85 had invalid e-mail addresses), and 143 were returned (response rate, 41 percent). Overall, 66 percent of respondents chose implant-based reconstruction, 25 percent chose autologous reconstruction, and 9 percent chose no reconstruction. Respondents selecting autologous reconstruction cited cosmetic outcome as the most important factor considered in 47 percent of cases, compared with 14 percent of those choosing implant-based breast reconstruction (p = 0.0001). Invasiveness of the procedure/recovery time was cited as the most important factor by 83 percent of those surgeons opting for no breast reconstruction and by 51 percent of those choosing implant-based breast reconstruction (p = 0.0175). Board-certified female plastic surgeons exhibit a strong desire to pursue implant-based breast reconstruction over autologous reconstruction. When it was chosen, autologous reconstruction was felt to offer improved aesthetic outcomes. When making such a decision, patients can use female plastic surgeons as a resource for information, thus helping them to make an informed decision.


Irradiance dependence of the He-Ne laser-induced protection against UVC radiation in E. coli strains.
He-Ne laser pre-irradiation-induced protection against UVC damage was investigated in wild-type E. coli K12 strain AB1157 and its isogenic DNA repair mutant strains. At a dose of 7 kJ/m(2), pre-irradiation was observed to induce protection in recA proficient strains (AB1157 and uvrA(-) AB1886) at both the irradiances investigated (2 and 100 W/m(2)). However, at the same dose (7 kJ/m(2)), while no protection was observed at 100 W/m(2) in the recA(-) strain, some protection appeared to be there at 2 W/m(2). Mechanistic studies carried out on these strains at the two irradiances suggest that, whereas the protection observed at 100 W/m(2) is mediated by singlet oxygen, that observed at 2 W/m(2) is not. Further, the fact that protection at 100 W/m(2) was observed only in recA proficient strains suggests that it may arise due to the induction of DNA repair processes controlled by the recA gene. The latter may arise due to the oxidative stress produced by singlet oxygen generated by He-Ne laser irradiation. In contrast, the protection observed at 2 W/m(2) appears to be independent of the DNA repair proficiency of the strain.


Structure of Physical Activity Opportunities Contribution to Children's Physical Activity Levels in After-School Programs.
The primary purpose of this study was to determine which physical activity (PA) opportunity elicits the most moderate to vigorous PA (MVPA) in after-school programs. This study used a 3-group cross-over design in which participants were exposed to 3 variations of activity structures: free play, organized, or a mixture. PA was measured using ActiGraph GT3X+ accelerometers. All data were transformed into percentage of time spent sedentary or in MVPA. Repeated-measures mixed-effects models were used to examine differences in MVPA and sedentary among the 3 activity sessions. Participants included 197 unique children, aged 5-12 years, and were 53% male and 55% white. Statistically significant differences were observed in the percentage of time boys spent in MVPA during free play and mixed compared with organized only sessions (35.8% and 34.8% vs 29.4%). No significant difference was observed in the percentage of time girls spent in MVPA during free play compared with organized or mixed (27.2% and 26.1% vs 26.1%). Both boys and girls experienced ∼10% less time sedentary during free play compared with the others. Offering free play during PA opportunities can help children attain as much if not more MVPA compared with only offering organized, adult-led games.


Complete versus incomplete coronary revascularization of patients with multivessel coronary artery disease.
The treatment of patients with multivessel coronary artery disease in need of a revascularization procedure is influenced by the clinical situation (stable vs. unstable), comorbid conditions (diabetes mellitus), and anatomical variables (proximal left anterior artery stenosis, left ventricular dysfunction). Given the invasive nature of coronary artery bypass graft (CABG) operations, surgeons have embraced the concept of complete anatomical revascularization in one procedural stage since the inception of CABG surgery. However, achieving complete coronary revascularization has been more challenging with percutaneous coronary intervention (PCI), and as a result, incomplete procedures are far more common than complete ones. Data continue to emerge on the potential benefits of complete revascularization on clinical outcomes and suggest that complete revascularization should be the goal of therapy whenever possible. The heart team should carefully review the degree to which each revascularization modality can achieve this goal during procedural planning.


Slow myosin in developing rat skeletal muscle.
Through S1 nuclease mapping using a specific cDNA probe, we demonstrate that the slow myosin heavy-chain (MHC) gene, characteristic of adult soleus, is expressed in bulk hind limb muscle obtained from the 18-d rat fetus. We support these results by use of a monoclonal antibody (mAb) which is highly specific to the adult slow MHC. Immunoblots of MHC peptide maps show the same peptides, uniquely recognized by this antibody in adult soleus, are also identified in 18-d fetal limb muscle. Thus synthesis of slow myosin is an early event in skeletal myogenesis and is expressed concurrently with embryonic myosin. By immunofluorescence we demonstrate that in the 16-d fetus all primary myotubes in future fast and future slow muscles homogeneously express slow as well as embryonic myosin. Fiber heterogeneity arises owing to a developmentally regulated inhibition of slow MHC accumulation as muscles are progressively assembled from successive orders of cells. Assembly involves addition of new, superficial areas of the anterior tibial muscle (AT) and extensor digitorum longus muscle (EDL) in which primary cells initially stain weakly or are unstained with the slow mAb. In the developing AT and EDL, expression of slow myosin is unstable and is progressively restricted as these muscles specialize more and more towards the fast phenotype. Slow fibers persisting in deep portions of the adult EDL and AT are interpreted as vestiges of the original muscle primordium. A comparable inhibition of slow MHC accumulation occurs in the developing soleus but involves secondary, not primary, cells. Our results show that the fate of secondary cells is flexible and is spatially determined. By RIA we show that the relative proportions of slow MHC are fivefold greater in the soleus than in the EDL or AT at birth. After neonatal denervation, concentrations of slow MHC in the soleus rapidly decline, and we hypothesize that, in this muscle, the nerve protects and amplifies initial programs of slow MHC synthesis. Conversely, the content of slow MHC rises in the neonatally denervated EDL. This suggests that as the nerve amplifies fast MHC accumulation in the developing EDL, accumulation of slow MHC is inhibited in an antithetic fashion. Studies with phenylthiouracil-induced hypothyroidism indicate that inhibition of slow MHC accumulation in the EDL and AT is not initially under thyroid regulation. At later stages, the development of thyroid function plays a role in inhibiting slow MHC accumulation in the differentiating EDL and AT.(ABSTRACT TRUNCATED AT 400 WORDS)


Homeopathic remedy for arsenic toxicity?: Evidence-based findings from a randomized placebo-controlled double blind human trial.
Millions of people are at risk of groundwater arsenic contamination, but supply of arsenic-free drinking water is grossly inadequate. The present study was intended to examine if a potentized homeopathic remedy reportedly showing ameliorating potentials in people inhabiting high-risk arsenic-contaminated areas but drinking arsenic-free water, can also ameliorate arsenic toxicity in subjects living in high-risk arsenic-contaminated areas, and drinking arsenic-contaminated water. This pilot study was conducted on 20 males and 19 females of village Dasdiya (arsenic contaminated) who initially agreed to act as volunteers; but as many as 14, mostly placebo-fed subjects, later dropped out. 18 volunteers, 14 males and 4 females, from a distant village, Padumbasan (arsenic-free), served as negative controls. In a double blind placebo-controlled study, a potentized remedy of homeopathic Arsenicum Album-30 and its placebo (Succussed Alcohol-30) were given randomly to volunteers. Arsenic contents in urine and blood and several widely accepted toxicity biomarkers and pathological parameters in blood were analyzed before and after 2 months of administration of either verum or placebo. Elevated levels of ESR, creatinine and eosinophils and increased activities of AST, ALT, LPO and GGT were recorded in arsenic exposed subjects. Decreased levels of hemoglobin, PCV, neutrophil percentages, and GSH content and low G-6-PD activity were also observed in the arsenic exposed people. The administration of "verum" appeared to make positive modulations of these parameters, suggestive of its ameliorative potentials. Most of the subjects reported better appetite and improvement in general health, thereby indicating possibility of its use in remote arsenic-contaminated areas as an interim health support measure to a large population at risk.


Somatostatin and its analog lanreotide inhibit the proliferation of dispersed human non-functioning pituitary adenoma cells in vitro.
Somatostatin is a powerful inhibitor of hormone secretion and cell proliferation. Treatment with somatostatin analogs in humans causes a reduction in size and secretory activity of some endocrine tumors, including somatotropic pituitary adenomas. Less studied are the effects of somatostatin agonists on non-functioning pituitary adenomas (NFPAs). In this study we characterized the effects of somatostatin and its analog lanreotide on the proliferation of NFPAs in vitro and the intracellular mechanisms involved. Twenty-three NFPA post-surgical specimens were analyzed for somatostatin receptor (SSTR) expression and 12 of them were cultured in vitro to study somatostatin's effects on cell proliferation, assessed by means of [(3)H]thymidine uptake, and the intracellular signaling. One or more SSTR subtypes were expressed in 90% of the adenomas tested. Somatostatin and lanreotide treatment inhibited phorbol myristate acetate (PMA)-induced cell proliferation. Vanadate pretreatment reversed somatostatin and lanreotide inhibition of PMA-induced DNA synthesis suggesting an involvement of tyrosine phosphatase in this effect. In the only adenoma tested, somatostatin directly induced a tyrosine phosphatase activity. Somatostatin and lanreotide caused also a significant inhibition of voltage-sensitive calcium channel activity induced by 40mmol/l K(+) depolarization in microfluorimetric analysis. These data show that somatostatin and lanreotide inhibit human NFPA cell proliferation in vitro, and suggest that activation of tyrosine phosphatases and inhibition of the activity of voltage-dependent calcium channels may represent intracellular signals mediating this effect.


Oxidative stress in breath-hold divers after repetitive dives.
Hyperoxia causes oxidative stress. Breath-hold diving is associated with transient hyperoxia followed by hypoxia and a build-up of carbon dioxide (CO₂), chest-wall compression and significant haemodynamic changes. This study analyses variations in plasma oxidative stress markers after a series of repetitive breath-hold dives. Thirteen breath-hold divers were asked to perform repetitive breath-hold dives to 20 metres' depth to a cumulative breath-hold time of approximately 20 minutes over an hour in the open sea. Plasma nitric oxide (NO), peroxinitrites (ONOO⁻) and thiols (R-SH) were measured before and after the dive sequence. Circulating NO significantly increased after successive breath-hold dives (169.1 ± 58.26% of pre-dive values; P = 0.0002). Peroxinitrites doubled after the dives (207.2 ± 78.31% of pre-dive values; P = 0.0012). Thiols were significantly reduced (69.88 ± 19.23% of pre-dive values; P = 0.0002). NO may be produced by physical effort during breath-hold diving. Physical exercise, the transient hyperoxia followed by hypoxia and CO₂ accumulation would all contribute to the increased levels of superoxide anions (O₂²⁻). Since interaction of O₂²⁻ with NO forms ONOO⁻, this reaction is favoured and the production of thiol groups is reduced. Oxidative stress is, thus, present in breath-hold diving.


The geographic distribution of trace elements in the environment: the REGARDS study.
Research on trace elements and the effects of their ingestion on human health is often seen in scientific literature. However, little research has been done on the distribution of trace elements in the environment and their impact on health. This paper examines what characteristics among participants in the Reasons for Geographic and Racial Differences in Stroke (REGARDS) study are associated with levels of environmental exposure to arsenic, magnesium, mercury, and selenium. Demographic information from REGARDS participants was combined with trace element concentration data from the US Geochemical Survey (USGS). Each trace element was characterized as either low (magnesium and selenium) or high (arsenic and mercury) exposure. Associations between demographic characteristics and trace element concentrations were analyzed with unadjusted and adjusted logistic regression models. Individuals who reside in the Stroke Belt have lower odds of high exposure (4th quartile) to arsenic (OR 0.33, CI 0.31, 0.35) and increased exposure to mercury (OR 0.65, CI 0.62, 0.70) than those living outside of these areas, while the odds of low exposure to trace element concentrations were increased for magnesium (OR 5.48, CI 5.05, 5.95) and selenium (OR 2.37, CI 2.22, 2.54). We found an association between levels of trace elements in the environment and geographic region of residence, among other factors. Future studies are needed to further examine this association and determine whether or not these differences may be related to geographic variation in disease.


A Compilation of Serum Concentrations of 12 Antipsychotic Drugs in a Therapeutic Drug Monitoring Setting.
No comprehensive collection of routine therapeutic drug monitoring data for antipsychotic drugs has been published. In this compilation, data on 12 antipsychotics are presented. The drugs included are amisulpride (n = 506), aripiprazole (n = 1610), clozapine (n = 1189), flupentixol (n = 215), haloperidol (n = 390), olanzapine (n = 10,268), perphenazine (n = 1065), quetiapine (n = 5853), risperidone (n = 3255), sertindole (n = 111), ziprasidone (n = 1235), and zuclopenthixol (n = 691). Because only one sample per patient is included, the number of patients equals the number of samples. For each drug, median serum concentrations as well as that of the 10th and 90th percentiles are given for a range of daily doses. Comparisons are made between males and females, between patients younger than 65 years and 65 years and older, and between those treated with a low and a high dose of each drug. The concentration-to-dose (C/D) ratio is the primary variable used in these comparisons. Coefficients of variation (CVs) for the serum concentrations of each drug within and between subjects are presented. In general, the C/D ratios were higher in females than in males, higher in those 65 years and older than in younger subjects, and lower in those treated with higher doses than in those treated with lower doses. CVs between individuals were larger than within subjects, and the CVs were highest for the drugs with short elimination half-lives. For each antipsychotic drug, the results presented can serve as a reference tool for pharmacokinetic interpretation of the individual patient's serum drug level. The compiled serum concentrations and the C/D ratios can support the physician's decision when individualizing dosing and determining treatment strategies for a specific patient.


Case Report: Rhinosporidiosis Literature Review.
Rhinosporidiosis is caused by Rhinosporidium seeberi, a pathogen currently considered a fungus-like parasite of the eukaryotic group Mesomycetozoea. It is usually a benign condition, with slow growth of polypoid lesions, with involvement of the nose, nasopharynx, or eyes. The clinical characteristics of a painless, friable, polypoid mass, usually unilateral, can guide the diagnosis, but the gold standard for diagnosis is histopathological findings. This article reviews the epidemiology, pathobiology, clinical manifestations, diagnostic strategies, and treatment approach for rhinosporidiosis.


Stability, growth, and decline in adult life span development of declarative memory: cross-sectional and longitudinal data from a population-based study.
Five-year changes in episodic and semantic memory were examined in a sample of 829 participants (35-80 years). A cohort-matched sample (N=967) was assessed to control for practice effects. For episodic memory, cross-sectional analyses indicated gradual age-related decrements, whereas the longitudinal data revealed no decrements before age 60, even when practice effects were adjusted for. Longitudinally, semantic memory showed minor increments until age 55, with smaller decrements in old age as compared with episodic memory. Cohort differences in educational attainment appear to account for the discrepancies between cross-sectional and longitudinal data. Collectively, the results show that age trajectories for episodic and semantic memory differ and underscore the need to control for cohort and retest effects in cross-sectional and longitudinal studies, respectively.


Telomere shortening and karyotypic alterations in hepatocytes in long-term transplanted human liver allografts.
The long-term fate of aged liver allografts in young recipients who received grafts from older donors is unknown. We evaluated graft aging by analyzing hepatocytic telomere length and karyotypic changes. Seventeen pediatric individuals who underwent living-donor liver transplantation for congenital biliary diseases were selected. At a median of 10.4 years post-transplant, ten had tolerated grafts with weaned off immunosuppressants, and seven had idiopathic post-transplantation hepatitis. Fluorescence in situ hybridization was used to evaluate the telomere signal intensity (TI) and karyotypic changes. First, we measured predictive age-dependent TI decline with regression analysis of donor livers. The mean TI at the earliest (within a year) and latest biopsies was significantly lower than the predicted TI of the studied allografts. With univariate analysis, a higher abnormal karyotype ratio in the donor liver was correlated with development of idiopathic post-transplantation hepatitis. With multivariate analysis that included clinical parameters, a greater TI decline at the earliest biopsy was correlated with the development of idiopathic post-transplantation hepatitis. In conclusion, graft aging as measured by TI decline and donor abnormal karyotype ratio was associated with idiopathic post-transplantation hepatitis of long-term transplanted liver allografts.


RnaPredict--an evolutionary algorithm for RNA secondary structure prediction.
This paper presents two in-depth studies on RnaPredict, an evolutionary algorithm for RNA secondary structure prediction. The first study is an analysis of the performance of two thermodynamic models, Individual Nearest Neighbor (INN) and Individual Nearest Neighbor Hydrogen Bond (INN-HB). The correlation between the free energy of predicted structures and the sensitivity is analyzed for 19 RNA sequences. Although some variance is shown, there is a clear trend between a lower free energy and an increase in true positive base pairs. With increasing sequence length, this correlation generally decreases. In the second experiment, the accuracy of the predicted structures for these 19 sequences are compared against the accuracy of the structures generated by the mfold dynamic programming algorithm (DPA) and also to known structures. RnaPredict is shown to outperform the minimum free energy structures produced by mfold and has comparable performance when compared to sub-optimal structures produced by mfold.


Hepatitis B virus infection is associated with younger median age at diagnosis and death in cancers.
Several non-hepatocellular cancers were linked with hepatitis B virus (HBV) infection. This study was aimed to quantify the potential associations between HBV infection and multiple non-hepatocellular cancers. Continuous cases, including 5,715 non-cancer and 40,963 cancer cases diagnosed from 2008 to 2014 in Sun Yat-sen University Cancer Center were analyzed. HBV DNA and hepatitis B core antigen (HBcAg) were examed in gastric cancer tissues by polymerase chain reaction and immunohistochemical staining. After adjusting for age, sex, year of diagnosis, smoking, drinking and family history of cancer, significant associations were found between serum HBsAg and frequently reported HBV-related non-hepatocellular cancers, including non-Hodgkin's lymphoma, cholangiocarcinoma and pancreatic cancer [adjusted odds ratio (AOR) and 95% confidence interval (CI): 1.89 (1.65-2.16)], as well as total other non-hepatocellular cancers [AOR and 95% CI: 1.12 (1.03-1.22)]. The median ages at diagnosis, all-cause death and cancer-specific death of serum HBsAg positive cancer patients were all significantly younger than those with serum HBsAg negative. HBV DNA was detected in 12.4% (34/275) gastric cancer tissues and HBcAg was most commonly detected in lymphocytes. This was the first report that HBV infection had a modest but significant nonspecific association with total non-hepatocellular cancers. Median age at diagnosis and death was significantly younger in serum HBsAg positive cancer patients. The underlying mechanism needs further investigation.


Development of antiangiogenin peptide using a phage-displayed peptide library.
Angiogenesis is essential for tumor growth and metastasis. Here, we have developed a peptide antagonist of human angiogenin, which is a potent and tumor-associated angiogenic factor. ANI-E peptide was derived from the phage clone, which binds to angiogenin via the disulfide-constrained octapeptide epitope that is displayed on its surface, and is displaced by actin. Disulfide-constrained ANI-E peptide inhibits the interaction of angiogenin with actin, which is regarded as the angiogenin-binding protein on the surface of endothelial cells, without any visible effect on the ribonucleolytic activity of angiogenin. The peptide also inhibits the neovascularization that is induced by angiogenin in the chick chorioallantoic membrane assay. The antiangiogenic activity of the peptide is specific for angiogenin because the peptide does not have any apparent effect on embryonic angiogenesis or the preexisting blood vessels. The disulfide bond and the glutamic acid inside the disulfide ring of ANI-E peptide are indispensable for its antiangiogenin activity. Furthermore, ANI-E peptide blocks the angiogenesis that is induced by the angiogenin-secreting PC3 human prostate adenocarcinoma cells, without any direct effect on the proliferation, as well as the adhesion of PC3 cells to angiogenin. Therefore, the inhibition of the tumor-induced angiogenesis by ANI-E peptide is most likely caused by the neutralization of the extracellular angiogenin that is secreted by PC3 cells. On the basis of our results, ANI-E peptide may be effective for the treatment of various human tumors that secrete angiogenin. Our results also strongly support the hypothesis that the interaction of angiogenin with the cell surface actin-like protein is essential for the biological action of angiogenin, and angiogenin has an essential role in tumor-induced angiogenesis.


A Cross Sectional Survey of International Horse-Racing Authorities on Injury Data Collection and Reporting Practices For Professional Jockeys.
Jockey injuries are common in professional horse-racing and can result in life-threatening or career-ending outcomes. Robust injury data are essential to understand the circumstances of injury occurrence and ultimately identify prevention opportunities. This study aimed to identify jockey injury surveillance practices of international horse-racing authorities (HRAs) and the specific data items collected and reported by each HRA. A cross-sectional survey of representatives (e.g. Chief Medical Officer) from international HRAs was conducted. An online and paper questionnaire was designed comprised of 32 questions. Questions considered the barriers and facilitators to data collection within each HRA, and where available, what data were collected and reported by HRAs. Representatives from 15 international racing jurisdictions were included, of which 12 reported collection of race day injuries or falls, using varied definitions of medical attention and time loss. Six HRAs did not have a definition for a jockey injury, and eight HRAs had no parameters for describing injury severity. Race day exposure was collected by two HRAs. Results were commonly presented by HRAs as the number of injuries (n = 9/15) or proportion of injured jockeys (n = 6/15). The lack of a designated role for collection, collation and reporting of data was the main barrier for injury surveillance. Twelve HRAs agreed that mandatory collection would be a strong facilitator to improving practice. Enhancement and standardization of international jockey injury surveillance is required to move forward with evidence informed prevention. Concurrent investigation of how reporting practices can be best supported within existing HRA structures is recommended.


Purification and characterization of a calcium-activated neutral protease from rabbit skeletal muscle which requires calcium ions of microM order concentration.
One form of calcium-activated neutral protease (CANP) highly sensitive to calcium ions was purified by column chromatographic procedures to homogeneity. The purified enzyme required microM order Ca2+ (mu CANP), and the half-maximum activity was attained at 50 microM Ca2+. The electrophoretic mobility in a non-denaturing buffer showed that this enzyme is less acidic than another CANP which required mM order Ca2+ (mCANP). On SDS-polyacrylamide gel electrophoresis, the enzyme separated into two components with molecular weights of 79,000 and 28,000, respectively. Of these, the former was slightly larger than the counterpart of mCANP (Mr 76,000). Thus, mu CANP cannot be derived from mCANP by limited autolysis.


[Aura without migraine or Alice in Wonderland syndrome: Pediatric clinical case].
Migraine with aura in children is often reported, typical aura without headache is reported sometimes, but persistent aura and Alice in Wonderland syndrome is exceptionally reported. We present a case corresponding to the last one mentioned in a 6-year-old patient who had at least two episodes with the typical characteristics of aura without migraine before developing more frequently the complete clinical picture of aura and subsequently headache. The complementary studies systematically done to the patient were normal and she improved with conventional therapy.It is important to emphasize that the syndrome should be suspected in any patient who presents the described manifestations in order to approach to the diagnosis of aura without migraine, to discard either organic pathology or another cause and to indicate adequate therapeutic measures.


Patterns, causes and consequences of defensive microbiome dynamics across multiple scales.
The microbiome can significantly impact host phenotypes and serve as an additional source of heritable genetic variation. While patterns across eukaryotes are consistent with a role for symbiotic microbes in host macroevolution, few studies have examined symbiont-driven host evolution or the ecological implications of a dynamic microbiome across temporal, spatial or ecological scales. The pea aphid, Acyrthosiphon pisum, and its eight heritable bacterial endosymbionts have served as a model for studies on symbiosis and its potential contributions to host ecology and evolution. But we know little about the natural dynamics or ecological impacts of the heritable microbiome of this cosmopolitan insect pest. Here we report seasonal shifts in the frequencies of heritable defensive bacteria from natural pea aphid populations across two host races and geographic regions. Microbiome dynamics were consistent with symbiont responses to host-level selection and findings from one population suggested symbiont-driven adaptation to seasonally changing parasitoid pressures. Conversely, symbiont levels were negatively correlated with enemy-driven mortality when measured across host races, suggesting important ecological impacts of host race microbiome divergence. Rapid drops in symbiont frequencies following seasonal peaks suggest microbiome instability in several populations, with potentially large costs of 'superinfection' under certain environmental conditions. In summary, the realization of several laboratory-derived, a priori expectations suggests important natural impacts of defensive symbionts in host-enemy eco-evolutionary feedbacks. Yet negative findings and unanticipated correlations suggest complexities within this system may limit or obscure symbiont-driven contemporary evolution, a finding of broad significance given the widespread nature of defensive microbes across plants and animals.


Clinical profile and outcomes of De novo posttransplant thrombotic microangiopathy.
Thrombotic microangiopathy (TMA) after kidney transplant is rather uncommon but an important reversible cause of graft loss. This retrospective study of biopsy-proven posttransplant TMA was done to identify the important etiological factors, clinical features, and outcomes of post transplant TMA in a tertiary care referral hospital in northern India. This retrospective study was conducted among all renal transplant recipients who presented with graft dysfunction between 1989 and 2015. All the cases were looked for their etiology, clinical course, treatment modalities, and renal outcomes. The study was conducted in accord with prevailing ethical principles and reviewed by our own institutional review board. Seventeen patients out of 2000 (0.008%) transplants done during the study period had posttransplant TMA, out of which all the patients had de novo TMA, and the median time of presentation after transplantation was four months. Systemic TMA was noted in only four patients. Biopsy revealed associated rejection in five patients and associated calcineurin inhibitor (CNI) toxicity in 12 patients. Patients with TMA due to CNI toxicity were managed with CNI reduction or switching to alternate CNI or mammalian target of rapamycin inhibitors. In addition, antithymocyte globulin and plasma exchange were used in rejection-associated TMA. While four out of 12 patients (33%) in CNI-related TMA developed end-stage renal disease (ESRD), all patients in rejection-associated TMA developed ESRD. The overall one-year graft survival was 47%, whereas five- and 10-year survival was 35%. There was no significant difference in graft survival between localized and systemic TMAs (P = 0.4). Posttransplant TMA should be suspected even if there are no systemic features of hemolysis and early graft biopsy and prompt action is needed. The occurrence of TMA in the setting of rejection is associated with grave prognosis.


Disruption of the RICTOR/mTORC2 complex enhances the response of head and neck squamous cell carcinoma cells to PI3K inhibition.
Phosphoinositide 3-kinase (PI3K) is aberrantly activated in head and neck squamous cell carcinomas (HNSCC) and plays a pivotal role in tumorigenesis by driving Akt signaling, leading to cell survival and proliferation. Phosphorylation of Akt Thr308 by PI3K-PDK1 and Akt Ser473 by mammalian target of rapamycin complex 2 (mTORC2) activates Akt. Targeted inhibition of PI3K is a major area of preclinical and clinical investigation as it reduces Akt Thr308 phosphorylation, suppressing downstream mTORC1 activity. However, inhibition of mTORC1 releases feedback inhibition of mTORC2, resulting in a resurgence of Akt activation mediated by mTORC2. While the role of PI3K-activated Akt signaling is well established in HNSCC, the significance of mTORC2-driven Akt signaling has not been thoroughly examined. Here we explore the expression and function of mTORC2 and its obligate subunit RICTOR in HNSCC primary tumors and cell lines. We find RICTOR to be overexpressed in a subset of HNSCC tumors, including those with PIK3CA or EGFR gene amplifications. Whereas overexpression of RICTOR reduced susceptibility of HNSCC tumor cells to PI3K inhibition, genetic ablation of RICTOR using CRISPR/Cas9 sensitized cells to PI3K inhibition, as well as to EGFR inhibition and cisplatin treatment. Further, mTORC2 disruption led to reduced viability and colony forming abilities of HNSCC cells relative to their parental lines and induced loss of both activating Akt phosphorylation modifications (Thr308 and Ser473). Taken together, our findings establish RICTOR/mTORC2 as a critical oncogenic complex in HNSCC and rationalize the development of an mTORC2-specific inhibitor for use in HNSCC, either combined with agents already under investigation, or as an independent therapy.


Immediate responses of multi-focal low level laser therapy on quadriceps in knee osteoarthritis patients.
Low-level laser therapy (LLLT) applying on knee osteoarthritis (OA) patients has shown positive outcomes in analgesic effect and functional recovery. However, few studies applied such therapy on large area of quadriceps muscle in these patients. The aim of this study was to evaluate immediate effect of multi-focal LLLT on quadriceps of knee OA patients in pain and functional performance. Fifty-one participants with knee OA were enrolled and evaluated before (T1) and immediately after intervention (T2) by knee joint pain in numeric rating scale (NRS), walking speed, timed five-chair stands, and quadriceps strength by isokinetic dynamometer. Intervention with two multi-focal Gallium-Aluminum-Arsenide laser devices, each device with 36 laser diodes (wavelength 808 ± 10 nm, continuous, mean power 50 mW, 30 minutes), applied simultaneously over bilateral quadriceps with a total dose of 180 J for each thigh. The multi-focal LLLT significantly improved knee joint pain as measured by the NRS (54% reduction), timed five-chair stands, and walking speed (P < .05). Knee extensor strength also increased in terms of peak torque and force of concentric and eccentric contraction as measured by isokinetic dynamometer (P < .05). In conclusion, single-session multi-focal LLLT on quadriceps in knee OA patients has immediate beneficial effect on knee pain reduction, quadriceps strengthening and functional performance recovery. Long-term effect requires further investigation. Multi-focal LLLT on quadriceps might serve as an alternative non-invasive treatment option in these patients.


Development and Implementation of a Community Paramedicine Program in Rural United States.
Community paramedicine (CP) is an innovative care model focused on medical management for patients suffering from chronic diseases or other conditions that result in over-utilization of healthcare services. Despite their value, CP care models are not widely used in United States healthcare settings. More research is needed to understand the feasibility and effectiveness of implementing CP programs. Our objective was to develop a CP program to better meet the needs of complex, high-utilizer patients in a rural setting. We conducted an observational descriptive case series in a community, 25-bed, critical access hospital and primary care clinic in a rural Wisconsin county. Multiple stakeholders from the local health system and associated ambulance service were active participants in program development and implementation. Eligible patients receiving the intervention were identified as complex or high need by a referring physician. Primary outcomes included measures of emergency department, hospital, and clinic utilization. Secondary measures included provider and patient satisfaction. We characterized 32 unique patients as high utilizers requiring assistance in medical management. These patients were enrolled into the program and categorized as high utilizers requiring assistance in medical management. The median age was 76 years, and 68.8% were female. After six months, we found a statistically significant decline in patient utilization for primary care (53.3%, p = .006) and ED visits (59.3%, p = .007), but not for hospitalizations (60%, p = .13, non-significant (NS), compared to the six months preceding enrollment. Overall, the total number of healthcare contacts was increased after implementation (623 before vs 790 after, + 167, +26.8%). Implementation of the CP program resulted in increased overall use of local healthcare resources in patients referred by physicians as high utilizers. The implementation of an in-home CP program targeting high users of healthcare resources resulted in a decrease in utilization in the hospital, ED, and primary care settings; however, it was balanced and exceeded by the number of CP visits. CP programs align well with population health strategies and could be better leveraged to fill gaps in care and promote appropriate access to healthcare services. Further study is required to determine whether the shift in type of healthcare access reduces or increases cost.


Lipopolysaccharide-dependent down-regulation of CD27 expression on T cells activated with superantigen.
To investigate the mechanisms underlying T-cell responses during superantigen (SAg) stimulation, we analysed the effects of SAg on CD27 expression with or without lipopolysaccharide (LPS) as a novel regulator of T-cell function. CD27 is expressed on the majority of resting peripheral blood T cells (CD27low). Activation of T cells by SAg induces high levels of CD27 surface expression (CD27high) accompanied with simultaneous CD30 receptor expression. After prolonged activation in vitro, the level of CD27 expression became intermediate. The effects of LPS on down-regulation of CD27high expression on CD30+ T cells were dose-dependent. Separating LPS-stimulated monocytes from T cells by mechanical dispersion abolished its inhibitory effect, indicating the requirement for direct interactions between monocytes and T cells. We also found that SAg up-regulated CD80 expression on CD14+ monocytes and LPS inhibited SAg-induced CD80 expression after 24 hr of stimulation. Up-regulation of CD152 (CTLA-4) was selective, since it was found to be preferentially expressed on the CD30+ population. Competitive experiments using soluble blocking peptides showed that addition of CD28 or CD80 peptide recovered LPS-induced down-regulation of CD27high expression on CD30+ T cells. These observations suggested that the presence of low levels of CD80 on monocytes may partially inhibit CD27 expression due to inefficient delivery of positive signals via CD28/CD80 interaction, and that the increased levels of CD80 enhance the inhibition through interactions with CD152 which is expressed at the highest levels after 48 hr of activation.


Neuroimaging techniques in epilepsy.
To review state-of-the-art neuroimaging modalities in epilepsy and their clinical applications. PubMed literature searches to March 2010, using the following key words: 'epilepsy', 'positron emission tomography (PET)', 'single photon emission computed tomography (SPECT)', 'MR volumetry', 'diffusion tensor imaging', and 'functional MR imaging'. All articles including neuroimaging techniques in epilepsy were included in the review. High-field magnetic resonance imaging is fundamental for high-resolution structural imaging. Functional radionuclide imaging (positron emission tomography/single-photon emission computed tomography) can provide additional information to improve overall accuracy, and show good results with high concordance rates in temporal lobe epilepsy. Magnetic resonance spectroscopy is a useful adjunct consistently demonstrating changing metabolites in the epileptogenic region. Magnetic resonance volumetric imaging shows excellent sensitivity and specificity for temporal lobe epilepsy but thus far it has been inconsistent for extratemporal epilepsy. Diffusion tensor imaging with tractography allows visualisation of specific tracts such as connections with the language and visual cortex to enhance preoperative evaluation. Functional magnetic resonance imaging using blood oxygen level-dependent activation techniques is mainly used in presurgical planning for the high-sensitivity mapping of the eloquent cortex. Both contrast-bolus and arterial spin labelling magnetic resonance perfusion imaging show good correlation with clinical lateralisation of seizure disorder. Structural imaging is essential in localisation and lateralisation of the seizure focus. Functional radionuclide imaging or advanced magnetic resonance imaging techniques can provide complementary information when an epileptogenic substrate is not identified or in the presence of non-concordant clinical and structural findings.


Elucidation of the relative and absolute stereochemistry of the kalimantacin/batumin antibiotics.
Kalimantacin A and batumin exhibit potent and selective antibiotic activity against Staphylococcus species including MRSA. Both compounds are formed via a hybrid polyketide synthase/non-ribosomal peptide synthetase (PKS-NRPS) biosynthetic pathway and from comparison of the gene clusters it is apparent that batumin from Pseudomonas batumici and kalimantacin from P. fluorescens are the same compound. The linear structure of this unsaturated acid was assigned by spectroscopic methods, but the relative and absolute stereochemistry of the five stereocentres remained unknown. Herein we describe isolation of kalimantacin A and two further metabolites 17,19-diol 2 and 27-descarbomyl hydroxyketone 3 from cultures of P. fluorescens. Their absolute and relative stereochemistries are rigorously determined using a multidisciplinary approach combining natural product degradation and fragment synthesis with bioinformatics and NMR spectroscopy. Diol 2 has the 5R, 15S, 17S, 19R, 26R, 27R configuration and is the immediate biosynthetic precursor of the bioactive kalimantacin A formed by oxidation of the 17-alcohol to the ketone.


[Congenital disorders of tyrosine metabolism and their correction in children with tumors].
The paper deals with the evaluation of quantitative indexes of urine excretion of tyrosine and its metabolites--highly carcinogenic p-hydroxyphenyllactic acid and homogentisic acid--in infants suffering various tumors and their parents. Also, the effect of ascorbic acid (100 mg/kg body weight daily) on tyrosine metabolism was studied. The sick children and their parents showed increased urine excretion of tyrosine and p-hydroxyphenyllactic acid matched by a decrease in that of homogentisic acid. The said changes suggest inhibition of p-hydroxyphenylpyruvate dioxygenase (CE 126.96.36.199). High doses of ascorbic acid were shown to arrest or significantly inhibit the excretion of carcinogenic metabolite of tyrosine in patients with hemoblastoses and nephroblastoma.


Antigenotoxic and antioxidant effects of the Mongolian medicinal plant Leptopyrum fumarioides (L): an in vitro study.
Leptopyrum fumarioides has been used in the traditional medicine of Mongolia for the treatment of various diseases, including drug intoxications. However, since there is only sparse information about its chemistry, active components, and pharmacological and toxicological effects, the major aim of the present study employing mouse lymphoma cells was to evaluate the genotoxic and antigenotoxic/antioxidative effects of extracts and components isolated from this plant. A crude methanol extract was separated into three different sub-extracts: dichloromethane, n-butanol, and water. The major constituent of the n-butanol extract, i.e., the flavone luteolin-7-O-glucoside and a mixture of the most abundant compounds in the dichloromethane sub-extract were then isolated. DNA damage was evaluated using the comet assay; the antioxidant activity was evaluated using the DPPH radical scavenging assay. The crude methanol extract, the dichloromethane sub-extract and the mixture of compounds isolated from the latter fraction, increased the level of DNA damage after three hours of exposure. In contrast, no increase in DNA damage was observed in the cells that had been exposed to the n-butanol and water sub-extracts, or to the pure flavone. When non-DNA damaging concentrations of extracts and compounds were tested together with the DNA damaging agent catechol, all sub-extracts were found to reduce the catechol-induced DNA damage (the flavone was then found to be the most effective protective agent). The n-butanol sub-extract and the flavone were also found to have the most prominent antioxidative effects. Based on the results from the present study, components in Leptopyrum fumarioides were found to protect the DNA damage induced by catechol, probably by acting as potent antioxidants.


Significance of adhesion molecules expression for estimation of serous ovarian cancer prognosis.
To study the adhesion molecules CD44 and E-cadherin expression in serous ovarian cancer (OC) and their relationship with clinicopathological peculiarities of tumor process and prognosis. The study was performed on OC samples from 72 patients with serous OC of stages I-III. Expression of CD44 and E-cadherin in tumor samples was evaluated with the use of immunohistochemical analysis. Immunohistochemical analysis has revealed that in 58.3% of OC patients CD44 expression was detected in more than 10% of cells with average level of 30.0В±5.6%. E-cadherin expression was present in 47.2% of tumors, and 12.2В±3.6% cells were immunopositive. CD44 and E-cadherin expression depends on degree of cytomorphological malignancy and cell differentiation. A reverse correlation between CD44 and E-cadherin expression in primary serous OC (r=-0.38, p < 0.05) has been found. Increased CD44 expression (mean index) was not observed in peritoneal metastases compared to primary tumors, but analysis of individual indexes showed increase of CD44 expression in 46.6% OC patients with metastases. The dependence between overall survival of OC patients and the molecular phenotype of tumor cells, in particular, poor prognosis for OC patients with CD44(+)/E-cadherin(-) and CD44(+)/budding(+) tumor cells phenotype, has been revealed. The results of morphological and immunohistochemical analysis has shown the increase of adhesive features of serous ovarian cancer cells what may be significant for estimation of ovarian cancer aggressiveness and development of metastatic process.


[Patho-physiology of renal dysfunction in Bardet-Biedl Syndrome].
Bardet-Biedl Syndrome (BBS) is a rare autosomal recessive disorder with renal and extra-renal involvement. The wide spectrum of clinical manifestations is associated to the high genetic heterogeneity. To date 21 genes have been identified in humans and the majority of them encode proteins located on the basal body of the primary cilium. For this reason the disease is has been included among the 'ciliopathies'. The renal involvement is extremely heterogeneous in BBS and is considered the main cause of morbidity and mortality. Recent evidences have suggested that mutations in BBS6, 10 and 12 are associated with a more severe renal dysfunction. The most common renal dysfunction is the urine concentrating defect, even though the underlying mechanism is not completely known. Recently we have demonstrated that hyposthenuria in BBS patients has a renal origin, and depends on desmopessin resistance. The majority of hyposthenuric BBS patients have a combined defect to both concentrate and dilute the urine. The combined defect is associated with a blunted increased urine Aquaproine-2 (u-AQP2) excretion in antidiuresis. A ccordingly, in vitro BBS10 silencing prevented AQP2 trafficking to the apical plasma membrane. However, after long term water restriction hyposthenuric BBS patients showed the same u-AQP2 excretion compared with controls, suggesting that other mechanisms are implicated into the pathogenesis of hyposhtenuria. The complete molecular mechanism underlying hyposhtenuria remains largely unknown in BBS. Whether this defect may represent a predictor factor for poor renal outcome remains to be elucidated.


Precision Cardiovascular Medicine: State of Genetic Testing.
In the 15 years following the release of the first complete human genome sequences, our understanding of rare and common genetic variation as determinants of cardiovascular disease susceptibility, prognosis, and therapeutic response has grown exponentially. As such, the use of genomics to enhance the care of patients with cardiovascular diseases has garnered increased attention from clinicians, researchers, and regulatory agencies eager to realize the promise of precision genomic medicine. However, owing to a large burden of "complex" common diseases, emphasis on evidence-based practice, and a degree of unfamiliarity/discomfort with the language of genomic medicine, the development and implementation of genomics-guided approaches designed to further individualize the clinical management of a variety of cardiovascular disorders remains a challenge. In this review, we detail a practical approach to genetic testing initiation and interpretation as well as review the current state of cardiovascular genetic and pharmacogenomic testing in the context of relevant society and regulatory agency recommendations/guidelines.