Structural insights into the evolution of the adaptive immune system.
The adaptive immune system, which is based on highly diverse antigen receptors that are generated by somatic recombination, arose approximately 500 Mya at the dawn of vertebrate evolution. In jawed vertebrates, adaptive immunity is mediated by antibodies and T cell receptors (TCRs), which are composed of immunoglobulin (Ig) domains containing hypervariable loops that bind antigen. In striking contrast, the adaptive immune receptors of jawless vertebrates, termed variable lymphocyte receptors (VLRs), are constructed from leucine-rich repeat (LRR) modules. Structural studies of VLRs have shown that these LRR-based receptors bind antigens though their concave surface, in addition to a unique hypervariable loop in the C-terminal LRR capping module. These studies have revealed a remarkable example of convergent evolution in which jawless vertebrates adopted the LRR scaffold to recognize as broad a spectrum of antigens as the Ig-based antibodies and TCRs of jawed vertebrates, with altogether comparable affinity and specificity.


The viral protein gp120 decreases the acetylation of neuronal tubulin: potential mechanism of neurotoxicity.
The human immunodeficiency virus (HIV) envelope protein gp120 promotes axonal damage and neurite pruning, similar to that observed in HIV-positive subjects with neurocognitive disorders. Thus, gp120 has been used to examine molecular and cellular pathways underlying HIV-mediated neuronal dysfunction. Gp120 binds to tubulin beta III, a component of neuronal microtubules. Microtubule function, which modulates the homeostasis of neurons, is regulated by polymerization and post-translational modifications. Based on these considerations, we tested the hypothesis that gp120 induces dynamic instability of neuronal microtubules. We first observed that gp120 prevents the normal polymerization of tubulin in vitro. We then tested whether gp120 alters the post-translational modifications in tubulin by examining the ability of gp120 to change the levels of acetylated tubulin in primary rat neuronal cultures. Gp120 elicited a time-dependent decrease in tubulin acetylation that was reversed by Helix-A peptide, a compound that competitively displaces the binding of gp120 to neuronal microtubules. To determine whether post-translational modifications in tubulin also occur in vivo, we measured acetylated tubulin in the cerebral cortex of HIV transgenic rats (HIV-tg). We observed a decrease in tubulin acetylation in 5- and 9-month-old HIV-tg rats when compared to age-matched wild type. Neither changes in microglia morphology nor alterations in mRNA levels for interleukin-1β and tumor necrosis factor α were detected in 5-month-old animals. Our findings propose neuronal microtubule instability as a novel mechanism of HIV neurotoxicity, without evidence of enhanced inflammation.


Edge-Guided Camouflaged Object Detection via Multi-Level Feature Integration.
Camouflaged object detection (COD) aims to segment those camouflaged objects that blend perfectly into their surroundings. Due to the low boundary contrast between camouflaged objects and their surroundings, their detection poses a significant challenge. Despite the numerous excellent camouflaged object detection methods developed in recent years, issues such as boundary refinement and multi-level feature extraction and fusion still need further exploration. In this paper, we propose a novel multi-level feature integration network (MFNet) for camouflaged object detection. Firstly, we design an edge guidance module (EGM) to improve the COD performance by providing additional boundary semantic information by combining high-level semantic information and low-level spatial details to model the edges of camouflaged objects. Additionally, we propose a multi-level feature integration module (MFIM), which leverages the fine local information of low-level features and the rich global information of high-level features in adjacent three-level features to provide a supplementary feature representation for the current-level features, effectively integrating the full context semantic information. Finally, we propose a context aggregation refinement module (CARM) to efficiently aggregate and refine the cross-level features to obtain clear prediction maps. Our extensive experiments on three benchmark datasets show that the MFNet model is an effective COD model and outperforms other state-of-the-art models in all four evaluation metrics (Sα, Eϕ, Fβw, and MAE).


Clp ATPases are required for stress tolerance, intracellular replication and biofilm formation in Staphylococcus aureus.
The Hsp100/Clp ATPases constitute a family of closely related proteins of which some members function solely as chaperones whereas others additionally can associate with the unrelated ClpP peptidase forming a Clp proteolytic complex. We have investigated the role of four Clp ATPases in the versatile pathogen, Staphylococcus aureus. Previously, we showed that ClpX is required for expression of major virulence factors and for virulence of S. aureus, but not for survival during heat shock. In the present study, we have inactivated clpC, clpB and clpL and, while none of these mutations affected toxin production, both ClpC and ClpB and to a minor extent ClpL were required for intracellular multiplication within bovine mammary epithelial cells. These defects were paralleled by an inability of the clpC mutant to grow at high temperature and of the clpB mutant to induce thermotolerance indicating that the protective functions of these proteins are required both at high temperature and during infection. By primer extension analysis and footprint studies, we show that expression of clpC and clpB is controlled by the negative heat-shock regulator, CtsR, and that ClpC is required for its repressor activity. Thus, ClpC is a likely sensor of stress encountered during both environmental stress and infection. In addition to virulence factor production the ability to form biofilms is of importance to S. aureus as a nosocomial pathogen. Interestingly, biofilm formation was reduced in the absence of ClpX or ClpC whereas it was enhanced in the absence of ClpP. Thus, our data show that Clp proteolytic complexes and the Clp ATPases control several key processes of importance to the success of S. aureus as a pathogen.


Effectiveness and safety of anti-TNF therapy for ankylosing spondylitis: a real-world study.
Aim: To evaluate the effectiveness and safety of anti-TNF drugs for ankylosing spondylitis. Materials & methods: A prospective cohort study was performed at a pharmacy in the Brazilian Public Health System. Effectiveness by Bath Ankylosing Spondylitis Disease Activity Index, functionality by Health Assessment Questionnaire Disability Index, quality of life by European Quality of Life Five-Dimensions and safety was assessed at 6 and 12 months of follow-up. Results: About 160 patients started the treatment with adalimumab, etanercept or infliximab. There was a statistically significant improvement in disease activity, functionality and quality of life at 6 and 12 months (p < 0.05). Conclusion: This real-world study has shown that anti-TNF drugs are effective and well tolerated for ankylosing spondylitis patients.


Repurposing metformin: an old drug with new tricks in its binding pockets.
Improvements in healthcare and nutrition have generated remarkable increases in life expectancy worldwide. This is one of the greatest achievements of the modern world yet it also presents a grave challenge: as more people survive into later life, more also experience the diseases of old age, including type 2 diabetes (T2D), cardiovascular disease (CVD) and cancer. Developing new ways to improve health in the elderly is therefore a top priority for biomedical research. Although our understanding of the molecular basis of these morbidities has advanced rapidly, effective novel treatments are still lacking. Alternative drug development strategies are now being explored, such as the repurposing of existing drugs used to treat other diseases. This can save a considerable amount of time and money since the pharmacokinetics, pharmacodynamics and safety profiles of these drugs are already established, effectively enabling preclinical studies to be bypassed. Metformin is one such drug currently being investigated for novel applications. The present review provides a thorough and detailed account of our current understanding of the molecular pharmacology and signalling mechanisms underlying biguanide-protein interactions. It also focuses on the key role of the microbiota in regulating age-associated morbidities and a potential role for metformin to modulate its function. Research in this area holds the key to solving many of the mysteries of our current understanding of drug action and concerted effects to provide sustained and long-life health.


Targeted panel sequencing in the routine diagnosis of mature T- and NK-cell lymphomas: report of 128 cases from two German reference centers.
Diagnosing any of the more than 30 types of T-cell lymphomas is considered a challenging task for many pathologists and currently requires morphological expertise as well as the integration of clinical data, immunophenotype, flow cytometry and clonality analyses. Even considering all available information, some margin of doubt might remain using the current diagnostic procedures. In recent times, the genetic landscape of most T-cell lymphomas has been elucidated, showing a number of diagnostically relevant mutations. In addition, recent data indicate that some of these genetic alterations might bear prognostic and predictive value. Extensive genetic analyses, such as whole exome or large panel sequencing are still expensive and time consuming, therefore limiting their application in routine diagnostic. We therefore devoted our effort to develop a lean approach for genetic analysis of T-cell lymphomas, focusing on maximum efficiency rather than exhaustively covering all possible targets. Here we report the results generated with our small amplicon-based panel that could be used routinely on paraffin-embedded and even decalcified samples, on a single sample basis in parallel with other NGS-panels used in our routine diagnostic lab, in a relatively short time and with limited costs. We tested 128 available samples from two German reference centers as part of our routine work up (among which 116 T-cell lymphomas), which is the largest routine diagnostic series reported to date. Our results showed that this assay had a very high rate of technical success (97%) and could detect mutations in the majority (79%) of tested T-cell lymphoma samples.


CD8+ T-cell exhaustion in cancer: mechanisms and new area for cancer immunotherapy.
Immunotherapies have emerged as the most promising area in cancer treatments in recent years. CD8+ T cells, as one of the primary effector cells of anticancer immunity, however, when infiltrating in cancer tissues, are generally in dysfunctional states termed T-cell exhaustion. Exhausted CD8+ T cells are characterized by impaired activity and proliferative ability, increased apoptotic rate and reduced production of effector cytokines. Such dysfunctional CD8+ T cells serve as a barrier in successful cancer elimination. Investigation on the mechanism of T-cell exhaustion was aiming to sustain or restore the efficiency of CD8+ T cells infiltrating in cancer, which may help to develop novel strategies to overcome cancer. Recent studies have found several vital mechanisms of CD8+ T-cell exhaustion and provided novel avenues through targeting CD8+ T-cell exhaustion to enhance anticancer immunity. Here, we review the recent progress in the study of CD8+ T-cell exhaustion to make a summary and to provide a framework for further researches.


Toward a resolution of the introns early/late debate: only phase zero introns are correlated with the structure of ancient proteins.
We present evidence that a well defined subset of intron positions shows a non-random distribution in ancient genes. We analyze a database of ancient conserved regions drawn from GenBank 101 to retest two predictions of the theory that the first genes were constructed by exon shuffling. These predictions are that there should be an excess of symmetric exons (and sets of exons) flanked by introns of the same phase (positions within the codon) and that intron positions in ancient proteins should correlate with the boundaries of compact protein modules. Both these predictions are supported by the data, with considerable statistical force (P values < 0.0001). Intron positions correlate to modules of diameters around 21, 27, and 33 A, and this correlation is due to phase zero introns. We suggest that 30-40% of present day intron positions in ancient genes correspond to phase zero introns originally present in the progenote, while almost all of the remaining intron positions correspond to introns added, or moved, appearing equally in all three intron phases. This proposal provides a resolution for many of the arguments of the introns-early/introns-late debate.


Ultraviolet-B-induced oxidative stress and responses of the ascorbate-glutathione cycle in a marine macroalga Ulva fasciata.
The regulation of the antioxidant defence system by ultraviolet-B (UV-B) was determined in a marine macroalga Ulva fasciata Delile exposed to low (0.5, 1 W m(-2)), medium (2.5, 5 W m(-2)), and high (10, 20 W m(-2)) UV-B irradiance. UV-B > or =2.5 W m(-2) increased H2O2 contents that are positively correlated with lipid peroxidation and total peroxide contents. Inhibition of the UV-B-induced H2O2 increase by a specific O2.- scavenger, 1,2-dihydroxy-benzene-3,5-disulphonic acid, shows that O2.- is the primary source of H2O2. Superoxide dismutase activity was increased by UV-B with a peak at 2.5 W m(-2), which did not match the H2O2 pattern. Alleviation of UV-B-induced oxidative damage by a H2O2 scavenger, dimethylthiourea, and a free radical scavenger, sodium benzoate, which inhibited UV-B-induced H2O2 accumulation, suggests that oxidative damage caused by UV-B > or = 2.5 W m(-2) is ascribed to accumulated H2O2. However, a decrease in growth rate and TTC reduction ability only at high UV-B doses indicates that the defence and repairing systems operate at low and medium UV-B doses. H2O2 not only can be excreted but can also be detoxified via the ascorbate-glutathione cycle. Increases in catalase, peroxidase, ascorbate peroxidase, and glutathione reductase activities and ascorbate (AsA) and glutathione pools, as well as AsA regeneration ability, function to keep the balance of cellular H2O2 under low UV-B doses. Dehydroascorbate reductase and monodehydroascorbate reductase are responsible for AsA regeneration under low and medium UV-B radiation, respectively. The appearance of oxidative damage in medium and high UV-B flux is attributable to a lower induction of the ascorbate-glutathione cycle as an antioxidant defence system. Overall, the availability of antioxidants and the induction of antioxidant enzyme activities for detoxifying reactive oxygen species (ROS) are regulated in U. fasciata against UV-B-induced oxidative stress, and experiments using ROS scavengers demonstrate that the antioxidant defence system is modulated by O2.- or H2O2.


The cysteine knot of platelet glycoprotein Ib beta (GPIb beta) is critical for the interaction of GPIb beta with GPIX.
The glycoprotein Ib (GPIb) complex is composed of GPIb alpha covalently attached to GPIb beta and noncovalently complexed with GPIX and GPV. Patients with Bernard-Soulier syndrome demonstrate that mutations in either GPIb beta or GPIX result in an absence of platelet GPIb alpha. This occurs through the interaction of GPIX with GPIb beta. The precise sites of interaction of GPIb beta with GPIX are not known. To characterize the interaction of GPIb beta and GPIX, we developed an anti-GPIb beta monoclonal antibody MBC 257.4, whose epitope was in the N-terminal region of GPIb beta. N-terminal truncations of GPIb beta were expressed in mammalian cells. N-terminal truncations of GPIb beta, missing the first 14, 26, or 31 amino acids, were surface-expressed but did not enable coexpressed GPIX to be surface expressed, suggesting that the site of interaction with GPIX was modified by these deletions. GPIb beta and GPIX chimeras corresponding to predicted boundaries were used to define the sites of interaction of GPIb beta with GPIX. Replacing the N-terminal disulfide loops of GPIb beta (amino acids 1-14) with the corresponding disulfide loops of GPIX (amino acids 1-22) resulted in surface expression of coexpressed wildtype GPIX. However, when the N terminus of GPIb beta was replaced to residue 32 with the N terminus of GPIX (amino acids 1-36), GPIX did not surface express with this chimera. These results suggest that the cysteine knot region of GPIb beta in the N terminus is critical for the conformation of GPIb beta that interacts with GPIX and further suggests that a critical interaction of GPIb beta with GPIX involve residues 15 through 32 of GPIb beta


The search for interstellar glycine.
Millimeter arrays can be used to identify hot young molecular cores which contain large, highly saturated interstellar molecules, including biomolecules. These cores are prime locations for searching for interstellar glycine. The current status of the interstellar glycine search is discussed.


[Bronchoalveolar lavage in childhood: technical aspects and reference values].
(BAL) has been increasingly used in recent years also in children. In paediatric pulmonary diseases, however, interpretation has proved difficult in the absence of standard reference values for BAL in childhood. It is the purpose of the present publication to discuss the methodological aspects of BAL in childhood and to proviced criteria by means of standard reference values for the assessment of BAL in paediatric patients. In a group of 50 children aged 3 to 15 years with healthy lungs, who were subjected to BAL during an elective surgical intervention, we examined both cellular and non-cellular components of the BAL. Whereas the percentage of macrophages and lymphocytes in the total population did not differ significantly from the standard values in adults, the absolute and relative granulocyte count was found to be elevated in children under 8 years of age. The CD4/CD8 quotient of BAL was 0.7 on the average, and hence clearly below the standard reference values defined for adults. In the non-cellular components we found an increase in the level of all proteins from plasma, whereas locally produced proteins did not differ from the findings in healthy adults. The total phospholipid content of BAL was markedly enhanced in children below 8 years of age and decreased with increasing age. These differences between children and adults should be taken into account when interpreting pathological findings. They are the basis for better defining the ranking of BAL in paediatric pulmonary diseases.


Methicillin-resistant Staphylococcus aureus arthritis of the wrist treated with arthrodesis using vascularized fibular transfer: a case report.
There have been few reports of vascularized fibular transfer for septic arthritis of the wrist. We report on a 52-year-old male who presented with methicillin-resistant Staphylococcus aureus (MRSA) arthritis of the wrist following a severe crush injury. We performed on this patient wrist arthrodesis using curettage of the lesion and vascularized fibular transfer with comparatively good results. Arthrodesis of the wrist using vascularized fibular transfer is useful for reconstructing bone destruction or defects accompanying intractable arthritis of the wrist such as caused by MRSA arthritis.


Mechanism of action of a new prostaglandin antihypertensive, viprostol [CL 115 347; (dl)-15-deoxy-16-hydroxy-16(alpha/beta)-vinyl-prostaglandin E2 methyl ester]: (I). Vasodilation.
Viprostol [(dl)-15-deoxy-16-hydroxy-16(alpha/beta)-vinyl-prostaglandin E2 methyl ester; CL 115 347], injected directly into coronary, renal, mesenteric, femoral and carotid arteries of the anaesthetized beagle dogs at doses which did not lower the systemic arterial blood pressure, increased blood flow of the vascular beds being studied. Viprostol was as potent as I-prostaglandin E2 (I-PGE2) in the renal bed, but less potent in the other vascular beds. Viprostol was as potent as I-PGE2 in relaxing smooth muscle of the perfused isolated central ear artery of the rabbit. Viprostol maintained its antihypertensive effect in spontaneously hypertensive rats (SHR) pretreated with indomethacin, chlorpheniramine plus cimetidine, atropine or propranolol, suggesting that the vasodilating effects of viprostol were not mediated through the endogenous prostaglandin, histamine, cholinergic nervous system or beta-adrenergic nervous system. The antihypertensive effects of viprostol were not altered in SHR with bilateral nephrectomy or bilaterally ligated ureters, suggesting that the action of viprostol was not dependent on the secretory or excretory functions of the kidneys. In conclusion, viprostol exerts its antihypertensive action mainly by vasodilation in a way similar to the natural PGE2.


Pregnancy with macroprolactinoma.
A macroprolactinoma recurred in a 25-year-old lady, who had initially presented with inability to conceive, secondary amenorrhea, galactorrhea and persistent headache. She was diagnosed as a patient of pituitary macroadenoma of 1.7 cm with elevated serum prolactin level. She was given Bromocriptine, which normalized her menstruation as well as the prolactin level followed by conception during treatment. Pregnancy remained uneventful till 27 weeks when she developed severe headache and total loss of vision from left eye and partial from right eye at 27 weeks. MRI showed enlargement of macroadenoma upto 2.5 cm with compression on optic chiasma. Transsphenoidal adenectomy was performed. After surgery visual field defect improved but plasma prolactin level remained elevated. She delivered vaginally at 39 weeks. Later, treatment with Bromocriptine (15 mg/day) failed to keep prolactin level normal and Lisuride hydrogen (0.8 mg/day) reduced the prolactin levels.


AACR-FDA-NCI cancer biomarkers collaborative.
The past few years have seen a frantic effort in an attempt to discover and validate novel biomarkers for the diagnosis and treatment of diseases such as cancer. Unfortunately, the rate at which biomarkers are being used in actual patient treatment is not increasing accordingly. There are a number of reasons for this discrepancy; one is the failure to recognize and appreciate which validated biomarkers have the greatest chance of being used in a clinical setting. In addition, there is a general lack of understanding within the basic research community of the intricacies involved in the regulatory process required to produce a clinically useful biomarker. Fortunately, a collaboration involving the American Association for Cancer Research, the National Cancer Institute and the US FDA hopes to expedite the process of bringing validated biomarkers to the point where they can have an impact on the diagnosis and treatment of disease.


[Clinical applications of beta-thromboglobulin and platelet factor 4 radioimmune assays (author's transl)].
beta-Thromboglobulin (beta-TG) and the anti-heparin activity platelet factor 4 (PF4) were simultaneously assayed by a radioimmune method in 108 subjects, most of whom had already be studied for platelet kinetics. The following data were obtained : while there was no technical cross-reaction, the two assays correlated closely, which indicates that they explore the same physiological phenomena : when controls were chosen at random in a population of wide age range, considerable differences were observed in the results, the values that could be considered excessive in a large percentage of cases, particularly among old, "normal" individuals ; there was no correlation between beta-TG and PF4 values on the one hand platelet destruction rate on the other ; plasma levels of beta-TG and PF4 were significantly higher in patients with vascular diseases and polycythaemia vera than in control but clearly overlapped with the values observed in controls of the same age group. The clinical significance of these data requires, to be assessed, a long-term prospective study aimed at determining whether patients with high beta-TG and PF4 levels present a high risk of thromboembolic complications and whether this risk can be reduced by correcting the biological abnormality with agents altering platelet function.


Does echocardiography allow the monitoring of the cardiac effects of nitrates?
Echocardiography is a very useful non-invasive method of cardiac diagnosis. It has been widely used in evaluating the effects of some drugs, such as nitrates, prazosin, and propranolol, on heart and seems very successful. In this paper, the echocardiographic studies of the effects of nitrates on left ventricular dimension and volume have been reviewed and an attempt has been made to answer whether or not echocardiography can allow to monitor the effects of nitrates in patient groups and further in individual patients.


Energy Landscapes and Heat Capacity Signatures for Monomers and Dimers of Amyloid-Forming Hexapeptides.
Amyloid formation is a hallmark of various neurodegenerative disorders. In this contribution, energy landscapes are explored for various hexapeptides that are known to form amyloids. Heat capacity (CV) analysis at low temperature for these hexapeptides reveals that the low energy structures contributing to the first heat capacity feature above a threshold temperature exhibit a variety of backbone conformations for amyloid-forming monomers. The corresponding control sequences do not exhibit such structural polymorphism, as diagnosed via end-to-end distance and a dihedral angle defined for the monomer. A similar heat capacity analysis for dimer conformations obtained using basin-hopping global optimisation shows clear features in end-to-end distance versus dihedral correlation plots, where amyloid-forming sequences exhibit a preference for larger end-to-end distances and larger positive dihedrals. These results hold true for sequences taken from tau, amylin, insulin A chain, a de novo designed peptide, and various control sequences. While there is a little overall correlation between the aggregation propensity and the temperature at which the low-temperature CV feature occurs, further analysis suggests that the amyloid-forming sequences exhibit the key CV feature at a lower temperature compared to control sequences derived from the same protein.


Impairments in glucose tolerance can have a negative impact on cognitive function: a systematic research review.
There is an increasing body of research investigating whether abnormal glucose tolerance is associated with cognitive impairments, the evidence from which is equivocal. A systematic search of the literature identified twenty-three studies which assessed either clinically defined impaired glucose tolerance (IGT) or variance in glucose tolerance within the clinically defined normal range (NGT). The findings suggest that poor glucose tolerance is associated with cognitive impairments, with decrements in verbal memory being most prevalent. However, the evidence for decrements in other domains was weak. The NGT studies report a stronger glucose tolerance-cognition association than the IGT studies, which is likely to be due to the greater number of glucose tolerance parameters and the more sensitive cognitive tests in the NGT studies compared to the IGT studies. It is also speculated that the negative cognitive impact of abnormalities in glucose tolerance increases with age, and that glucose consumption is most beneficial to individuals with poor glucose tolerance compared to individuals with normal glucose tolerance. The role of potential mechanisms are discussed.


Role of nitric oxide-dependent and -independent mechanisms in peristalsis and accommodation in the rabbit distal colon.
Previous findings, which have been confirmed in this study, indicate that nitric oxide (NO)-dependent and -independent (apamin-sensitive) mechanisms underlie the electrically induced non-adrenergic, noncholinergic (NANC) relaxation in the circular muscle of rabbit distal colon. Based on this evidence, we further investigated whether, and to what extent, these separate NANC components participate in the maintenance of circular muscle tone, reflex relaxations evoked by localized balloon distension or during peristalsis (descending inhibition) and in the accommodation of colonic wall in response to graded intraluminal fluid delivery. NG-nitro-L-arginine (L-NNA) (10-300 microM), apamin (100 nM) and tetrodotoxin (60 nM), enhanced the spontaneous low tone and phasic activity in circular muscle strips. In experiments on peristalsis, L-NNA (30 and 300 microM) shortened the latency of peristaltic wave initiation and increased the velocity of propulsion of an intraluminally distended balloon (range, 0.1-1 ml). The latter effect was mimicked by the NO scavenger oxyhemoglobin (30 microM). Velocity of propulsion was enhanced by apamin (100 nM) at low balloon distension (0.1 and 0.2 ml), whereas it was reduced at high distension volumes (1 ml), due to disruption of descending inhibition. A combination of L-NNA (300 microM) and apamin (100 nM) blocked peristalsis, due to persisting spasms of the circular muscle. L-NNA (300 microM) did not affect the amplitude of distension-evoked ascending reflex contraction and slightly inhibited the descending reflex relaxation. By contrast, the latter reflex was virtually abolished by apamin (100 nM).(ABSTRACT TRUNCATED AT 250 WORDS)


[Corona virus disease 2019 lesion segmentation network based on an adaptive joint loss function].
Corona virus disease 2019 (COVID-19) is an acute respiratory infectious disease with strong contagiousness, strong variability, and long incubation period. The probability of misdiagnosis and missed diagnosis can be significantly decreased with the use of automatic segmentation of COVID-19 lesions based on computed tomography images, which helps doctors in rapid diagnosis and precise treatment. This paper introduced the level set generalized Dice loss function (LGDL) in conjunction with the level set segmentation method based on COVID-19 lesion segmentation network and proposed a dual-path COVID-19 lesion segmentation network (Dual-SAUNet++) to address the pain points such as the complex symptoms of COVID-19 and the blurred boundaries that are challenging to segment. LGDL is an adaptive weight joint loss obtained by combining the generalized Dice loss of the mask path and the mean square error of the level set path. On the test set, the model achieved Dice similarity coefficient of (87.81 ± 10.86)%, intersection over union of (79.20 ± 14.58)%, sensitivity of (94.18 ± 13.56)%, specificity of (99.83 ± 0.43)% and Hausdorff distance of 18.29 ± 31.48 mm. Studies indicated that Dual-SAUNet++ has a great anti-noise capability and it can segment multi-scale lesions while simultaneously focusing on their area and border information. The method proposed in this paper assists doctors in judging the severity of COVID-19 infection by accurately segmenting the lesion, and provides a reliable basis for subsequent clinical treatment.


Response of a seagrass fish assemblage to improved wastewater treatment.
We compared the structure of a seagrass fish assemblage near a sewage outlet before and after improvements to wastewater treatment. To determine whether responses by the fish assemblage were due to changes in water quality or to other factors, comparisons were made with the structure of a fish assemblage from a nearby site unaffected by sewage effluent. Total species richness, density and biomass of fish, decreased at both sites over the 30-year period. An increase in mean trophic level near the sewage outlet following improvements in water quality indicated that wastewater treatment had another important effect. This result is consistent with the reductions in food webs supporting pelagic and benthic fishes that typically accompany decreases in nutrient inputs. Although improvements to wastewater treatment explained much of the variation in the structure of the fish assemblage at PC, our results also suggest that fishing and climate change, at both sites.


Synergistic antitumor activity of triple-regulated oncolytic adenovirus with VSTM1 and daunorubicin in leukemic cells.
V-set and transmembrane domain-containing 1 (VSTM1), which is downregulated in bone marrow cells from leukemia patients, may provide a diagnostic and treatment target. Here, a triple-regulated oncolytic adenovirus was constructed to carry a VSTM1 gene expression cassette, SG611-VSTM1, and contained the E1a gene with a 24-nucleotide deletion within the CR2 region under control of the human telomerase reverse transcriptase promoter, E1b gene directed by the hypoxia response element, and VSTM1 gene controlled by the cytomegalovirus promoter. Real-time quantitative PCR and Western blot analyses showed that SG611-VSTM1 expressed VSTM1 highly efficiently in the human leukemic cell line K562 compared with SG611. In Cell Counting Kit-8 and flow cytometric assays, SG611-VSTM1 exhibited more potent anti-proliferative and pro-apoptotic effects in leukemic cells compared with SG611 and exerted synergistic cytotoxicity with low-dose daunorubicin (DNR) in vitro. In xenograft models, SG611-VSTM1 intratumorally injected at a dose of 1 × 10(9) plaque forming units combined with intraperitoneally injected low-dose DNR displayed significantly stronger antitumor effects than either treatment alone. Histopathologic examination revealed that SG611-VSTM1 induced apoptosis of leukemic cells. These results implicate an important role for VSTM1 in the pathogenesis of leukemia, and SG611-VSTM1 may be a promising agent for enhancing chemosensitivity in leukemia therapy.


Viloxazine extended-release capsules for the treatment of attention-deficit/ hyperactivity disorder in adult patients.
Attention-deficit/hyperactivity disorder (ADHD) is a common neurobehavioral disorder with symptoms that may persist in up to 90% of adults diagnosed during childhood and continue to cause significant impairment throughout the lifespan. In the United States (US), amphetamine and methylphenidate formulations have been available to treat ADHD for several decades. Only one nonstimulant, atomoxetine, was available for the treatment of ADHD in adults until recently. In April 2022, a second nonstimulant, viloxazine extended-release (VLX-ER), became available in the US for the treatment of adult ADHD. Efficacy was previously established in placebo-controlled trials in children and adolescents. VLX-ER is a norepinephrine reuptake inhibitor with serotonin activity. The efficacy in adults, adverse event profile, pharmacokinetics, drug-drug interactions, and metabolism of VLX-ER are reviewed. Despite the availability of effective pharmacological treatments for ADHD, many patients discontinue treatment in less than 1 year. Stimulants are effective in more than 80% of patients; however, some may have difficulty tolerating them. Although there were no head-to-head studies, the effect size of VLX-ER in an adult efficacy trial was lower than has been shown for stimulants. Nevertheless, the approval of VLX-ER adds another effective ADHD treatment option for adults.


[Ocular penetration of dexamethasone sodium phosphate via soft contact lenses].
The ocular penetration of dexamethasone sodium phosphate was measured by HPLC and RIA, and the concentrations in the aqueous humor and iris of a soft contact lens (SCL) group and eyedrop-only group of white rabbits were compared. The efficacy of dexamethasone phosphate eyedrops in eyes of white rabbits with SCL was also investigated using protein in the aqueous humor of the eyes of rabbits with experimental endophthalmitis induced by endotoxin and prostaglandin E2 as an index. The concentration of dexamethasone sodium phosphate transferred to the aqueous humor and irises in the SCL group was significantly higher than that in the eyedrop-only group. SCLs containing dexamethasone sodium phosphate was effective against experimental endophthalmitis, and the protein in the aqueous humor was significantly lower in the SCL group than in the controls.


Relationship Between Objective Financial Burden and the Health-Related Quality of Life and Mental Health of Patients With Cancer.
The high and increasing costs of cancer care can lead to financial burden for patients and their families. However, no study has specifically examined the association between objective measures of financial burden and the health-related quality of life (HRQOL) and psychological health of patients with cancer. Data on patients with cancer were obtained from the 2010 to 2014 Medical Expenditure Panel Survey. High financial burden was defined as a ratio of annual family out-of-pocket health care expenditures to family income exceeding either 10% or 20%. Multivariable linear regressions were used to estimate the relationship between high financial burden and patient HRQOL, nonspecific psychological distress, and depressed mood. Of 6,799 patients with cancer, 15% and 6% experienced high financial burden exceeding 10% and 20% of family income, respectively. Compared with those without high financial burden, patients with cancer with high financial burden were more likely to have significantly lower HRQOL and a greater tendency toward nonspecific psychological distress. Higher levels of financial burden were associated with lower HRQOL, with a stronger relationship observed with physical health than with mental health and a greater tendency toward nonspecific psychological distress. Depressed mood was not significantly associated with high financial burden. High financial burden among patients with cancer was significantly associated with lower HRQOL and poor mental health. Along with efforts to reduce health care costs for cancer survivors, additional interventions are necessary to ensure the HRQOL and psychological health of cancer survivors.


Efficacy assessment of sustained intraperitoneal paclitaxel therapy in a murine model of ovarian cancer using bioluminescent imaging.
We evaluated the pre-clinical efficacy of a novel intraperitoneal (i.p.) sustained-release paclitaxel formulation (PTX(ePC)) using bioluminescent imaging (BLI) in the treatment of ovarian cancer. Human ovarian carcinoma cells stably expressing the firefly luciferase gene (SKOV3(Luc)) were injected i.p. into SCID mice. Tumour growth was evaluated during sustained or intermittent courses of i.p. treatment with paclitaxel (PTX). In vitro bioluminescence strongly correlated with cell survival and cytotoxicity. Bioluminescent imaging detected tumours before their macroscopic appearance and strongly correlated with tumour weight and survival. As compared with intermittent therapy with Taxol, sustained PTX(ePC) therapy resulted in significant reduction of tumour proliferation, weight and BLI signal intensity, enhanced apoptosis and increased survival times. Our results demonstrate that BLI is a useful tool in the pre-clinical evaluation of therapeutic interventions for ovarian cancer. Moreover, these results provide evidence of enhanced therapeutic efficacy with the sustained PTX(ePC) implant system, which could potentially translate into successful clinical outcomes.


Frontal sinus cementifying ossifying fibroma.
Cementifying ossifying fibroma is a fibro-osseous lesion which produces cementum. Cementifying ossifying fibroma is a mesodermal, non-odontogenic tumor of ectopic multipotential periodontal membrane blast cells. Although, it is a benign bone tumor, it has an aggressive destructive behavior in patients under 20 years of age with high tendency for recurrence. Cementifying ossifying fibroma can be recognized microscopically by the presence of cementicles. This is a case of frontal sinus cementifying ossifying fibroma presenting with proptosis in a 39-year-old male.


Isoflurane is a suitable alternative to ether for anesthetizing rats prior to euthanasia for gene expression analysis.
Diethyl ether (ether) had been widely used in Japan for anesthesia, despite its explosive properties and toxicity to both humans and animals. We also had used ether as an anesthetic for euthanizing rats for research in the Toxicogenomics Project (TGP). Because the use of ether for these purposes will likely cease, it is required to select an alternative anesthetic which is validated for consistency with existing TGP data acquired under ether anesthesia. We therefore compared two alternative anesthetic candidates, isoflurane and pentobarbital, with ether in terms of hematological findings, serum biochemical parameters, and gene expressions. As a result, few differences among the three agents were observed. In hematological and serum biochemistry analysis, no significant changes were found. In gene expression analysis, four known genes were extracted as differentially expressed genes in the liver of rats anesthetized with ether, isoflurane, or pentobarbital. However, no significant relationships were detected using gene ontology, pathway, or gene enrichment analyses by DAVID and TargetMine. Surprisingly, although it was expected that the lung would be affected by administration via inhalation, only one differentially expressed gene was extracted in the lung. Taken together, our data indicate that there are no significant differences among ether, isoflurane, and pentobarbital with respect to effects on hematological parameters, serum biochemistry parameters, and gene expression. Based on its smallest affect to existing data and its safety profile for humans and animals, we suggest isoflurane as a suitable alternative anesthetic for use in rat euthanasia in toxicogenomics analysis.


Determination of the liver cytosolic proteins that bind to p-hydroxyacetophenone.
The purpose of the present study was to determine the proteins that bind to acetophenones in the liver. Immobilized p-hydroxyacetophenone (p-HAP) was used as a ligand of affinity chromatography. Analysis using sodium dodesyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) demonstrated that five polypeptides in the liver cytosolic fraction specifically bound to the p-HAP matrix. These polypeptides were digested with Lys-specific protease and used to generate peptide maps by reversed-phase high-performance liquid chromatography. Consequently, identification from a data base of protein sequences revealed that the five polypeptides were glycogen phosphorylase, cytosolic aldehyde dehydrogenase, adenosine kinase, class I alcohol dehydrogenase and glutathione S-transferase A2. In addition to p-HAP, acetylsalicylic acid also displayed a prominent ability to elute these five enzymes from the p-HAP affinity column loaded with the cytosolic fraction of the liver. Thus, p-HAP has affinities to the above liver enzymes and is a useful ligand for analysis of them.


[Cancer in the elderly. Clinico-epidemiologic considerations].
The elderly population is rapidly increasing in Western countries. Old age indirectly represents one of the most important risk factors for the development of neoplasias. In Italy in 1991 a total of 14,572 deaths from tumours were recorded, equivalent to 27% of all deaths; of these over 66% affected individuals aged over 65 years old. In the 65-74 age bracket mortality caused by tumour is the prime cause of death in Italy and is undergoing an exponential increase over time. The most frequently found tumours and above all the cause of the greatest number of deaths in the elderly population are lung cancer, prostate cancer, breast cancer, colorectal cancer, stomach cancer. To date the only weapon that has proved efficacious in some tumours (breast, colon-rectal, prostate, melanoma) is early diagnosis through screening and adequate treatment. The elderly are an extremely heterogeneous population and it is therefore not easy to provide guidelines that are applicable to the entire population. Firstly, it is important to provide more information in order to permit effective and targeted prevention. In order to fulfil this objective, structures offering the necessary equipment and skills will have to be create; training must be provided for personnel in this type of preventive medicine; lastly, the awareness of doctors towards this type of medicine must be increased by providing the correct information.


Localization, characterization, and second messenger coupling of pituitary adenylate cyclase-activating polypeptide receptors in the fetal human adrenal gland during the second trimester of gestation.
The distribution and pharmacological properties of pituitary adenylate cyclase-activating polypeptide (PACAP) receptors were studied in the fetal human adrenal gland during the second trimester of gestation. Autoradiographic studies, using [125I]PACAP27 as a radioligand, revealed that PACAP-binding sites are exclusively located on chromaffin cells of adrenals from fetuses 14-20 weeks old. Biochemical characterization of binding revealed the occurrence of a single class of PACAP-binding sites with a dissociation constant value of 0.32-0.74 nmol/L and a binding capacity of 0.30-0.81 pmol/mg wet tissue. PACAP27 and PACAP38 were equipotent in competing for [125I]PACAP27 binding (IC50 = 0.28-0.64 nmol/L and 0.15-0.81 nmol/L, respectively), and the Hill coefficients were close to 1. In contrast, vasoactive intestinal polypeptide was much less efficient in displacing the tracer (IC50 = 4-362 nmol/L), and the Hill coefficients were less than 0.6. PACAP38 induced a dose-dependent increase in cAMP production in fetal human adrenal cell suspension (ED50 = 0.07 +/- 0.02 nmol/L), as well as in cells maintained in culture for 5 days (5.4 +/- 1.8 nmol/L). In contrast, PACAP38 induced a modest increase in inositol phosphate formation. These data indicate that type I PACAP receptors are present in the early stages of the human medulla organization during the process of migration of chromaffin cells from the periphery to the central part of the gland. The present results suggest that PACAP could be involved in the regulation of the human adrenochromaffin cells during ontogenesis.


Distinct mutations in two patients with leukocyte adhesion deficiency and their functional correlates.
Two patients with leukocyte adhesion deficiency (LAD), one with a moderate phenotype (patient 14) and one with a severe phenotype (patient 2) who had been shown to have a normal sized beta subunit protein precursor, were analyzed in an attempt to determine the molecular basis for their disease. RNase mapping located possible mutations to two distinct but adjacent regions of the beta subunit cDNA. Sequencing of patient-derived cDNA clones in this region revealed a C for T difference at amino acid 149 in patient 14 which resulted in the substitution of a leucine for a proline, and an A for G substitution at amino acid 169 in patient 2 which mutated a glycine to an arginine. The mutated amino acids are in a region of the cDNA that is highly conserved between the beta subunits of the integrin family and are identical in all known integrin beta subunits. Co-transfection of the beta subunit cDNA containing the patient 2 mutation with the wild-type alpha subunit of LFA-1 in a mammalian expression system resulted in no expression of LFA-1. In the case of the mutation in patient 14 there was markedly diminished expression of LFA-1 with loss of function and loss of the epitope for a number of anti-beta mAbs. Normal half-life of the mutant beta subunits, and previous demonstration of a lack of alpha/beta complex formation during biosynthesis in patient cells, suggest a defect in association with the alpha subunit. Association with beta is required for expression of the alpha subunit of LFA-1. Loss of functional expression with both of these beta subunit mutations suggests that they lie in a site critical for association with the alpha subunit.


Adult respiratory distress syndrome and sepsis.
In the 25 yrs since the original description of adult respiratory distress syndrome, much has been learned concerning the pathology and pathophysiology of the syndrome. Investigations into the humoral mediator of the syndrome are proceeding. Despite these advances and the advances in intensive care medicine, adult respiratory distress syndrome remains a considerable clinical challenge, especially when associated with sepsis.


Oleodendrimers: a novel class of multicephalous heterolipids as chemical penetration enhancers for transdermal drug delivery.
This paper reports synthesis and evaluation of Janus type generation G-1 and G-2 dendrimers. The dendrimers have been constructed by linking two building blocks, dendrons and oleic acid, through ester and amide bonds and were well characterized by Fourier-transform infrared (FT-IR), (1)H NMR, (13)C NMR and electrospray ionization mass spectrometry (ESI-MS). The dendrimers have been evaluated for in vitro cytotoxicity using sulforhodamine B assay (SRB assay) and in vivo skin irritation potential. The ester linked dendrimers did not exhibit any cytotoxicity even up to 80 μg/ml while G-1 and G-2 generations dendrimers with amide linkage exhibited toxicity above 70 μg/ml and 21 μg/ml, respectively, none of the dendrimers showed any skin irritation. All the dendrimers, tested for their skin permeation enhancement potential using diclofenac sodium (DS) as a model drug at a concentration of 1% in gels, showed significant increase in steady-state flux (ERflux) of the drug as compared to control (without enhancer), and oleic acid. Amongst the dendrimers, the ester linked G-1 and G-2 dendrimers showed highest ERflux, 3.33 ± 0.31 and 3.39 ± 0.21, respectively.


Cryopreservation of equine embryos: current state-of-the-art.
During the past 15 years, embryo transfer (ET) has become increasingly widespread within the sport-horse breeding industry. At present, however, the vast majority (>95%) of horse embryos are transferred fresh or after chilled storage for up to 24 h, whereas cryopreservation is rarely employed despite its obvious potential for simplifying recipient mare management and facilitating long-term storage and international transport of embryos. A number of inter-related factors have contributed to the slow development and implementation of equine embryo cryopreservation, and these include the following: (i) the absence of commercially available products for reliably stimulating superovulation; (ii) very poor pregnancy rates following cryopreservation of embryos >300 μm in diameter; (iii) difficulty in recovering embryos at early developmental stages amenable to cryopreservation; and (iv) interembryo variation in susceptibility to cryodamage. However, acceptable success rates (>55% pregnancy) have been reported for both slow-frozen and vitrified small embryos (<300 μm), and there is renewed interest in cryopreservation, not only in the context of standard ET programmes, but also because it would facilitate pre-implantation genetic testing and allow wider access to techniques for producing embryos in vitro, such as intracytoplasmic sperm injection and nuclear transfer. This article will review the current status of equine embryo cryopreservation.


The dexamethasone suppression test for major depression among opiate addicts.
The authors gave the dexamethasone suppression test (DST) to 42 opiate addicts 2 weeks after detoxification. The DST provided a sensitivity of 80% and a specificity of 93% for major depression diagnosed according to the Research Diagnostic Criteria.


Activation of the alternative complement pathway in ascitic fluid during spontaneous bacterial peritonitis.
We studied complement and immunoglobulin profiles on the serum and ascitic fluid of a patient before and during gram-negative spontaneous bacterial peritonitis (SBP). During the infection, activation of the alternative complement pathway in ascitic fluid was manifested by a 35% reduction in functional activity and depression of both properdin and factor B concentrations to nondetectable levels. Activation of the complement cascade was also demonstrated by a 50% reduction in the C3 concentration and depression of total hemolytic complement. There was no evidence of complement activation of a functionally intact complement system in the ascitic fluid of cirrhotic patients. Complement consumption in ascitic fluid may predispose the cirrhotic to SBP.


EVIDENCE THAT AN OUTCROSSING POPULATION IS A DERIVED LINEAGE IN A HERMAPHRODITIC FISH (RIVULUS MARMORATUS).
Rivulus marmoratus is the only known vertebrate with obligate, synchronous hermaphroditic fertilization. Males can be experimentally induced in the laboratory and are rare or absent in most populations, but at the isolated Twin Cays, Belize, locality, males are relatively abundant. At this locality, evidence of outcrossing has been documented in this otherwise automictic cloning species. Phylogenetic analysis of restriction sites and sequence characters revealed that all Florida and Belize western Caribbean populations (including Twin Cays) are phyletically indistinguishable yet divergent from eastern populations in Brazil and the Bahamas. Further, these western lineages shared a common ancestor more recently than all other populations. Therefore, the Twin Cays population is not a remnant ancestral outcrossing population. Outcrossing is suspected to have evolved as a phenotypically plastic character, and its expression in R. marmoratus may be dormant unless triggered by some ecological factor that is not well understood.


Manganese exposure and neurotoxic effects in children.
Manganese (Mn) is the fifth most abundant metal on earth. Although it is a well understood essential trace element, in excess, Mn is neurotoxic. Initial toxic symptoms associated with Mn are of psychiatric nature and are clinically defined as locura manganica. Neurological signs of Mn toxicity include dystonia, progressive bradykinesia, and disturbance of gait, slurring, and stuttering of speech with diminished volume. Studies indicate that children who ingested Mn in the drinking water (WMn) at or above a level of 0.241mg/L for a minimum of three years performed more poorly in school as measured by mastery of language, mathematics, and in their overall grade average. The Mn-exposed children also performed more poorly on a battery of neurobehavioral tests. It was also found a significant association between higher WMn and lower cognitive performance, verbal function, and full-scale intelligence quotient (IQ) scores. Young children appear to make up a vulnerable group in exposed populations. Toxicity of WMn is a problem particularly in areas of industrial waste or where Mn is leaching from the soil into public drinking water. Practical and cost-effective approaches are available to remove Mn from drinking water. It is crucial to protect developing brains against Mn toxicity.


2-Thiopheneacetic acid directed synthesis of Au nanorosette as an SERS-active substrate.
Rosette-like nanoscale Au materials were simply prepared through one-pot reduction of the AuCl₄⁻ precursor by 2-thiopheneacetic acid (2-TAA) without extra surface capping ligands at room temperature. 2-TAA underwent polymerization into polythiophene derivatives while the AuCl₄⁻ precursor was simultaneously reduced into various Au nanostructures. In situ generated polythiophene derivatives played a significant role of surface passivation in guiding the shape of Au nanostructures. The morphology of Au nanostructures was strongly dependent on the molar ratio of 2-TAA to AuCl₄⁻. At lower [2-TAA]/[AuCl₄⁻] ratios, uniform rosette-like Au microparticles consisting of 30-nm-thick Au nanoplates were observed. The Au nanoplates had either triangular prismatic or hexagonal geometry with many defects. Uniform Au nanorosettes could be easily deposited on the Si substrate by drop-casting and were subsequently used as highly active SERS substrates for the detection of methylene blue and crystal violet by Raman spectroscopy. Upon increasing the ratio of [2-TAA]/[AuCl₄⁻], Au nanoparticles or nanorods heavily surrounded with polythiophene polymers were obtained.


Low Buffer Capacity and Alternating Motility along the Human Gastrointestinal Tract: Implications for in Vivo Dissolution and Absorption of Ionizable Drugs.
In this study, we determined the pH and buffer capacity of human gastrointestinal (GI) fluids (aspirated from the stomach, duodenum, proximal jejunum, and mid/distal jejunum) as a function of time, from 37 healthy subjects after oral administration of an 800 mg immediate-release tablet of ibuprofen (reference listed drug; RLD) under typical prescribed bioequivalence (BE) study protocol conditions in both fasted and fed states (simulated by ingestion of a liquid meal). Simultaneously, motility was continuously monitored using water-perfused manometry. The time to appearance of phase III contractions (i.e., housekeeper wave) was monitored following administration of the ibuprofen tablet. Our results clearly demonstrated the dynamic change in pH as a function of time and, most significantly, the extremely low buffer capacity along the GI tract. The buffer capacity on average was 2.26 μmol/mL/ΔpH in fasted state (range: 0.26 and 6.32 μmol/mL/ΔpH) and 2.66 μmol/mL/ΔpH in fed state (range: 0.78 and 5.98 μmol/mL/ΔpH) throughout the entire upper GI tract (stomach, duodenum, and proximal and mid/distal jejunum). The implication of this very low buffer capacity of the human GI tract is profound for the oral delivery of both acidic and basic active pharmaceutical ingredients (APIs). An in vivo predictive dissolution method would require not only a bicarbonate buffer but also, more significantly, a low buffer capacity of dissolution media to reflect in vivo dissolution conditions.


DCDS: A Real-time Data Capture and Personalized Decision Support System for Heart Failure Patients in Skilled Nursing Facilities.
Heart disease is the leading cause of death in the United States. Heart failure disease management can improve health outcomes for elderly community dwelling patients with heart failure. This paper describes DCDS, a real-time data capture and personalized decision support system for a Randomized Controlled Trial Investigating the Effect of a Heart Failure Disease Management Program (HF-DMP) in Skilled Nursing Facilities (SNF). SNF is a study funded by the NIH National Heart, Lung, and Blood Institute (NHLBI). The HF-DMP involves proactive weekly monitoring, evaluation, and management, following National HF Guidelines. DCDS collects a wide variety of data including 7 elements considered standard of care for patients with heart failure: documentation of left ventricular function, tracking of weight and symptoms, medication titration, discharge instructions, 7 day follow up appointment post SNF discharge and patient education. We present the design and implementation of DCDS and describe our preliminary testing results.


Identification of N epsilon-carboxymethyllysine as a degradation product of fructoselysine in glycated protein.
The chemistry of Maillard or browning reactions of glycated proteins was studied using the model compound, N alpha-formyl-N epsilon-fructoselysine (fFL), an analog of glycated lysine residues in protein. Incubation of fFL (15 mM) at physiological pH and temperature in 0.2 M phosphate buffer resulted in formation of N epsilon-carboxymethyllysine (CML) in about 40% yield after 15 days. CML was formed by oxidative cleavage of fFL between C-2 and C-3 of the carbohydrate chain and erythronic acid (EA) was identified as the split product formed in the reaction. Neither CML nor EA was formed from fFL under a nitrogen atmosphere. The rate of formation of CML was dependent on phosphate concentration in the incubation mixture and the reaction was shown to occur by a free radical mechanism. CML was also identified by amino acid analysis in hydrolysates of both poly-L-lysine and bovine pancreatic ribonuclease glycated in phosphate buffer under air. CML was also detected in human lens proteins and tissue collagens by HPLC and the identification was confirmed by gas chromatography/mass spectroscopy. The presence of both CML and EA in human urine suggests that they are formed by degradation of glycated proteins in vivo. The browning of fFL incubation mixtures proceeded to a greater extent under a nitrogen versus an air atmosphere, suggesting that oxidative degradation of Amadori adducts to form CML may limit the browning reactions of glycated proteins. Since the reaction products, CML and EA, are relatively inert, both chemically and metabolically, oxidative cleavage of Amadori adducts may have a role in limiting the consequences of protein glycation in the body.


[Treatment of unstable thoracolumbar spinal injuries--determination of its present status and a preliminary report on transpedicular stabilization].
Follow-up checks were made on 19 patients with instabile injuries of the thoracolumbar spine. Poor late results were recorded from eleven of them who had undergone conservative or inadequate surgical treatment. Axial malposition of severe clinical disorders were among the findings. Transpedicular osteosynthesis of instable fractures of thoracic or lumbar regions of the spine has been practised by the authors since 1986, with plate fixation of only two vertebral segments. Pathoneurological disorders are handled by laminectomy or hemilaminectomy followed by H-shaped interspine bone grafting. Posterior rim fragments, burst off the spine, are either reduced or removed. Intraoperative perimyelography is possible. Therapeutic exercises are initiated immediately after the operation. After wound healing, the patient is allowed to get out of bed, supported by a plaster corset. Eleven patients have so far undergone the above surgical treatment. Four of them have resumed their original jobs, whereas three patients with paraplegia have been transferred to a rehabilitation centre.


Protective effect of olopatadine hydrochloride against LPS-induced acute lung injury: via targeting NF-κB signaling pathway.
Morbidity and mortality rates associated with acute lung injury/acute respiratory distress syndrome (ALI/ARDS) are high (30-40%). Nuclear factor-kappa B (NF-κB) is a transcription factor, associated with transcription of numerous cytokines leading to cytokine storm, and thereby, plays a major role in ALI/ARDS and in advanced COVID-19 syndrome. Considering the role of NF-κB in ALI, cost-effective in silico approaches were utilized in the study to identify potential NF-κB inhibitor based on the docking and pharmacokinetic results. The identified compound was then pharmacologically validated in lipopolysaccharide (LPS) rodent model of acute lung injury. LPS induces ALI by altering alveolar membrane permeability, recruiting activated neutrophils and macrophages to the lungs, and compromising the alveolar membrane integrity and ultimately impairs the gaseous exchange. Furthermore, LPS exposure is associated with exaggerated production of various proinflammatory cytokines in lungs. Based on in silico studies Olopatadine Hydrochloride (Olo), an FDA-approved drug was found as a potential NF-κB inhibitor which has been reported for the first time, and considered further for the pharmacological validation. Intraperitoneal LPS administration resulted in ALI/ARDS by fulfilling 3 out of the 4 criteria described by ATS committee (2011) published workshop report. However, treatment with Olo attenuated LPS-induced elevation of proinflammatory markers (IL-6 and NF-κB), oxidative stress, neutrophil infiltration, edema, and damage in lungs. Histopathological studies also revealed that Olo treatment significantly ameliorated LPS-induced lung injury, thus conferring improvement in survival. Especially, the effects produced by Olo medium dose (1 mg/kg) were comparable to dexamethasone standard. In nutshell, inhibition of NF-κB pathway by Olo resulted in protection and reduced mortality in LPS- induced ALI and thus has potential to be used clinically to arrest disease progression in ALI/ARDS, since the drug is already in the market. However, the findings warrant further extensive studies, and also future studies can be planned to elucidate its role in COVID-19-associated ARDS or cytokine storm.


Consumer perspectives on a pericoital contraceptive pill in India and Uganda.
Studies suggest that women in some countries have adopted emergency contraceptive pills as a routine method of family planning. This practice indicates there may be latent demand for a pericoital contraceptive pill taken only when a woman has sexual intercourse, and labeled and marketed for use as a regular contraceptive method. To understand the appeal and potential market for a pericoital contraceptive pill, 39 focus groups and 23 in-depth interviews were conducted with women and men in Lucknow, Uttar Pradesh, India, and Kampala, Uganda. A total of 281 individuals participated in this qualitative study. In general, women embraced the idea of a female-controlled method that would be easier than taking a daily oral contraceptive pill and that could be taken either before or after sexual intercourse; in Uganda, especially, women approved of the fact that the method could be taken without a partner's knowledge. Although we do not yet know the extent of side effects for this method, women expressed some concerns about the level and nature of potential side effects. The results suggest that a pericoital contraceptive pill would be well-received by consumers in both country settings. If its efficacy and side effects are acceptable, a pericoital contraceptive pill could fill a gap for female-controlled, discreet, coitus-related contraception, particularly among women who do not have sex very frequently.


Granulation tissue polyposis associated with carcinoid tumours of the small intestine.
We report two patients with ileal carcinoid tumours which were associated with polyps due to mucosal granulation tissue proliferation. In both cases the tumours had extensively infiltrated the small bowel wall and mesentery, and one had hepatic metastases. The mucosal surface of each specimen showed numerous, pale brown, sessile polyps which were restricted to the intestinal segment involved by carcinoid tumour, although not always closely related to neoplastic cells. The polyps were formed by the proliferation of capillaries, smooth muscle cells and myofibroblasts as demonstrated by immunohistochemistry and electronmicroscopy.


Nano-conjugate fluorescence probe for the discrimination of phosphate and pyrophosphate.
We describe a pyrophosphate (PPi) probe that is based on a fluorescent dicarboxylate-substituted poly(para-phenyleneethynylene) (PPE) and 10 nm cobalt-iron spinel nanoparticles (NPs) in aqueous media. The spinel NPs efficiently quench the fluorescence of the PPE at a concentration of 20-30 pmol. Addition of phosphate anions to the PPE-NP construct displaces the quenched PPE to give rise to a fluorescent response; we found that PPi and phosphate (Pi) have significantly different binding affinities for the self-assembled materials. We can discern >40 nM PPi in the presence of 0.1 mM Pi at pH 7, which suggests that these assemblies may be useful in bio-analytical applications. This displacement assay was used to effectively determine the ability of pyrophosphatase to hydrolyze PPi to Pi.


Newborn Health and Child Mortality Across England.
Although the immediate impact of neonatal illness is well recognized, its wider and longer term outcomes on childhood mortality and the role of specific illnesses across childhood are unclear. To investigate how many deaths in childhood are associated with neonatal illness and the underlying conditions of the children who died. This population-based cohort study of children who died before age 10 years in England between April 1, 2019, and March 31, 2021, used data from the National Child Mortality Database. Data analysis was performed from September 2022 to May 2023. Children who received care in a neonatal unit after birth plus those who died in the first day of life, before admission to a neonatal unit, were considered to have likely neonatal illness. The primary outcome was the relative risk (RR) of dying, stratified by likely neonatal illness and specific neonatal conditions. Comparisons were made using the χ2 or likelihood ratio test, as appropriate. A total of 4829 children were included (median [IQR] age at death, 28 [2-274] days; 2606 boys [54.8%]; 2690 White children [64.0%]). Overall, 3456 children who died (71.6%) had evidence of likely neonatal illness. Children with neonatal illness were more likely to die before their tenth birthday than those without evidence of neonatal illness (RR, 13.82; 95% CI, 13.00-14.71). The estimated population-attributable risk fraction for neonatal illness among all deaths before age 10 years was 66.4% (95% CI, 64.9%-67.9%). Children with preceding neonatal illness who died were more likely to have underlying behavioral or developmental disorders (odds ratio [OR], 3.31; 95% CI, 2.47-4.42), chronic neurological disease (OR, 3.00; 95% CI, 2.51-3.58), and chronic respiratory disease (OR, 3.01; 95% CI, 2.43-3.73) than children without neonatal illness. In this cohort study, most children who died before age 10 years had some evidence of neonatal illness, and they died of a range of causes, including infections and sudden, unexpected, unexplained death. These findings suggest that improvements to perinatal morbidity, an area with an existing evidence base for improvement, may have important impacts on child health across the next decade.


Structure of catalase determined by MicroED.
MicroED is a recently developed method that uses electron diffraction for structure determination from very small three-dimensional crystals of biological material. Previously we used a series of still diffraction patterns to determine the structure of lysozyme at 2.9 Å resolution with MicroED (Shi et al., 2013). Here we present the structure of bovine liver catalase determined from a single crystal at 3.2 Å resolution by MicroED. The data were collected by continuous rotation of the sample under constant exposure and were processed and refined using standard programs for X-ray crystallography. The ability of MicroED to determine the structure of bovine liver catalase, a protein that has long resisted atomic analysis by traditional electron crystallography, demonstrates the potential of this method for structure determination.


Di-iodothyronine as part of the oestradiol and catechol oestrogen receptor--the role of iodine, thyroid hormones and melatonin in the aetiology of breast cancer.
Hypothyroidism and low iodine intake may be important aetiological factors in oestrogen dependent tumours of the breast, uterus and ovary. They are preventable risk factors. Iodine supplementation will hopefully lead to a decreased incidence of these cancers in future generations. The present author proposes that the tyrosyl residue in the hydrophobic oestrogen binding site of the oestrogen receptor is post translationally modified to monoiodotyrosine and hence 3,3' di-iodothyronine monoamine (T2) by peroxidase activity. He has previously proposed that various monoamine receptors are also T2 based. The densities of these receptors are increased in hypothyroidism and they exert control over release of prolactin and other hormones, including melatonin at multiple sites in the hypothalamic--pituitary axis. Melatonin is a metabolite of serotonin and hence melatonin receptors may be T2 or rT3 based as well. These factors could be significant in the aetiology of breast cancer as high prolactin and melatonin levels may be protective. Oestrogen receptor density may be increased in hypothyroidism as is certain monoamine receptor density. This would amplify the effect of high circulation oestrogen levels in hypothyroidism and may help explain why hypothyroidism and low iodine intake are risk factors for breast, uterine and ovarian cancer.


Enhanced Raman scattering in slow-light photonic crystals for chip-scale frequency conversion and optical amplification.
We present measurements of enhanced Raman scattering in silicon slow-light photonic crystal waveguides. By utilizing both the Bragg gap edge dispersion of TM-like modes for pump enhancement and the TE-like fundamental mode onset for Stokes enhancement, a six-fold increase in the spontaneous Raman scattering was observed in the double slow-light regime. Both forward and backward Stokes signals are examined, with continuous-wave measurements, in our low-loss photonic crystal membranes. The measured nonlinear enhancement matches well with our numerical model and simulations, and are described in detail in this paper. These observations support the development of chip-scale frequency conversion and optical amplification in silicon nanophotonics.


[Leptospirillum-like bacteria and their role in pyrite oxidation].
Two strains of Leptospirillum-like bacteria isolated from dumps of Alaverdi and Akhtala sulfide ore deposits in Armenia were studied. The optimum and maximum temperatures for the growth of both strains were 37 and 40 degrees C, respectively. The pH optimum was 2.0-2.3. Bacterial growth and ferrous iron oxidation were inhibited by yeast extract. The pyrite-leaching activity of the Leptospirillum-like bacteria under mesophilic conditions was close to that of Acidithiobacillus ferroxidans and exceeded by 2.0-2.7 times the activity of these moderately thermophilic bacteria at 37 degrees C. The leaching of pyrite by Leptospirillum-like bacteria increased in the presence of sulfur-oxidizing bacteria, particularly, in their association with a thermotolerant sulfur-oxidizing bacterium.


Usefulness of Virtual Expiratory CT Images to Compensate for Respiratory Liver Motion in Ultrasound/CT Image Fusion: A Prospective Study in Patients with Focal Hepatic Lesions.
To assess whether virtual expiratory (VE)-computed tomography (CT)/ultrasound (US) fusion imaging is more effective than conventional inspiratory (CI)-CT/US fusion imaging for hepatic interventional procedures. This prospective study was approved by the Institutional Review Board, and informed consent was obtained from each patient. In total, 62 patients with focal hepatic lesions referred for hepatic interventional procedures were enrolled. VE-CT images were generated from CI-CT images to reduce the effects of respiration-induced liver motion. The two types of CT images were fused with real-time US images for each patient. The operators scored the visual similarity with the liver anatomy upon initial image fusion and the summative usability of complete image fusion using the respective five-point scales. The time required for complete image fusion and the number of point locks used were also compared. In comparison with CI-CT/US fusion imaging, VE-CT/US fusion imaging showed significantly higher visual similarity with the liver anatomy on the initial image fusion (mean score, 3.9 vs. 1.7; p < 0.001) and higher summative usability for complete image fusion (mean score, 4.0 vs. 1.9; p < 0.001). The required time (mean, 11.1 seconds vs. 22.5 seconds; p < 0.001) and the number of point locks (mean, 1.6 vs. 3.0; p < 0.001) needed for complete image fusion using VE-CT/US fusion imaging were significantly lower than those needed for CI-CT/US fusion imaging. VE-CT/US fusion imaging is more effective than CI-CT/US fusion imaging for hepatic interventional procedures.


Phospholipase A₂ activity in first episode schizophrenia: associations with symptom severity and outcome at week 12.
Intracellular phospholipases A₂ (inPLA₂) are activated during monoaminergic neurotranismision and act as key enzymes in cell membrane repair and remodelling, neuroplasticity, neurodevelopment, apoptosis, synaptic pruning, neurodegenerative processes and neuroinflammation. Several independent studies found increased inPLA₂ activity in drug-naïve first episode and chronic schizophrenia. This study investigates if inPLA₂ activity is associated with symptoms severity and treatment response in first episode schizophrenia (FES). InPLA₂ activity was measured in serum of 35 young FES patients (mean age: 19.36 ± 3.32, mean duration of illness: 7.53 ± 6.28 months, 16 neuroleptic-naïve) before and after 12 weeks of treatment with second-generation antipsychotic medications (olanzapine, quetiapine or risperidone), as well as in 22 healthy controls matched for age. Psychopathology and social functioning were assessed at the same time points. Baseline inPLA₂ activity was significantly increased in drug-naïve and treated FES patients compared to healthy controls. Baseline inPLA₂ activity was also associated with severity of negative symptoms and lower functioning at baseline. Furthermore, baseline inPLA₂ activity was associated with improvement in negative symptoms and functioning within the first 12 weeks of treatment. Intracellular PLA₂ activity is increased in first episode schizophrenia and associated with symptom severity and outcome after 12 weeks of treatment. Future studies should investigate the implications of inPLA₂ activity as a potential predictor of treatment response for different antipsychotic agents.


[Cloning of the gene of Saccharomyces cerevisiae yeasts that determines resistance to the toxic action of cadmium ions].
A gene conferring resistance to cadmium in Saccharomyces cerevisiae was isolated from a yeast gene library created on the basis of the pL3 vector. The phenotype of resistance is only expressed in the yeast cells with cloned DNA inserted into a multicopy plasmid. Integration of the plasmid into chromosome or introduction of the centromeric region into the plasmid decreases the level of cadmium resistance. The cloned Sau3A I fragment of the yeast chromosome is 3.5 kbp in size. Restriction analysis and subcloning experiments showed the gene to be located within 1.6 kbp of the XhoI-Sau3A I fragment of DNA. Instability was observed in the vicinity of the XhoI-Sau3A I fragment of the yeast DNA in Escherichia coli.


[Sensitive noncompetitive measurement of small molecules by open sandwich immunoassay].
Proteins having multiple epitopes can be usually measured with sandwich ELISA employing two kinds of antibodies, which permits high sensitivity as well as a wide working range of more than three orders of magnitude. On the other hand, so-called monovalent antigens with MW less than 1000 are not susceptible to sandwich assays due to their small size and have almost always been measured in competitive assays. However, while a competitive assay needs only one antibody due to the principle of ratiometric measurement, optimization of the reaction conditions is necessary to attain suitable sensitivity and working range, which are often inferior to those of sandwich assays. As an alternative immunoassay for small antigens, here we propose a noncompetitive "open sandwich" immunoassay, which is based on the principle of stabilization of the antibody variable region Fv upon binding with antigen. Using ELISA to detect labeled V(H) fragments bound to immobilized V(L) in the presence of sample in microplate wells, various small molecules with MW around 200-300 can be measured with a superior detection limit and working range compared with those achieved with the corresponding competitive assays. The results indicate a common antigen recognition mode of anti-hapten antibody and wide applicability of the assay to the sensitive and handy analysis of low molecular-weight substances in areas such as environmental analysis and clinical diagnostics.


Groundwater chemistry determines the prokaryotic community structure of waterworks sand filters.
Rapid sand filtration is essential at most waterworks that treat anaerobic groundwater. Often the filtration depends on microbiological processes, but the microbial communities of the filters are largely unknown. We determined the prokaryotic community structures of 11 waterworks receiving groundwater from different geological settings by 16S rRNA gene-based 454 pyrosequencing and explored their relationships to filtration technology and raw water chemistry. Most of the variation in microbial diversity observed between different waterworks sand filters could be explained by the geochemistry of the inlet water. In addition, our findings suggested four features of particular interest: (1) Nitrospira dominated over Nitrobacter at all waterworks, suggesting that Nitrospira is a key nitrifying bacterium in groundwater-treating sand filters. (2) Hyphomicrobiaceae species were abundant at all waterworks, where they may be involved in manganese oxidation. (3) Six of 11 waterworks had significant concentrations of methane in their raw water and very high abundance of the methanotrophic Methylococcaceae. (4) The iron-oxidizing bacteria Gallionella was present at all waterworks suggesting that biological iron oxidation is occurring in addition to abiotic iron oxidation. Elucidation of key members of the microbial community in groundwater-treating sand filters has practical potential, for example, when methods are needed to improve filter function.


Neoplasms of urinary bladders of hamsters treated with 2-acetylaminofluorene and indole.
Tested in hamsters was the effect of neonatal treatment with 2-acetylaminofluorene (AAF) followed by prolonged feeding in combination with indole on the development of bladder tumors. Neonatal males and females were given intraperitoneal injections of AAF, 5 mg/100 g body weight, 3 times weekly until weaning. They were then fed a synthetic diet containing 0.06% AAF and 1.6% indole. The 26 hamsters surviving 10-12 months developed transitional cell carcinomas of the bladder, all but 2 of which were invasive. No hamsters had malignant tumors in the liver. Seven hamsters developed peliosis of the spleen.


Interaction of atomic hydrogen with single-walled carbon nanotubes: a density functional theory study.
We have studied the interaction of atomic hydrogen with (5,5) and (10,0) single-walled carbon nanotubes (SWNT) using density functional theory. These calculations use Gaussian orbitals and periodic boundary conditions. We compare results from the local spin density approximation, generalized gradient approximation (GGA), and hybrid density functionals. We have first kept the SWNT geometric structure fixed while a single H atom approaches the tube on top of a carbon atom. In that case, a weakly bound state with binding energies from -0.8 to -0.4 eV was found. Full geometry relaxation leads to a strong SWNT deformation, weakening the nearest C-C bonds and increasing the binding energy by about 1 eV. Full hydrogen coverage of the (5,5) SWNT converts this metallic nanotube into an insulator with a band gap of 3.4 eV for the GGA functional and 4.8 eV for the hybrid functional. Hybrid functionals perform similar to pure density functional theory functionals for the calculation of binding energies while band gaps critically depend on the functional choice.


Treatment of prostatitis in the rat.
A spontaneous, nonacute, age-dependent prostatitis was found in a high incidence in the lateral prostatic lobes of Lewis rats. Such rats were treated with methylprednisolone, indometacin , testosterone, hexyloxyphenylproprionate , polyestradiol phosphate, various antibiotics, or were caged together with female rats. The effect of the different treatment modalities was evaluated microscopically by blind observation of the degree of inflammatory reaction in the lateral prostate. Methylprednisolone and the testosterone ester caused a reduction of the inflammatory reactions and so did caging with female rats. The similarity of the rat prostatitis to the human condition may suggest the possibility of using corticosteroids or androgens for the treatment of patients with nonacute prostatitis.


Biology and efficacy of Aprostocetus (Eulophidae: Hymenoptera) as a parasitoid of the blueberry gall midge complex: Dasineura oxycoccana and Prodiplosis vaccinii (Diptera: Cecidomyiidae).
In the southeastern United States, bud-infesting larvae of two gall midge species, Dasineura oxycoccana (Johnson) and Prodiplosis vaccinii (Felt), destroy from 20 to 80% of the rabbiteye blueberry crop, Vaccinium virgatum Aiton (syn. V. ashei Reade). These midge larvae are attacked by five species of parasitoid wasps. The most effective of these is the bivoltine eulophid Aprostocetus sp. nr. marylandensis (Eulophidae), whose adults constitute one-third of the gall midge parasitoids, active in both conventional and organic blueberry fields. Broods of Aprostocetus use several reproductive strategies to keep sole possession of their larval hosts. As solitary endoparasitoids as well as facultative hyperparasitoids, precocial larvae of Aprostocetus devour hosts organs along with any younger siblings and rival parasitoid broods. Although larger hosts are preferred, any sized larvae can be parasitized, which reduces brood congestion and infanticide. An Aprostocetus female spends an hour or more in a systematic hunt for hosts, during which time 40 to 100% of midge larvae encountered are parasitized. Aprostocetus females could have located hosts more quickly had they recognized host-feeding scars as cues. Even so, high rates of larval parasitism achieved by Aprostocetus may kill as many midges as insecticides do.


Syntheses, structures, and magnetic properties of unusual nonlinear polynuclear copper(II) complexes containing derivatives of 1,2,4-triazole and pivalate ligands.
Novel polynuclear Cu(II) complexes containing derivatives of 1,2,4-trizaole and pivalate ligands, [Cu(3)(mu(3)-OH)(mu-adetrz)(2)(piv)(5)(H(2)O)].6.5H(2)O (1) (adetrz = 4-amino-3,5-diethyl-1,2,4-triazole, piv = pivalate), [Cu(4)(mu(3)-OH)(2)(mu-atrz)(2)(mu-piv)(4)(piv)(2)].2MeOH.H(2)O (2) (atrz = 4-amino-1,2,4-triazole), [Cu(4)(mu(3)-OH)(2)(mu-tbtrz)(2)(mu-piv)(2)(piv)(4)].4H(2)O (3) (tbtrz = 4-tert-butyl-1,2,4-trizaole), and [Cu(4)(mu(3)-O)(2)(mu-admtrz)(4)(admtrz)(2)(mu-piv)(2)(piv)(2)].2[Cu(2)(mu-H(2)O)(mu-admtrz)(piv)(4)].13H(2)O [4 = 4a.2(4b).13H(2)O; admtrz = 4-amino-3,5-dimethyl-1,2,4-triazole], have been prepared and structurally characterized. 1 is an asymmetrical triangular complex containing a [Cu(3)(mu(3)-OH)] core with two Cu---Cu edges spanned by bridging adetrz ligands. 2, 3, and 4a are novel tetranuclear compounds containing a [Cu(4)(mu(3)-O)(2)] or [Cu(4)(mu(3)-OH)(2)] core with Cu---Cu edges spanned by bridging 1,2,4-triazole or pivalate ligands. 4b is a dinuclear compound with one admtrz and one water bridge, and it is the first dinuclear Cu(II) triazole complex with one bridging water molecule. 1 is one of few reported triangular Cu(II) complexes with derivatives of 1,2,4-triazole, while 2, 3, and 4a are the first group of the nonlinear tetranuclear Cu(II) compounds with derivatives of 1,2,4-triazole. Variable-temperature magnetic susceptibility studies on the powder samples of 1-3 reveal the overall antiferromagnetic coupling between Cu(II) ions with J values of -55.6 to -12.8 cm(-1) (1), -216.4 to 0 cm(-1) (2), and -259.8 to 4.8 cm(-1) (3).


Prediction of performance reduction in self-paced exercise as modulated by the rating of perceived exertion.
Rating of perceived exertion (RPE) is a scale of exercise difficulty and has been hypothesized to be a regulator of work rate during self-pacing. The goal of this work was to develop a dynamic prediction of RPE and to characterize the control strategy employed to reduce work rate during self-paced exercise using RPE as feedback. Training and test data were acquired from the literature to develop a linear regression of RPE as a function of four physiological variables: core temperature, mean-weighted skin temperature, metabolic rate, and integral of relative oxygen consumption (R (2) = 0.85). A thermoregulatory model was used to predict core and mean-weighted skin temperature. Utilizing self-paced cycling and running data from the literature, we characterized reductions in work rate with a proportional-derivative control algorithm with RPE as feedback. Bland-Altman analysis revealed the necessity to parameterize RPE equations for untrained and endurance-trained individuals. Afterwards, dynamic predictions of RPE were accurate for a wide range of activity levels and air temperatures for walking, running, and cycling (LoA and bias of 2.3 and -0.03, respectively). For self-paced exercise, the control algorithm characterized the trend and magnitude of work rate reductions for cycling and running, and showed regulated RPE to be less conservative for shorter vs. longer duration exercise. A novel methodology to characterize self-paced work intensity, based upon dynamic physiologic response, is provided. The complete model is a useful tool that estimates performance decrements during self-paced exercise and predicts tolerance time for exhaustive fixed-rate exercise.


On re-excitation of feline motoneurones: its mechanism and consequences.
Conditions required for re-excitation of lumbosacral motoneurones, i.e. for double impulses in the motor axons associated with a single soma-dendritic action potential, were examined in cats anaesthetized with pentobarbitone and paralysed with gallamine triethiodide. Simultaneous recording from a motoneurone (intracellular, and in some experiments also extracellular), and from its axon in a ventral root, was used to assess the relations between the soma and the double axonal action potentials. Action potentials (greater than 70 mV) evoked by brief depolarizing current pulses applied intracellularly were never observed to cause re-excitation. Re-excitation could, however, be regularly induced by procedures which increased the delay between the initial segment and soma-dendritic components of these potentials. Re-excitation could be evoked (i) when brief hyperpolarizing pulses were applied before the onset of the soma-dendritic spikes, (ii) when the depolarizing pulses were applied on a background of long hyperpolarizing pulses or (iii) when two action potentials were evoked in a quick succession (by two brief depolarizing pulses). No relationship was found between the presence of re-excitation of motor axons and the presence of the delayed depolarization which follows the soma-dendritic spikes. Neither re-excitation nor delayed depolarization were found to be dependent upon re-excitation of the initial segment. These observations are thus at variance with previous suggestions that the initial segment spikes induce the re-excitation of motor axons and that the initial segment spikes cause the delayed depolarization following soma-dendritic spikes. Since re-excitation of a motor axon occurred without any signs of a second initial segment spike, it is concluded that it is initiated at the level of the axon, most likely at the first node of Ranvier. Re-excitation of motor axons was also observed during repetitive firing induced by intracellular current injection. However, it occurred then only occasionally, and only under strong depolarizing drive. It is thus not expected to be a common phenomenon under natural conditions of repetitive firing.


Comparative effectiveness of membrane bioreactors, conventional secondary treatment, and chlorine and UV disinfection to remove microorganisms from municipal wastewaters.
Log removals of bacterial indicators, coliphage, and enteric viruses were studied in three membrane bioreactor (MBR) activated-sludge and two conventional secondary activated-sludge municipal wastewater treatment plants during three recreational seasons (May-Oct.) when disinfection of effluents is required. In total, 73 regular samples were collected from key locations throughout treatment processes: post-preliminary, post-MBR, post-secondary, post-tertiary, and post-disinfection (UV or chlorine). Out of 19 post-preliminary samples, adenovirus by quantitative polymerase chain reaction (qPCR) was detected in all 19, enterovirus by quantitative reverse transcription polymerase chain reaction (qRT-PCR) was detected in 15, and norovirus GI by qRT-PCR was detected in 11. Norovirus GII and Hepatitis A virus were not detected in any samples, and rotavirus was detected in one sample but could not be quantified. Although culturable viruses were found in 12 out of 19 post-preliminary samples, they were not detected in any post-secondary, post-MBR, post-ultraviolet, or post-chlorine samples. Median log removals for all organisms were higher for MBR secondary treatment (3.02 to >6.73) than for conventional secondary (1.53-4.19) treatment. Ultraviolet disinfection after MBR treatment provided little additional log removal of any organism except for somatic coliphage (>2.18), whereas ultraviolet or chlorine disinfection after conventional secondary treatment provided significant log removals (above the analytical variability) of all bacterial indicators (1.18-3.89) and somatic and F-specific coliphage (0.71 and >2.98). Median log removals of adenovirus across disinfection were low in both MBR and conventional secondary plants (no removal detected and 0.24), and few removals of individual samples were near or above the analytical variability of 1.2 log genomic copies per liter. Based on qualitative examinations of plots showing reductions of organisms throughout treatment processes, somatic coliphage may best represent the removal of viruses across secondary treatment in both MBR and conventional secondary plants. F-specific coliphage and Escherichia coli may best represent the removal of viruses across the disinfection process in MBR facilities, but none of the indicators represented the removal of viruses across disinfection in conventional secondary plants.


Obesity is a disease.
The American Medical Association House of Delegates' vote in June to classify obesity as a "disease state with multiple pathophysiological aspects requiring a range of interventions to advance obesity treatment and prevention" sparked national debate. Critics worry calling obesity a disease will take focus off prevention and remove personal responsibility from the equation. Supporters say the classification is bound to spur innovative policy, research, and treatment approaches to obesity. Physicians by and large hope that the new label will persuade private health plans and Medicaid to pay for treatment of obese patients.


Why does a protein's evolutionary rate vary over time?
The sequences of different proteins evolve at different rates. The relative evolutionary rate (ER) of a single protein also changes over evolutionary time. The cause of this ER fluctuation remains uncertain, and study of this phenomenon may shed light on protein evolution more broadly. We have characterized ER fluctuation in mammals and Drosophila. We found little correlation between the amount of rate variation observed for a protein and such factors as its expression level or phylogenetic distribution. Perhaps more surprisingly, we found little correlation between our measure of rate variation and ER itself. We also investigated the extent to which the ERs of different domains of a protein vary independently. We found that rates of different domains do tend to vary together. In fact, rates at positions in different domains are coupled just as strongly as rates at equally distant positions in the same domain. These findings provide clues to the protein evolutionary process.


Erythropoietin is not a risk factor for severe retinopathy of prematurity among high risk preterm infants.
Retinopathy of prematurity (ROP) is a developmental retinal vaso-proliferative disease and a leading cause of blindness in children. Early gestational age, low birth weight and unregulated oxygen exposure are the main risk factors for the development of ROP. There are conflicting reports of a possible association between recombinant Erythropoietin (rhEPO) use and an increased risk for the development of ROP. To determine whether rhEPO is an independent risk factor for the development of severe ROP among preterm infants with a gestational age of 23 to 32 weeks and a birth weight <1500 g. We performed a retrospective study of risk factors for ROP on a cohort of 1762 premature infants born between 2009 and 2014, half of whom received rhEPO. To examine the association between treated ROP and rhEPO, a propensity score (PS) analysis was performed using the inverse probability of treatment weighted (IPTW) approach. The incidence of treated ROP was 7.3% (129/1762). PS analysis did not show an association between rhEPO and severe ROP needing treatment or ROP stage 2 or higher, in either the whole population or in the subgroup of babies born at 23 to 28 weeks gestation, in whom the incidence of severe ROP was the highest. Of 117 patients treated for Type 1 or worsening stage 3 ROP, 17 were first diagnosed after NICU discharge. Our study showed no association between Erythropoietin use and severe ROP and highlights the importance of Ophthalmology follow up after hospital discharge.


Surgical treatment of compensatory head position in congenital nystagmus.
Records of 32 patients with congenital nystagmus who underwent surgery for a significant head turn were reviewed to assess the effectiveness of current surgical approaches. Eighteen patients (Group I) had single binocular vision and had bilateral recess/resect procedures either according to Parks' 5-6-7-8 scheme (nine patients) or "augmented" from 10% to 40% for large head turns (nine patients). Fourteen (78%) had a reduction of the turn to 15 degrees or less, six (33%) being 5 degrees or less. One was overcorrected . Fourteen patients had a heterotropia in addition to head turn. Seven (Group II) had bilateral surgery to attempt to correct the head turn and reduce the tropia to less than 10 prism diopters. In three cases (43%) there was both successful treatment of the tropia and reduction of the turn to 15 degrees or less. The tropia was successfully treated in six patients (86%). In seven patients (Group III) monocular recess/resect surgery was done to attempt to correct both abnormalities. In four (57%) the turn was reduced to 15 degrees or less with successful treatment of the tropia at the same time. Five (71%) had a tropia of less than or equal to 10 diopters postoperatively. Overall, the head turns were reduced to 15 degrees or less in 23 (72%) of the 32 patients including 11 (35%) with 5 degrees or less residual turn. Three (9%) were overcorrected at least 10 degrees. The frequency of undercorrections indicates that larger amounts of surgery have to be done, especially for large turns, although ductions will be limited in some gaze positions, at times significantly, to achieve this goal.