A perfusion chamber for high-resolution light microscopy of cultured cells.
A perfusion chamber suitable for general microscopic work on upright and inverted microscopes with cultured cells is described. The device features a minimal internal volume and large field of view within small outer dimensions. In particular the chamber body is very flat to accommodate large-aperture condenser and objective lenses. It is constructed of innocuous, sterilizable materials, incorporates permanent seals for the windows, is robust and simple to use.


[Autologous blood transfusion in elective heart surgery: prospective study of 189 consecutive patients].
Blood autotransfusion has entered a new phase in blood transfusion technique, since it represents an important alternative in eliminating the risks connected with blood transfusion: viral hepatitis, AIDS, blood transfusion reactions, and alloimmunization. Transfusion requirements during cardiac surgical procedures have steadily decreased; nowadays most adult patients require no transfusion during surgery. Patients (pts) receiving bank-blood may develop infectious diseases (hepatitis, AIDS, etc.). We have studied how to avoid the risk of infections with homologous blood transfusions. We present our experience of day-hospital pre-operative autologous blood collection. One-hundred-eighty-nine patients undergoing primary myocardial revascularization or valvular replacement were submitted to the drainage of 350 ml of blood three times every four days before surgery. The blood was centrifuged at once, to separate red cells from plasma. Surgeries were performed 21 days after the first drainage; iron therapy was recommended. After surgery pts received blood only if haematocrit was lower than 28%. The following data were recorded: no. of pts who received homologous blood; blood loss and homologous total blood volume used for each pt. Average blood loss was 1230 cc for ischemic pts and 701 cc for valvular pts. Non-A non B hepatitis occurred in 3/189 pts (1.5%). All of them had received homologous blood transfusions. Our data show clearly that autotransfused pts had a better post-operative period; less bank-blood and fewer transfusions have been used. No pt had collateral effects such as angina or hypotension from blood drawing. Our data show that severe cardiac diseases do not represent an absolute contraindication to heavy blood drainage.(ABSTRACT TRUNCATED AT 250 WORDS)


Usefulness of Early Plate Removal in Patients With Occlusal Discrepancies After Sagittal Split Ramus Osteotomy.
The aim of this study was to evaluate the effects of early removal of fixed plates in patients with occlusal discrepancies after sagittal split ramus osteotomy (SSRO) with a focus on the positional relationship of the temporomandibular joint (TMJ). Sagittal split ramus osteotomy with/without Le Fort I osteotomy was performed on 98 patients with mandibular prognathism. Of these 98 patients, 15 with occlusal discrepancies and/or TMJ symptoms underwent early plate removal after SSRO. Finally, 12 consecutive patients were evaluated in this study: 7 underwent bilateral SSRO, 1 underwent unilateral SSRO, and 4 underwent bilateral SSRO with maxillary advancement. The axiolateral TMJ Schuller method was used to evaluate the TMJ position. The authors measured 3 spaces (anterior, superior, and posterior joint spaces) between the condyle and glenoid fossa in the sagittal plane. The anterior and superior joint spaces were significantly larger immediately after the operation than before the operation. After early plate removal, these spaces significantly decreased in size. The posterior joint space increased, but with no significant difference. Three months after SSRO, the size of each of the 3 spaces was closely related to its preoperative size. In conclusion, these results suggest that early removal of fixed plates is 1 treatment option for postoperative occlusal discrepancies after SSRO.


Increasing the capacity of health surveillance assistants in community mental health care in a developing country, Malawi.
Mental health services in Malawi are centralized in the three tertiary units which are located one in each of the three regions of Malawi and this means that most people with mental health problems do not get help. With severe shortages of mental health professionals in the country, integration of mental health into existing primary and community health services is the most feasible way of increasing access to services for people with mental health problems. This paper discusses a pilot program of integrating mental health in the activities of Health Surveillance Assistants (HSAs) who are community health workers in Malawi.


Effects of toxic β-glucosides on carbohydrate metabolism in cotton bollworm, Helicoverpa armigera (Hübner).
The purpose of this study was to evaluate the effects of three toxic β-glucosides, phlorizin, santonin, and amygdalin, on carbohydrate metabolism in the cotton bollworm, Helicoverpa armigera (Hübner), when diets mixed with β-glucosides were fed to third-instar larvae. The growth of the larvae was significantly inhibited by exposure to santonin after 96 hr but not obviously affected by phlorizin and amygdalin. The midgut trehalase activities were only 51.7%, 32%, and 42.5% of that of the control after treatment with phlorizin, santonin and amygdalin at 2 mg/ml, respectively. In the hemolymph and fat body, the amount of trehalose decreased in all cases. However, the effects of santonin on the alteration of the glycogen and glucose levels as well as the activities of glycogen phosphorylase, were different than those of the other two β-glucosides. It appears that the three β-glucosides have different influences on the carbohydrate metabolism of cotton bollworm.


Replacement of Vena Cava up to the Right Atrium during Living Donor Liver Transplantation for Echinococcus alveolaris.
Management of advanced stage of Echinococcus alveolaris is a very difficult procedure. Surgical treatment like resection and liver transplantation is accepted procedure nowadays. Here we presented a case report of Echinococcus alveolaris which invaded the inferior vena cava up to the right atrium and surrounding tissues. This patient underwent living donor liver transplantation with replacement of inferior vena cava up to the right atrium with cryopreserved cadaveric aortic graft. This procedure is very difficult but it is a life-saving chance for patients in advanced cases of Echinococcus alveolaris.


Development and implementation of a local government survey to measure community supports for healthy eating and active living.
The ability to make healthy choices is influenced by where one lives, works, shops, and plays. Locally enacted policies and standards can influence these surroundings but little is known about the prevalence of such policies and standards that support healthier behaviors. In this paper, we describe the development of a survey questionnaire designed to capture local level policy supports for healthy eating and active living and findings and lessons learned from a 2012 pilot in two states, Minnesota and California, including respondent burden, survey sampling and administration methods, and survey item feasibility issues. A 38-item, web-based, self-administered survey and sampling frame were developed to assess the prevalence of 22 types of healthy eating and active living policies in a representative sample of local governments in the two states. The majority of respondents indicated the survey required minimal effort to complete with half taking < 20 min to complete the survey. A non-response follow-up plan including emails and phone calls was required to achieve a 68% response rate (versus a 37% response rate for email only reminders). Local governments with larger residential populations reported having healthy eating and active living policies and standards more often than smaller governments. Policies that support active living were more common than those that support healthy eating and varied within the two states. The methods we developed are a feasible data collection tool for estimating the prevalence of municipal healthy eating and active living policies and standards at the state and national level.


Beta-globin gene cluster haplotypes in an admixed Venezuelan population.
Haplotype frequencies for the beta-globin gene cluster were determined in 96 chromosomes from a Venezuelan mixed population. Twelve haplotypes were observed, the most frequent of which were 2, 3, and 4; all others showed prevalences below 10%. When the results were compared with those of 13 other groups using a phylogenetic tree, they clustered in an intermediate position somewhat nearer to those of four sub-Saharan African populations. This is in accordance with historical and other genetic data which indicate an important contribution of people of African ancestry to this population. The high heterozygosity (80%) and the intermediate position in the tree, on the other hand, possibly reflect past intermixture of these African-derived individuals with Europeans and Amerindians. Am. J. Hum. Biol. 9:323-327, 1997. © 1997 Wiley-Liss, Inc.


Unphosphorylated H1 is enriched in a specific region of the promoter when CDC2 is down-regulated during starvation.
Tetrahymena thermophila macronuclear histone H1 is phosphorylated by a cdc2 kinase, and H1 phosphorylation regulates CDC2 expression by a positive feedback mechanism. In starved wild-type cells, decreased expression of the CDC2 gene is correlated with a global reduction in the phosphorylation of H1 and reduced phosphorylation of H1 in the region upstream of the CDC2 gene. To determine whether the reduced H1 phosphorylation upstream of the CDC2 gene is merely a reflection of global dephosphorylation or is due to specific targeting of dephosphorylation of H1 to the CDC2 promoter during starvation, the CDC2 promoter was mapped, and the distributions of phosphorylated and unphosphorylated H1 across the CDC2 gene were determined using chromatin immunoprecipitation. Unphosphorylated H1 is specifically enriched in a region of the CDC2 promoter when the gene's expression is reduced during starvation but not when CDC2 is highly active in growing cells. The region of unphosphorylated H1 coincides with a region that is essential for CDC2 expression. These studies are the first in vivo demonstration that the phosphorylation of H1 is being regulated at a fine level and that unphosphorylated H1 can be specifically targeted to a promoter, where it likely regulates transcription in a gene-specific manner.


Uptalk in L2 English: the phonetic identity and perception of final declarative rises in Serbian EFL.
Uptalk has been increasingly documented in different L1 English varieties and communicative contexts, but is rarely recognized in formal L2 educational contexts, where it is still attributed to learners' inadequate mastery of intonation. This study is a cross-sectional corpus-based exploration of the phonetic realization of uptalk in Serbian EFL students' semi-spontaneous expository speech, and its perception as a sentence-finality signal. The corpus comprised all rising intonation units (IU) produced by 14 female and 9 male participants, classified by structural clues as syntactic continuation, listing, polar questions, or uptalk, to explore the relatedness of the phonetic properties to structural position and gender. Next, 100 EFL students rated selected phrases, illustrating continuation rises, uptalk, and final falls, as possibly sentence final, on a 5-point scale. The findings showed that uptalk was consistently produced as a phonetically distinct signal, characterized by a larger pitch excursion, a steeper rise slope, a higher rise peak, and a longer post-IU pause. Females produced wider pitch excursions and steeper slopes. Uptalk examples were ranked high as possible sentence-finality signals. The study suggests that EFL speakers' uptalk should be recognized as a novel socio-pragmatic prosodic device, deliberately used even in more formal academic contexts.


[Inter-fascial dissection in laparoscopic colorectal cancer surgery: principles of retraction and exposure and techniques in fascial separation].
Colorectal surgery for malignancies has evolved into an era of careful and precise dissection along mesorectal or mesocolic fascia to achieve the so-called total mesorectal excision or complete mesocolic excision. The wide use of laparoscopic technique prompted more anatomical, histological, and embryological studies. This leads to a deeper and more precise understanding of fascial anatomy concerning colorectal surgery, though controversies exist. The complicated anatomy of multilayer parietal fasciae and dense adhesion between fasciae at specific sites still represent a major hindrance to perform a precise inter-fascial dissection. Colorectal surgeons should be familiar with the onion-like arrangement of the visceral and parietal fasciae. The dedicated assistants should provide three-directional traction and adjust the direction of forces timely in a manner that the resultant forces are always in a direction perpendicular to the fasciae that are to be dissected. The fixation of the mesorectum and the mesocolon to the pelvic and abdominal wall can also be exploited as a natural counter-retraction. To separate loosely attached visceral and parietal fasciae, the application of splitting forces on opposite fasciae or sliding the forceps along the interface will provide quick separation and maintenance of the integrity of the fasciae. In summary, careful attention to the direction and strength of three directional retractions on parietal and visceral fasciae will help stretch and open up the areolar surgical tissue plane, skillful maneuver in separation and dividing of the attachment of two fasciae will ensure a precise inter-fascial dissection and help achieve total mesorectal excision or complete mesocolic excision, reducing the risk of the residual of the mesentery and inadvertent injuries to adjacent tissues and autonomic nerves.


The legal status of cats in New Zealand: a perspective on the welfare of companion, stray, and feral domestic cats (Felis catus).
Pinpointing and safeguarding the welfare status of domestic cats is problematic, especially in New Zealand where cats are introduced predators with significant impact on indigenous fauna. Usually the identification of welfare status depends on conservational, legal, and public attitudes that are often contrasting. Cats may rapidly transgress definitions placed on them, confounding attempts to categorize them. In 1 generation, cats can move from a human-dependent state ("stray" or "companion") to wild ("feral"). Often this categorization uses arbitrary behavioral and or situational parameters; consequent treatment and welfare protection for these cats are similarly affected. Terminology used to describe cats is not equitable across research. However, the New Zealand Animal Welfare (Companion Cats) Code of Welfare 2007 seeks to create a new definition of the terms companion, stray, and feral. It distinguishes between cats who live within and without human social constructs. This legislation mandates that cats in human environments or indirectly dependent on humans cannot be classified as feral. Such definitions may prove vital when safeguarding the welfare of free-living domestic cats and cat colonies.


Expression of Krüppel-like factor 4 in breast cancer tissues and its effects on the proliferation of breast cancer MDA-MB-231 cells.
The aim of the present study was to detect the expression of Krüppel-like factor 4 (KLF4) in breast cancer tissues and to evaluate the effect on the proliferation of breast cancer MDA-MB-231 cells. The expression of KLF4 protein in 239 breast cancer tissues and 40 paracancerous tissues were detected by an immunohistochemical assay, and its correlation with clinical pathological parameters was analyzed. A eukaryotic expression vector, pcDNA3.1-KLF4, was constructed by transient transfection of breast cancer MDA-MB-231 cells with liposomes (experimental group). The untransfected cells and those transfected with empty plasmid pcDNA3.1 were used as the blank and negative control groups, respectively. The expression of the KLF4 gene and protein in the three groups were detected by reverse transcription polymerase chain reaction and western blotting, respectively. Furthermore, the cell proliferative capacity was detected by an MTT assay. The positive expression rate of KLF4 protein in breast cancer tissues (39.0%, 93/239) was significantly lower than that of paracancerous tissues (77.5%, 31/40) (P<0.05). In addition, KLF4 protein expression in breast cancer tissues was correlated with pathological type, histological grade and lymphatic metastasis (P<0.05). KLF4 mRNA and protein were both expressed by the experimental group, but not by the two control groups. Meanwhile, the proliferative capacity of the experimental group was also significantly decreased. A significant decrease in the positive expression rate of KLF4 protein in breast cancer tissues was correlated with several clinical pathological parameters. In addition, transfection of the KLF4 gene inhibited the proliferation of breast cancer cells, suggesting that this gene is important in the onset and progression of this type of cancer.


Rapid diagnosis of the economically important fruit fly, Bactrocera correcta (Diptera: Tephritidae) based on a species-specific barcoding cytochrome oxidase I marker.
The guava fruit fly, Bactrocera correcta (Bezzi) (Diptera: Tephritidae), is an invasive pest of fruit and vegetable crops that primarily inhabits Southeast Asia and which has the potential to become a major threat within both the Oriental and Australian oceanic regions as well as California and Florida. In light of the threat posed, it is important to develop a rapid, accurate and reliable method to identify B. correcta in quarantine work in order to provide an early warning to prevent its widespread invasion. In the present study, we describe a species-specific polymerase chain reaction assay for the diagnosis of B. correcta using mitochondrial DNA cytochrome oxidase I (mtDNA COI) barcoding genes. A B. correcta-specific primer pair was designed according to variations in the mtDNA COI barcode sequences among 14 fruit fly species. The specificity and sensitivity of the B. correcta-specific primer pair was tested based on the presence or absence of a band in the gel profile. A pair of species-specific B. correcta primers was successfully designed and named BCOR-F/BCOR-R. An ∼280 bp fragment was amplified from specimens belonging to 17 geographical populations and four life stages of B. correcta, while no such diagnostic bands were present in any of the 14 other related fruit fly species examined. Sensitivity test results demonstrated that successful amplification can be obtained with as little as 1 ng μl⁻¹ of template DNA. The species-specific PCR analysis was able to successfully diagnose B. correcta, even in immature life stages, and from adult body parts. This method proved to be a robust single-step molecular technique for the diagnosis of B. correcta with respect to potential plant quarantine.


Chromosome pairing in inter-racial hybrids of the house musk shrew (Suncus murinus, Insectivora, Soricidae).
Two chromosome races of the house shrew Suncus murinus that differ from each other for five Robertsonian translocations (8.17, 9.13, 10.12, 11.16, and 14.15), heterochromatic insertions in chromosomes 7 and X, and multiple rearrangements in the Y chromosome were crossed and then intercrossed in captivity to produce a hybrid stock. Electron-microscopic analysis of synaptonemal complexes in fertile and sterile hybrid males was carried out. Meiosis in sterile males did not progress beyond pachytene and was severely disrupted. Meiotic arrest was not determined by structural heterozygosity: heterozygotes for all variant chromosomes distinguishing two parental races were found in both sterile and fertile male hybrids. Fertile hybrids demonstrated an orderly pairing of all chromosomes. In heterozygotes for Robertsonian fusions, completely paired trivalents were formed between the Robertsonian metacentrics and homologous acrocentrics. In heterozygotes for chromosome 7, bivalents with a small buckle were observed in a small fraction of pachytene cells. No differences were found in the morphology and pairing pattern of sex bivalents, composed of the X and Y chromosomes derived from the same or different parental races. Univalents, multivalents, and associations between X and Y chromosomes and autosomal trivalents, as well as associations of autosomal trivalents with each other, were observed in a small fraction of the pachytene cells of fertile males. Our results indicate that the system controlling male sterility in interracial hybrids of S. murinus is of genic rather than of chromosomal type.


The roles of geography and founder effects in promoting host-associated differentiation in the generalist bogus yucca moth Prodoxus decipiens.
There is ample evidence that host shifts in plant-feeding insects have been instrumental in generating the enormous diversity of insects. Changes in host use can cause host-associated differentiation (HAD) among populations that may lead to reproductive isolation and eventual speciation. The importance of geography in facilitating this process remains controversial. We examined the geographic context of HAD in the wide-ranging generalist yucca moth Prodoxus decipiens. Previous work demonstrated HAD among sympatric moth populations feeding on two different Yucca species occurring on the barrier islands of North Carolina, USA. We assessed the genetic structure of P. decipiens across its entire geographic and host range to determine whether HAD is widespread in this generalist herbivore. Population genetic analyses of microsatellite and mtDNA sequence data across the entire range showed genetic structuring with respect to host use and geography. In particular, genetic differentiation was relatively strong between mainland populations and those on the barrier islands of North Carolina. Finer scale analyses, however, among sympatric populations using different host plant species only showed significant clustering based on host use for populations on the barrier islands. Mainland populations did not form population clusters based on host plant use. Reduced genetic diversity in the barrier island populations, especially on the derived host, suggests that founder effects may have been instrumental in facilitating HAD. In general, results suggest that the interplay of local adaptation, geography and demography can determine the tempo of HAD. We argue that future studies should include comprehensive surveys across a wide range of environmental and geographic conditions to elucidate the contribution of various processes to HAD.


Mirror-Image Dependence: Targeting Enantiomeric G-Quadruplex DNA Using Triplex Metallohelices.
Natural d-DNA and l-DNA are mirror-image counterparts. However, because of the inherent flexibility and conformation diversity of DNA, it is still not clear how enantiomeric compounds recognize d-DNA and l-DNA. Herein, taking G-quadruplex (G4) DNA as an example that has diverse conformations and distinct biofunctions, the binding of ten pairs of iron triplex metallohelices to d- and l-G4 DNA were evaluated. The Δ-enantiomer binds to d-DNA and the Λ-enantiomer binds to l-DNA, exhibiting almost the same stabilization effect and binding affinity. The binding affinity of the Δ-metallohelix with d-G4 is nearly 70-fold higher than that of Λ-metallohelix binding d-G4. Δ-Metallohelix binding to d-G4 follows a two-step binding process driven by a favorable enthalpy contribution to compensate for the associated unfavorable entropy.


Cyclofenil treatment of scleroderma--a controlled study.
A double-blind cross-over study of cyclofenil and placebo treatments was conducted in 11 scleroderma patients. Each treatment period was for four months. A range of subjective and objective measurements failed to reveal any consistent or obvious response to cyclofenil. One patient died of his disease during active treatment. Four were withdrawn because of liver dysfunction which was attributed to cyclofenil. It remains to be seen whether treatment over longer periods will exert a clinically significant effect.


Risk factors affecting amputation in diabetic foot.
A diabetic foot is the most common cause of non-traumatic lower extremity amputations (LEA). The study seeks to assess the risk factors of amputation in patients with diabetic foot ulcers (DFU). The study was conducted on 351 patients with DFUs from January 2010 to December 2018. Their demographic characteristics, disease history, laboratory data, ankle-brachial index, Wagner classification, osteomyelitis, sarcopenia index, and ulcer sizes were considered as variables to predict outcome. A chi-square test and multivariate logistic regression analysis were performed to test the relationship of the data gathered. Additionally, the subjects were divided into two groups based on their amputation surgery. Out of the 351 subjects, 170 required LEA. The mean age of the subjects was 61 years and the mean duration of diabetes was 15 years; there was no significant difference between the two groups in terms of these averages. Osteomyelitis (hazard ratio [HR], 6.164; 95% confidence interval [CI], 3.561-10.671), lesion on percutaneous transluminal angioplasty (HR, 2.494; 95% CI, 1.087-5.721), estimated glomerular filtration rate (eGFR; HR, 0.99; 95% CI, 0.981-0.999), ulcer size (HR, 1.247; 95% CI, 1.107-1.405), and forefoot ulcer location (HR, 2.475; 95% CI, 0.224-0.73) were associated with risk of amputation. Osteomyelitis, peripheral artery disease, chronic kidney disease, ulcer size, and forefoot ulcer location were risk factors for amputation in diabetic foot patients. Further investigation would contribute to the establishment of a diabetic foot risk stratification system for Koreans, allowing for optimal individualized treatment.


Clinico-haematological profile of pure red cell aplasia in children.
Pure red cell aplasia (PRCA) is a rare disorder, characterized by isolated failure of erythropoiesis. The clinico-haematological profile of 16 patients with PRCA is presented in this communication. Fourteen patients had Diamond-Blackfan anaemia (DBA), one had transient erythroblastopenia of childhood, and one patient had PRCA secondary to carbamazepine. Physical abnormalities were observed in 50 per cent of patients with DBA. Of the nine patients with DBA who were administered prednisolone and had a regular follow-up, four (44.4 per cent) had no response, three (33.3 per cent) responded fully, and two (22.2 per cent) were steroid dependent.


Regression analysis of networked data.
This paper concerns regression methodology for assessing relationships between multi-dimensional response variables and covariates that are correlated within a network. To address analytical challenges associated with the integration of network topology into the regression analysis, we propose a hybrid quadratic inference method that uses both prior and data-driven correlations among network nodes. A Godambe information-based tuning strategy is developed to allocate weights between the prior and data-driven network structures, so the estimator is efficient. The proposed method is conceptually simple and computationally fast, and has appealing large-sample properties. It is evaluated by simulation, and its application is illustrated using neuroimaging data from an association study of the effects of iron deficiency on auditory recognition memory in infants.


Glucocorticoid regulation of corticotropin-releasing factor1 receptor expression in pituitary-derived AtT-20 cells.
Corticotropin-releasing factor (CRF) receptors represent one of the primary sites for negative feedback of the pituitary by adrenocortical glucocorticoid hormones; however, the molecular mechanisms involved have yet to be elucidated. The present study examines the mechanisms by which glucocorticoids regulate CRF-R1 expression in the pituitary cell line, AtT-20. Treatment of these cells with dexamethasone resulted in a concentration- and time-dependent inhibition of CRF-R1 mRNA that was significant by 1 hr and maximal after 4 hr. Levels of CRF-R1 mRNA then returned to control levels after 24 hr. Similar changes were observed when the cells were treated with corticosterone. Pro-opiomelanocortin mRNA was also decreased after dexamethasone pretreatment; however, the time course was much slower with a significant effect only detected after 6 hr. Further analysis of the mechanisms that mediate glucocorticoid regulation of CRF-R1 mRNA was conducted These studies demonstrated that glucocorticoid incubation significantly decreases the rate of CRF-R1 gene transcription, as determined by nuclear run-on analysis. In addition, the result demonstrate that glucocorticoid incubation significantly decreases CRF-R1 mRNA stability by approximately 50%. The down-regulation of CRF-R1 mRNA was dependent on de novo protein synthesis, as it was blocked by pretreatment with cycloheximide. This represents a novel mechanism for glucocorticoid negative feedback regulation of CRF-R1 expression.


"Naming and Framing": The Impact of Labeling on Health State Values for Multiple Sclerosis.
Health state valuation is a key input in many economic evaluations that inform resource allocation across competing healthcare interventions. Empirical evidence has shown that, in preference elicitation surveys, respondents may value a health state differently if they are aware of the condition causing it ('labeling effects'). This study investigates the impact of including a multiple sclerosis (MS) label for valuation of MS health states. Health state values for MS were elicited using two internet-based surveys in representative samples of the UK population ( n = 1702; n = 1788). In one survey respondents were not informed that health states were caused by MS. The second survey included a condition label for MS. Surveys were identical in all other ways. Health states were described using a MS-specific eight-dimensional classification system (MSIS-8D), and the time trade-off valuation technique was used. Differences between values for labeled and unlabeled states were assessed using descriptive statistics and multivariate regression methods. Adding a MS condition label had a statistically significant effect on mean health state values, resulting in lower values for labeled MS states v. unlabeled states. The data suggest that the MS label had a more significant effect on values for less severe states, and no significant effect on values for the most severe states. The inclusion of the MS label had a differential impact across the dimensions of the MSIS-8D. Across the MSIS-8D, predicted values ranged from 0.079 to 0.883 for unlabeled states, and 0.066 to 0.861 for labeled states. Differences reported in health state values, using labeled and unlabeled states, demonstrate that condition labels affect the results of valuation studies, and can have important implications in decision-analytic modelling and in economic evaluations.


Constipation is not a risk factor for hemorrhoids: a case-control study of potential etiological agents.
Constipation is widely believed to cause hemorrhoids. However, recent evidence questions the importance of constipation in the development of this common disorder. In the present study, the association of hemorrhoids with constipation and other potential risk factors was examined. The analysis consisted of 325 consecutive patients who underwent proctoscopy at the Milwaukee VA Medical Center during 1989. Each participant was questioned regarding hemorrhoid symptoms, bowel habits, and associated medical conditions. At the time of proctoscopy, the presence or absence of hemorrhoids was recorded. Hemorrhoids were identified in 168 subjects; the remaining 157 served as controls. No significant association was observed between constipation and hemorrhoids. Increasing age, cirrhosis, and varicose veins likewise were not associated with hemorrhoids. In contrast, the subjective complaint of diarrhea [odds ratio 2.1; 95% confidence limits (CL) 1.2-3.7] and obesity (odds ratio 1.7; 95% CL 1.1-2.7) were significantly associated with the presence of hemorrhoids. The results of this analysis suggest that diarrhea but not constipation may represent a risk factor for the development of hemorrhoids.


[Asparaginase mediated acid adaptation of mycobacteria].
Mycobacterium tuberculosis can survive inside host cells for a long time. To elucidate the acid adaptation mechanism mediated by the asparagine (Asn) metabolic pathway, the activity of asparaginase (AnsA) and growth of mycobacteria were characterized. The nonpathogenic M. bovis BCG was used as a model strain to characterize AnsA function. The recombinant AnsA was further expressed in E. coli, purified by Co2+ charged resin, and analyzed by enzymatic method. M. bovis BCG mutant (DansA) was constructed by homologous recombination. The growth and ammonia generation by M. bovis BCG wild type and mutant were evaluated in acidic medium. Ammonia was detected in the presence of AnsA and Asn. Deletion of ansA significantly delayed the growth of M. bovis BCG for 10 days in acidic medium. Asn was used by M. bovis BCG to produce ammonia, increasing pH. The acid adaptation of M. bovis BCG was mediated by ammonia, the metabolic product of asparagine. This provides new clue to uncover the survival mechanism of M. tuberculosis inside the acidic niche environment of host macrophage.


Proton therapy in the management of lymphoma.
Hodgkin lymphoma and non-Hodgkin lymphoma are common hematologic malignancies with such a favorable prognosis that many survivors live decades after combined chemotherapy and radiotherapy. These survivors, however, are at risk of developing late treatment adverse effects from collateral damage, including secondary malignancies, cardiac toxicities, and ischemic heart diseases. Recent efforts have successfully reduced the radiation dose and treatment field without compromising cure rates. Proton therapy has the potential of further lowering the treatment-related toxicity owing to its ability to deposit a high dose only at the target. Although this finding has been supported by several current dosimetric studies, its utilization in the management of lymphoma has been limited during the past 5 years because of the paucity of facilities and the difficulty of obtaining insurance coverage. With diligent follow-up, the clinical impact of proton therapy will be established to improve the therapeutic ratio and to reduce late treatment-related morbidity.


[Evaluation of four antigens for the detection of anti-Mycobacterium bovis antibodies by enzyme immunoassay].
An enzyme-linked immunosorbent assay (ELISA) for the diagnosis of bovine tuberculosis through the detection of specific seric antibodies has recently been developed in our laboratory. In order to assess its reproducibility and select the most adequate antigen, four bovine PPDs from different sources were evaluated in parallel: PPD M. bovis strain AN5, CEPANZO standard (CPZ), PPD M. bovis strain AN5, European Economic Community standard (EEC), PPD M. bovis strain AN5, prepared from non heated bacilli, killed by phenol (P) and PPD. M. bovis BCG strain prepared at the Pasteur Institute, Paris (BCG). Sera from 22 healthy cattle from tuberculosis free area and 20 bacteriologically confirmed tuberculous animals were employed in simultaneous assays. Antibody mean and standard deviations from healthy cattle expressed as optical density (OD) values were 45 +/- 22 when CPZ was used as antigen, 24 +/- 10 with EEC, 103 +/- 56 with P and 56 +/- 20 with BCG. Mean O.D. from tuberculous cattle were 588 +/- 158, 510 +/- 234, 782 +/- 138 and 441 +/- 189 with antigens CPZ, EEC, P and BCG respectively. A close correlation was observed when results obtained with EEC and P were compared with that of CPZ (r: 0.97 and 0.94 respectively). A lower specificity was achieved when BCG was used as antigen being also lower its correlation with the results obtained with CPZ (r: 0.87). It is concluded that our ELISA would achieve similar sensitivity and specificity if CPZ, EEC and P were used as antigens. On the other hand, BCG would not be suitable for this assay.


Rat salivary gland function after fractionated irradiation.
The aim of this study was to investigate longitudinal effects of fractionated irradiation, with various total doses, on salivary gland function in the rat. Female Sprague-Dawley rats were irradiated with 4, 5, 6, 7 or 8 Gy per day on five consecutive days. Irradiation was given to the head and neck region. Whole saliva was collected before and 2, 15 and 26 weeks after irradiation. In general the effects of irradiation on salivary gland function were found to be related to dose and time after exposure. Secretion rates were significantly decreased two weeks after irradiation with doses of 30 Gy or higher, after 15 weeks with 25 Gy or higher, and after 26 weeks with 20 Gy or higher. Response patterns to irradiation differed between the salivary constituents. Thus, the conclusions from this study are that early and late effects display different patterns and that the model used to study variations in salivary gland function after fractionated irradiation must be adjusted to the question addressed.


Calcium absorption--a paradigm for mineral absorption.
Intestinal calcium absorption proceeds by two mechanisms, an active transcellular process that takes place in the duodenum and a passive paracellular process throughout the small intestine. This article characterizes the three steps of transcellular calcium movement-entry, intracellular diffusion and extrusion-and identifies conditions that must be satisfied for other mineral ions to move transcellularly as part of a transepithelial transport process. Passive calcium movement is down a chemical gradient with the amount absorbed by this pathway determined in large measure by the sojourn time, most of which is spent in the ileum. Because transcellular movement of most mineral ions other than calcium, where measured, is either small or negligible, passive transport is likely to be the major route of intestinal absorption, the nature of which, however, has not been well established experimentally.


Monoclonal antibodies directed to leukocyte differentiation antigens for therapeutic use.
During the past years monoclonal antibodies directed to human leukocyte differentiation antigens have resulted not only in fundamental new knowledge of function and differentiation of leukocyte-populations, but have also turned out to be excellent and new tools for the treatment of lymphoid malignancies. Most widely and successfully monoclonal antibodies directed to T-cell differentiation antigens have been used for immunosuppression in organ transplantation. In the present paper, experimental and clinical data are analyzed which may give information about the criteria for clinical effectivity of distinct monoclonal antibodies. The conclusions clearly show, that not only specificity of an antibody, e.g. recognition of a distinct antigen, but also the recognized epitope, affinity and isotype of an antibody play a crucial role for the biological effectivity of a monoclonal antibody. Clinical success in antibody-therapy will therefore be dependent on the possibilities to alter antibody molecules according to the special clinical situation.


[A case of paraneoplastic limbic encephalitis associated with neck neuroendocrine carcinoma].
A 69-year-old man presented with a history of personality change for several years. He was admitted to our hospital due to partial seizure. A cerebrospinal fluid test and an electroencephalogram showed no specific abnormalities, but brain magnetic resonance imaging revealed abnormal findings in the right temporal pole, bilateral amygdala to hippocampus, and insular cortex. He was diagnosed with limbic encephalitis accompanied by partial seizure, and received infusion of an antiepileptic agent and acyclovir. Additional examinations for malignancy and autoimmune disease were performed, and neck CT and MRI revealed a neck tumor. Neck lymph node biopsy suggested lymph node metastasis of a neuroendocrine neoplasm derived from other organs. He did not want aggressive treatment involving surgical resection and chemotherapy, and thus, conservative treatment was chosen by an otorhinolaryngologist and immunotherapy was not used. After discharge, the neck tumor grew gradually. To manage the focal mass effect, chemotherapy and surgical resection followed by chemoradiotherapy were performed by the otorhinolaryngologist on days 244 and 325 of the disease course, respectively. Histology of resected tissues disclosed neck neuroendocrine carcinoma derived from a submandibular gland. His personality change improved temporarily after surgical resection, but then worsened again with regrowth of the tumor. He died on day 723. After death, a blood test revealed the presence of anti-amphiphysin antibody. This case suggests that neck neuroendocrine carcinoma can induce paraneoplastic limbic encephalitis, and in such cases, early surgical resection of the neck tumor with suspected lymph node metastasis is necessary both to control symptoms associated with encephalitis and to exclude carcinoma derived from the neck itself.


An Investigation on the Quantitative Structure-Activity Relationships of the Anti-Inflammatory Activity of Diterpenoid Alkaloids.
Diterpenoid alkaloids are extracted from plants. These compounds have broad biological activities, including effects on the cardiovascular system, anti-inflammatory and analgesic actions, and anti-tumor activity. The anti-inflammatory activity was determined by carrageenan-induced rat paw edema and experimental trauma in rats. The number of studies focused on the determination, quantitation and pharmacological properties of these alkaloids has increased dramatically during the past few years. In this work we built a dataset composed of 15 diterpenoid alkaloid compounds with diverse structures, of which 11 compounds were included in the training set and the remaining compounds were included in the test set. The quantitative chemistry parameters of the 15 diterpenoid alkaloids compound were calculated using the HyperChem software, and the quantitative structure-activity relationship (QSAR) of these diterpenoid alkaloid compounds were assessed in an anti-inflammation model based on half maximal effective concentration (EC50) measurements obtained from rat paw edema data. The QSAR prediction model is as follows: log ( E C 50 ) = - 0.0260 × SAA + 0.0086 × SAG + 0.0011 × VOL - 0.0641 × HE - 0.2628 × LogP - 0.5594 × REF - 0.2211 × POL - 0.1964 × MASS + 0.088 × BE + 0.1398 × HF (R² = 0.981, Q² = 0.92). The validated consensus EC50 for the QSAR model, developed from the rat paw edema anti-inflammation model used in this study, indicate that this model was capable of effective prediction and can be used as a reliable computational predictor of diterpenoid alkaloid activity.


Fellow As Teacher Curriculum: Improving Rheumatology Fellows' Teaching Skills During Inpatient Consultation.
Enhancing rheumatology fellows' teaching skills in the setting of inpatient consultation may have a broad positive impact. Such efforts may improve fellows' clinical skills and overall patient care. Most importantly, effective resident-fellow teaching interactions may not only increase residents' knowledge of rheumatology but may influence their career choice. However, a number of barriers to the resident-fellow teaching interaction have been identified, including fellows' teaching skills. We developed the Fellow As Clinical Teacher (FACT) curriculum in order to enhance fellows' teaching skills during inpatient consultation. The FACT curriculum was delivered in two 45-minute workshops during the 3-day Winter Symposium of the Carolinas Fellows Collaborative. We evaluated its effect with self-assessment surveys and fellow performance on the objective structured teaching exercise (OSTE) before and after participation in the curriculum. Nineteen fellows from 4 rheumatology training programs participated in the pre- and post-curriculum OSTEs and 18 fellows completed pre- and post-curriculum surveys. OSTE scores improved on 5 of the 8 items assessed, and the total OSTE score improved as well (34.7 versus 29.5; P < 0.01) after the FACT curriculum. Fellows' self-assessment of their teaching skills and intent to teach during consultation also increased after participation in the curriculum. The FACT curriculum, focused on teaching during consultation, improved fellows' teaching skills and attitudes toward teaching. Improving and increasing fellow teaching, particularly in the consultation setting, may impact patient care, resident and fellow learning, and teaching skills of future faculty, and could potentially influence residents' career choice.


Resilience anchors for children in an out-of-home care institution during and after COVID-19.
Growing numbers of children of all ages grow up in out-of-home care institutions due to personal and socioecological risk variables that destabilized their families of origin. In the aftermath of the COVID-19 pandemic which disrupted lives and development, there is particular interest in how children who grow up in out-of-home care institutions cope and develop. This paper reports the findings of a study that sought to document anchors of resilience in children who resided in a care institution run by a non-governmental, church-based welfare organization in one of the central provinces of South Africa. In line with recent developments in childcare, the organization mainly functions via smaller child and youth group homes across the province (compared to bigger children's homes in the past). In our qualitative, phenomenological study, we used the participatory, child-friendly, and less intrusive draw-and-write technique to generate data. We asked the 20 participating children of one of these group homes to make drawings that mirror their lives, and to write paragraphs in which they described their drawings. All were school-going children in care, aged from 12 to 19. There were 11 girls and 9 boys in the study, and one of these identified as "other." The grades ranged from 7 to 12 and they spoke African languages, namely Afrikaans, Sesotho, Setswana and IsiXhosa. We used inductive content analysis to process the data, and the findings indicate that, notwithstanding personal and socioecological risks during and after the COVID-19 pandemic, the resilience of the participants was anchored by a number of universal personal strengths as well as socioecological resources.


Obstacles to successful antiretroviral treatment of HIV-1 infection: problems & perspectives.
Mutations in human immunodeficiency virus type 1 (HIV-1) are a major impediment to successful highly active antiretroviral therapy (HAART) and the design of anti-HIV vaccines. Although HAART has made long-term suppression of HIV a reality, drug resistance, drug toxicity, drug penetration, adherence to therapy, low levels of continued viral replication in cellular reservoirs and augmentation of host immune responses are some of the most important challenges that remain to be sorted out. Continuing viral replication in the face of HAART leads to the accumulation of drug resistance mutations, increase in viral loads and eventual disease progression. Patients who fail therapy have minimal options for their clinical management. Therefore, a clear understanding of the pathogenesis of drug-resistant HIV-1, and all of the issues that influence the success of HAART is urgently needed. In the present article, we discuss various obstacles to HIV therapy, and provide perspectives relating to these issues that are critical in determining the success or failure of HAART.


How patchy is patchy villous atrophy?: distribution pattern of histological lesions in the duodenum of children with celiac disease.
In celiac disease (CD) the degree of histological damage in the duodenum may vary, but there is some controversy about the coexistence of villous atrophy and normal mucosa in different biopsy sites, i.e., patchy villous atrophy. We prospectively evaluated the degree, frequency, and distribution of histological lesions among different duodenal sites as well as within each duodenal biopsy. Over the last 4 years, in each patient with suspected CD (positive anti-transglutaminase antibodies), four to five endoscopic biopsies were taken from the duodeno-jejunal flexure/distal duodenum (D3), intermediate duodenum (D2), proximal duodenum (D1), and duodenal bulb (B). Biopsies were subjected to hematoxylin/eosin staining and immunostaining with anti-CD3 monoclonal antibodies for intraepithelial lymphocyte (IEL) count. Duodenal lesions were classified according to Marsh-Oberhuber, and CD was diagnosed according to the European Society of Pediatric Gastroenterology, Hepatology, and Nutrition criteria. Six hundred and eighty-six children did have CD. A degree of villous atrophy was found in 660/686 patients (96.2%), total villous atrophy was present in 550/686 (80.1%), and 320/686 (46.6%) had different lesions at different sites, but none of these patients had entirely normal biopsies. In all, 116 of 686 (16.9%) had variable lesions within the same biopsy, with grade 2+3A being the most frequent association (43%), followed by 2+3A+3B (27%) and 2+3A+3B+3C (22%). All these 116 patients also had histologically normal areas within the same biopsy, but anti-CD3 immunostaining showed that IELs were always increased in such areas. In all the cases, the severity of duodenal lesions significantly increased in an aborad manner (chi(2)=52.38 with alpha=0.01 and d.f.=12; P<0.0001). No correlation was found between type and distribution of histologic lesions and clinical presentation of CD. In newly diagnosed CD, some variability of histological lesions can be found, even within the same duodenal biopsy, in which areas of apparently normal mucosa with increased IEL number often exist. We also confirm our previous findings that duodenal lesions may vary among different biopsies; lesion severity has a proximal-to-distal gradient, but no patient has entirely normal duodenal biopsies. The awareness of such histological variability may help establish a correct diagnosis of CD.


Profile of Women receiving Second-trimester Safe Abortion Service at Paropakar Maternity and Women's Hospital.
Background Second trimester abortions account for a small percentage of abortions globally. Abortion is one of the safest medical procedures. Accessibility of safe and high quality second-trimester abortion care can decrease severe complications associated with it. Objective The profile of women undergoing second-trimester abortion was assessed during the study. The profile includes demographic and clinical aspects. The study also aims to find out the rate of surgical and medical methods of abortion and to understand its indications. Method It is a study based on retrospective data collected from charts available at the comprehensive abortion care center at the Paropakar Maternity and Women's Hospital. The data from Shrawan to Magh 2077 B.S. was collected and calculations were done on MS Excel. Result Higher proportion of women receiving second-trimester abortion were of the age between 21-25 years (n=31; 31%). Most women were literate (n=69; 69%) and n=38(38%) were nulliparous. The most common indication for abortion was maternal mental issues (n=32; 32%). The rate of medical abortion was higher (n=71; 71%) than surgical method of abortion (n=29; 29%). Conclusion There exists both demographic and clinical variations in women undergoing secondtrimester abortion. Though few women undergo second trimester abortion than first-trimester the variation in profile is significant with most women referring to second trimester abortion due to later diagnosis of pregnancy, women facing financial and logistic barriers to health care, victims of violence and women with pregnancy complications or fetal anomalies.


[Development of External Stimuli-responsive Organic π Molecules and the Creation of Novel Photo-functions].
We have found that the spiro form of aminobenzopyranoxanthene (ABPX) exhibits dual solvatochromic and nanoaggregate fluorescence in organic solvents by spectrophotometric and theoretical analyses. The dual fluorescence properties of ABPX were adjustable in response to water content, and served as a new detection principal for naked-eye visualization (above 0.5 wt%) and quantification (0.010-0.125 wt%) of water in tetrahydrofuran. In investigating the optical properties of a dicationic form of ABPX, ABPX containing linear n-alkyl chains at amino groups were then synthesized. These ABPX exhibited fluorescence emission in the far-red and NIR wavelength regions, and we noted an increased fluorescence quantum efficiency with increasing n-alkyl chain length. To further improve the fluorescence quantum yields, we have designed and synthesized ABPX with different nitrogen-containing fused rings. It was kinetically demonstrated that the structurally rigid conjugation of the xanthene moiety is an effective molecular design for the drastic enhancement of fluorescence emission efficiency.


Mechanisms of Cx43 and Cx26 transport to the plasma membrane and gap junction regeneration.
Previous reports have suggested that Cx26 exhibits unique intracellular transport pathways en route to the cell surface compared with other members of the connexin family. To directly examine and compare nascent and steady-state delivery of Cx43 and Cx26 to the plasma membrane and gap junction biogenesis we expressed fluorescent-protein-tagged Cx43 and Cx26 in BICR-M1Rk and NRK cells. Static and time-lapse imaging revealed that both connexins were routed through the Golgi apparatus prior to being transported to the cell surface, a process inhibited in the presence of brefeldin A (BFA) or the expression of a dominant-negative form of Sar1 GTPase. During recovery from BFA, time-lapse imaging of nascent connexin Golgi-to-plasma membrane delivery revealed many dynamic post-Golgi carriers (PGCs) originating from the distal side of the Golgi apparatus consisting of heterogeneous vesicles and long, tubular-like extensions. Vesicles and tubular extensions were also observed in HBL-100 cells expressing a human, disease-linked, Golgi-localized Cx26 mutant, D66H-GFP. A diffuse cell surface rim of fluorescent-protein-tagged wild-type connexins was observed prior to the appearance of punctate gap junctions, which suggests that random fusion of PGCs occurred with the plasma membrane followed by lateral diffusion of connexins into clusters. Fluorescence recovery after photobleaching studies revealed that Cx26-YFP was more mobile within gap junction plaques compared with Cx43-GFP. Intriguingly, Cx43-GFP delivery and gap junction regeneration was inhibited by BFA and nocodazole, whereas Cx26-GFP delivery was prevented by BFA but not nocodazole. Collectively, these studies suggest that during gap junction biogenesis two phylogenetically distinct members of the connexin family, Cx43 and Cx26, share common secretory pathways, types of transport intermediates and turnover dynamics but differ in their microtubule-dependence and mobility within the plasma membrane, which might reflect differences in binding to protein scaffolds.


New 1H-pyrazole-4-carboxamides with antiplatelet activity.
Nine title compounds were synthesized and investigated in the Born test for their antiplatelet activities against collagen, ADP, adrenaline, and platelet activating factor (PAF) as inducers of the aggregation. Using collagen three compounds with IC50 values below 100 microM were found (3b, 3e, 3i). Activities in nanomolar concentrations were observed against ADP (3b, IC50 = 9.4 nM), adrenaline (3i, IC50 = 5.8 nM), and platelet activating factor (3e, IC50 = 0.45 nM).


Spatially gradated hydrogel platform as a 3D engineered tumor microenvironment.
There is an acute need for biomaterial tools that recreate the heterogeneous brain-tumor microenvironment. A microfluidic mixing tool is reported to encapsulate glioblastoma multiforme cells within miniaturized gelatin hydrogels containing overlapping patterns of tumor-inspired matrix signals. This approach permits in situ analysis of glioma cells at the molecular and genomic level as well as the potential for clinical insight.


Acute hepatitis in recurrent hereditary polyserositis (familial Mediterranean fever).
The clinical features of two cases of Hereditary Recurrent Polyserositis HRP (Familial Mediterranean Fever) in related Yemeni children resident in Saudi Arabia are described. One presented with recurrent acute hepatitis, which has not been previously documented. These are the first cases of HRP reported in Arabs originating from the Arabian Peninsula.


<Editors' Choice> Treatment strategy for upper cervical epidural abscess: a literature review.
We aimed to determine available evidences in the literature regarding surgical approaches and methods, timing of surgical interventions, duration of perioperative antibiotics, and duration of nonsurgical treatments (antibiotics administration) in patients with upper cervical (occiput-C2) epidural abscess (UCEA). We performed a literature review of the articles on surgical interventions and antibiotic therapy to treat UCEA, searching the PubMed database for relevant articles published in the English language (as of March 2020). In total, 53 patients with UCEA were identified. Permanent limb paralysis or death was observed in 1/15 (6.7%) patients who received the transoral approach and 2/15 (13.3%) patients who received the transcervical approach, 1/26 (3.8%) patients who underwent surgery before the onset of paralysis, and 2/4 (50.0%) patients who underwent surgery after the onset of paralysis. In 85%-89% of cases, antibiotic administration was continued for 6-12 weeks, which was determined by the confirmation of reduced inflammatory response and/or abscess disappearance on imaging. Differences in surgical approaches may not be associated with the incidence of permanent limb paralysis or death. Surgical interventions before limb paralysis onset are recommended in UCEA patients. In perioperative and nonoperative treatments, antibiotic administration for 6-12 weeks may be supported based on the confirmation of reduced inflammatory response and/or abscess disappearance on imaging. Further investigations are needed.


Aryl ketones from Acronychia pedunculata with cyclooxygenase-2 inhibitory effects.
1-[2,4-Dihydroxy-6-methoxy-3,5-bis(3-methylbut-2-en-1-yl)phenyl]ethanone (1), and a new aryl ketone, named acrovestenol (2), were isolated as cyclooxygenase-2 (COX-2) inhibitory principles from a CH2Cl2 extract of the bark of Acronychia pedunculata by a bioassay-guided fractionation procedure. Compound 2 inhibited COX-2 with an IC50 value of 142.0+/-2.15 microM, compared to the COX-2 inhibitory reference compound NS-398 with an IC50 value of 11.3+/-1.12 microM. Compound 1 inhibited COX-2-catalyzed PG biosynthesis with 68% at a concentration of 500 microM. The structures were determined by UV, IR, and 1D- and 2D-NMR, including TOCSY, HSQC-DEPT, and HMBC, and MS investigations.


The production of tissue-specific histone complements during development.
At least two mechanisms generate tissue differences in the histone subtype composition during development: subtype dilution and subtype replacement. Subtype dilution, which occurs when cells continue dividing after having ceased to synthesize one more histone subtypes, allows the elimination of stable subtypes. It is the major mechanism generating cell differences in histone composition in sea urchin embryogenesis. Subtype replacement has been observed in mammalian tissues, both in the intact animal and in cultured cells. It is most evident in nondividing cells but occurs to some extent in dividing cells as well. Examples of the two mechanisms are presented and their possible biological significance, as well as that of the differences they produce, is discussed.


Wind profiling based on the optical beam intensity statistics in a turbulent atmosphere.
Reconstruction of the wind profile from the statistics of intensity fluctuations of an optical beam propagating in a turbulent atmosphere is considered. The equations for the spatiotemporal correlation function and the spectrum of weak intensity fluctuations of a Gaussian beam are obtained. The algorithms of wind profile retrieval from the spatiotemporal intensity spectrum are described and the results of end-to-end computer experiments on wind profiling based on the developed algorithms are presented. It is shown that the developed algorithms allow retrieval of the wind profile from the turbulent optical beam intensity fluctuations with acceptable accuracy in many practically feasible laser measurements set up in the atmosphere.


Homocysteine induced impairment of nitric oxide-dependent vasorelaxation is reversible by the superoxide dismutase mimetic TEMPOL.
Elevated plasma homocysteine concentrations in renal patients are associated with accelerated cardiovascular disease. The mechanism(s) by which homocysteine acts remains unclear however, evidence implicates a role involving endothelial dysfunction. Rat femoral arteries after acute or 4-h pre-incubation with racemic D,L-homocysteine (100 microM) were mounted on a myograph, pre-constricted with phenylephrine (10 microM) and responses to acetylcholine-dependent vasorelaxation examined. The incubations were repeated in the presence of indomethacin (10 microM), omega-nitro-L-arginine methyl ester (100 microM), L-arginine (100 microM), tetrahydrobiopterin (1 microM), catalase (1200 U/ml), ebselen, a peroxynitrite chelator (20 microM) and TEMPOL, a superoxide dismutase mimetic (1 mM). Results are shown as means+/-standard error, expressed as per cent relaxation to acetylcholine added (nmol/l). Increasing concentrations of homocysteine had no affect when added directly to basally relaxed or pre-constricted freshly isolated vessels. However, 4-h pre-incubation with or without homocysteine significantly shifted the acetylcholine EC(50) (EC(50) was defined as the concentration of acetylcholine that caused relaxation of the phenylephrine contracted tissue by 50%), control((4 h)) = 74.7 nmol/l+/-10.5 vs 100 microM D,L-homocysteine((4 h)) = 159.9 nmol/l+/-20.6; P<0.05) without affecting maximal relaxation. Response to endothelial independent relaxation was unaffected. Indomethacin, indomethacin and omega-nitro-L-arginine methyl ester, l-arginine and tetrahydrobiopterin, catalase and ebselen had no effect on the EC(50) in homocysteine-exposed arteries. However, TEMPOL normalized vasorelaxation in homocysteine-treated arteries (75.2 nmol/l+/-14.6) but had no effect on the 4-h control group. Moreover, washing TEMPOL from the treated vessels restored endothelial dysfunction in D,L-homocysteine-treated vessels (163.9 nmol/l+/-34.1). We conclude that homocysteine causes endothelial dysfunction by up-regulating a potential superoxide generating system resulting in reduced nitric oxide bio-availability.


Re-examining poikilocytosis in goats: prevalence, type and association with age and disease.
Domestic goats (Capra aegagrus hircus) are a food, fiber and companion animal. Abnormal erythrocyte shapes (poikilocytes) are considered normal in young goats, but their association with disease is not well described. Likewise, there is little information on the significance of poikilocytosis in adult goats. The objective of this study was to investigate the prevalence, severity and type of poikilocytosis in young and adult goats and its association with age, sex, breed, laboratory results, and underlying disease. We retrospectively examined clinical and laboratory data from 1254 goats presented at the University of California-Davis Veterinary Medical Teaching Hospital from 1997 to 2019. We analyzed 313 blood smears from goats with moderate or marked (MOD-MKD) poikilocytosis on initial blood smear evaluation. Number and type of poikilocytes per 1000 red blood cells (RBCs) were enumerated. Laboratory values and primary disease categories were compared with the severity and type of poikilocytosis. Kids (<6 mos) and juveniles (>6 mos to <1 year) had a higher prevalence of MOD-MKD poikilocytosis (95/210, 45.2% kids; 27/59, 45.8% juveniles) than adult goats (≥1 year; 190/982, 19.3%) (p < 0.001). Kids had a higher percentage of elliptocytes, dacryocytes, and schistocytes and a lower percentage of polygonal and spiculated RBCs than juvenile and adult goats (p < 0.001). Of goats with MOD-MKD (vs NONE-SLIGHT) poikilocytosis, kids had lower HGB and MCH, and higher RDW (p ≤ 0.02); juveniles and adult goats had lower HCT, MCV, MCH, and albumin concentration (p ≤ 0.01), and all age groups had lower total CO2 concentration and higher PLT counts (p < 0.03). Adult goats with MOD-MKD poikilocytosis also had higher BUN:Cr ratios (p = 0.005). Gastrointestinal parasitism, Johne's disease, diarrhea/enteritis, lice, hepatic disease and renal disease (but not urolithiasis) were significantly associated with MOD-MKD poikilocytosis (p < 0.001). Goats with hepatic and renal disease had a higher prevalence and percentage of spiculated cells (p = 0.001). Goats with Johne's disease had a higher prevalence of polygonal cells (93.3%) and dacryocytes (66.7%) than other diseases, and elliptocytes predominated in a higher proportion (36.0%) of adult goats with GI parasitism vs other diseases (p < 0.05). These findings suggest that iron deficiency is an important pathophysiologic mechanism of poikilocytosis in juvenile and adult goats, and possibly in kids, whether due to iron-restricted erythropoiesis, chronic hemorrhage, functional iron deficiency, or a combination of these mechanisms. Further investigation into the detection and monitoring of iron deficiency and the value of poikilocytosis as a diagnostic marker of iron status in goats is warranted.


Insight into the Endocrine System and the Immune System: A Review of the Inflammatory Role of Prolactin in Rheumatoid Arthritis and Psoriatic Arthritis.
Rheumatoid arthritis (RA) is a chronic autoimmune disease that affects females three times more frequently than males. A potential role for hormones, such as prolactin (PRL), may in part explain this phenomenon. The risk of developing RA is increased in women who are lactating after the first pregnancy, which might be related to breastfeeding and the release of PRL. Other studies found a protective effect of PRL on RA development. Some studies have reported that hyperprolactinemia is more common in RA and serum PRL levels are correlated with several disease parameters, although others could not confirm these findings. Overall the plasma PRL levels are on average not elevated in RA. Previously, a small number of open-label clinical trials using bromocriptine, which indirectly decreases PRL levels, were performed in RA patients and showed clinical benefit, although others found the opposite effect. Locally produced PRL at the site of inflammation may have a crucial role in RA as well, as it has been shown that PRL can be produced by synovial macrophages. Locally produced PRL has both pro-inflammatory and anti-inflammatory effects in arthritis. Psoriatic arthritis (PsA) is also an autoinflammatory disease, in which the prolactin receptor is also expressed in macrophages. The aim of this review is to provide an overview of the potential role of PRL signaling in inflammatory joint diseases (RA and PsA) and its potential as a therapeutic target.


Low-Grade Myxofibrosarcoma of the Rectus Abdominus Muscle Infiltrating into Abdominal Cavity: A Case Report.
Objective: Myxofibrosarcoma (MFS) is a relatively rare tumor that is histologically characterized by myxoid stroma and spindle cell proliferation. This tumor most commonly arises as a slow growing, enlarging painless mass in the extremities of elderly patients. Methods: We report a case of a primary, low-grade MFS in the rectus abdominis muscle infiltrating the abdominal cavity of a 75-year-old man. Results: The patient underwent a wide excision of the right abdominal wall mass with a 3-cm surgical margin from the scar due to a biopsy. The tumor infiltrated the urinary bladder, peritoneum, and external iliac vessels. Twenty-six months after the initial operation, he had recurrences in his abdominal wall, urinary bladder, and right iliac vessels. Conclusions: To our knowledge, primary MFS of the muscle in the abdomen has not been documented previously. Although this case was histopathologically classified as a low-grade tumor, it infiltrated the abdominal cavity. The tumor is suspected to have penetrated the abdominal cavity below the linea arcuata, which lacks the posterior sheath of the rectus abdominis muscle; from there, it could easily spread without being blocked by any biological barriers.


The Yin and Yang of Invariant Natural Killer T Cells in Tumor Immunity-Suppression of Tumor Immunity in the Intestine.
CD1d-restricted invariant natural killer T (iNKT) cells are known as early responding, potent regulatory cells of immune responses. Besides their established role in the regulation of inflammation and autoimmune disease, numerous studies have shown that iNKT cells have important functions in tumor immunosurveillance and control of tumor metastasis. Tumor-infiltrating T helper 1 (TH1)/cytotoxic T lymphocytes have been associated with a positive prognosis. However, inflammation has a dual role in cancer and chronic inflammation is believed to be a driving force in many cancers as exemplified in patients with inflammatory bowel disease that have an increased risk of colorectal cancer. Indeed, NKT cells promote intestinal inflammation in human ulcerative colitis, and the associated animal model, indicating that NKT cells may favor tumor development in intestinal tissue. In contrast to other cancers, recent data from animal models suggest that iNKT cells promote tumor formation in the intestine by supporting an immunoregulatory tumor microenvironment and suppressing TH1 antitumor immunity. Here, we review the role of iNKT cells in suppression of tumor immunity in light of iNKT-cell regulation of intestinal inflammation. We also discuss suppression of immunity in other situations as well as factors that may influence whether iNKT cells have a protective or an immunosuppressive and tumor-promoting role in tumor immunity.


Cystic Lymphatic Malformation with Lymphedema Treated by Lymphaticovenular Anastomosis Combined with Ethanol Sclerotherapy.
The incidence of cystic lymphatic malformation (CL) in an extremity is very rare. CL can be a cause of lymphedema in a lower limb. The most effective treatment for CL is sclerotherapy or excision; however, these treatments have the potential to cause fibrosis and obliteration of ruptured lymphatic vessels, which impairs lymphatic drainage and increases the risk of lymphedema. Lymphaticovenular anastomosis (LVA) combined with sclerotherapy may be a minimally invasive treatment option for CL in a lower limb. In this report, we describe a patient with CL complicated by lymphedema in a lower extremity, who we treated using LVA and ethanol sclerotherapy with satisfactory results. The patient was a 60-year-old man with a CL measuring 16 cm in diameter in the thigh region above the knee with lower limb lymphedema. The percentage of excess volume of the affected lower leg was 24.7%, preoperatively. Combined treatment using LVA with sclerotherapy was performed under general anesthesia. At 2 years after surgery, the reduction of excess volume of affected lower leg between preoperative and postoperative was 85.4%. In our previous report, the same combination therapy was used to treat CL and prevent lymphedema; however, in the present case, it was used to treat CL and lymphedema at one stage. LVA combined with sclerotherapy is a complementary minimally invasive treatment for CL accompanied by lymphedema.


Impact of the time interval between lymph node recurrence and lymphadenectomy on melanoma patient survival.
Lymphadenectomy is currently the standard treatment for melanoma patients with palpable lymph node (LN) metastasis. There is no recommendation as to the time when surgery should be performed in France. The aim of this retrospective study was to assess the impact of the time interval preceding lymphadenectomy on patient outcomes. Patients who underwent lymphadenectomy for LN macrometastasis (Stage IIIB/C AJCC) between 2005 and 2012 were included. Both the time interval between the first suspicion of LN recurrence (physical examination and imaging) and lymphadenectomy and the time interval between the multidisciplinary team meeting and lymphadenectomy were recorded. The impact of these time intervals on patient relapse-free survival (RFS) and overall survival (OS) were analysed. The regression optimized (ROP) model was used to identify ghost factors for the Cox model. A total of 154 patients were included. The median time interval between the multidisciplinary team meeting and lymphadenectomy was 22 days (IQR: 6 to 66). The median time interval between the first suspicion of LN recurrence and lymphadenectomy was 59 days (IQR: 15 to 676). Taking into account the effect identified by the regression optimized (ROP) model, these times were associated with an increased risk of recurrence and mortality (p<0.001 and p = 0.01, respectively). Our study demonstrates that increasing the time interval preceding lymphadenectomy significantly reduces patient RFS and OS.


Altered drug metabolism during pregnancy: hormonal regulation of drug-metabolizing enzymes.
Medication use during pregnancy is prevalent, but pharmacokinetic information of most drugs used during pregnancy is lacking in spite of known effects of pregnancy on drug disposition. Accurate pharmacokinetic information is essential for optimal drug therapy in mother and fetus. Thus, understanding how pregnancy influences drug disposition is important for better prediction of pharmacokinetic changes of drugs in pregnant women. Pregnancy is known to affect hepatic drug metabolism, but the underlying mechanisms remain unknown. Physiological changes accompanying pregnancy are probably responsible for the reported alteration in drug metabolism during pregnancy. These include elevated concentrations of various hormones such as estrogen, progesterone, placental growth hormones and prolactin. This review covers how these hormones influence expression of drug-metabolizing enzymes (DMEs), thus potentially responsible for altered drug metabolism during pregnancy. The reader will gain a greater understanding of the altered drug metabolism in pregnant women and the regulatory effects of pregnancy hormones on expression of DMEs. In-depth studies in hormonal regulatory mechanisms as well as confirmatory studies in pregnant women are warranted for systematic understanding and prediction of the changes in hepatic drug metabolism during pregnancy.


Functional properties and physiological roles of organic solute channels.
Membrane channels provide routes for the rapid, passive movement of solutes across plasma and intracellular membranes. It is generally assumed that the major physiological role of membrane channels is to transport inorganic ions for processes such as transepithelial salt absorption and secretion, cell volume regulation, signal transduction, and control of membrane electrical properties. Increasing evidence indicates, however, that channels play an important role in organic solute transport in a wide variety of cell types and organisms. Some of the major physiological roles of organic solute channels include uptake of nutrients, excretion of metabolic waste products, volume-regulatory organic osmolyte transport, and control of mitochondrial metabolism. This article reviews the functions and characteristics of channels that participate in the transport and regulation of both charged and electroneutral organic solutes.


What can we gain from subpopulation universal pulses? A simulation-based study.
The aim of the study was to explore a novel methodology for designing universal pulses (UPs) that balances the benefits of a calibration-free approach with subject-specific online pulse design. The proposed method involves segmenting the population into subpopulations with variability in anatomical shapes and positions reduced to 75%, 50%, and 25% of their original values while keeping the mean values unchanged. An additional 25% extreme case with a large volume of interest and shifted position was included. For each group, a 5kT-points universal inversion pulse was designed and assessed by the normalized root mean square error (NRMSE) on the target longitudinal magnetization profile. The performance was compared to the conventional one-size-fits-all approach. A total of 132 electromagnetic simulations were executed to generate representative anatomies and specific absorption rate (SAR) distributions in a three-dimensional parameter space comprised of head breadth, head length, and Y-shift. The 99.9th percentile on the peak local SAR distribution was utilized to establish an intersubject variability safety margin. UPs designed for subpopulations with decreased head shape and position variability reduced the anatomical safety margin by up to 20%. Furthermore, when a head was significantly different to the average case, the proposed approach improved the inversion homogeneity by up to 24%, compared to the conventional one-size-fits-all approach. Subpopulation UPs present an opportunity to improve the B 1 + $$ {\mathrm{B}}_1^{+} $$ homogeneity and reduce anatomical SAR safety margins at 7T without additional acquisition time for calibration.


Association between anthropometric measures of regional fat mass and heart rate variability in obese women.
Impaired autonomic function could be the mechanism for the development of cardiometabolic diseases in obesity. Hence, simple anthropometric measures of overall and central obesity could be screening markers for autonomic imbalance. We analysed the relationship between heart rate variability (HRV) parameters and obesity indicators. Sixty-three obese women underwent blood pressure, lipids and anthropometric measurements, body composition assessment, HOMA (homeostasis model assessment) index calculation and short-term HRV analysis. The correlation between obesity indicators and HRV parameters was influenced by age. In the multiple regression model, sagittal abdominal diameter (SAD) was a significant negative predictor of lnLF/HF (logarithmically transformed ratio of low to high frequencies) and lnLFnorm, and positive predictor of HFnorm (normalized high frequencies); the significant relationship remained even after adjustment for age, HOMA, blood pressure, lipid profile, menopause, body mass index (BMI) and body fat percentage (FAT). Anterior forearm skinfold showed inverse association with HRV. Correlation between waist circumference and waist-to-height ratio (WHtR) with lnLF/HF, as well as between anterior thigh skinfold with lnLF/HF, LFnorm (normalised low frequencies) and HFnorm was lost after further adjustments. Among all anthropometric measures, SAD and anterior forearm skinfold thickness showed the highest predictive ability for HRV. Markers of lower sympathetic and higher cardiac parasympathetic activity corresponded with indicator of central obesity, while indicators of peripheral obesity showed completely opposite relationship with markers of cardiac autonomic activity.


Effects of Astragalus membranaceus and its main components on the acute phase endothelial dysfunction induced by homocysteine.
This study was designed to investigate the effects of Astragalus membranaceus (AM) and its main components, astragalus saponin (ASP), astragalus polysaccharide (APS) and aminobutyric acid (GABA), on homocysteine (Hcy) induced acute impairment of vascular tone and to explore whether the antioxidant mechanism was involved in AM protective effect. Inhibitory effects of Hcy and protective effects of AM and its main components on endothelium-dependent relaxation of aortic rings were determined by isometric tension recordings and nitric oxide signaling was assayed with 125I-cGMP RIA Kit. Furthermore, generation of reactive oxygen species (ROS) in endothelial cells was detected using 5-(6)-chloromethyl-2',7'-dichlorodihydrofluorescein diacetate (CM-H2DCF-DA). Hcy significantly inhibited endothelium-dependent relaxation to acetylcholine (ACh) in a dose-dependent manner, and decreased cGMP levels increased by ACh in aorta. Furthermore, superoxide dismutase (SOD), AM, and ASP markedly attenuated inhibition of vasorelaxation and downregulation of cGMP level by Hcy, and APS exerted a tendency to reverse both of the depressive responses, while GABA had no similar effects. Additionally, partially impaired relaxation by Hcy was completely blocked due to the presence of N(omega)-nitro-L-arginine-methyl ester (L-NAME), which could not be further altered by treatment with AM, ASP, APS or GABA. Finally, Hcy significantly increased intracellular ROS levels in endothelial cells as measured by CM-H2DCF-DA fluorescence. SOD, AM, ASP, and APS, but not GABA, inhibited Hcy-stimulated ROS generation. This study demonstrated that AM and ASP, potently protected endothelium-dependent relaxation against the acute injury from Hcy through nitric oxide regulatory pathways, in which antioxidation played a key role.


Root and Shoot Biomass Growth of Constructed Floating Wetlands Plants in Saline Environments.
Constructed Floating Wetlands (CFWs) are increasingly being used globally in freshwater environments such as urban lakes and ponds to remove pollutants from urban stormwater runoff. However, to date there has been limited research into the use and performance of these systems in saline environments. This study compared the root and shoot biomass growth and nutrient uptake of five different plant species, Chrysopogon zizanioides, Baumea juncea, Isolepis nodosa, Phragmites australis and Sarcocornia quinqueflora, in three different saltwater treatments over a 12-week period. The aim of the study was to identify which of the plant species may be most suitable for use in CFWs in saline environments. Plant nutrient uptake testing revealed that Phragmites australis had the greatest percentage increase (1473⁻2477%) of Nitrogen mass in the shoots in all treatments. Sarcocornia quinqueflora also had impressive Nitrogen mass increase in saltwater showing an increase of 966% (0.208 ± 0.134 g). This suggests that the use of Phragmites australis and Sarcocornia quinqueflora plants in CFWs installed in saline water bodies, with regular harvesting of the shoot mass, may significantly reduce Nitrogen concentrations in the water. Isolepis nodosa had the greatest percentage increase (112% or 0.018 ± 0.020 g) of Phosphorous mass in the shoots in the saltwater treatment. Baumea juncea had the greatest percentage increase (315% or 0.026 ± 0.012 g) of Phosphorous mass in the roots in the saltwater treatment. This suggests that the use of Isolepis nodosa and Baumea juncea plants in CFWs installed in saline water bodies may significantly reduce Phosphorous concentrations in the water if there was a way to harvest both the shoots above and the roots below the CFWs. The study is continuing, and it is anticipated that more information will be available on CFW plants installed in saline environments in the near future.


A neutralization-inhibition enzyme immunoassay for anti-idiotypic antibodies that block monoclonal antibodies neutralizing Semliki Forest virus.
In this paper we compare a solid-phase enzyme immunoassay (EIA) with a neutralization-inhibition enzyme immunoassay (NI-EIA) for the determination of anti-idiotypic antibodies against Semliki Forest virus (SFV)-neutralizing monoclonal antibodies (MAs) UM 5.1 (IgG2a) and UM 1.4 (IgG2a). Against these MAs strong immune sera were induced in female BALB/c mice by two subcutaneous injections, 3 weeks apart, with keyhole limpet hemocyanin coupled MA mixed with the saponin Quil A. Rabbit immune sera were prepared by intracutaneous injections of purified MA mixed with either FCA (first immunization) or IFA (second and third immunization). In the NI-EIA serum is preincubated with neutralizing MA, in wells of 96-well plates, before SFV is added. Binding of anti-idiotypic antibodies to MA results in a diminished capacity of that MA to neutralize SFV. After 1 h incubation with SFV L cells are added and residual infectious virus is allowed to multiply for 6 h at 37 degrees C. Then the monolayers are fixed with glutaraldehyde and subsequently SFV is quantified with a horseradish peroxidase-labelled SFV-specific MA. Low absorbance values indicate that the neutralizing capacity of MA is intact and that blocking antibodies were not present in serum. In contrast high absorbance values indicate that blocking (anti-idiotypic) antibodies had abrogated the neutralizing capacity of MA. With the strongly neutralizing MA UM 5.1 as idiotypic antigen the NI-EIA proved to be at least as sensitive as the solid-phase EIA. Furthermore both normal mouse serum-absorbed rabbit immune sera and mouse immune sera were not cross-reactive in both solid-phase EIA and NI-EIA.


Cancer antigen-125 plasma level as a biomarker of new-onset atrial fibrillation in postmenopausal women.
Plasma cancer antigen (CA)-125 is a tumour marker recently shown to be associated with systolic heart failure and new-onset atrial fibrillation (AF) after myocardial infarction. However, no reports have described the relationship between CA-125 and new-onset AF in healthy postmenopausal women. The aim of the present study was to evaluate the relationship between CA-125 and new-onset AF in postmenopausal women. Between 2005 and 2015, 2086 women, including 1012 postmenopausal women, visited our hospital for annual health check-ups. We excluded patients with systolic dysfunction, chronic inflammatory disease, chronic obstructive pulmonary disease, histories of AF or neoplastic diseases. A total of 746 postmenopausal women underwent thorough physical examinations, including those for biomarkers such as brain natriuretic peptide, high-sensitivity C-reactive protein (hs-CRP) and CA-125. During the 10-year observation period, AF was documented in 31 participants (4.2%). The mean age of participants developing AF (75±6 years) was higher than that of those without AF (68±8 years). Participants developing AF showed significantly higher CA-125 (11.4±6.3 U/mL) and hs-CRP (0.10±0.11 mg/dL) levels than did those without AF (7.7±3.2 U/mL, p<0.01; 0.07±0.08 mg/dL, p<0.05). Cox regression analyses revealed ageing (HR 1.3; 95% CI 1.08 to 1.57; p<0.01) and plasma CA-125 levels (HR 1.29; 95% CI 1.10 to 1.51; p=0.02) as independent predictors of AF. High CA-125 levels might be associated with new-onset AF in healthy postmenopausal women.


Anesthetic implications for patients with rate-responsive pacemakers.
One hundred thousand adults and children in the United States receive pacemakers each year, of which 85% are rate-responsive pacemakers (RRPs). Recent advances in the fields of computer programming and computer chip technology have led to the myriad development of RRPs, which contain sensors that automatically adjust the pacing rate to match the physiologic changes that occur during physical exertion. Because patients with RRPs may experience heart rate changes in the operating room due to ''normal'' sensor function, anesthesiologists must be aware of the new developments in RRP sensor technology to properly manage these patients. Increases in respiratory rate and tidal volume as well as the use of electrocautery have been reported to accelerate the paced rate of pacemakers with minute ventilation sensors. Likewise, patient movement and saws that produce vibrations can accelerate the paced rate of patients with piezoelectric crystal sensors. This paper discusses the history of pacemaker development, reviews the currently used RRP sensors, and recommends procedures for the perioperative management of these patients. Knowledge of sensor type and factors that stimulate them will help the anesthesiologist understand the cause of these changes so that he will be able to manage clinically significant hemodynamic changes due to RRP sensor activation.


The effects of fetal adrenalectomy at 110 days gestational age on AVP and CRH mRNA expression in the hypothalamic paraventricular nucleus of the ovine fetus.
AVP and CRH produced in the parvocellular neurons of the paraventricular nucleus (PVN) have both been implicated in the regulation of anterior pituitary ACTH synthesis and secretion. In sheep, fetal ACTH secretion increases around 120 days gestational age (dGA). Little is known about adrenal regulation of AVP and CRH immediately prior to this critical period. We investigated the effects of adrenalectomy and subsequent cortisol (F) administration on PVN AVP and CRH mRNA in the fetal sheep PVN at 109-125 dGA. At 109-113 dGA, fetal sheep adrenals were removed (ADX)(n = 8); or sham surgery performed (CONT)(n = 4). From day 6 post ADX, maternal plasma cortisol and fetal plasma ACTH and cortisol levels were determined daily by radioimmunoassays. From day 7 post ADX, cortisol (4 micrograms/min) was continuously infused intravenously to four ADX fetuses (ADX + F). Fetal hypothalami were collected at 123-125 dGA, and studied by in-situ hybridization and quantitative autoradiography for AVP and CRH mRNA. Plasma cortisol levels remained low in CONT and ADX fetuses (< 4.9 ng/ml), while during cortisol infusion to ADX + F fetuses, plasma F increased to 16.4 +/- 2.2 and 22.3 +/- 3.2 ng/ml (mean +/- S.E.M.) on day 10 and 13, respectively. Plasma ACTH levels increased significantly in ADX compared with CONT fetuses. This ACTH increase was completely suppressed in ADX + F fetuses. AVP mRNA abundance in the whole PVN was the same in all three groups, however, a separate analysis of AVP mRNA abundance in parvocellular and magnocellular regions of the PVN revealed that AVP mRNA in the parvocellular PVN showed a significant increase in ADX and suppression in ADX + F fetuses when compared to CONT. AVP mRNA in the magnocellular PVN remained unchanged. PVN CRH mRNA expression was augmented in ADX and suppressed in ADX + F when compared to CONT fetuses. We conclude that in fetal sheep at 109-125 dGA: AVP and CRH mRNA abundance in the parvocellular region of the PVN are increased by adrenalectomy and that cortisol inhibits this increase.


Nucleic acid-based therapeutics for the treatment of central nervous system disorders.
Nucleic acid-based therapeutics (NBTs) are an emerging class of drugs with potential for the treatment of a wide range of central nervous system conditions. To date, pertaining to CNS indications, there are two commercially available NBTs and a large number of ongoing clinical trials. However, these NBTs are applied directly to the brain due to very low blood brain barrier permeability. In this review, we outline recent advances in chemical modifications of NBTs and NBT delivery techniques intended to promote brain exposure, efficacy, and possible future systemic application.


Utilization of human hematopoietic cell lines for the propagation and characterization of HBLV (human herpesvirus 6).
Details of the productive infection of established human cell lines of diverse origin by HBLV (also designated Human Herpesvirus 6) are described in this report. The infection and replication of HBLV in several T and B lymphoid and other cell lines was observed by electron microscopic examination, by the detection of viral antigen expression by indirect immunofluorescence assay (IFA) and by the presence of HBLV DNA by Southern blot hybridization. Several of these cell lines produced large amounts of virus. For this reason and because of the absence of other human herpesviruses, these lines have provided a valuable resource for the preparation of reagents and the development of assays for the detection and characterization of HBLV. The isolation and characterization of new HBLV isolates from patients with chronic fatigue syndrome were also facilitated by using some of the cell lines reported here. The host range of HBLV in established cell lines, therefore, does not appear to be limited to the B lymphocytes, as initially suggested by in vivo studies. The infection of T and B lymphocytes, megakaryocytes and neuronal cells in vitro suggests a need for the evaluation of diverse hematological and neurological disorders to shed light on a possible HBLV involvement.


Ionic Liquid-Based Colloidal Formulations for the Synthesis of Nano-MOFs: Applications in Gas Adsorption and Water Desalination.
Microemulsions (MEs) comprising choline dioctylsulfosuccinate [Cho][AOT], a biobased ionic liquid (IL) surfactant as an emulsifier, (R)-(+)-limonene (RL) as a nonpolar phase, and ethylene glycol (EG)/ethanolammonium formate (EOAF) as an organic solvent/low-viscosity IL polar component were constructed. Spontaneous aggregation of [Cho][AOT] was observed with a negative ΔH form using isothermal titration calorimetry. The aggregates of [Cho][AOT] in RL showed a critical micellar concentration (cmc) of ∼5.49 mM, EG (cmc ∼3.99 mM), and EOAF (cmc ∼1.56 mM), and these are further characterized by various techniques. These novel IL-based MEs have been used as nanoreactors for the sustainable synthesis of uniform nanosized metal-organic frameworks (N-MOFs), such as MIL-53(Al), HKUST-1, UIO-66-NH2, and ZIF-8, with a precise control over size and morphology at room temperature. Characterization of N-MOFs has been performed using scanning electron microscopy, powder X-ray diffraction, and Fourier transform infrared spectroscopy. The synthesized N-MOFs have been used to prepare stable and uniform thin film nanocomposite nanofiltration membranes, suitable for desalination of brackish water with excellent flux (31.8 LMH/bar) and rejection (99.0%) of divalent salts.


Piperazin incorporated Schiff Base derivatives: Assessment of in vitro biological activities, metabolic enzyme inhibition properties, and molecular docking calculations.
The cytotoxic activities of the compounds were determined by the 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) method in human breast cancer (MCF-7), human cervical cancer (HeLa), and mouse fibroblast (L929) cell lines. The compounds MAAS-5 and four modified the supercoiled tertiary structure of pBR322 plasmid DNA. MAAS-5 showed the highest cytotoxic activity in HeLa, MCF-7, and L929 cells with IC50 values of 16.76 ± 3.22, 28.83 ± 5.61, and 2.18 ± 1.22 µM, respectively. MAAS-3 was found to have almost the lowest cytotoxic activities with the IC50 values of 93.17 ± 9.28, 181.07 ± 11.54, and 16.86 ± 6.42 µM in HeLa, MCF-7, and L929 cells respectively at 24 h. Moreover, the antiepileptic potentials of these compounds were investigated in this study. To this end, the effect of newly synthesized Schiff base derivatives on the enzyme activities of carbonic anhydrase I and II isozymes (human carbonic anhydrase [hCA] I and hCA II) was evaluated spectrophotometrically. The target compounds demonstrated high inhibitory activities compared with standard inhibitors with Ki values in the range of 4.54 ± 0.86-15.46 ± 8.65 nM for hCA I (Ki value for standard inhibitor = 12.08 ± 2.00 nM), 1.09 ± 0.32-29.94 ± 0.82 nM for hCA II (Ki value for standard inhibitor = 18.22 ± 4.90 nM). Finally, the activities of the compounds were compared with the Gaussian programme in the B3lyp, HF, M062X base sets with 6-31++G (d,p) levels. In addition, the activities of five compounds against various breast cancer proteins and hCA I and II were compared with molecular docking calculations. Also, absorption, distribution, metabolism, excretion, and toxicity analysis was performed to investigate the possibility of using five compounds as drug candidates.


[Enantioseparation of tropicamide by capillary electrophoresis with square wave amperometric detection].
An enantioseparation method for tropicamide by high performance capillary electrophoresis with square wave amperometric detection (SWAD) was developed. The enantiomers of tropicamide were baseline separated in 16 min with an uncoated fused-silica capillary (75 microm i.d. x 50 cm) under the optimum conditions: 7 mmol/L Tris-10 mmol/L citric acid-2 mmol/L H3BO(3)-15 mmol/L beta-cyclodextrin (beta-CD) (pH 3.0) as background electrolyte, SWAD balance potential (E(b)) + 0.80 V (vs. Ag/AgCl), separation voltage 15 kV, injection height 20 cm, and injection time 10 s. The calibration curve of the enantiomers showed good linearity in the range from 5 micromol/L to 750 micromol/L with detection limit of 2 micromol/L. The average recoveries of added standards were 96% - 103%. The effects of the concentrations of beta-CD and the boric acid, the pH of the background electrolyte on resolutions (Rs) of the enantiomers were discussed in details. The proposed method was applied to the determination of a commercial tropicamide eye-drops sample without pre-treatment, and satisfactory results were obtained.


CD30+ large cell transformation of mycosis fungoides after psoralen plus ultraviolet A photochemotherapy.
Psoralen plus ultraviolet A (PUVA) photochemotherapy is widely used for the therapy of mycosis fungoides (MF). Clinical progression of MF is often associated with an increase in the size of tumour cells known as transformation. We report two patients with CD30+ large cell transformation that appeared after low-dose PUVA therapy for MF. Clinical data, histopathology, immunohistopathology and T-cell receptor gene rearrangement were studied. Nodules consisted of atypical large cells that expressed CD30. Monoclonal rearrangement of T-cell receptors was observed in one case. Low-dose PUVA therapy may be associated with CD30+ large cell transformation in patients with MF.