Production of recombinant proteins in Mycobacterium smegmatis for structural and functional studies.
Protein production using recombinant DNA technology has a fundamental impact on our understanding of biology through providing proteins for structural and functional studies. Escherichia coli (E. coli) has been traditionally used as the default expression host to over-express and purify proteins from many different organisms. E. coli does, however, have known shortcomings for obtaining soluble, properly folded proteins suitable for downstream studies. These shortcomings are even more pronounced for the mycobacterial pathogen Mycobacterium tuberculosis, the bacterium that causes tuberculosis, with typically only one third of proteins expressed in E. coli produced as soluble proteins. Mycobacterium smegmatis (M. smegmatis) is a closely related and non-pathogenic species that has been successfully used as an expression host for production of proteins from various mycobacterial species. In this review, we describe the early attempts to produce mycobacterial proteins in alternative expression hosts and then focus on available expression systems in M. smegmatis. The advantages of using M. smegmatis as an expression host, its application in structural biology and some practical aspects of protein production are also discussed. M. smegmatis provides an effective expression platform for enhanced understanding of mycobacterial biology and pathogenesis and for developing novel and better therapeutics and diagnostics.


Papillary Thyroid Carcinoma: Ectopic Malignancy versus Metastatic Disease.
Papillary thyroid carcinoma frequently metastasizes to regional lymph nodes. However, cervical lymph node metastasis as a sole manifestation of occult papillary thyroid carcinoma is rarely observed. Ectopic thyroid is an uncommon condition defined as the presence of thyroid tissue at a site other than pretracheal area. Approximately 1-3% of all ectopic thyroid tissue is located in the lateral neck. This entity may represent the only functional thyroid tissue in the body. Malignant transformation of ectopic thyroid is uncommon; but even rarer is the development of papillary carcinoma on it. We present a case of a 33-year-old man with an incidental lateral neck mass diagnosed after a motor vehicle accident. Total thyroidectomy and lymph node resection were completed without evidence of papillary thyroid carcinoma. Malignant transformation of heterotopic thyroid tissue was the final diagnosis. The possibility of an ectopic thyroid cancer should be considered in the differential diagnosis of a pathological mass in the neck. The uniqueness of this case strives in the rarity that the thyroid gland was free of malignancy, despite ectopic tissue being positive for thyroid carcinoma. Management strategies, including performance of total thyroidectomy, neck dissection, and treatment with radioiodine, should be based on individualized risk assessment.


Protective phenotypes of club cells and alveolar macrophages are favored as part of endotoxin-mediated prevention of asthma.
Atopic asthma is a chronic allergic disease that involves T-helper type 2 (Th2)-inflammation and airway remodeling. Bronchiolar club cells (CC) and alveolar macrophages (AM) are sentinel cells of airway barrier against inhaled injuries, where allergy induces mucous metaplasia of CC and the alternative activation of AM, which compromise host defense mechanisms and amplify Th2-inflammation. As there is evidence that high levels of environmental endotoxin modulates asthma, the goal of this study was to evaluate if the activation of local host defenses by Lipopolysaccharide (LPS) previous to allergy development can contribute to preserving CC and AM protective phenotypes. Endotoxin stimulus before allergen exposition reduced hallmarks of allergic inflammation including eosinophil influx, Interleukin-4 and airway hyperreactivity, while the T-helper type 1 related cytokines IL-12 and Interferon-γ were enhanced. This response was accompanied by the preservation of the normal CC phenotype and the anti-allergic proteins Club Cell Secretory Protein (CCSP) and Surfactant-D, thereby leading to lower levels of CC metaplasia and preventing the increase of the pro-Th2 cytokine Thymic stromal lymphopoietin. In addition, classically activated alveolar macrophages expressing nitric oxide were promoted over the alternatively activated ones that expressed arginase-1. We verified that LPS induced a long-term overexpression of CCSP and the innate immune markers Toll-like receptor 4, and Tumor Necrosis Factor-α, changes that were preserved in spite of the allergen challenge. These results demonstrate that LPS pre-exposition modifies the local bronchioalveolar microenvironment by inducing natural anti-allergic mechanisms while reducing local factors that drive Th2 type responses, thus modulating allergic inflammation.


[Intra-articular injections of hyaluronic acid for anterior disc displacement of temporomandibular joint].
Anterior disc displacement (ADD) of temporomandibular joint (TMJ) is regarded as one of the major findings in temporomandibular disorders (TMD). It is related to joint noise, pain, mandibular dysfunction, degenerative change and osteoarthritis. In the mean time, the pathological changes were found in synovial membrane and synovial fluid. Hyaluronic acid is a principal component of the synovial fluid which plays an important role in nutrition, lubrication, anti-inflammation and cartilage repair. The synthesis, molecule weight, and concentration of hyaluronic acid are decreased during TMD and cause TMJ degenerative changes. The clinical conditions, pathological changes, the mechanism of action for hyaluronic acid and the treatment of anterior disc displacement of TMJ are discussed in this article.


Nuclear inclusions of tonofilaments and keratohyalin granules in the condyloma acuminatum.
Electron-microscopic investigations of the condyloma acuminatum show different nuclear inclusions in keratinocytes of the granular layer or the layer immediately below. Fibrous structures were indentified as tonofilaments and irregularly shaped electron-dense substances as keratohyalin. However the question remains as to whether there is a connection between the viral infection and the occurrence of such intranuclear inclusions.


Clinical management of canine seizures.
Client education is the most important step in seizure management. The owner should be well informed about canine epilepsy and the present limitations regarding treatment. Regular follow-up should be performed for drug monitoring, evaluation of the metabolic profile, and assessment of seizure frequency. Other methods of treatment may become available in the future to treat the dog with epilepsy. Potassium bromide, the first anticonvulsant discovered, is being reevaluated as an anticonvulsant in the dog. The dose advocated is 50 mg per kg once daily. This author has no experience with this drug. GABA-mimetic drugs have been shown to exert potent anticonvulsant effects against chemically and electrically induced seizures in rats. Surgical treatment of intractable epilepsy, such as division of the corpus callosum, is being performed in man. Also in man, cerebellar stimulation is effective in controlling intractable epilepsy. Recently, acupuncture treatment has been shown to reduce seizure frequency in a few epileptic dogs. Unfortunately, the ideal anticonvulsant drug still does not exist. Every patient should be evaluated as an individual. Administration of multiple drugs should be avoided whenever possible to minimize side effects and drug interactions.


Italian guidelines for the diagnosis and treatment of patients with haemophilia and inhibitors.
The Italian Association of Haemophilia Centres reviewed and finally approved in November 2004 the new Italian Guidelines for the diagnosis and treatment of patients with clotting factor inhibitors. The recommendations have been based on the identification of levels of clinical evidence derived from the systematic review carried out in 2003 by the School of Health and Related Research, the University of Sheffield, UK, and further integrated by clinical studies published from 2003 to 2004. The Italian guidelines consist of six major domains concerning inhibitor definition, epidemiology, risk factors, diagnosis, inhibitor eradication, management of bleeding episodes, in patients with congenital and acquired coagulation disorders, with 121 statements, 59 synthesis and 54 recommendations. We report here recommendations and open issues concerning the diagnosis and monitoring of inhibitors, inhibitor eradication and the management of bleeding in patients with haemophilia A and B.


Prevention of Congenital Syphilis Through Antenatal Screenings in Lusaka, Zambia: A Systematic Review.
Congenital syphilis is one of the preventable diseases caused by the gram-negative bacteria Treponema pallidum; yet, it imposes a serious global health and economic burden, with more than half of the cases resulting in serious adverse outcomes, including infant mortality. Mother-to-child transmission (MTCT) of syphilis is estimated at 3.6 million adjusted life years (DALYs) and around $309 million in medical costs. In 2006, an estimated 9.7 million children of age less than five years died in developing countries; almost four million were neonatal deaths. There were 3.2 million stillbirths globally, among whom 95% were in the developing countries. In sub-Saharan Africa, there is an estimated 2.7% (0.1%-10.3%) of pregnant women infected with syphilis, representing more than 900,000 pregnancies at risk each year. There were many non-specific and specific diagnostic tests used in the past, which required laboratory equipment and electricity, but there are many newer tests available now that provide rapid results with high sensitivity and specificity, e.g., the immunochromatographic strip (ICS) and rapid syphilis tests (RST). Early syphilis can be completely eliminated with a single injection of penicillin, which is readily available, cheap, and highly effective, and treating pregnant women with penicillin is 98% effective at preventing congenital syphilis. Targeting women at a high risk of having syphilis makes universal screening in antenatal programs the most efficacious way to prevent syphilis-associated morbidity and mortality. The potential for a program to prevent congenital syphilis in the perinatal, neonatal, and postnatal periods is evident. While considering resource allocation to child survival programs in areas where the prevalence of syphilis is high, officials need to include antenatal syphilis screening, using rapid tests and treatment at the first contact of the mother with the health care system. In countries like Zambia and other resource-limited settings, a same-day test and treatment with penicillin should be prioritized to achieve the goal of eliminating congenital syphilis. Eliminating MTCT of syphilis through screening and treatment in antenatal care (ANC) is highly cost-effective in a wide range of settings, especially in countries with a high prevalence.


Identification of Kashmir bee virus in France using a new RT-PCR method which distinguishes closely related viruses.
A new RT-PCR protocol has been developed, avoiding potential misdiagnosis of Kashmir bee virus (KBV) linked to the use of KBV primers designed originally. The PCR assay validation was realised taking into account the analytical specificity and the PCR detection limit. KBV was detected in a bee sample collected in France from an apparently healthy apiary in 2012. The specificity of the primers was confirmed by sequencing the PCR product. This French sequence clustered into the KBV genotype by phylogenetic analysis, while previous French sequence isolates collected in 2002 belong to the IAPV genotype. These data represent the first detection of KBV in France.


[Epidemiological Clinical Characteristics of Acute Cardiac Insufficiency in a Tertiary Hospital in Lima, Peru].
The objective of this study was to describe the clinical and epidemiological characteristics of acute heart failure (AHF) in 1,075 patients from a hospital in Lima, Peru. The average patient age was 74 years and 55% of study subjects were men. Moreover, 39% of patients in the sample had AHF with low ejection fraction, 15% had intermediate ejection fraction, and 46% had preserved ejection fraction. The most common comorbidities were hypertension (52.6%) and coronary disease (51%). Moreover, 29.2% of patients had previous hospitalizations due to AHF. The median hospital stay was 3 days. The rate of in- hospital mortality from all causes was 7.2%. Hospitalization was more common in elderly patients with multiple comorbidities. The low use of prescription drugs together with the high rate of previous hospitalizations may explain the high mortality rate reported in this study.


Inflammatory pseudotumor of the lung. Ultrastructural and light microscopic study of a myxomatous variant.
A case of a myxomatous variant of an inflammatory pseudotumor of the lung is reported with clinicopathologic findings including its ultrastructure. The pathologic features of inflammatory pseudotumor of the lung are compared with similar tumors arising elsewhere and bear a close resemblance to granulation tissue, which suggests that inflammatory pseudotumors represent a reparative process rather than a neoplastic process. Recognition of this entity is imperative for a correct diagnosis and in order to differentiate it from malignant neoplasm, since only conservative management is necessary for its treatment.


Structure-function constraints of tracheid-based xylem: a comparison of conifers and ferns.
The ferns comprise one of the most ancient tracheophytic plant lineages, and occupy habitats ranging from tundra to deserts and the equatorial tropics. Like their nearest relatives the conifers, modern ferns possess tracheid-based xylem but the structure-function relationships of fern xylem are poorly understood. Here, we sampled the fronds (megaphylls) of 16 species across the fern phylogeny, and examined the relationships among hydraulic transport, drought-induced cavitation resistance, the xylem anatomy of the stipe, and the gas-exchange response of the pinnae. For comparison, the results are presented alongside a similar suite of conifer data. Fern xylem is as resistant to cavitation as conifer xylem, but exhibits none of the hydraulic or structural trade-offs associated with resistance to cavitation. On a conduit diameter basis, fern xylem can exhibit greater hydraulic efficiency than conifer and angiosperm xylem. In ferns, wide and long tracheids compensate in part for the lack of secondary xylem and allow ferns to exhibit transport rates on a par with those of conifers. We suspect that it is the arrangement of the primary xylem, in addition to the intrinsic traits of the conduits themselves, that may help explain the broad range of cavitation resistance in ferns.


Urinary Angiotensinogen Predicts Renal Disease Activity in Lupus Nephritis.
Aims: A noninvasive indicator of renal histological lesions and disease activity in lupus nephritis (LN) is needed for timely and targeted treatment before overt renal injury. Here, we tested the utility of urinary angiotensinogen (UAGT) to predict renal disease activity in LN. Results: A prospective, three-stage study was performed in patients with LN. In stage I, UAGT was measured in 140 newly diagnosed LN patients. UAGT significantly increased in LN patients, correlating well with kidney angiotensinogen expression and histological activity. Patients with LN class IV exhibited the highest UAGT compared with other histopathological classes of LN. For identifying LN class IV, a particularly aggressive type of LN, UAGT outperformed the conventional clinical measures and improved their performance. In stage II, UAGT was monitored in 61 subjects from stage I for up to 12 months. UAGT decreased after induction therapy and remained low in patients with LN remission during follow-up. For predicting therapy success at month 12, the area under the receiver operating characteristics curve of UAGT reduction at month 4 was 0.83, outperforming that of 24-h proteinuria. In stage III, UAGT was monitored in 12 LN patients before, during, and after the onset of renal flares. An elevation in UAGT predicted recurrence of LN, and a decline in UAGT after a renal flare heralded the remission of disease before conventional clinical measures. Innovation and Conclusion: UAGT in LN is a promising indicator for dynamic surveillance of renal disease activity and prediction of renal flares. Antioxid. Redox Signal. 31, 1289-1301.


Perceptions of empathic accuracy and assumed similarity in the coach-athlete relationship.
Drawing upon the methodology developed by Kenny and Acitelli (2001), this study examined empathic accuracy and assumed similarity in both parties' perceptions of coach-athlete relationships. One hundred and twenty-one coaches and athletes reported on their direct-perceptions and meta-perceptions of closeness, commitment and complementarity (3 Cs; Jowett & Cockerill, 2002), and of satisfaction with instruction, performance and external agents. There was evidence of both empathic accuracy and assumed similarity in coaches' and athletes' perceptions. Athletes were more accurate in identifying the specific content of their coaches' feelings in terms of closeness. Athletes and coaches from newly developed relationships displayed higher levels of empathic accuracy, whereas female athletes displayed higher levels of assumed similarity. Moreover, evidence suggested that athletes' and coaches' assumed similarity led to more accurate perceptions. Implications and future research directions are discussed.


Localization of the inosine triphosphatase locus (Itp) on chromosome 2 of the mouse.
An electrophoretic variant of the enzyme inosine triphosphatase was found by screening inbred strains of mice. Strains with the slower-migrating variant include BALB/cJ, DBA/1J, and PL/J. The Itp locus was mapped between the beta-2-microglobulin (B2m) and the agouti (a) loci on chromosome 2. The mapping of Itp on chromosome 2 identifies a chromosomal segment that has been conserved since the divergence of lineages leading to mouse and man.


Children's views on postsurgical pain in recovery units in Norway: A qualitative study.
To explore children's postsurgical experiences with pain and pain management in the recovery unit. Children's pain is underestimated and undertreated. Untreated pain can cause unnecessary suffering, increased complication risks and may lead to chronic pain. Research exploring children's experiences with postoperative pain and pain management is limited. A qualitative, exploratory study. The study complied with the Consolidated Criteria for Reporting Qualitative Research (COREQ). Children (N = 20), 8-16 years old, took part in semi-structured interviews about their experiences with pain and postoperative pain management while they were in a recovery unit. Data were collected at two university hospitals in Norway. Content analysis was used to analyse the data. Three themes emerged from the interviews: "children's experiences of what felt unpleasant and painful," "children's experiences with pain management" and "children's recommendations for future pain management". About half of the children reported moderate to severe pain while in the recovery unit and they did not always tell their nurses when they had pain. They also reported experiencing pain in places other than their surgical wounds and stated that nausea and vomiting felt unpleasant and painful. The children indicated that pain medications and the use of nonpharmacological methods helped them cope with their pain and provided several recommendations about how to improve pain management. Paediatric postoperative pain management remains suboptimal. The children in our study provided useful information about their pain experiences, how to improve pain management and explained why they did not tell their nurses when they were in pain. These findings should direct further improvements in paediatric postoperative pain management, such as increased use of pain assessment tools and preparatory information, as well as more appropriate administration of pain medications.


Tumor necrosis factor receptor/tumor necrosis factor family members in antiviral CD8 T-cell immunity.
CD8 memory T cells can play a critical role in protection against repeated exposure to infectious agents such as viruses, yet can also contribute to the immunopathology associated with these pathogens. Understanding the mechanisms that control effective memory responses has important ramifications for vaccine design and in the management of adverse immune reactions. Recent studies have implicated several members of the tumor necrosis factor receptor (TNFR) family as key stimulatory and inhibitory molecules involved in the regulation of CD8 T cells. In this review, we discuss their control of the generation, persistence, and reactivation of CD8 T cells during virus infection.


Inactivation of ibeA and ibeT results in decreased expression of type 1 fimbriae in extraintestinal pathogenic Escherichia coli strain BEN2908.
IbeA in extraintestinal pathogenic Escherichia coli (ExPEC) strains was previously described for its role in invasion. Here we investigated the role of IbeA and IbeT, encoded by a gene located downstream of ibeA, in the adhesion of the avian ExPEC strain BEN2908 to human brain microvascular endothelial cells (HBMEC). The DeltaibeA mutant was less adhesive to HBMEC than the wild-type strain BEN2908 was. Because strain BEN2908 also expresses type 1 fimbriae, we measured the adhesion specifically due to IbeA by comparing the adhesive properties of a Deltafim derivative of strain BEN2908 to those of a double Deltafim DeltaibeA mutant. No differences were observed, indicating that the reduction of adhesion in BEN2908 DeltaibeA could be due to a decrease in type 1 fimbria expression. We indeed showed that the decreased adhesion of BEN2908 DeltaibeA was correlated with a decrease in type 1 fimbria expression. Accordingly, more bacteria had a fim promoter orientated in the off position in a culture of BEN2908 DeltaibeA than in a culture of BEN2908. Expression of fimB and fimE, two genes encoding recombinases participating in controlling the orientation of the fim promoter, was decreased in BEN2908 DeltaibeA. A reduction of type 1 fimbria expression due to a preferential orientation of the fim promoter in the off position was also seen in an ibeT mutant of strain BEN2908. We finally suggest a role for IbeA and IbeT in modulating the expression of type 1 fimbriae through an as yet unknown mechanism.


A simple analytical model for a fast 3D assessment of peripheral photon dose during coplanar isocentric photon radiotherapy.
Considering that cancer survival rates have been growing and that nearly two-thirds of those survivors were exposed to clinical radiation during its treatment, the study of long-term radiation effects, especially secondary cancer induction, has become increasingly important. To correctly assess this risk, knowing the dose to out-of-field organs is essential. As it has been reported, commercial treatment planning systems do not accurately calculate the dose far away from the border of the field; analytical dose estimation models may help this purpose. In this work, the development and validation of a new three-dimensional (3D) analytical model to assess the photon peripheral dose during radiotherapy is presented. It needs only two treatment-specific input parameter values, plus information about the linac-specific leakage, when available. It is easy to use and generates 3D whole-body dose distributions and, particularly, the dose to out-of-field organs (as dose-volume histograms) outside the 5% isodose for any isocentric treatment using coplanar beams [including intensity modulated radiotherapy and volumetric modulated arc therapy (VMAT)]. The model was configured with the corresponding Monte Carlo simulation of the peripheral absorbed dose for a 6 MV abdomen treatment on the International Comission on Radiological Protection (ICRP) 110 computational phantom. It was then validated with experimental measurements using thermoluminescent dosimeters in the male ATOM anthropomorphic phantom irradiated with a VMAT treatment for prostate cancer. Additionally, its performance was challenged by applying it to a lung radiotherapy treatment very different from the one used for training. The model agreed well with measurements and simulated dose values. A graphical user interface was developed as a first step to making this work more approachable to a daily clinical application.


Detection of molecular oxygen using nanosecond-laser-induced plasma.
Molecular oxygen (O2) concentration is measured by employing nanosecond laser-induced plasmas (ns-LIP) over a broad temperature spectrum ranging from 300 K to 1000 K, in the presence of an additional oxygen-containing molecule, CO2. Typically, emission spectra emanating from ns-LIP are devoid of molecular information, as the ns-LIP causes the dissociation of molecular species within the plasma. However, atomic oxygen absorption lines that momentarily appear at 777 nm in the broadband emission from the early-stage plasma are determined to be highly sensitive to the O2 mole fraction but negligibly affected by the CO2 mole fraction. The atomic O absorbing the plasma emission originates from the O2 adjacent to the plasma: robust UV radiation from the early-stage plasma selectively dissociates adjacent O2, exhibiting a relatively low photodissociation threshold, thus generating the specific meta-stable oxygen capable of absorbing photons at 777 nm. A theoretical model is introduced, explicating the formation of the meta-stable O atom from adjacent O2. To sustain the UV radiation from the plasma under high-temperature and low-density ambient conditions, a preceding breakdown is triggered by a split laser pulse (532 nm). This breakdown acts as a precursor, seeding electrons to intensify the inverse-Bremsstrahlung photon absorption of the subsequent laser pulse (1064 nm). Techniques such as proper orthogonal decomposition (POD) and support vector regression (SVR) are employed to precisely evaluate the O2 mole fraction (<1% uncertainty), by analyzing the short-lived (<10 ns) O2-indicator depicted in the early-stage plasma.


Association of single-nucleotide polymorphism on chromosome 9 and ischemic stroke in Heilongjiang province in China.
There is a significant correlation between ischemic stroke (IS) and chromosome 9. However, its status was uncertain in China's cold regions. 1920 IS patients, and 1920 healthy individuals were included in the study. Blood samples were collected. The association of SNPs with IS was evaluated by Sequenom, and logistic regression models adjusted for known risk factors of IS were constructed to assess the SNPs' associations in cases and controls. We found rs1333040 and rs2383207 were associated with IS, compared with primitive genotypes. The genotype CT of rs7027526 has a protective role during IS development, while the effect of the genotype TT is still not clear. These results changed after stratification by age and sex. In conclusion, rs1333040 and rs2383207 SNPs in CDKN2BAS are associated with ischemic stroke in the Chinese Han population. This study confirms the association between 9p21.3 and IS.


Mean platelet volume and neutrophil to lymphocyte ratio as parameters to indicate the severity of suicide attempt.
This study was designed to compare the complete blood count (CBC) values of 38 violent suicidal attempt (VSA) and 38 nonviolent suicidal attempt (NVSA) patients and 38 healthy controls, particularly mean platelet volume (MPV) and neutrophil to lymphocyte ratio (NLR). MPV was found to be significantly higher (P = 0.000), and PLT was found to be significantly lower in the VSA group (P = 0.001). NLR was found to be significantly higher (P = 0.005) in the VSA group. The area under the receiver operating characteristic curve of MPV levels for VSA was 0.78 and 0.68 for NLR in the VSA group when compared with the NVSA group.


Cardiopulmonary function as related to intestinal mucosal lesions in experimental septic shock.
The purpose of the present investigation was to explore a possible relationship between cardiopulmonary function and small intestinal mucosal lesions in experimental septic shock. The filling pressure of the left ventricle of the heart (LVEDP) and the maximal pressure change during systole (max dP/dt) were monitored continuously by a tip-transducer catheter. The pulmonary and femoral arterial pressures and pressure changes in the trachea were recorded. The aortic blood flow was measured electromagnetically. Stroke volume, external cardiac work, and the pulmonary vascular resistance could be calculated. Twenty-two cats were made septic by i.v. infusion of live E. coli bacteria. Controls received saline i.v. After 2 hours of bacteremia, preload was increased by a rapid infusion of dextran to unmask cardiac dysfunction. Small intestinal specimens were taken for microscopical examination. The occurrence and the degree of mucosal damage was established. Thirteen cats had virtually normal mucosal appearances, whereas nine disclosed severe lesions. The loading procedure revealed a more pronounced cardiac dysfunction in cats with severe mucosal damage, compared to those without. Pulmonary vasoconstriction, bronchoconstriction, and impaired ventilation-perfusion ratio were also demonstrated, but these signs of pulmonary dysfunction did not differ when comparing animals with and without mucosal damage. A correlation between impaired cardiac function on one hand and hypotension--intestinal mucosal lesions on the other, suggests cardiotoxic material released from the damaged intestine as one contributing mechanism.


Impression cytology with transfer: an easy method for detection of vitamin A deficiency.
We described a new method to stain epithelial cells harvested by ocular impression. The cells are immediately transferred on a glass slide after the sampling; this transfer permits a very simple staining and easier reading by light microscopy. The results are compared with those obtained with the previous techniques and confirm ocular impression as a good help for vitamin A status determination.


Short-term and long-term role of platelet activating factor as a mediator of in vivo platelet aggregation.
Platelet activating factor (PAF) is a phospholipid released upon stimulation by a variety of cells and has been implicated in several pathophysiological events such as asthma and inflammatory diseases. However, although the ability to aggregate platelets in vitro was the first biological activity ascribed to PAF, its role in contributing to the in vivo formation of arterial thrombi has not been thoroughly clarified. Intravascular platelet aggregation was initiated in two different animal models of arterial stenosis and endothelial injury. An external constrictor was positioned around rabbit carotid arteries and canine coronary arteries. After placement of the constrictor, a typical pattern of flow developed in the stenotic vessels. This pattern of flow, characterized by progressive reductions of carotid or coronary blood flow followed by spontaneous or induced restorations of flow (cyclic flow variations, CFVs), is related to recurrent platelet aggregation at the site of the stenosis followed by dislodgment of the thrombus. After observing CFVs for 30 minutes, BN52021 (up to 1.2 mg/kg), a potent and selective PAF antagonist, was given intravenously to rabbits (n = 12) and dogs (n = 10). BN52021 completely inhibited CFVs in 10 of 12 rabbits, whereas it was relatively ineffective in abolishing CFVs in dogs (only 2 of 10 animals inhibited). This different effect of BN52021 was not explained by too small a dose of the drug to achieve a complete blockade of PAF receptors in dogs, since ex vivo platelet aggregation was completely inhibited in both rabbits and dogs in response to exogenous PAF at concentrations up to 10(-5) mol/L. In a second group of 10 dogs, the hypothesis that PAF may become an important mediator of CFVs in dogs only several hours after endothelial injury was tested. After 30 minutes of baseline CFVs, these animals received a bolus of BN52021 up to 1.2 mg/kg. After this treatment, CFVs were completely abolished in 2 of 10 animals. The remaining 8 dogs were followed for an additional 8-hour period, at the end of which a second bolus of BN52021 was given. At this time, BN52021 was effective, as CFVs were abolished in 6 of 8 animals. These effects of BN52021 at 8 hours were not the consequence of a cumulative dose of the compound, since ex vivo platelet aggregation in response to PAF returned to baseline values immediately before administering the second dose. To identify possible sources of PAF other than aggregating platelets at the site of arterial stenosis, dogs in a third group were killed after 30 minutes (n = 7) and after 8 hours (n = 8) of CFVs. Histological sections of the stenotic coronary artery showed a marked leukocyte infiltration in these arterial segments after 8 hours of CFVs, whereas sections from dogs killed after 30 minutes showed only moderate or no infiltration. These data demonstrate that PAF plays an important role as a mediator of platelet aggregation in vivo in rabbits and dogs. In the canine model, PAF appears to become more important after leukocyte infiltration of the arterial wall, as it may contribute to initiating enough platelet activation to lead to cyclic flow variations at sites of arterial stenosis and endothelial injury. Data from the present study suggest that PAF antagonists may be used as antiplatelet agents.


Determination of interleukin-6 in lung transplantation: association with primary graft dysfunction.
Primary graft dysfunction (PGD) secondary to damage caused by ischemia/reperfusion is responsible for significant morbidity. It constitutes the main cause of early death following implantation. Our objective was to verify the association between PGD and activation of the inflammatory cascade by measuring interleukin-6 (IL-6) in the blood and the bronchoalveolar lavage (BAL) of the recipient. The 31 patients, including 22 bipulmonary and 9 unipulmonary cases, had severe PGD (ISHLT grade II) defined by: (1) radiographic infiltrates during the first 72 hours after transplantation, (2) PO2/FiO2 ratio <200 in the first 72 hours after the operation, and (3) no other cause of dysfunction. BAL and peripheral arterial blood samples were extracted prior to implantation (baseline level) and at 12, 24, and 48 hours after reperfusion. Samples were frozen to -80 degrees C until determination of IL-6 using an immunoassay technique (ELISA). In the 31 transplants (100%), there were elevated IL-6 contents in the BAL and blood compared with the baseline level (P < .0001). Among 11 patients with severe PGD (38.70%) In the BAL samples the concentration of IL-6 was significantly elevated (P < .05) compared with patients without PGD (P < .031). These finding were also observed in blood (P < .016) obtained at 12 hours. The analyses at 24 and 48 hours showed higher levels of IL-6 in the PGD group, although they were not significant. There was a significant elevation of IL-6 in blood and BAL during the first few hours after reperfusion of the graft, which was directly related to the development of PGD.


CD101, a long-acting echinocandin, and comparator antifungal agents tested against a global collection of invasive fungal isolates in the SENTRY 2015 Antifungal Surveillance Program.
CD101 is a novel echinocandin with exceptional chemical stability and long-acting pharmacokinetics. The activity of CD101 and comparators was evaluated using CLSI broth microdilution methods against 713 invasive fungal isolates, including 589 Candida spp. (6 species), 14 C. neoformans, 97 A. fumigatus and 13 A. flavus species complex collected worldwide during 2015. All C. tropicalis, C. krusei and C. dubliniensis, 99.7% of C. albicans and 98.3% of C. glabrata were inhibited by ≤0.12 µg/mL of CD101, and these isolates were susceptible/wild type to other echinocandins using CLSI clinical breakpoint and epidemiological cutoff value (ECV) interpretive criteria. C. parapsilosis displayed higher MIC values (range 0.25-2 µg/mL), but similar results were observed for other echinocandins. One C. glabrata and one C. albicans with CD101 MIC value at 1 and 0.25 µg/mL possessed F625S and S645P alterations on FKS1, respectively. These isolates also displayed elevated MIC values for at least one clinically available echinocandin. Fluconazole resistance was noted for 6.6% of C. glabrata and 3.6% C. parapsilosis. Echinocandins had limited activity against C. neoformans. CD101 activity against A. fumigatus and A. flavus (MEC ≤0.03 µg/mL) was comparable to other echinocandins (MEC ≤0.03 µg/mL). These moulds had MIC values below ECVs for the mould-active azoles. CD101 was as active as other echinocandins against common fungal organisms recovered from invasive fungal infections. The extended half-life profile is very desirable as less frequent dosing of this agent should facilitate shorter and more cost-effective hospital stays, improve compliance for outpatients, and provide more convenient outpatient prophylaxis.


Nanoporous alumina capsules for cellular macroencapsulation: transport and biocompatibility.
Currently, the most widely used treatment for diabetes is the daily subcutaneous injection of recombinant human insulin. Daily injections, however, cannot match the physiological biphasic behavior of normal insulin release, nor can they precisely meet the demands of food intake, exercise, and stress. Cellular encapsulation, or immunoisolation, is a possible solution to this problem. This cell-based therapy allows patients to receive the benefits of more physiological insulin and blood glucose regulation, without the need for immunosuppressants that are associated with organ or cell transplantation. Immunoisolation capsules were fabricated out of aluminum and aluminum oxide using a two-step anodization procedure. The diffusion behavior of glucose, immunoglobulin G (IgG), and insulin were measured. Furthermore, the functionality and viability of encapsulated MIN6 cells were tested. Finally, live cells were stained and imaged using confocal microscopy. Data indicated that this device is effective in allowing the transport of relevant molecules such as glucose and insulin, while at the same time significantly impeding the transport of IgG, suggesting that it would be efficacious in protecting cell grafts in vivo. Furthermore, encapsulated cells were able to respond dynamically to glucose input signals. Finally, cell staining showed that the viability of encapsulated cells is maintained after 24 h, although the cells appear to be more heavily concentrated at the area that is closest to the membrane. This study has shown that nanoporous alumina membranes, with well-controlled pore sizes, can be used for the encapsulation of therapeutic cells and may provide an alternative encapsulation strategy for the treatment of diabetes.


Diagnostic imaging findings and endocrine test results in dogs with pituitary-dependent hyperadrenocorticism that did or did not have neurologic abnormalities: 157 cases (1989-2005).
To compare imaging findings in dogs with pituitary-dependent hyperadrenocorticism (PDH) that did or did not have neurologic abnormalities. Design-Retrospective case series. 157 dogs with PDH that did (n = 73) or did not (84) have neurologic abnormalities. Medical records were reviewed for the presence and nature of clinical signs of CNS disease, and computed tomographic and magnetic resonance images were reviewed for evidence of a pituitary tumor. 60 of the 84 (71%) dogs without neurologic abnormalities and 48 of the 73 (66%) dogs with neurologic abnormalities had a detectable pituitary tumor. However, 17 of the 84 (20%) dogs without neurologic abnormalities had a pituitary macrotumor (ie, a tumor > or = 10 mm in height), and 41 of the 73 (56%) dogs with neurologic abnormalities did not have a detectable pituitary tumor or had a pituitary microtumor. Vague signs of CNS dysfunction (ie, lethargy, inappetence, and mental dullness) were more specific for detection of pituitary macrotumors than were CNS-specific signs (ie, seizure or blindness). Results suggested that there was no apparent relationship between a pituitary tumor and development of neurologic abnormalities in dogs with PDH. In addition, neurologic abnormalities in dogs with pituitary macrotumors were often vague (ie, lethargy, inappetence, and mental dullness).


Rhein inhibits the growth of Propionibacterium acnes by blocking NADH dehydrogenase-2 activity.
Introduction. Rhein (4, 5-dihydroxyanthraquinone-2-carboxylic acid) has various properties, including anti-inflammatory, antioxidant and anticancer activities. However, the mechanism underlying the role of rhein in antimicrobial activity remains largely unknown.Aim. This study aims to identify potential natural compounds of rhein that are capable of inhibiting Cutibacterium acnes and elucidate the effects of rhein on NADH dehydrogenase-2 activity in C. acnes.Methodology. The anti-C. acnes activity of compounds was analysed using minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), the paper disc diffusion test and the checkerboard dilution test. To check whether rhein was inhibitory, putative type II NADH dehydrogenase (NDH-2) of C. acnes was analysed, cloned and expressed in Escherichia coli, and then NDH-2 purification was assessed with Ni-NTA before rhein inhibition of NADH dehydrogenase-2 activity was checked with ferricyanide [K3Fe(CN)6] as a substrate.Results. The results showed that the MIC of rhein against C. acnes was 6.25 µg ml-1, while the MBC was 12.5 µg ml-1, and there was a 38 mm inhibition zone in the paper disc diffusion test. Rhein showed an additive two- to fourfold reduction of the MIC value with four antibiotics on the checkerboard dilution test. The purified NADH dehydrogenase gene product showed a size of approximately 51 kDa and had a V max of 23 µmol and a K m of 280 µm. The inhibitory effect of rhein against NADH dehydrogenase-2 activity was non-competitive with ferricyanide [K3Fe(CN)6] with a K i value of 3.5-4.5 µm.Conclusion. This study provided evidence of the inhibitory effects of rhein on the growth of C. acnes by blocking of NADH dehydrogenase-2 activity. This mechanism of inhibitory activity in the reduction of ROS formation and ATP productivity should be further tested in C. acnes and the question of whether rhein inhibits the natural growth of C. acnes should be investigated.


Carnitine/Organic Cation Transporter OCTN1 Negatively Regulates Activation in Murine Cultured Microglial Cells.
Brain immune cells, i.e., microglia, play an important role in the maintenance of brain homeostasis, whereas chronic overactivation of microglia is involved in the development of various neurodegenerative disorders. Therefore, the regulation of microglial activation may contribute to their treatment. The aim of the present study was to clarify the functional expression of carnitine/organic cation transporter OCTN1/SLC22A4, which recognizes the naturally occurring food-derived antioxidant ergothioneine (ERGO) as a substrate in vivo, in microglia and its role in regulation of microglial activation. Primary cultured microglia derived from wild-type mice (WT-microglia) and mouse microglial cell line BV2 exhibited time-dependent uptake of [3H]- or d9-labeled ERGO. The uptake was markedly decreased in cultured microglia from octn1 gene knockout mice (octn1 -/--microglia) and BV2 cells transfected with small interfering RNA targeting the mouse octn1 gene (siOCTN1). These results demonstrate that OCTN1 is functionally expressed in murine microglial cells. Exposure of WT-microglia to ERGO led to a significant decrease in cellular hypertrophy by LPS-stimulation with concomitant attenuation of intracellular reactive oxygen species (ROS), suggesting that OCTN1-mediated ERGO uptake may suppress cellular hypertrophy via the inhibition of ROS production with microglial activation. The expression of mRNA for interleukin-1β (IL-1β) after LPS-treatment was significantly increased in octn1 -/--microglia and siOCTN1-treated BV2 cells compared to the control cells. Meanwhile, treatment of ERGO minimally affected the induction of IL-1β mRNA by LPS-stimulation in cultured microglia and BV2 cells. Thus, OCTN1 negatively regulated the induction of inflammatory cytokine IL-1β, at least in part, via the transport of unidentified substrates other than ERGO in microglial cells.


Zinc excess impairs Mycobacterium bovis growth through triggering a Zur-IdeR-iron homeostasis signal pathway.
Zinc excess is toxic to bacteria and, thus, represents an important innate defense mechanism of host cells, especially against mycobacterial infections. However, the signaling pathway triggered by zinc excess and its relationship with iron homeostasis remain poorly understood in mycobacteria. Here, we characterize a novel Zur-IdeR-iron homeostasis signaling pathway that modulates the growth of Mycobacterium bovis under zinc toxicity. We found that the regulator Zur interacts with the iron-homeostasis regulator IdeR, enhancing the DNA-binding ability of IdeR. Excess zinc disrupts this interaction and represses ideR transcription through Zur, which promotes the expression of iron uptake genes and leads to the accumulation of intracellular iron in M. bovis. The elevated iron levels lower the bacterial survival ability under excess zinc stress. Consistently, deleting zur hinders intracellular iron accumulation of M. bovis and enhances bacterial growth under stress, while silencing ideR impairs the growth of the wild-type and zur-deleted strains under the same conditions. Interestingly, both Zur and IdeR are conserved in bacteria facing zinc toxicity. Overall, our work uncovers a novel antimicrobial signal pathway whereby zinc excess disrupts iron homeostasis, which may deepen our understanding of the crosstalk mechanism between iron and zinc homeostasis in bacteria.IMPORTANCEAs a catalytic and structural cofactor of proteins, zinc is essential for almost all living organisms. However, zinc excess is toxic and represents a vital innate immunity strategy of macrophages to combat intracellular pathogens, especially against mycobacterial pathogens such as Mycobacterium tuberculosis, the causative agent of tuberculosis. Here, we first characterize an antibacterial signaling pathway of zinc excess and its relationship with iron homeostasis in M. bovis. We found that excess zinc inhibits the transcription of ideR and its DNA-binding activity through Zur, which, in turn, promotes the expression of iron uptake genes, causes intracellular iron accumulation, and finally impairs the bacterial growth. This study reveals the existence of the Zur-IdeR-iron homeostasis pathway triggered by zinc excess in M. bovis, which will shed light on the crosstalk mechanisms between zinc and iron homeostasis in bacteria and the antimicrobial mechanisms of host-mediated zinc toxicity.


A plant-based diet differentially affects the global hepatic methylome in rainbow trout depending on genetic background.
Replacing fish meal and oil in trout diets with plant-derived ingredients is a contemporary challenge to move towards more sustainable aquaculture practices. However, such dietary replacement causes hepatic metabolic changes that have not yet been elucidated. Here, we aimed to decipher the effect of a 100% plant-based diet on the hepatic global DNA methylation landscape in trout and assess whether changes depend on fish genetic background. We analysed the global methylome and the expression of DNA (de)methylation-related genes of three isogenic lines that exhibit similar growth when fed a marine resource-based diet (M diet), but differ in their responses to a plant-based diet (V diet). Our results revealed that the V diet induced a decrease in 5-cytosine combined with an increase in 5-hydroxymethylcytosine in two of the three analysed lines. For one of these 2 affected lines, when fed the M diet but at the same feed intake of the V diet (MR), no methylome differences were highlighted between M and MR or between MR and V-fed trout whereas for the other affected line, M fed trout displayed a divergent methylome profile from MR and V fed fish. DNA (de)methylation-related genes were also affected by the V or MR diets. Our findings showed that the global hepatic methylome of trout is affected by a V diet, depending on genetic background. This latter effect seems to be due to either a decreased feed intake alone or combined with the effect of the dietary composition per se.


Actinomycin V Suppresses Human Non-Small-Cell Lung Carcinoma A549 Cells by Inducing G2/M Phase Arrest and Apoptosis via the p53-Dependent Pathway.
Actinomycin V, extracted and separated from marine-derived actinomycete Streptomyces sp., as the superior potential replacement of actinomycin D (which showed defect for its hepatotoxicity) has revealed an ideal effect in the suppression of migration and invasion in human breast cancer cells as referred to in our previous study. In this study, the involvement of p53 in the cell cycle arrest and pro-apoptotic action of actinomycin V was investigated in human non-small-cell lung carcinoma A549 cells. Results from the 3-(4,5-dimethylthiazol)-2,5-diphenyltetrazolium bromide assay showed that cytotoxic activity of actinomycin V on A549 cells (with wild-type p53) was stronger than the NCI-H1299 cells (p53-deficient). Actinomycin V upregulated both of the protein and mRNA expression levels of p53, p21Waf1/Cip1 and Bax in A549 cells. For this situation, actinomycin V decreased the M-phase related proteins (Cdc2, Cdc25A and Cyclin B1) expression, arrested cells in G2/M phase and subsequently triggered apoptosis by mediating the Bcl-2 family proteins' expression (Bax and Bcl-2). Furthermore, the effects of cell cycle arrest and apoptosis in A549 cells which were induced by actinomycin V could be reversed by the pifithrin-α, a specific inhibitor of p53 transcriptional activity. Collectively, our results suggest that actinomycin V causes up-regulation of p53 by which the growth of A549 cells is suppressed for cell cycle arrest and apoptosis.


Integrated Magnetic Bead-Quantum Dot Immunoassay for Malaria Detection.
Malaria persists as a disease of high morbidity and mortality due to improper diagnosis, overuse of drugs, rapidly evolving drug resistant parasites, and poor disease monitoring. The two common tests used in developing countries, microscopic examination of Glemsa slides and rapid diagnostic tests (RDTs), have limitations associated with variability in specificity and sensitivity, and qualitative outcome. Here we report on an immunoassay using magnetic beads for capture and quantum dots for detection of histidine-rich protein 2 (HRP2). Conventional immunoassays, such as ELISA, and molecular analysis tools, such as PCR, are difficult to implement in low resource settings. Therefore, to provide a proof-of-principle of translation of this assay to low resource settings, we demonstrate HRP2 detection in an automated droplet-based microfluidic device. Droplet-based platforms have the potential to allow translation of molecular detection assays to point-of-care use in low resource settings.


Effect of peppermint-lemon aromatherapy on nausea-vomiting and quality of life in pediatric patients with leukemia: A randomized controlled trial.
Nausea and vomiting, frequently induced by chemotherapy, can delay treatment protocols and the healing process. The aim of this study is to determine how aromatherapy inhalation with peppermint and lemon using a diffuser affects nausea-vomiting management and quality of life in 2-12-year-old children undergoing chemotherapy. The study utilized a pretest-posttest control group experimental design with randomized groups. A total of 90 children who met the inclusion criteria were included in the study. The experimental group received Mentha Piperita and Citrus Lemon essential oils through a diffuser, while the placebo group received water through a diffuser. The control group did not receive any intervention. Pulse and respiratory rates of children treated with aromatherapy were found to be significantly lower than the other groups. After aromatherapy application, quality of life of the children in the experimental group was significantly higher than the other groups. The change in the Index of Nausea, Vomiting, and Retching scores of the experimental group on the 4th chemotherapy cycle compared to the 1st chemotherapy cycle was significantly higher than the change in the other groups. Consequently, it was determined that inhalation aromatherapy with peppermint-lemon was effective in the management of chemotherapy-induced nausea-vomiting symptoms and quality of life compared to the placebo and control groups. Inhalation aromatherapy with mint-lemon can be used as an alternative method to improve the quality of life in children with leukemia who suffer from chemotherapy-induced nausea and vomiting.


Measuring Explicit Word Learning of Preschool Children: A Development Study.
The purpose of this article is to present preliminary results related to the development of a new measure of explicit word learning. The measure incorporated elements of explicit vocabulary instruction and dynamic assessment and was designed to be sensitive to differences in word learning skill and to be feasible for use in clinical settings. The explicit word learning measure included brief teaching trials and repeated fine-grained measurement of semantic knowledge and production of 3 novel words (2 verbs and 1 adjective). Preschool children (N = 23) completed the measure of explicit word learning; standardized, norm-referenced measures of expressive and receptive vocabulary; and an incidental word learning task. The measure of explicit word learning provided meaningful information about word learning. Performance on the explicit measure was related to existing vocabulary knowledge and incidental word learning. Findings from this development study indicate that further examination of the measure of explicit word learning is warranted. The measure may have the potential to identify children who are poor word learners. https://doi.org/10.23641/asha.5170738.


Effects of ground, steam-flaked, and steam-rolled corn grains on performance of lactating cows.
Five types of processed corn were compared to determine the effects of processing on lactational performance and nutrient digestibilities in dairy cows. Forty lactating Holstein cows averaging 160 +/- 60 d in milk were randomly divided into five groups on the basis of pretreatment milk yields and were fed diets containing 40% corn grain for 56 d. Treatments were 1) finely ground corn, 2) coarsely ground corn, 3) steam-flaked corn at a low density, 4) steam-flaked corn at a medium density, and 5) steam-rolled corn. Cows fed the diet containing steam-flaked corn at a medium density had a higher milk yield (37.1 kg/d) than did cows fed the diets containing coarsely ground corn, steam-flaked corn at a low density, or steam-rolled corn; cows fed the diet containing finely ground corn had an intermediate milk yield (35.5 kg/d). Efficiency of feed utilization was greater, and dry matter intake was lower, for cows fed the diet containing finely ground corn than for cows fed the other diets. The fat content of milk was higher for cows fed the diets containing coarsely ground corn and steam-rolled corn than for cows fed the diet containing steam-flaked corn at a medium density. Milk protein and SNF contents and yields of protein, lactose, and SNF did not differ among diets. Apparent starch digestibilities in the total digestive tract were lower for diets containing coarsely ground corn (87.4%) and steam-rolled corn (91.3%) than for the other diets (X = 96.3%). Milk yield was highest for cows fed the diet containing steam-flaked corn with a medium density. Fine grinding resulted in the greatest efficiency of feed utilization.


[Effect of MLL-AF9 fusion gene silence of acute monocytic leukemia cell line THP-1 on cyclin-dependent kinase inhibitor p27 expression].
To explore the effect of MLL-AF9 fusion gene silence on p27 expression and transcription regulation in THP-1 cells. Small interference RNA (siRNA) fragments targeting THP-1 cells specific MLL-AF9 fusion gene were designed and constructed, and transfected into THP-1 by lipofectamine. Flow cytometry was used to detect siRNA transfection efficiency. The level of MLL-AF9 mRNA expression was examined by RT-PCR and the expression of MLL-AF9 and p27 protein was detected by Western blot. Chromatin immunoprecipitation (ChIP) assay was used to confirm whether MLL-AF9 binds to the p27 promoter in THP-1 cell. SiRNA transfection efficiency was (69.1 +/- 1.8)%. The level of p27 expression was up-regulated at both mRNA [(0.84 +/- 0.12) vs (0.35 +/- 0.03) of control group] and protein levels after MLL-AF9 expression was significantly inhibited in siRNA-transfected cells (0.31 +/- 0.07) compared with that in the controls (1.25 +/- 0.13) (P<0.01). MLL-AF9 fusion protein bond to DNA fragment of p27 gene promoter region in THP-1 cell. MLL-AF9 fusion gene silence up-regulates p27 gene expression, and the mechanism maybe the recovery of p27 gene expression due to MLL-AF9 fusion protein binding to p27 promoter.


The one-year results of half-dose photodynamic therapy with verteporfin in chronic or recurrent central serous chorioretinopathy.
To evaluate the one-year results of half-dose photodynamic therapy with verteporfin in chronic or recurrent central serous chorioretinopathy (CSC). A Retrospective, consecutive case series. Twenty-seven eyes of 27 patients with chronic symptomatic CSC or recurrent CSC underwent photodynamic therapy (PDT) with half-dose (3 mg/m2) verteporfin. The demographic data such as age, side, gender spot sizes of laser photodynamic therapy (PDT) were recorded. The primary outcomes were the best correct visual acuity (BCVA), central foveal thickness using the optical coherence tomography (OCT) and complication were recorded as secondary outcome at baseline, month 1,3,6 and 12 post PDT RESULTS: At 12 months after half-dose PDT the mean logMAR BCVA improved from 0.32 to 0.18 (p = 0.001), the mean central foveal thickness decreased from 375.52 microm to 186.52 mirom (p < 0.001). The results also showed significant improvement of logMAR BCVA and decreased central foveal thickness after 1 month (0.32 to 0.22, p = 0.003 and 375.52 microm to 175.41 microm, p < 0.001) and maintained the results until one-year follow-up. Twenty-five eyes (92.59%) showed complete resolution of subretinal fluid at 1 month, 27 patients (100%) showed complete resolution at 3 month and all sustained the complete resolution until the last visit. No serious complications were recorded during and after the treatment. The half-dose PDT in area of fluorescein leakage is one of the effective treatment options for chronic or recurrent CSC, especially in patients who cannot be performed by focal laser photocoagulation. The treatment sustained the good visual results and has no serious complications up to one-year.


Repair and care of chest wall defects.
The formidable challenge of chest wall repair calls upon a multidisciplinary health care approach. The vital organs housed within the chest must be protected and the vascularity of available tissues used for repair must be maintained. Once chest wall reconstruction is complete, the patient must be closely monitored for stability and tissues must be protected from internal and external assaults. The following discussion will review the options for repair and the care of the patient undergoing chest wall defect reconstruction.


Display rule "regulators": the relationship between supervisors and worker emotional exhaustion.
This field study examined the effect of supervisory regulation of display rules--the rules about what kind of emotion to express on the job (R. Ekman, 1992; A. Rafaeli & R. I. Sutton, 1987)--on the emotional exhaustion of subordinates. On the basis of a sample of 940 call center employees, the authors found that worker emotional exhaustion varied across supervisors within jobs, suggesting that emotion work is influenced at the supervisory, rather than job, level. Moreover, the authors found that the importance supervisors place on interpersonal job demands of their workers was positively related to worker emotional exhaustion. Worker career identity moderated the interpersonal-job-demands--emotional-exhaustion relationship, but self-efficacy did not. Study conclusions and suggestions for future research are provided.


Pain management autobiographies and reluctance to use opioids for cancer pain management.
Although pain management education results in improved pain control for some patients, it does not work for all patients because some patients remain reluctant or unwilling to use prescribed analgesics to their optimal effect. In a randomized clinical trial that tested the effectiveness of the PRO-SELF Pain Control Program, 11 patients declined to increase their analgesic use despite moderate to severe pain. These patients were selected for a qualitative analysis of their audiotaped discussions about pain management with their intervention nurses. This analysis revealed that these patients often spontaneously provided detailed explanations about why they were reluctant or unwilling to take analgesics in general or opioids in particular. We termed these explanatory accounts pain management autobiographies because of their narrative character and multilayered, richly detailed quality. Pain management autobiographies included stories about (1) previous experience with chronic pain management, including stigmatizing interactions with clinicians and family members; (2) bad experiences with cancer pain management, including severe constipation; and 3) strongly held conventions about medication use, including the belief that all medications are "toxins" that should be avoided. The study findings suggest that a small subset of patients with cancer pain may need interventions such as individual or family counseling or alternative pain management strategies to augment education about opioids.


The Lived Experience of a Newly Diagnosed Older Person With HIV in Ukraine.
Low- and middle-income countries are home to 80% of older people with HIV (OPWH). Ukrainian OPWH experience higher mortality and decreased antiretroviral therapy (ART) initiation than younger patients, but there is little data examining OPWH's perspectives around new diagnosis and impact on care. In this study, we examined accounts of 30 newly diagnosed OPWH in Ukraine, exploring challenges faced in the peri-diagnosis period. Themes emerged representing the longitudinal coping process: OPWH (1) viewed themselves as low risk before diagnosis; (2) experienced HIV diagnosis as a traumatic event challenging their self-image; (3) used disclosure to seek support among a small circle of family, friends, or health care providers; (4) avoided disclosure to outsiders including primary care providers for fears of stigma and breaches in confidentiality; (5) viewed age as an asset; and (6) used HIV diagnosis as starting point for growth. These findings highlight the need for age-specific programming to increase HIV knowledge and coping, increase screening, and improve long-term planning.


The role of CT, PET-CT, and MRI in ovarian cancer.
New treatment developments in ovarian cancer have led to a renewed interest in staging advanced ovarian cancer. The treatment of females with ovarian cancer patients has a strong multidisciplinary character with an essential role for the radiologist. This review aims to provide an overview of the current position of CT, positron emission tomography-CT, and MRI in ovarian cancer and how imaging can be used to guide multidisciplinary team discussions.


General Signal Amplification Strategy for Nonfaradic Impedimetric Sensing: Trastuzumab Detection Employing a Peptide Immunosensor.
A label-free and reagent-free peptide mimotope capacitive biosensor has been developed for cancer drug (trastuzumab) quantification based on nonfaradic readout. The low sensitivity issue of capacitive biosensors was overcome with two innovations: peptide mimotope mixed self-assembled monolayer (SAM) biointerface and dilution of the analysis buffer. Signal amplification was achieved through dilution of phosphate-buffered saline (PBS) to tune Cdl to dominate the overall capacitance change upon target binding, which contribution is often negligible without dilution. After 1000× dilution, the limit of detection was lowered 500-fold (0.22 μg/mL) and the sensitivity was increased 20-fold [0.04192 (μg/mL)-1] in comparison with undiluted PBS. The proposed signal amplification strategy is more straightforward and practical compared to biorecognition element engineering and other strategies. The proposed method was further applied to planar electrodes for optimizing sensing response time to less than 1 min.


Evaluation of anti-HIF and anti-angiogenic properties of honokiol for the treatment of ocular neovascular diseases.
Pathological activation of the hypoxia-inducible-factor (HIF) pathway leading to expression of pro-angiogenic genes, such as vascular endothelial growth factor (VEGF), is the fundamental cause of neovascularization in ocular ischemic diseases and cancers. We have shown that pure honokiol inhibits the HIF pathway and hypoxia-mediated expression of pro-angiogenic genes in a number of cancer and retinal pigment epithelial (RPE) cell lines. The crude extracts, containing honokiol, from Magnolia plants have been used for thousands of years in the traditional oriental medicine for a number of health benefits. We have recently demonstrated that daily intraperitoneal injection of honokiol starting at postnatal day (P) 12 in an oxygen induced retinopathy mouse model significantly reduced retinal neovascularization at P17. Here, we evaluate the mechanism of HIF inhibition by honokiol in RPE cells. Using chromatin immunoprecipitation experiments, we demonstrate that honokiol inhibits binding of HIF to hypoxia-response elements present on VEGF promoter. We further show using a number of in vitro angiogenesis assays that, in addition to anti-HIF effect, honokiol manifests potent anti-angiogenic effect on human retinal micro vascular endothelial cells. Our results suggest that honokiol possesses potent anti-HIF and anti-angiogenic properties. These properties of honokiol make it an ideal therapeutic agent for the treatment of ocular neovascular diseases and solid tumors.


[Coordination of individualized diagnosis and therapy of prostatic cancer].
The individualisation of the diagnostics and the therapy of the carcinoma of the prostate renders necessary the coordination between medical check-up, statement and early forms of the carcinoma of the prostate, histological ascertainment, basis diagnostics and enlarged diagnostics as well as a highly specialised therapy. In the paper a model is demonstrated with the help of which it is possible to comprehend curable patients with carcinoma of the prostate and to use curative methods of treatment in a differentiated way. The author enters the possibilities of an exact TNM-determination. The continuation of curative methods of treatment, the intensifying of medical check-ups and the scientific optimization of palliative forms of treatment are decisive factors for the improvement of the survival rates in the carcinoma of the prostate.


Catalytic asymmetric rearrangement of allylic trichloroacetimidates. A practical method for preparing allylic amines and congeners of high enantiomeric purity.
COP-Cl catalyzes the rearrangement of (E)-allylic trichloroacetimidates to provide transposed allylic trichloroacetamides of high enantiopurity, a transformation that underlies the first truly practical method for transforming prochiral allylic alcohols to enantioenriched allylic amines and congeners. The high functional group compatibility of this asymmetric rearrangement and the demonstrated broad utility in synthesis of the allylic trichloroacetimidate to allylic trichloroacetamide conversion are singular features of this new catalytic asymmetric reaction.


Investigating the role of the ROS/CncC signaling pathway in the response to xenobiotics in Spodoptera frugiperda using Sf9 cells.
Spodoptera frugiperda (fall armyworm, FAW) is an invasive polyphagous lepidopteran pest that has developed sophisticated resistance mechanisms involving detoxification enzymes to eliminate toxic compounds it encounters in its diet including insecticides. Although its inventory of detoxification enzymes is known, the mechanisms that enable an adapted response depending on the toxic compound remain largely unexplored. Sf9 cells were used to investigate the role of the transcription factors, Cap n' collar isoform C (CncC) and musculoaponeurotic fibrosarcoma (Maf) in the regulation of the detoxification response. We overexpressed CncC, Maf or both genes, and knocked out (KO) CncC or its repressor Kelch-like ECH associated protein 1 (Keap1). Joint overexpression of CncC and Maf is required to confer increased tolerance to indole 3-carbinol (I3C), a plant secondary metabolite, and to methoprene, an insecticide. Both molecules induce reactive oxygen species (ROS) pulses in the different cell lines. The use of an antioxidant reversed ROS pulses and restored the tolerance to I3C and methoprene. The activity of detoxification enzymes varied according to the cell line. Suppression of Keap1 significantly increased the activity of cytochrome P450s, carboxylesterases and glutathione S-transferases. RNAseq experiments showed that CncC mainly regulates the expression of detoxification genes but is also at the crossroads of several signaling pathways (reproduction and immunity) maintaining homeostasis. We present new data in Sf9 cell lines suggesting that the CncC:Maf pathway plays a central role in FAW response to natural and synthetic xenobiotics. This knowledge helps to better understand detoxification gene expression and may help to design next-generation pest insect control measures.


Myofasciitis in the domestic ferret.
Since late 2003, an inflammatory disease of muscle and fascia has been diagnosed in several ferrets at Northwest ZooPath, and this report describes the condition in 17 ferrets. It is a disease of young ferrets, characterized by rapid onset of clinical signs, high fever, neutrophilic leukocytosis, treatment failure, and death (or euthanasia). Gross lesions include atrophy of skeletal muscle; red and white mottling and dilatation of the esophagus; and splenomegaly. Histologically, moderate to severe suppurative to pyogranulomatous inflammation is in the skeletal muscle and the fascia at multiple sites, including esophagus, heart, limbs, body wall, head, and lumbar regions. Myeloid hyperplasia of spleen and/or bone marrow also is a prominent feature. Ultrastructural lesions include mitochondrial swelling, intracellular edema, disruption of myofibrils and Z bands. Bacterial and viral cultures, electron microscopy, immunohistochemistry, and polymerase chain reaction were negative for a variety of infectious agents. The clinical presentation and distribution of lesions suggests that polymyositis in domestic ferrets is likely a distinct entity. The etiopathogenesis if this condition is not known.


A peptide from ICAM-2 binds to the leukocyte integrin CD11a/CD18 and inhibits endothelial cell adhesion.
Numerous leukocyte functions depend on adhesive intercellular interactions. The leukocyte-specific integrins CD11a/CD18 (lymphocyte function-associated antigen-1 (LFA-1)) and CD11b/CD18 (complement type 3 receptor (Mac-1)), which bind to the intercellular adhesion molecules ICAM-1 and ICAM-2, play a key role in adhesion. Little is known about the binding in molecular detail. We have now defined a peptide region from the first immunoglobulin domain of ICAM-2 that is specifically involved in binding to CD11a/CD18. A synthetic peptide from this part of ICAM-2, covering residues 21-42, bound to purified CD11a/CD18 and inhibited the adhesion of endothelial cells to this integrin. It also inhibited the binding of B lymphoblastoid cells to endothelial cells. Leukocytes bound to the peptide coated on plastic. Several shorter peptides from the same region showed less or no activity.


Association of primary glaucomas with retinal vein occlusion.
To determine the association of Primary glaucoma with retinal vein occlusion. A prospective cross-sectional study was carried out in B.P. Koirala Lions Centre for Ophthalmic Studies from April 2002-July 2003. All diagnosed cases of retinal vein occlusion were evaluated in detail for the presence of primary glaucoma. Detailed history was taken. Best corrected visual acuity was recorded, slit-lamp evaluation of anterior segment was performed, intraocular pressure was assessed with the help of applanation tonometer, gonioscopy was done and fundus evaluation was done under full mydriasis. Out of 50 patients, 19 (38%) were central retinal vein occlusion and 31(62%) were branch retinal vein occlusion. Majority of the retinal vein occlusion patients were in the age group of (61-70 years) 12 out of 50 patients (24%). There were 12 patients of primary glaucoma in 50 patients of retinal vein occlusion. Out of which 11 patients had primary open angle glaucoma and 1 patient had primary angle closure glaucoma. This study shows association of primary glaucoma as a risk factor of retinal vein occlusion. Evaluation of retinal vein occlusion patients for primary glaucoma would be worthwhile for early detection and prevention of blindness.


Food and consumers: Where are we heading?
The translation of recent advances in nutrition sciences into enhanced population health and well-being depends on the development of a deeper understanding of human food consumption patterns and the factors which influence them. Food consumption patterns are dynamic and are influenced by complex, interrelated biological, social, cultural and psychological processes. These are evident in recent attempts to discriminate nutrition and health-related dietary patterns in terms of consumer lifestyles and belief systems. Consumers' pursuit of health and well-being through food consumption will be illustrated through reference to recent Australian studies. Some of the effects of societal changes associated with globalization: gender, work and family roles; materialism; information technology; and increasing longevity, on food consumption trends will be explored. Finally, the implications of these developments for the activities of health professionals, food companies and other agencies will be raised.


The pComb3 Phagemid Family of Phage Display Vectors.
A phagemid is a plasmid that contains the origin of replication and packaging signal of a filamentous phage. Following bacterial transformation, a phagemid can be replicated and amplified as a plasmid, using a double-stranded DNA origin of replication, or it can be replicated as single-stranded DNA for packaging into filamentous phage particles. The use of phagemids enables phage display of large proteins, such as antibody fragments. Phagemid pComb3 was among the first phage display vectors used for the generation and selection of antibody libraries in the 50-kDa Fab format, a monovalent proxy of natural antibodies. Affording a robust and versatile tool for more than three decades, phage display vectors of the pComb3 phagemid family have been widely used for the discovery, affinity maturation, and humanization of antibodies in Fab, scFv, and single-domain formats from naive, immune, and synthetic antibody repertoires. In addition, they have been used for broadening phage display to the mining of nonimmunoglobulin repertoires. This review examines conceptual, functional, and molecular features of the first-generation phage display vector pComb3 and its successors, pComb3H, pComb3X, and pC3C.


Characterization of the cytochrome system of a nitrogen-fixing strain of a sulfate-reducing bacterium: Desulfovibrio desulfuricans strain Berre-Eau.
Two c-type cytochromes were purified and characterized by electron paramagnetic resonance (EPR) and nuclear magnetic resonance (NMR) spectroscopic techniques, from the sulfate-reducer nitrogen-fixing organism, Desulfovibrio desulfuricans strain Berre-Eau (NCIB 8387). The purification procedures included several chromatographic steps on alumina, carboxymethylcellulose and gel filtration. A tetrahaem and a monohaem cytochrome were identified. The multihaem cytochrome has visible, EPR and NMR spectra with general properties similar to other low-potential bis-histidinyl axially bound haem proteins, belonging to the class of tetrahaem cytochrome c3 isolated from other Desulfovibrio species. The monohaem cytochrome c553 is ascorbate-reducible and its EPR and NMR data are characteristic of a cytochrome with methionine-histidine ligation. Their properties are compared with other homologous proteins isolated from sulfate-reducing bacteria.


Tree proximity, soil pathways and common mycorrhizal networks: their influence on the utilization of redistributed water by understory seedlings.
Hydraulic redistribution (HR) is a process by which water moves through plant roots from moist to dry soils. An experiment was conducted to quantify the influence of common mycorrhizal networks (CMNs) and proximity to mature HR-source trees on the water relations of surrounding seedlings. Douglas-fir (Pseudotsuga menziesii var glauca (Mirb.) Franco) seedlings were planted at four distances (0.5, 1, 2.5, and 5 m) from six mature Douglas-fir trees, either directly into soil (soil plus CMN pathway) or inside 0.5 microm mesh bags (soil-only pathway). Deuterated water was used to irrigate soil beside mature trees in order to identify different HR water pathways to surrounding seedlings. This was followed by measurements of seedling deuterium enrichment, seedling water potential, soil water potential, gravimetric soil water content, and tree root density surrounding the seedlings. There was no significantly detectable difference in the quantity of HR water transferred to seedlings having access to soil and CMN pathways or soil-only pathways of water movement. Water from the irrigation plot contributed up to 1.4% of the water of Douglas-fir seedlings. Based on the assumption that the only pathway through which seedlings could access irrigation water was through the mature trees, we estimate that as much as 21.6% of the seedling water was supplied by the nearby tree. Seedling water potential was not significantly affected either by proximity to mature trees or pathway, suggesting HR may have compensated for increasing tree competitive effects with proximity. It is also possible that the lack of difference was due to a relatively moist summer. Our results suggest that residual mature trees are potentially important for hydraulic redistribution to regenerating seedlings in harvested dry interior Douglas-fir forests.


Virulence Factors and Azole-Resistant Mechanism of Candida Tropicalis Isolated from Candidemia.
Limited knowledge exists on the virulence factors of Candida tropicalis and the mechanisms of azole resistance that lead to an intensified pathogenicity and treatment failure. We aimed to evaluate the virulence factors and molecular mechanisms of azole resistance among C. tropicalis isolated from patients with candidemia. Several virulence factors, including extracellular enzymatic activities, cell surface hydrophobicity (CSH), and biofilm formation, were evaluated. Antifungal susceptibility pattern and expression level of ERG11, UPC2, MDR1, and CDR1 genes of eight (4 fluconazole resistance and 4 fluconazole susceptible) clinical C. tropicalis isolates were assessed. The correlation between the virulence factors and antifungal susceptibility patterns was analyzed. During a 4 year study, forty-five C. tropicalis isolates were recovered from candidemia patients. The isolates expressed different frequencies of virulence determinants as follows: coagulase 4 (8.9%), phospholipase 5 (11.1%), proteinase 31 (68.9%), esterase 43 (95.6%), hemolysin 44 (97.8%), biofilm formation 45 (100%) and CSH 45(100%). All the isolates were susceptible to amphotericin B and showed the highest resistance to voriconazole. There was a significant positive correlation between micafungin minimum inhibitory concentrations (MICs) and hemolysin production (rs = 0.316). However, we found a negative correlation between fluconazole MICs and esterase production (rs = -0.383). We observed the high expression of ERG11 and UPC2 genes in fluconazole-resistant C. tropicalis isolates. C. tropicalis isolated from candidemia patients extensively displayed capacities for biofilm formation, hemolysis, esterase activity, and hydrophobicity. In addition, the overexpression of ERG11 and UPC2 genes was considered one of the possible mechanisms of azole resistance.


The Intensive Care Medicine research agenda on critically ill oncology and hematology patients.
Over the coming years, accelerating progress against cancer will be associated with an increased number of patients who require life-sustaining therapies for infectious or toxic chemotherapy-related events. Major changes include increased number of cancer patients admitted to the ICU with full-code status or for time-limited trials, increased survival and quality of life in ICU survivors, changing prognostic factors, early ICU admission for optimal monitoring, and use of noninvasive diagnostic and therapeutic strategies. In this review, experts in the management of critically ill cancer patients highlight recent changes in the use and the results of intensive care in patients with malignancies. They seek to put forward a standard of care for the management of these patients and highlight important updates that are required to care for them. The research agenda they suggest includes important studies to be conducted in the next few years to increase our understanding of organ dysfunction in this population and to improve our ability to appropriately use life-saving therapies or select new therapeutic approaches that are likely to improve outcomes. This review aims to provide more guidance for the daily management of patients with cancer, in whom outcomes are constantly improving, as is our global ability to fight against what is becoming the leading cause of mortality in industrialized and non-industrialized countries.


Effects of Fas-ligand overexpression on alveolar type II cell growth kinetics in perinatal murine lungs.
We determined the time-specific effects of FasL overexpression on perinatal alveolar type II cell growth kinetics. To achieve temporal overexpression of respiratory epithelium-specific FasL expression, tetracycline inducible CCSP-rtTA/FasL-TetOp transgenic mice were given doxycycline (Dox) from gestational d 14 (E14) to E19 (antenatal treatment group), from postnatal d 1 (P1) to P7 (postnatal group), or from E14 to P7 (combined antenatal and postnatal group). Antenatal Dox administration induced an increase of pulmonary FasL mRNA levels in double transgenic animals up to >300-fold over single transgenic littermate controls, associated with massive fetal respiratory epithelial apoptosis and excessive postnatal lethality. Although animals from the combined antenatal/postnatal Dox treatment group continued to display evidence of increased apoptosis, there was a paradoxical increase in alveolar type II cell proliferation, resulting in a net increase in type II cell density, elevated pulmonary surfactant protein C levels and improved postnatal survival. Postnatal Dox administration was also associated with increased type II cell density, although FasL up-regulation was more variable. In conclusion, these results, and our previous studies, suggest that FasL signaling has dual timing-dependent proapoptotic and proproliferative effects on postcanalicular type II cell kinetics.


Diagnosis and treatment of depression in the elderly.
Depression and suicide are significant problems in the elderly, both in terms of their severity and their prevalence. It is particularly difficult to distinguish depression from early dementia, since elderly depressed patients often deny mood disorder and focus on their memory problems. This differential diagnostic dilemma is further complicated by the fact that 20 percent of Alzheimer-type dementia patients have moderate to severe depression. An even higher prevalence of depression can be seen in elderly patients with stroke or Parkinson's disease. Most all of the depressive disorders of the elderly are amenable to one form or combination of therapies: pharmacologic, electro-convulsive, or psychotherapy. Tricyclic antidepressants are often associated with adverse drug reactions in the elderly, so alternatives such as MAO inhibitors, alprazolam, bupropion and psychostimulants are currently being explored in this patient population.


Molecular Markers for Sensitive Detection of Plasmodium falciparum Asexual Stage Parasites and their Application in a Malaria Clinical Trial.
Plasmodium falciparum parasite life stages respond differently to antimalarial drugs. Sensitive stage-specific molecular assays may help to examine parasite dynamics at microscopically detectable and submicroscopic parasite densities in epidemiological and clinical studies. In this study, we compared the performance of skeleton-binding protein 1 (SBP1), ring-infected erythrocyte surface antigen, Hyp8, ring-exported protein 1 (REX1), and PHISTb mRNA for detecting ring-stage trophozoite-specific transcripts using quantitative reverse transcriptase polymerase chain reaction. Markers were tested on tightly synchronized in vitro parasites and clinical trial samples alongside established markers of parasite density (18S DNA and rRNA) and gametocyte density (Pfs25 mRNA). SBP1 was the most sensitive marker but showed low-level expression in mature gametocytes. Novel markers REX1 and PHISTb showed lower sensitivity but higher specificity for ring-stage trophozoites. Using in vivo clinical trial samples from gametocyte-negative patients, we observed evidence of persisting trophozoite transcripts for at least 14 days postinitiation of treatment. It is currently not clear if these transcripts represent viable parasites that may have implications for clinical treatment outcome or transmission potential.


Innovation in dermatology: where are the dermatologists? A retrospective review of the Pitchbook Database.
To investigate the extent to which dermatologists are involved in innovative dermatology start-up companies, we analyzed the Pitchbook database which covers private capital markets. From January 1, 2010 to June 23, 2021, there were 105 dermatology companies, of which six (5.7%) had a dermatologist chief executive officer (CEO), 16 (15.2%) had a dermatologist founder, and 19 (18.1%) had a dermatologist advisor or board member. There were 98 dermatologists with leadership positions, including six (6.1%) CEOs, 21 (21.4%) founders, 11 (11.2%) chief medical officers (CMOs), and 60 (61.2%) advisors. Our findings highlight the underrepresentation of dermatologists in start-up ventures relating to dermatology.


Periodontal status and treatment needs among Spanish military personnel.
Problems arising from poor gingival and periodontal health in military personnel account for around 10% of the emergencies of oral origin during deployments and manoeuvres. Consequently, it is important to ascertain the prior oral health situation among Spanish military personnel. To assess the periodontal health status and treatment needs of a population of Spanish Army personnel at the General Almirante Base at Marines, in the province of Valencia, and calculate the time required to cover the treatment needs of this population. Observational cross-sectional study of periodontal health status in a representative sample of the population in question. The community periodontal index (CPI) was used in accordance with WHO criteria. The sample comprised 387 subjects, selected by systematically sampling those attending the sick bay at the base for different reasons. The examinations were carried out by a single examiner; the intra-observer Kappa index was 0.83. Data collection began in December 2003 and ended in July 2004. All sextants were healthy in 7,2% of the sample. The most prevalent condition was the presence of calculus, especially in the younger population. 7.8% had 4-5 mm periodontal pockets and 2.3% had pockets of 6 mm or more, all in the over-25 age group. The women below 25 years of age were significantly more healthy than the men. The healthy sextants mean was 2.38, without differences by rank. Privates presented a significantly higher mean figure for sextants with bleeding, while officers and non-commissioned officers presented a higher average number of sextants with pockets. Practically every subject needed oral hygiene instructions and scaling and root planing but only 2.3%, all over 25 years old, required complex treatments. The estimated treatment time need is 1 hour per person per year.


Negative specific heat, the thermodynamic limit, and ergodicity.
Thermodynamic stability, in particular, the positivity of the specific heat in the microcanonical ensemble, is not an automatic consequence of the thermodynamic limit. But it holds under special circumstances such as for the most important case of quantum-mechanical Coulomb systems. Therefore, it is surprising that there are experimental indications to the contrary. In this Letter we study a simple model for which the microcanonical specific heat is positive, if the system is ergodic. However, if the system is not ergodic, the energy shell in phase space has some ergodic components with a negative specific heat. This provides another possible general pathway for a negative specific heat in addition to the commonly accepted, the small number of particles.


Age-specific hysterectomy and oophorectomy prevalence rates and the risks for cancer of the reproductive system.
A sample survey was conducted in 1982 to determine the prevalence of hysterectomy and oophorectomy among upstate New York women, ages 25-74. The effects of this surgery on age-specific estimates of the risk for cancer of the uterus, cervix, and ovary were calculated. Overall, 16.9 per cent of the women reported having had a hysterectomy and 9.9 per cent reported a bilateral oophorectomy. The adjustment for age-specific hysterectomy increased the 1977-1979 average annual incidence rate of cervical and uterine cancer by 21 per cent. In several five-year age categories, the increase reached 54 per cent. The 1977-1979 average annual incidence rate of ovarian cancer increased by 12 per cent after adjusting for age-specific bilateral oophorectomy. The increase reached 29 per cent in one five-year age group. The sample results show a lower prevalence of hysterectomy among women 25 to 40 years old and among women 70 to 74 than estimates based on the application of mathematical models to data on surgical incidence.


Living with a leaky skin: upregulation of ion transport proteins during sloughing.
Amphibian skin is a multifunctional organ providing protection from the external environment and facilitating the physiological exchange of gases, water and salts with the environment. In order to maintain these functions, the outer layer of skin is regularly replaced in a process called sloughing. During sloughing, the outermost layer of the skin is removed in its entirety, which has the potential to interfere with skin permeability and ion transport, disrupting homeostasis. In this study, we measured, in vivo, the effects of sloughing on the cutaneous efflux of ions in toads Rhinella marina kept in freshwater conditions. We also measured transepithelial potential, cutaneous resistance, active ion transport and the distribution, abundance and gene expression of the key ion transport proteins sodium-potassium ATPase (NKA) and epithelial sodium channel (ENaC) during sloughing. We hypothesised that the increase in transepithelial efflux of ions during sloughing is a consequence of increased permeability and/or a reduction in the abundance or expression of cutaneous ion transport proteins, resulting in disruption of internal ion homeostasis. There was a significant increase in sodium and chloride efflux during sloughing in R. marina However, although in vitro skin resistance decreased after sloughing, active sodium transport increased commensurate with an increase in NKA and ENaC protein abundance in the skin. These changes in skin function associated with sloughing did not affect the maintenance of internal electrolyte homeostasis. These results suggest that during sloughing, amphibians actively maintain internal homeostasis by increasing cutaneous rates of ion uptake.


Laser-induced micropore formation and modification of cartilage structure in osteoarthritis healing.
Pores are vital for functioning of avascular tissues. Laser-induced pores play an important role in the process of cartilage regeneration. The aim of any treatment for osteoarthritis is to repair hyaline-type cartilage. The aims of this study are to answer two questions: (1) How do laser-assisted pores affect the cartilaginous cells to synthesize hyaline cartilage (HC)? and (2) How can the size distribution of pores arising in the course of laser radiation be controlled? We have shown that in cartilage, the pores arise predominately near chondrocytes, which promote nutrition of cells and signal molecular transfer that activates regeneration of cartilage. In vivo laser treatment of damaged cartilage of miniature pig joints provides cellular transformation and formation of HC. We propose a simple model of pore formation in biopolymers that paves the way for going beyond the trial-and-error approach when choosing an optimal laser treatment regime. Our findings support the approach toward laser healing of osteoarthritis.


An experimental assessment of algal calcification as a potential source of atmospheric CO2.
Marine vegetated ecosystems such as seagrass meadows are increasingly acknowledged as important carbon sinks based on their ability to capture and store atmospheric carbon dioxide, thereby contributing to climate change mitigation. Most studies on carbon storage in marine ecosystems have focused on organic carbon, leaving inorganic carbon processes such as calcification unaccounted for, despite of their critical role in the global carbon budget. This is probably because of uncertainties regarding the role of calcification in marine carbon budgets as either atmospheric CO2 source or sink. Here, we conducted a laboratory experiment to investigate the influence of a calcifying alga (Corallina officinalis L.) on seawater carbon content, using a non-calcifying alga (Ulva lactuca L.) as a control. In a first part, algae were incubated separately while measuring changes in seawater pH, total alkalinity (TA) and total dissolved inorganic carbon (DIC). The amount of carbon used in photosynthetic uptake and production of CaCO3 was then calculated. In a second, directly following, part the algae were removed and DIC levels were allowed to equilibrate with air until the pH stabilized and the loss of CO2 to air was calculated as the difference in total DIC from the start of part one, to the end of the second part. The results showed that C. officinalis caused a significant and persistent reduction in total dissolved inorganic carbon (DIC), TA and seawater pH, while no such permanent changes were caused by U. lactuca. These findings indicate that calcification can release a significant amount of CO2 to the atmosphere and thereby possibly counteract the carbon sequestration in marine vegetated ecosystems if this CO2 is not re-fixed in the system. Our research emphasises the importance of considering algal calcification in future assessments on carbon storage in coastal areas.


Biological proton pumping in an oscillating electric field.
Time-dependent external perturbations provide powerful probes of the function of molecular machines. Here we study biological proton pumping in an oscillating electric field. The protein cytochrome c oxidase is the main energy transducer in aerobic life, converting chemical energy into an electric potential by pumping protons across a membrane. With the help of master-equation descriptions that recover the key thermodynamic and kinetic properties of this biological "fuel cell," we show that the proton pumping efficiency and the electronic currents in steady state depend significantly on the frequency and amplitude of the applied field, allowing us to distinguish between different microscopic mechanisms of the machine. A spectral analysis reveals dominant reaction steps consistent with an electron-gated pumping mechanism.


Reexamination of the difference in susceptibility to adjuvant-induced arthritis among LEW/Crj, Slc/Wistar/ST and Slc/SD rats.
The present investigations were performed to assess the differences among rat colonies commonly used for neurophysiological research regarding the development of complete Freund's adjuvant (CFA)-induced arthritis. Inflammatory signs including edema in the paw fluctuated remarkably among individual Wistar (Slc/Wistar/ST) and Sprague-Dawley (Slc/SD) rats, while the inflammatory signs of Lewis (LEW/Crj) rats appeared earlier and was severer and more consistent than Slc/Wistar/ST and Slc/SD rats. Edema in the hind paw developed in 100% of LEW/Crj rats with the lowest dose of CFA (0.6 mg/rat) used as compared with 64% of Slc/Wistar/ST (CFA 1 mg/rat) and 38% of Slc/SD rats (CFA 1.2 mg/rat). Retardation of weight gain was observed in Slc/Wistar/ST and Slc/SD rats in contrast to a severe weight decrease in inflamed LEW/Crj rats after the development of arthritis.