Respiratory and other hazards of isocyanates.
Isocyanates are increasingly being used for manufacturing polyurethane foam, elastomers, adhesives, paints, coatings, insecticides, and many other products. At present, they are regarded as one of the main causes of occupational asthma. The large number of workers who are exposed to these chemicals have a concentration-dependent risk of developing chronic airway disorders, especially bronchial asthma. Different pathophysiologic mechanisms are involved. Immunoglobulin E (IgE)-mediated sensitization and irritative effects have been clearly demonstrated in both exposed subjects and animals. Presumably, neural inflammation due to neuropeptide release of capsaicin-sensitive afferent nerves is crucial. We collected data on 1780 isocyanate workers who had been examined by our groups. Of them 1095 (including subjects from outpatient departments) had work-related symptoms, predominantly of the respiratory tract. Specific IgE antibodies were found in 14% of the 1095 subjects. The methacholine challenge test was shown to be an inadequate predictor of the results of inhalative isocyanate provocation tests in workers and in asthmatic controls. Isocyanate (toluene diisocyanate TDI) air concentrations of 10 ppb (0.07 mg/m3) and 20 ppb (0.14 mg/m3), respectively, did not cause significant bronchial obstruction in the majority of previously unexposed asthmatics with bronchial hyperreactivity. IgG-mediated allergic alveolitis, a rare disease among isocyanate workers, was found in approximately 1% of the symptomatic subjects. Experimental studies exhibit dose-dependent toxic effects and give evidence for tachykinin-mediated bronchial hyperreactivity after exposure to isocyanates. The clinical role of genotoxic effects of isocyanates and their by-products demonstrated here in vitro and in vivo has yet to be clarified.


Impact of early diagnosis on functional disability in rheumatoid arthritis.
To determine whether early diagnosis is beneficial for functional status of various disease durations in rheumatoid arthritis (RA) patients. A total of 4,540 RA patients were enrolled as part of the Korean Observational Study Network for Arthritis (KORONA). We defined early diagnosis as a lag time between symptom onset and RA diagnosis of ≤ 12 months, whereas patients with a longer lag time comprised the delayed diagnosis group. Demographic characteristics and outcomes were compared between early and delayed diagnosis groups. Logistic regression analyses were performed to identify the impact of early diagnosis on the development of functional disability in RA patients. A total of 2,597 patients (57.2%) were included in the early diagnosis group. The average Health Assessment Questionnaire-Disability Index (HAQ-DI) score was higher in the delayed diagnosis group (0.64 ± 0.63 vs. 0.70 ± 0.66, p < 0.01), and the proportion of patients with no functional disability (HAQ = 0) was higher in the early diagnosis group (22.9% vs. 20.0%, p = 0.02). In multivariable analyses, early diagnosis was independently associated with no functional disability (odds ratio [OR], 1.19; 95% confidence interval [CI], 1.01 to 1.40). In a subgroup analysis according to disease duration, early diagnosis was associated with no functional disability in patients with disease duration < 5 years (OR, 1.37; 95% CI, 1.09 to 1.72) but not in patients with longer disease duration (for 5 to 10 years: OR, 1.07; 95% CI, 0.75 to 1.52; for ≥ 10 years: OR, 0.92; 95% CI, 0.65 to 1.28). Early diagnosis is associated with no functional disability, especially in patients with shorter disease duration.


Milk Transmission of HTLV-1 and the Need for Innovative Prevention Strategies.
Breastfeeding is recommended by the World Health Organization for at least 6 months up to 2 years of age, and breast milk protects against several diseases and infections. Intriguingly, few viruses are transmitted via breastfeeding including Human T-cell leukemia virus Type 1 (HTLV-1). HTLV-1 is a highly oncogenic yet neglected retrovirus, which primarily infects CD4+ T-cells in vivo and causes incurable diseases like HTLV-1-associated inflammatory conditions or Adult T-cell leukemia/lymphoma (ATLL) after lifelong viral persistence. Worldwide, at least 5-10 million people are HTLV-1-infected and most of them are unaware of their infection posing the risk of silent transmissions. HTLV-1 is transmitted via cell-containing body fluids such as blood products, semen, and breast milk, which constitutes the major route of mother-to-child transmission (MTCT). Risk of transmission increases with the duration of breastfeeding, however, abstinence from breastfeeding as it is recommended in some endemic countries is not an option in resource-limited settings or underrepresented areas and populations. Despite significant progress in understanding details of HTLV-1 cell-to-cell transmission, it is still not fully understood, which cells in which organs get infected via the oral route, how these cells get infected, how breast milk affects this route of infection and how to inhibit oral transmission despite breastfeeding, which is an urgent need especially in underrepresented areas of the world. Here, we review these questions and provide an outlook how future research could help to uncover prevention strategies that might ultimately allow infants to benefit from breastfeeding while reducing the risk of HTLV-1 transmission.


Broadband CARS spectral phase retrieval using a time-domain Kramers-Kronig transform.
We describe a closed-form approach for performing a Kramers-Kronig (KK) transform that can be used to rapidly and reliably retrieve the phase, and thus the resonant imaginary component, from a broadband coherent anti-Stokes Raman scattering (CARS) spectrum with a nonflat background. In this approach we transform the frequency-domain data to the time domain, perform an operation that ensures a causality criterion is met, then transform back to the frequency domain. The fact that this method handles causality in the time domain allows us to conveniently account for spectrally varying nonresonant background from CARS as a response function with a finite rise time. A phase error accompanies KK transform of data with finite frequency range. In examples shown here, that phase error leads to small (<1%) errors in the retrieved resonant spectra.


Facile and green synthesis of pullulan derivative-stabilized Au nanoparticles as drug carriers for enhancing anticancer activity.
In this work, we report for the first time AuNPs reduced/stabilized/capped with modified para-aminobenzoic acid-quat188-pullulan (PABA-QP) as excellent nanocarriers for delivery of doxorubicin to enhance the activity and safety of these systems. Spherical AuNPs@PABA-QP obtained by facile and green synthesis under optimum conditions were characterized by UV-VIS, TEM, EDS, SAED, XRD, ATR-FTIR and zeta-potential analyses and showed a narrow size distribution of 13.7 ± 1.9 nm. DOX was successfully loaded onto AuNPs@PABA-QP via intermolecular interactions with high drug loading. DOX-AuNPs@PABA-QP (IC50 = 0.39μM) showed a 2.1-fold higher cytotoxicity against Chago cells than DOX alone (IC50 = 0.82μM), while exhibiting less cytotoxicity against normal cells (Wi-38). Moreover, DOX-AuNPs@PABA-QP also demonstrated high intracellular uptake by endocytosis, arrested in S and G2-M phases of the cell cycle (total S/G2-M increased to approximately 18.0%), induced excellent cytotoxicity, and increased the fraction of late-apoptotic cells (18.6%). Consequently, it is suggested that the novel combination of DOX-AuNPs@PABA-QP has the potential to be developed for human cancer treatment.


Inhibition of Jv by ANF in rat proximal straight tubules requires angiotensin.
The effects of atrial natriuretic factor (ANF) on fluid absorption (Jv) by isolated perfused proximal straight tubules of rats were investigated. ANF alone (10(-8) M) added to the bath had no significant effect on absorption. In contrast, when tubules were first treated with 1.6 X 10(-10) M angiotensin II, this same concentration of ANF lowered fluid absorption from 0.99 +/- 0.03 to 0.69 +/- 0.02 nl.mm-1.min-1. A lower dose of ANF, 2 X 10(-10) M, reduced fluid absorption in the presence of angiotensin II from 1.13 +/- 0.06 to 0.65 +/- 0.05 nl.mm-1.min-1, an inhibition of 40%. Since guanosine 3',5'-cyclic monophosphate (cGMP) is reportedly part of the second messenger system of ANF, the effects of dibutyryl-cGMP (DBcGMP) on fluid absorption were studied. This membrane-permeant form of cGMP mimicked the effects of ANF, reducing fluid absorption from 1.15 +/- 0.18 to 0.54 +/- 0.08 nl.mm-1.min-1. These studies suggested the following: 1) ANF can regulate fluid absorption in the proximal nephron; 2) this inhibition occurs only in the presence of angiotensin; and 3) cGMP is part of the second messenger system of ANF in the rat proximal straight tubule, as it is in other tissues.


Osteomyelitis as a cause of brachial plexus neuropathy.
Two infants, younger than 2 months of age, had brachial plexus involvement and true paralysis secondary to osteomyelitis of the proximal humerus. To date, no previous cases of this association have been reported. Electromyography (EMG) demonstrated significant denervation in muscles innervated by the upper trunk, with scattered involvement of other muscles. Nerve conduction velocities were normal in both patients. Both patients had significant residual weakness after appropriate antibiotic therapy. One patient, 12 months after therapy for osteomyelitis, continued to demonstrate both clinical and EMG evidence of a lower motor neuron neuropathy. The pathophysiologic cause was thought to be ischemic nerve damage secondary to occlusion of vasa nervorum.


Beta-cell replacement for type I diabetes.
The ability to achieve insulin independence with either solid-organ pancreas or islet transplantation has increased the number of patients seeking beta-cell replacement as an alternative to insulin therapy. Despite dramatic improvements in the ability to achieve insulin independence following solid-organ pancreas transplantation, the secondary complications of long-standing diabetes are frequently irreversible by the time surgical intervention is justified based on the risk of this procedure. Pancreatic islet transplantation provides a safer and less invasive alternative for beta-cell replacement that could be justified earlier in the course of diabetes to prevent the development of secondary complications. Recent advances in the technology of islet isolation, as well as the ability to prevent the alloimmune and recurrent autoimmune response following islet transplantation with immunosuppressive regimens that are not toxic to beta cells, have rekindled an interest in this field. Widespread application of islet transplantation will depend on further improvements in selective immunosuppression, development of immunologic tolerance, and finding new sources of beta cells.


Excess TPX2 Interferes with Microtubule Disassembly and Nuclei Reformation at Mitotic Exit.
The microtubule-associated protein TPX2 is a key mitotic regulator that contributes through distinct pathways to spindle assembly. A well-characterised function of TPX2 is the activation, stabilisation and spindle localisation of the Aurora-A kinase. High levels of TPX2 are reported in tumours and the effects of its overexpression have been investigated in cancer cell lines, while little is known in non-transformed cells. Here we studied TPX2 overexpression in hTERT RPE-1 cells, using either the full length TPX2 or a truncated form unable to bind Aurora-A, to identify effects that are dependent-or independent-on its interaction with the kinase. We observe significant defects in mitotic spindle assembly and progression through mitosis that are more severe when overexpressed TPX2 is able to interact with Aurora-A. Furthermore, we describe a peculiar, and Aurora-A-interaction-independent, phenotype in telophase cells, with aberrantly stable microtubules interfering with nuclear reconstitution and the assembly of a continuous lamin B1 network, resulting in daughter cells displaying doughnut-shaped nuclei. Our results using non-transformed cells thus reveal a previously uncharacterised consequence of abnormally high TPX2 levels on the correct microtubule cytoskeleton remodelling and G1 nuclei reformation, at the mitosis-to-interphase transition.


Statistical, spectral and non-linear analysis of the heart rate variability during wakefulness and sleep.
As a first step in a program designed to study the central control of the heart rate variability (HRV) during sleep, we conducted polysomnographic and electrocardiogram recordings on chronically-prepared cats during semi- restricted conditions. We found that the tachogram, i.e. the pattern of heart beat intervals (RR intervals) was deeply modified on passing from alert wakefulness through quiet wakefulness (QW) to sleep. While the tachogram showed a rhythmical pattern coupled with respiratory activity during non-REM sleep (NREM), it turned chaotic during REM sleep. Statistical analyses of the RR intervals showed that the mean duration increased during sleep. HRV measured by the standard deviation of normal RR intervals (SDNN) and by the square root of the mean squared difference of successive intervals (rMSSD) were larger during REM and NREM sleep than during QW. SD-1 (a marker of short- term variability) and SD-2 (a marker of long-term variability) measured by means of Poincaré plots increased during both REM and NREM sleep compared to QW. Furthermore, in the spectral analysis of RR intervals, the band of high frequency (HF) was larger in NREM and REM sleep in comparison to QW, whereas the band of low frequency (LF) was larger only during REM sleep in comparison to QW. The LF/HF ratio was larger during QW compared either with REM or NREM sleep. Finally, sample entropy analysis used as a measure of complexity, was higher during NREM in comparison to REM sleep. In conclusion, HRV parameters, including complexity, are deeply modified across behavioral states.


The unique red cell heterogeneity of SC disease: crystal formation, dense reticulocytes, and unusual morphology.
Knowledge concerning SS (homozygous for the beta s gene) red blood cell (RBC) heterogeneity has been useful for understanding the pathophysiology of sickle cell anemia. No equivalent information exists for RBCs of the compound heterozygote for the beta s and beta c genes (SC) RBCs. These RBCs are known to be denser than most cells in normal blood and even most cells in SS blood (Fabry et al, J Clin Invest 70:1284, 1981). We have analyzed the characteristics of SC RBC heterogeneity and find that: (1) SC cells exhibit unusual morphologic features, particularly the tendency for membrane "folding" (multifolded, unifolded, and triangular shapes are all common); (2) SC RBCs containing crystals and some containing round hemoglobin (Hb) aggregates (billiard-ball cells) are detectable in circulating SC blood; (3) in contrast to normal reticulocytes, which are found mainly in a low-density RBC fraction, SC reticulocytes are found in the densest SC RBC fraction; and (4) both deoxygenation and replacement of extracellular Cl- by NO3- (both inhibitors of K:Cl cotransport) led to moderate depopulation of the dense fraction and a dramatic shift of the reticulocytes to lower density fractions. We conclude that the RBC heterogeneity of SC disease is very different from that of SS disease. The major contributions of properties introduced by HbC are "folded" RBCs, intracellular crystal formation in circulating SC cells, and apparently a very active K:Cl cotransporter that leads to unusually dense reticulocytes.


Attentional modulation in the detection of irrelevant deviance: a simultaneous ERP/fMRI study.
Little is known about the neural mechanisms that control attentional modulation of deviance detection in the auditory modality. In this study, we manipulated the difficulty of a primary task to test the relation between task difficulty and the detection of infrequent, task-irrelevant deviant (D) tones (1,300 Hz) presented among repetitive standard (S) tones (1,000 Hz). Simultaneous functional magnetic resonance imaging (fMRI)/event-related potentials (ERPs) were recorded from 21 subjects performing a two-alternative forced-choice duration discrimination task (short and long tones of equal probability). The duration of the short tone was always 50 msec. The duration of the long tone was 100 msec in the easy task and 60 msec in the difficult task. As expected, response accuracy decreased and response time (RT) increased in the difficult compared with the easy task. Performance was also poorer for D than for S tones, indicating distraction by task-irrelevant frequency information on trials involving D tones. In the difficult task, an amplitude increase was observed in the difference waves for N1 and P3a, ERP components associated with increased attention to deviant sounds. The mismatch negativity (MMN) response, associated with passive deviant detection, was larger in the easy task, demonstrating the susceptibility of this component to attentional manipulations. The fMRI contrast D > S in the difficult task revealed activation on the right superior temporal gyrus (STG) and extending ventrally into the superior temporal sulcus, suggesting this region's involvement in involuntary attention shifting toward unattended, infrequent sounds. Conversely, passive deviance detection, as reflected by the MMN, was associated with more dorsal activation on the STG. These results are consistent with the view that the dorsal STG region is responsive to mismatches between the memory trace of the standard and the incoming deviant sound, whereas the ventral STG region is activated by involuntary shifts of attention to task-irrelevant auditory features.


Detecting time-specific differences between temporal nonlinear curves: Analyzing data from the visual world paradigm.
In multiple fields of study, time series measured at high frequencies are used to estimate population curves that describe the temporal evolution of some characteristic of interest. These curves are typically nonlinear, and the deviations of each series from the corresponding curve are highly autocorrelated. In this scenario, we propose a procedure to compare the response curves for different groups at specific points in time. The method involves fitting the curves, performing potentially hundreds of serially correlated tests, and appropriately adjusting the overall alpha level of the tests. Our motivating application comes from psycholinguistics and the visual world paradigm. We describe how the proposed technique can be adapted to compare fixation curves within subjects as well as between groups. Our results lead to conclusions beyond the scope of previous analyses.


DAB2IP Downregulation Enhances the Proliferation and Metastasis of Human Gastric Cancer Cells by Derepressing the ERK1/2 Pathway.
DAB2IP (DOC2/DAB2 interactive protein) is downregulated in several cancer types, and its downregulation is involved in tumor cell proliferation, apoptosis, metastasis, and epithelial-mesenchymal transition (EMT). We aimed to investigate the potential role of DAB2IP in the development and progression of gastric cancer. DAB2IP levels were analyzed in human gastric cancer and adjacent normal tissues by Western blots and immunohistochemistry. Potential roles of DAB2IP in regulating gastric cancer cell growth and metastasis were examined by genetic manipulation in vitro. The molecular signaling was determined to understand the mechanisms of observed DAB2IP effects. DAB2IP level is lower in gastric cancer tissues as compared to paired normal tissues. Knockdown of DAB2IP enhanced gastric cancer cell growth and metastasis in vitro and promoted EMT progress at both protein and mRNA levels. Silencing DAB2IP activated extracellular signal-regulated kinase 1/2 (ERK1/2) pathway, and the enhanced proliferation and migration ability induced by DAB2IP knockdown were reduced after incubation with U0126 in SGC7901 gastric cancer cells. Inhibition of DAB2IP enhances gastric cancer cell growth and metastasis through targeting the ERK1/2 signaling, indicating that it may serve as a potential target for treatment of gastric cancer.


Risk of liver injury after α-glucosidase inhibitor therapy in advanced chronic kidney disease patients.
Although α-glucosidase inhibitors (AGIs) are commonly used for controlling postprandial blood glucose, AGIs-induced liver injuries have been reported. However, the relationship between AGIs and liver injuries in advanced chronic kidney disease (CKD) patients remains unexplored. In this nationwide case-control study, we recruited 1765 advanced diabetic CKD patients, who received AGIs therapy from January 1, 2000 to December 31, 2010 as the study sample and 5295 matched controls. Recent and former AGIs users were defined as patients who received the AGIs prescription for 30-60 d and 30-210 d before the event of liver injury. The risk of AGIs-induced liver injury was examined using time-dependent Cox proportional hazards model. Liver injury occurred in 3.9% of patients in the study group and 3.3% of patients in the control group. AGIs use did not increase the risk of liver injury in advanced CKD patients (P = 0.19). The stratified analysis indicated no increased risk of liver injury in all AGIs-using subgroups (all P > 0.05). The available evidence supports extending the use of AGIs without increasing the risk of liver injury in patients with advanced CKD. Additional randomized controlled trials are warranted to confirm our results.


Coronavirus JHM: tryptic peptide fingerprinting of virion proteins and intracellular polypeptides.
The virion proteins and intracellular polypeptides of the murine coronavirus MHV-JHM have been analysed by two-dimensional fingerprinting of their [35S]methionine-containing tryptic peptides. The analysis shows that the virion proteins gp98, gp65, pp60 and p23 are distinct. Virion protein gp25 has the same polypeptide component as p23, and virion protein gp170 has a polypeptide component related to gp98. The six virus polypeptides synthesized in infected cells, 150K, 65K, 60K, 30K, 23K and 14K are also distinct. The 170K and 98K species, which are produced by processing, are related to 150K. The 25K species is a processed form of 23K. The identity of corresponding species in the cell and in the virion has been shown and a model describing the genesis of coronavirus JHM proteins can now be proposed.


Transcriptomic Signatures in Sepsis and a Differential Response to Steroids. From the VANISH Randomized Trial.
There remains uncertainty about the role of corticosteroids in sepsis with clear beneficial effects on shock duration, but conflicting survival effects. Two transcriptomic sepsis response signatures (SRSs) have been identified. SRS1 is relatively immunosuppressed, whereas SRS2 is relatively immunocompetent. We aimed to categorize patients based on SRS endotypes to determine if these profiles influenced response to either norepinephrine or vasopressin, or to corticosteroids in septic shock. A post hoc analysis was performed of a double-blind, randomized clinical trial in septic shock (VANISH [Vasopressin vs. Norepinephrine as Initial Therapy in Septic Shock]). Patients were included within 6 hours of onset of shock and were randomized to receive norepinephrine or vasopressin followed by hydrocortisone or placebo. Genome-wide gene expression profiling was performed and SRS endotype was determined by a previously established model using seven discriminant genes. Samples were available from 176 patients: 83 SRS1 and 93 SRS2. There was no significant interaction between SRS group and vasopressor assignment (P = 0.50). However, there was an interaction between assignment to hydrocortisone or placebo, and SRS endotype (P = 0.02). Hydrocortisone use was associated with increased mortality in those with an SRS2 phenotype (odds ratio = 7.9; 95% confidence interval = 1.6-39.9). Transcriptomic profile at onset of septic shock was associated with response to corticosteroids. Those with the immunocompetent SRS2 endotype had significantly higher mortality when given corticosteroids compared with placebo. Clinical trial registered with www.clinicaltrials.gov (ISRCTN 20769191).


Chronotype is differentially associated with lifetime mood and panic-agoraphobic spectrum symptoms in patients with bipolar disorder and healthy controls.
Although the association between chronotype and mood disorders has been consistently reported, conversely, attempts to measure the association between chronotype and anxiety symptoms have generated inconsistent results. We aimed at evaluating whether chronotype (assessed through subjective and objective measures) is associated with lifetime mood and panic-agoraphobic spectrum symptoms in healthy controls (HCs) and in patients with bipolar disorder (BD). Overall, 173 subjects, patients with BD in euthymic phase (n = 76) and HC (n = 97), were evaluated through the reduced Morningness-Eveningness Questionnaire (rMEQ), actigraphy monitoring and mood and panic-agoraphobic spectrum self-report (MOODS-SR and PAS-SR). The discrepancy between objective (actigraphic-based) versus subjective (rMEQ-based) circadian typology was estimated through the Circadian Classification Discrepancy Index (CCDI). rMEQ-based evening chronotype (ET) was associated with higher scores in MOODS-SR depressive and rhythmicity and vegetative functions domains in HC and BD.Both ET and morning chronotypes (MT) were associated with higher PAS-SR scores in BD only. Actigraphic-based MT was associated with higher MOODS-SR depressive scores in HC. Likewise, the discrepancy between actigraphic-based and rMEQ-based circadian typology was associated with depressive symptoms in HC only. Self-reported ET was consistently associated with mood symptoms, while associations with panic-agoraphobic symptoms only emerged in BD and involved both extreme chronotypes. The discrepancy between the preferred circadian typology (rMEQ-based) and the actual one (actigraphic-based) could contribute to depressive symptoms in HC. These results pave the way for interventional studies targeting circadian typology in an attempt to prevent or treat mental health disorders.


Vitamin D level and its relation to muscle and fat mass in adult male Arabs.
Low levels of vitamin D have been linked with increased adiposity and diminished muscle strength. Whether it is also related to fat deposition in muscle tissues is not studied well. This study explored the associations between circulating 25-hydroxyvitamin D (25(OH)D) and fat deposition in muscle tissues of adult Arab males. A total 465 adult Saudi males were included in this cross-sectional study. Anthropometrics, body composition and muscle strength were assessed. Serum 25(OH)D was determined and quantified enzymatically. They were grouped according to vitamin D status: deficient (25(OH)D < 50 nmol/l) N = 325 (69.9%) and sufficient (25(OH)D > 50 nmol/l)140 (30.1%). Mean level of lean/height2, lean-arm-legs and lean-arms-legs/height2 were significantly higher in 25(OH)D deficient participants (p-values 0.03; 0.05 and 0.01 respectively). Thigh strength was significantly higher in 25(OH)D sufficient participants than their deficient counterparts (p = 0.02). In all participants, a significant correlation between 25(OH)D was observed with age and thigh-strength (p-values < 0.05), while a significant inverse correlation between 25(OH)D and lean/height2, lean-arms-legs, lean-arms-legs/height2, fat (%) region, fat arms, fat legs, fat trunk, lean legs were noted. In conclusion, low circulating 25(OH)D is associated with enhanced fat infiltration in muscle tissues of adult Arab males.


In vitro activity of ceftaroline and ceftobiprole against methicillin-resistant Staphylococcus aureus with decreased susceptibility to vancomycin isolated in paediatric patients.
Minimal inhibitory concentrations (MIC, mg/l) of ceftaroline and ceftobiprole were evaluated over 70 methicillin-resistant Staphylococcus aureus (MRSA) strains with vancomycin MIC ≥1 isolated in a paediatric hospital. The proportion of non-wild-type strains (MIC > epidemiological cut off) was 18% for ceftobiprole and 64% for ceftaroline. Only 1.4% of strains was resistant to ceftobiprole, and none to ceftaroline. These results are worrisome, since show the presence of non-negligible proportions of MRSA strains with high MIC values for ceftaroline and ceftobiprole in a setting where both drugs were never used.


SARS-CoV-2-reactive interferon-γ-producing CD8+ T cells in patients hospitalized with coronavirus disease 2019.
There is limited information on severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) T-cell immune responses in patients with coronavirus disease 2019 (COVID-19). Both CD4+ and CD8+ T cells may be instrumental in resolution of and protection from SARS-CoV-2 infection. Here, we tested 25 hospitalized patients either with microbiologically documented COVID-19 (n = 19) or highly suspected of having the disease (n = 6) for presence of SARS-CoV-2-reactive CD69+ expressing interferon-γ (IFN-γ) producing CD8+ T cells using flow-cytometry for intracellular cytokine staining assay. Two sets of overlapping peptides encompassing the SARS-CoV-2 Spike glycoprotein N-terminal 1 to 643 amino acid sequence and the entire sequence of SARS-CoV-2 M protein were used simultaneously as antigenic stimulus. Ten patients (40%) had detectable responses, displaying frequencies ranging from 0.15 to 2.7% (median of 0.57 cells/µL; range, 0.43-9.98 cells/µL). The detection rate of SARS-CoV-2-reactive IFN-γ CD8+ T cells in patients admitted to intensive care was comparable (P = .28) to the rate in patients hospitalized in other medical wards. No correlation was found between SARS-CoV-2-reactive IFN-γ CD8+ T-cell counts and SARS-CoV-2 S-specific antibody levels. Likewise, no correlation was observed between either SARS-CoV-2-reactive IFN-γ CD8+ T cells or S-specific immunoglobulin G-antibody titers and blood cell count or levels of inflammatory biomarkers. In summary, in this descriptive, preliminary study we showed that SARS-CoV-2-reactive IFN-γ CD8+ T cells can be detected in a non-negligible percentage of patients with moderate to severe forms of COVID-19. Further studies are warranted to determine whether quantitation of these T-cell subsets may provide prognostic information on the clinical course of COVID-19.


Disseminated infection with Neospora caninum in a ten-year-old dog.
Naturally-occurring neosporosis with multiple organ involvement was identified in a 10-year-old neutered male Basset hound dog. Clinical signs were first noticed 3 weeks prior to referral and consisted of crouched stance and mild pelvic limb ataxia. Dexamethasone administration had provided transient improvement. On presentation to the teaching hospital, clinical signs included depression, pelvic limb ataxia, inability to stand without assistance, and pain on deep palpation of the cervical and lumbar vertebral column. Lesions were found in the myocardium, liver, spleen, adrenal glands, brain, and spinal cord. Tachyzoites of Neospora caninum were found in the myocardium and adrenal glands. Organisms stained with anti-Neospora caninum, but not to anti-Toxoplasma gondii serum in an immunohistochemical test.


High-affinity chelator thiols for switchable and oriented immobilization of histidine-tagged proteins: a generic platform for protein chip technologies.
Protein micro-/nanoarrays are becoming increasingly important in systematic approaches for the exploration of protein-protein interactions and dynamic protein networks, so there is a high demand for specific, generic, stable, uniform, and locally addressable protein immobilization on solid supports. Here we present multivalent metal-chelating thiols that are suitable for stable binding of histidine-tagged proteins on biocompatible self-assembled monolayers (SAMs). The architectures and physicochemical properties of these SAMs have been probed by various surface-sensitive techniques such as contact angle goniometry, ellipsometry, and infrared reflection-absorption spectroscopy. The specific molecular organization of proteins and protein complexes was demonstrated by surface plasmon resonance, confocal laser scanning, and atomic force microscopy. In contrast to the mono-NTA/His6 tag interaction, which has major drawbacks because of its low affinity and fast dissociation, drastically improved stability of protein binding by these multivalent chelator surfaces was observed. The immobilized histidine-tagged proteins are uniformly oriented and retain their function. At the same time, proteins can be removed from the chip surface under mild conditions (switchability). This new platform for switchable and oriented immobilization should assist proteome-wide wide analyses of protein-protein interactions as well as structural and single-molecule studies.


Neonatal Schwartz-Jampel syndrome: a common autosomal recessive syndrome in the United Arab Emirates.
Schwartz-Jampel syndrome (SJS) is a heterogeneous autosomal recessive syndrome of myotonia and bone dysplasia. Two types have been recognised: the classical type with late infantile or childhood manifestation and a rarer form with neonatal manifestation. We report five families with a total of 11 children affected with severe neonatal SJS. All presented after birth with skeletal abnormalities and feeding difficulties. Five had the typical pursed appearance of the mouth. Nine died from respiratory complications (five in the neonatal period and four before 2 years of age). One (4 months old) remains hospitalised since birth requiring continuous oxygen supplementation and one (5 months old) requires nasogastric tube feeding and has repeated attacks of aspiration. Only seven of the 17 previously reported neonatal SJS cases had a similar course to the patients in this report. We suggest that within neonatal SJS there is a subgroup which manifests severe respiratory and feeding problems and has a poor prognosis. This report brings the total number of children with neonatal SJS reported from the UAE to 14. This represents the largest review of this syndrome to date from one centre and indicates that this syndrome is fairly common in the population of the UAE.


Bone morphogenetic proteins in soft-tissue reconstruction.
Different options are reviewed in the field of musculoskeletal tissue reconstruction, from the addition of biological actors (cells, growth factors, biological or artificial scaffolds) to the application of gene therapy or tissue engineering. Growth factors can enable innovative solutions to treat such disease if we can extrapolate to soft tissue the promising results obtained in bone reconstruction with bone morphogenetic proteins. However, as in bone reconstruction, soft-tissue regeneration will depend on the drug delivery carrier, the scaffold for the newly formed tissue, the dose of growth factor and the animal model, which must all be explored before extrapolation to clinical problems.


Non-invasive proximal adhesive restoration of natural non-cavitated proximal lesions.
To investigate the infiltration potential of different self-etch adhesives into natural non-cavitated proximal lesions and the effect of dehydration protocol on the infiltration of a self-etch adhesive. 29 extracted molars and premolars with natural proximal lesions (ICDAS 1-2) were sectioned through the lesion providing two samples from each lesion. To compare the different adhesives, three groups of eight lesions were abraded with fine metallic strips and then etched with 37% H3PO4 acid for 120 seconds. All teeth were stained with rhodamine isothiocyanate. After drying with compressed air and ethanol application, lesions were infiltrated with Scotchbond Universal, Clearfil SE Protect or OneCoat 7 Universal for 180 seconds and then coated with a thin layer of flowable composite (Tetric Flow). To compare the effect of dehydration protocol on infiltration, two groups of nine paired lesions were pretreated as described above. One group was dried using compressed air alone and the second group was dried using compressed air and ethanol, both groups were then infiltrated with Scotchbond Universal then coated with a thin film of flowable composite. After light curing, un-encapsulated dye was bleached by immersion in hydrogen peroxide. Remaining lesion pores were stained with sodium fluorescein solution. Thin cuts of the teeth were observed with confocal microscopy and computer image analysis was performed (ImageJ). ANOVA and Duncan post-hoc tests showed no significant differences of the infiltrated area between the three adhesives (P= 0.835), no significant difference was found between the group dried with air compared to the one dried with air and ethanol. It can be concluded that the tested adhesives may be used for infiltration of natural lesions following the described pretreatment. Enamel pretreatment with metallic strip and 37% H3PO4 acid promotes the infiltration of different adhesives into natural non-cavitated caries lesions.


The Pathway to Achieving Universal Health Coverage in the Democratic Republic of Congo: Obstacles and Prospects.
This paper explores the complexities surrounding achieving universal health coverage (UHC) in the Democratic Republic of Congo (DRC) and proposes viable strategies to overcome the obstacles. The study's findings contribute to the global discourse on UHC in resource-limited settings and hold significant implications for policy formulation and implementation in both DRC and similar contexts. The introduction emphasises the importance of UHC in promoting equitable access to quality healthcare services for all individuals. Nevertheless, the DRC faces numerous challenges on its path to UHC. This paper identifies four key challenges: Firstly, the fragile healthcare infrastructure in the DRC necessitates the establishment of better-equipped facilities, an adequate healthcare workforce, and improved access to essential medical supplies. These factors hinder the provision of comprehensive health services and impede progress towards UHC. Secondly, socio-economic barriers such as persistent poverty, income disparities, and regional variations pose significant obstacles to achieving UHC in the DRC. Limited financial resources and widespread poverty prevent individuals from accessing healthcare services, exacerbating health inequities. Thirdly, weak health governance, inadequate policy implementation, and limited coordination among stakeholders impede the effective delivery of healthcare services in the DRC. Thus, strengthening governance structures and enhancing policy implementation are essential for UHC. Lastly, the absence of comprehensive health information systems and poor data management hinder evidence-based decision-making and resource allocation. Addressing these deficiencies is vital for monitoring progress and guiding policy formulation towards UHC. Given these challenges, this paper proposes potential solutions and future perspectives for achieving UHC in the DRC. These include strengthening health systems, implementing social protection mechanisms, enacting policy reforms, enhancing governance structures, and strengthening health information systems. Investments in robust health information systems, data collection and management improvements and the enhancement of capacity for health research and surveillance facilitate evidence-based decision-making and progress towards UHC. In conclusion, the DRC faces obstacles related to healthcare infrastructure, socio-economic factors, governance issues, and deficiencies in health information systems in its pursuit of UHC. However, by addressing these challenges through targeted interventions, policy reforms, and improved governance, the DRC can make strides towards ensuring equitable access to high-quality healthcare for all its citizens. Collaboration between national and international stakeholders is crucial for sustaining progress towards UHC and promoting health equity within the country.


Omega-3 fatty acids and cognitive function in women.
Omega-3 fatty acids (FAs) could play an important role in maintaining cognitive function in aging individuals. The omega-3 FA docosahexaenoic acid is a major constituent of neuronal membranes and, along with the other long-chain omega-3 FAs from fish such as eicosapentaentoic acid, has been shown to have a wide variety of beneficial effects on neuronal functioning, inflammation, oxidation and cell death, as well as on the development of the characteristic pathology of Alzheimer's disease. Omega-3 FAs may prevent vascular dementia via salutary effects on lipids, inflammation, thrombosis and vascular function. Epidemiologic studies have generally supported a protective association between fish and omega-3 FA levels and cognitive decline. Some of the small, short-term, randomized trials of docosahexaenoic acid and/or eicosapentaentoic acid supplementation have found positive effects on some aspects of cognition in older adults who were cognitively intact or had mild cognitive impairment, although little effect was found in participants with Alzheimer's disease. Large, long-term trials in this area are needed.


Enhanced phosphate removal and potential recovery from wastewater by thermo-chemically calcinated shell adsorbents.
Shell from the seafood processing industry is an under-utilised waste resource worldwide. Calcite, the major component of shell is commonly used in wastewater treatment for the removal of phosphorus (P). Here, mussel and oyster shell-based adsorbents (MSB and OSB) were used for removal of P as phosphate (PO43-) from aqueous solution and secondary wastewater, following preparation through chemical calcination at 700 °C. Batch adsorption experiments were carried out to identify the effects of various operating parameters (e.g., pH, dosage, contact time, initial concentration of P ions, co-existing ions), while a desorption study helped to understand the availability of the bonded P. The optimal contact time for PO43- removal was 120 min using both adsorbents with the dose at 200 mg. Characterisation of the adsorbent was performed using SEM-EDX, pHpzc, BET, FTIR and XRD. The XRD analysis showed that both calcite and lime were present on the surface of the shell particles. P was adsorbed effectively through inner-sphere complexation and surface microprecipitation mechanisms, while an enhanced maximum P adsorption capacity of 12.44 mg/g for MSB and 8.25 mg/g for OSB was reached. The Redlich-Peterson isotherm model fitted well with the equilibrium isotherm data (R2 ≥ 0.97) which also suggested a heterogenic surface. The desorption study (on the saturated adsorbent) found that ~97% of bonded P could be plant available in soil. These results suggest that a shell-based adsorbent can serve as a promising material for P removal from real wastewater effluent and subsequently could be used as a soil conditioner.


[Perspectives on basic biotechnology research in Mexico].
The main fields of basic research in biotechnology in Mexico are reviewed, with emphasis in the problems relevant to public health, such as environmental pollution and vaccine development and production. The institutions that are responsible for these studies are analyzed in terms of their relative participation.


Detection of potato mop-top virus capsid readthrough protein in virus particles.
Potato mop-top furovirus (PMTV) RNA 3 encodes the 20 kDa coat protein and a larger readthrough protein of 67 kDa. The readthrough protein is expressed by suppression of the amber stop codon which terminates the coat protein gene. A 21 kDa C-terminal fragment of the readthrough protein was doned, fused to glutathione S-transferase and expressed in E. coli. An antiserum prepared against purified fusion protein was used in ELISA to detect the readthrough protein in extracts of PMTV-infected leaves. Immunogold labelling studies showed that the readthrough protein was located near one extremity of some of the virus particles.


Environmental triggers for connective tissue disease: the case of COVID-19 associated with dermatomyositis-specific autoantibodies.
The aim of the present review is to analyze the link between autoimmune diseases and environmental factors, in particular severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection (COVID-19) as it shares numerous features with the interstitial lung disease associated with connective tissue diseases positive for rare autoantibodies directed at highly specific autoantigens (i.e., MDA5 and RIG1) among the intracellular sensors of SARS-CoV-2 in the innate response against viruses. As shown in recent publications and in our original data, specific autoantibodies may be functionally relevant to COVID-19 infection. We evaluated sera from 35 hospitalized patients with COVID-19 to identify antinuclear antibodies and autoantibodies directed against specific antigenic targets, and we identified anti-nuclear antibodies (ANA) in 20/35 of patients with COVID-19 (57%), in patients with need for supplemental oxygen (90% vs. 20% in ANA-negative cases; P < 0.0001). In 7/35 COVID-19 sera, we detected anti-MJ/NXP2 (n = 3), anti-RIG1 (n = 2), anti-Scl-70/TOPO1 (n = 1), and anti-MDA5 (n = 1), overall associated with a significantly worse pulmonary involvement at lung computerized tomography scans. Eleven (31%) patients were positive for antibodies against the E2/E3 subunits of mitochondrial pyruvate dehydrogenase complex. Viral infections such as COVID-19 are associated with ANA and autoantibodies directed toward antiviral signaling antigens in particular in patients with worse pulmonary involvement.


Beyond the learning curve: incidence of bile duct injuries following laparoscopic cholecystectomy normalize to open in the modern era.
Early reports of higher complication rates, specifically bile duct injuries, raised concerns over the safety of laparoscopy over open cholecystectomy. This study aims to ascertain the rate, management, and perioperative outcomes of bile duct injury in an era beyond the laparoscopic learning curve. The New York State (NYS) Planning and Research Cooperative System longitudinal administrative database was used to identify patients. From 2005 to 2010, 156,315 patients were identified who had undergone laparoscopic cholecystectomy for symptomatic cholelithiasis or acute or chronic cholecystitis. Patients were then tracked with unique identifiers for common bile duct injury. Common bile duct injury was identified by ICD-9 and CPT diagnosis and procedure codes for patients who subsequently underwent hepatectomy, hepaticojejunostomy, or other bile duct surgery. From 2005 to 2010, 156,958 patients were identified who had undergone laparoscopic cholecystectomy for symptomatic cholelithiasis or acute or chronic cholecystitis. Of the total patients, 149 patients underwent a biliary duct procedure within a year. Twenty-four of them were diagnosed with gallbladder cancer and excluded, leaving 125 for further analysis. The biliary injuries were identified at a rate of 0.080 %. Thirty-one of those patients (24.8 %) underwent hepatectomy, 40 patients (32.0 %) underwent hepaticoenterostomy, and 54 patients (43.2 %) underwent primary repair of the bile duct. Thirty-two (26 %) patients were repaired on the same day of their initial procedure. Of the remaining 93 patients, 38 (30 %) were repaired within 10 days, seven (6 %) repaired between 11 and 20 days, and 48 (38 %) patients over 21 days from injury. In NYS, the rate of bile duct injury has now decreased to 0.08 % and mirrors the historical figures quoted for open cholecystectomy. This improvement likely reflects increased experience, improved instrumentation, and movement beyond the "learning curve."


Purpose Formulation, Coalition Building, and Evidence Use in Public-Academic Partnerships: Web-Based Survey Study.
Partnerships between academic institutions and public care agencies (public-academic partnerships [PAPs]) can promote effective policy making and care delivery. Public care agencies are often engaged in PAPs for evidence-informed policy making in health care. Previous research has reported essential partnership contextual factors and mechanisms that promote evidence-based policy making and practice in health care. However, the studies have not yet informed whether public care agency leaders' and academic researchers' perceptions of partnership purpose formulation and coalition building evolve through the PAP life cycle and whether public care agency leaders' use of research evidence differs through life cycle stages. This exploratory study aims to focus on PAPs designed to improve youth mental health and well-being outcomes. This study also aims to identify public care agency leaders' and academic researchers' perceptions of PAP purpose formulation (structure, goals, primary function, and agenda-setting process) and coalition building (mutual benefits, trust, convener's role, member role clarity, and conflict management) by PAP life cycle stage and examine whether public care agency leaders' use of research evidence differs according to the perception of PAP purpose formulation and coalition building through the PAP life cycle. A web-based survey of PAP experience was conducted by recruiting academic researchers (n=40) and public care agency leaders (n=26) who were engaged in PAPs for the past 10 years. Public care agency leaders additionally participated in the survey of the Structured Interview for Evidence Use scale (n=48). Most public care agency leaders and academic researchers in PAPs formed, matured, and sustained perceived their PAP as having purpose formulation context well aligned with their organizational purpose formulation context, pursuing mutual benefits, having leadership representation and role clarity, having a higher level of trust, and knowing how to handle conflicts. Most PAPs across all life cycle stages crystallized another issue to focus, but not all PAPs with issue crystallization had purpose reformulation. Public care agency leaders who trusted academic researchers in their PAP had greater use of research evidence. Public care agency leaders in PAPs that had gone through new issue crystallization also showed greater use of research evidence compared with those that had not. To promote public care agency leaders' use of research evidence, focusing on developing trusting partnerships and continuously crystallizing PAP issues are important. RR2-10.2196/14382.


Hydroxylapatite with substituted magnesium, zinc, cadmium, and yttrium. II. Mechanisms of osteoblast adhesion.
The present in vitro study investigated osteoblast adhesion on hydroxylapatite (HA) doped with either cadmium (Cd), zinc (Zn), magnesium (Mg), or yttrium (Y). Compared with any other dopant tested in the present study, osteoblast adhesion was significantly (p < 0.05) greater on HA doped with Y after 4 h; in addition, osteoblast adhesion increased with concentration (2-7 mol%) of Y in HA. The findings that HA doped with greater amounts of Y adsorbed higher concentrations of calcium and, subsequently, of vitronectin and collagen (proteins known to mediate osteoblast adhesion), but not of albumin, laminin, and fibronectin, may explain the observed enhanced adhesion of osteoblasts on these substrates. Interactions (i.e., adsorption and configuration/bioactivity) of vitronectin and collagen may have been promoted by increased porosity of doped HA. Through doping with Y, the present study provided the first evidence that HA can be synthesized and processed with improved cytocompatibility properties for osteoblast adhesion, and thus offered essential information for the design of novel proactive bioceramics. Proactive bioceramics which elicit specific, timely, and desired responses from surrounding cells and tissues are necessary for improving bonding of orthopaedic/dental implants to juxtaposed bone; such osseointegration will, undoubtedly, enhance implant efficacy.


Growth and Stress Axis Responses to Dietary Cholesterol in Nile Tilapia (Oreochromis niloticus) in Brackish Water.
Six isonitrogenous and isocaloric diets were formulated to contain 0% (control), 0.4, 0.8, 1.2, 1.6, or 2.4% dietary cholesterol and fed to juvenile Nile tilapia (Oreochromis niloticus) (2.20 ± 0.12 g) twice daily to apparent satiation for 8 weeks in triplicate at a salinity of 16. Fish fed 0.4% cholesterol showed a higher weight gain and specific growth rate and a lower feed coefficient ratio than fish fed other diets. No difference was found in the survival of Nile tilapia fed various levels of cholesterol. Cholesterol in the serum and liver and low-density lipoprotein cholesterol in the serum increased with the increase in the dietary cholesterol content. Relative to the control, no significant difference was found in the expression of head kidney P450scc mRNA between treatment groups. The expression of head kidney 11β-HSD2 mRNA was the highest in the control group, and it decreased significantly with increasing levels of diet cholesterol. Fish fed 0.4 or 1.2% cholesterol had a higher 20β-HSD2 mRNA expression in the head kidney than those fed other diets. Fish fed 0.8% cholesterol had higher expressions of GR1 and GR2B mRNA in the liver than other groups. Fish fed 0.4% cholesterol had the highest activity of gill Na+/K+-ATPase. Fish fed 0.8 to 2.4% cholesterol had higher serum cortisol contents than the fish in the control group and the fish fed 0.4% cholesterol. This study suggests that dietary cholesterol is not essential for Nile tilapia survival in brackish water, but 0.4% cholesterol supplementation in the Nile tilapia diet contributes to the improvement of hyperosmotic adaptation and increases in gill Na+/K+-ATPase activity and serum cortisol content by regulating the hypothalamic-pituitary-interrenal stress axis.


Glucose shortens the life span of C. elegans by downregulating DAF-16/FOXO activity and aquaporin gene expression.
Many studies have addressed the effect of dietary glycemic index on obesity and diabetes, but little is known about its effect on life span itself. We found that adding a small amount of glucose to the medium (2%) shortened the life span of C. elegans by inhibiting the activities of life span-extending transcription factors that are also inhibited by insulin signaling: the FOXO family member DAF-16 and the heat shock factor HSF-1. This effect involved the downregulation of an aquaporin glycerol channel, aqp-1. We show that changes in glycerol metabolism are likely to underlie the life span-shortening effect of glucose and that aqp-1 may act cell nonautonomously as a feedback regulator in the insulin/IGF-1-signaling pathway. Insulin downregulates similar glycerol channels in mammals, suggesting that this glucose-responsive pathway might be conserved evolutionarily. Together, these findings raise the possibility that a low-sugar diet might have beneficial effects on life span in higher organisms.


Current Diagnosis and Management of Hypersensitivity Pneumonitis.
Hypersensitivity Pneumonitis (HP) one of the most common interstitial lung diseases (ILDs) is characterized by exposure to an inhaled inciting antigen that leads to a host immunologic reaction determining interstitial inflammation and architectural distortion. The underlying pathogenetic mechanisms are unclear. The absence of international shared diagnostic guidelines and the lack of a "gold-standard" test for HP combined with the presence of several clinical and radiologic overlapping features makes it particularly challenging to differentiate HP from other ILDs, also in expert contests. Radiology is playing a more crucial role in this process; recently the headcheese sign was recognized as a more specific for chronic-HP than the extensive mosaic attenuation. Several classification proposals and diagnostic models have been advanced by different groups, with no prospective validation. Therapeutic options for HP have been limited to antigen avoidance and immunosuppressant drugs over the last decades. Several questions about this condition remain unanswered and there is a need for more studies.


The acquisition of task-specific productions and modifications of declarative representations in spatial-precuing tasks.
The stimulus-response translation stage of human information processing plays a mediating role of relating stimuli to assigned responses. The translation stage has been implicated as the locus of a pattern of differential precuing benefits obtained in spatial-choice tasks (Proctor & Reeve, 1986; Reeve & Proctor, 1985): When pairs of finger responses from the middle and index fingers of each hand are precued, the two leftmost and two rightmost responses show the greatest benefit. This pattern of differential benefits, which occurs regardless of whether the hand placement is adjacent or overlapped, has been attributed to spatially coded representations of the stimulus and response sets in the translation stage. Experiment 1 evaluated whether the mediating role of the translation stage changes with practice. All precued pairs of responses showed equivalent benefits in the last of three sessions. This result indicates that the spatial representations used initially to translate between stimuli and responses have been altered to be more efficient or have been replaced by productions that directly specify fingers. Experiment 2 used a fourth session in which subjects were transferred from the overlapped hand placement to the adjacent placement, or vice versa. For subjects in the former condition, the pattern of differential precuing benefits reappeared in the transfer session. This lack of transfer is consistent with the hypothesis that task-specific productions develop with practice that directly relate stimuli to fingers. For subjects who practiced with the adjacent placement and switched to the overlapped placement, only a nonsignificant tendency existed for the pattern of differential precuing benefits to reappear. This failure of the pattern to reappear could indicate that spatial representations continue to be used to translate between stimuli and responses. Alternatively, as occurs with the overlapped placement, task-specific productions could be acquired that relate stimuli to fingers. If so, the failure of the pattern of differential precuing benefits to reappear would reflect a modification in the representations that are used for translation in the transfer session. Specifically, if subjects were coding the stimulus and response sets on the basis of the distinction between the two hands, as well as the spatial distinction, the differential benefits would be minimized because hand coding should benefit different responses from those benefitted by spatial coding. These alternative explanations were evaluated in Experiment 3 by having subjects who practiced with the adjacent placement switch to a placement in which the hands were crossed completely.(ABSTRACT TRUNCATED AT 400 WORDS)


Novel Models to Identify Census Tracts for Hepatitis C Screening Interventions.
Increased screening efforts and the development of effective antiviral treatments have led to marked improvement in hepatitis C (HCV) patient outcomes. However, many people in the United States are still believed to have undiagnosed HCV. Geospatial modeling using variables representing at-risk populations in need of screening for HCV and social determinants of health (SDOH) provide opportunities to identify populations at risk of HCV. A literature review was conducted to identify variables associated with patients at risk for HCV infection. Two sets of variables were collected: HCV Transmission Risk and SDOH Level of Need. The variables were combined into indices for each group and then mapped at the census tract level (n = 233). Multiple linear regression analysis and the Pearson correlation coefficient were used to validate the models. A total of 4 HCV Transmission Risk variables and 12 SDOH Level of Need variables were identified. Between the 2 indexes, 21 high-risk census tracts were identified that scored at least 2 standard deviations above the mean. The regression analysis showed a significant relationship with HCV infection rate and prevalence of drug use (B = 0.78, P < .001). A significant relationship also existed with the HCV infection rate for households with no/limited English use (B = -0.24, P = .001), no car use (B = 0.036, P < .001), living below the poverty line (B = 0.014, P = .009), and median household income (B = -0.00, P = .009). Geospatial models identified high-priority census tracts that can be used to map high-risk HCV populations that may otherwise be unrecognized. This will allow future targeted screening and linkage-to-care interventions for patients at high risk of HCV.


Cysteine-rich secretory protein 2 binds to mitogen-activated protein kinase kinase kinase 11 in mouse sperm.
Cysteine-rich secretory protein (CRISP) 2 (previously TPX1) is a testis-enriched member of the CRISP family, and has been localized to both the sperm acrosome and tail. Like all members of the mammalian CRISP family, its expression pattern is strongly suggestive of a role in male fertility, but functional support for this hypothesis remains limited. In order to determine the biochemical pathways within which CRISP2 is a component, the putative mature form of CRISP2 was used as bait in a yeast two-hybrid screen of a mouse testis expression library. One of the most frequently identified interacting partners was mitogen-activated protein kinase kinase kinase 11 (MAP3K11). Sequencing and deletion experiments showed that the carboxyl-most 20 amino acids of MAP3K11 interacted with the CRISP domain of CRISP2. This interaction was confirmed using pull-down experiments and the cellular context was supported by the localization of CRISP2 and MAP3K11 to the acrosome of the developing spermatids and epididymal spermatozoa. Interestingly, mouse epididymal sperm contained an approximately 60-kDa variant of MAP3K11, which may have been a result of proteolytic cleavage of the longer 93-kDa form seen in many tissues. These data raise the possibility that CRISP2 is a MAP3K11-modifying protein or, alternatively, that MAP3K11 acts to phosphorylate CRISP2 during acrosome development.


Impact of Tai Chi on CRP, TNF-alpha and IL-6 in inflammation: a systematic review and meta-analysis.
Previous studies have reported that C reactive protein (CRP), tumor necrosis factor-alpha (TNF-alpha), and interleukin-6 (IL-6) in Coronavirus disease 2019 (COVID-19) patients are significantly increased, and their progressive increases are clinical warning indicators of severe and critical severity. The purpose of this meta-analysis is to evaluate the efficacy of Tai Chi on CRP, TNF-alpha and IL-6, and provide a basis for complementary treatment of COVID-19. Five English databases (PubMed, Web of Science, Physiotherapy Evidence Database, Cochrane Library and Embase) and four Chinese electronic databases (CNKI, Wanfang, China Science and Technology Journal Database and SinoMed) were searched from inception to April 1st, 2020. Combination MeSH and free text terms were used to make up search strategy. Interventions in RCTs were Tai Chi with or without comparison (usual care, health education, drug therapy, psychosocial therapy). Revman version 5.3 was used to analyze the extracted data. Continuous outcomes were described by SMD, and the I2 test was used to assess heterogeneity. Revised Physiotherapy Evidence Database scale was used to assess methodological quality. Nine RCTs involving 571 participants met the inclusion criteria, and the sample size ranged from 19 to 100 per study. Tai Chi can significantly reduce TNF-alpha (Tai Chi intervention: SMD =-0.92, 95% CI: -1.32 to -0.53; Tai Chi plus drug treatment intervention: SMD =-0.63, 95% CI: -1.15 to -0.11), moreover, it could reduce the amount of IL-6 (Tai Chi intervention: SMD =-0.62, 95% CI: -1.00 to -0.23; Tai Chi plus drug treatment intervention: SMD =-2.17, 95% CI: -3.69 to -0.64) and CRP (Tai Chi plus drug treatment intervention: SMD =-1.98, 95% CI: -2.47 to -1.50) while with a high exercise amount. A low exercise amount of Tai Chi showed poor efficacy on CRP (Tai Chi intervention: SMD =-0.18, 95% CI: -0.61 to 0.25; Tai Chi plus drug treatment intervention: SMD =-0.15, 95% CI: -0.47 to 0.16) and IL-6 (Tai Chi intervention: SMD =0.15, 95% CI: -0.24 to 0.55). The strength of evidence might be limited due to relatively low methodological quality, heterogeneity and indirectness. The overall results elucidate that Tai Chi could significantly reduce TNF-alpha while it did not show the same effects in IL-6 and CRP. After subgroup analysis, Tai Chi with a high exercise amount can reduce IL-6 and CRP. Tai Chi with a high exercise amount could be suggested as a complementary intervention for people with COVID-19. PROSPERO CRD42020177655.


Myelin-Independent Therapeutic Potential of Canine Glial-Restricted Progenitors Transplanted in Mouse Model of Dysmyelinating Disease.
Dysfunction of glia contributes to the deterioration of the central nervous system in a wide array of neurological disorders, thus global replacement of glia is very attractive. Human glial-restricted precursors (hGRPs) transplanted intraventricularly into neonatal mice extensively migrated and rescued the lifespan in half of the studied mice, whereas mouse GRPs (mGRPs) presented no therapeutic benefit. We studied in the same experimental setting canine GRPs (cGRP) to determine whether their therapeutic potential falls between hGRPs and mGRPs. Additional motivation for the selection of cGRPs was a potential for use in veterinary medicine. cGRPs were extracted from the brain of dog fetuses. The cells were transplanted into the anterior or posterior aspect of the lateral ventricle (LV) of neonatal, immunodeficient, dysmyelinated mice (Mbpshi, Rag2 KO; shiv/rag2). Outcome measures included early cell biodistribution, animal survival and myelination assessed with MRI, immunohistochemistry and electron microscopy. Grafting of cGRP into posterior LV significantly extended animal survival, whereas no benefit was observed after anterior LV transplantation. In contrast, myelination of the corpus callosum was more prominent in anteriorly transplanted animals. The extended survival of animals after transplantation of cGRPs could be explained by the vicinity of the transplant near the brain stem.


Perceptions of government knowledge and control over contributions of aid organizations and INGOs to health in Nepal: a qualitative study.
Almost 50% of the Nepali health budget is made up of international aid. International Non-Governmental Organizations working in the field of health are able to channel their funds directly to grass root level. During a 2010 conference, the Secretary of Population stated that the government has full knowledge and control over all funds and projects coming to Nepal. However, there are no documents to support this. The study aims to assess government and partner perceptions on whether Government of Nepal currently has full knowledge of contributions of international aid organizations and International Non-Governmental Organizations to health in Nepal and to assess if the government is able to control all foreign contributions to fit the objectives of Second Long Term Health Plan (1997-2017). A qualitative study was performed along with available literature review. Judgmental and snowball sampling led to 26 in depth interviews with key informants from the government, External Development Partners and International Non-Governmental Organizations. Results were triangulated based on source of data. Representatives of the Department of Health Services declined to be interviewed. Data collection was done until researchers felt data saturation had been reached with each group of key informants. While Ministry of Health and Population leads the sector wide approach that aims to integrate all donor and International Non-Governmental Organization contributions to health and direct them to the government's priority areas, questions were raised around its capacity to do so. Similarly, informants questioned the extent to which Social Welfare Council was able to control all International Non-Governmental Organizations contributions. Political tumult, corruption in the government, lack of human resources in the government, lack of coordination between government bodies, convoluted bureaucracy, and unreliability of donor and International Non-Governmental Organization contributions were identified as the main reasons for difficulties in aid integration. Despite its commitment to coordinate and control development assistance to the health sector, and its leadership position of the Sector Wide Approach, complete knowledge and effective coordination of all international contributions remains a challenge and is hampered by issues within the government as well as among External Development Partners and International Non-Governmental Organizations.


[Anatomy and histology of salivary glands of Triatominae].
Histological studies upon the salivary glands of ten species of triatomine bugs were performed looking for their number and structural organization in different genera. It was possible to evaluate the celular epithelium type of each gland, as well as the merocrine and apocrine secretions of the glands. Secretion run until the hilo and after to salivary pump and hypofaringe. The glandular components, D1, D2 and D3 are always present in the Triatoma, Panstrongylus and Diptelogaster but in Rhodnius there are only the first two pairs of glands. The salivary channels and the hilo are analyzed by histology. The whole pair D3 has a clear valve that regularizes the exit of the secretions to the hilo. According to the genus the valves appear in different locations. They have low and dense epithelium, and their nucleus are rich in chromatin. The secondary channels leaving these valves, are very different, with clear chitinous ringer, low level of chromatin in the nucleus and homogeneous cytoplasm.


[alpha-Hemolysin of E. coli].
The activity of alpha-hemolysin increased at the log growth phase in the culture of E. coli P678 Hly+ hemolytic strain; this activity diminished with the change into the stationary phase, and then fell sharply. Replacement of the culture medium in the stationary growth phase by fresh one led to restoration of the hemolytic activity of the culture. The culture fluid separated from the cells at the stationary growth phase produced an inhibitory action on the alpha-hemolysin Ca ions activated and stabilized the alpha-hemolysin. Sodium citrate and sucrose served as hemolysis inhibitors. The action of alpha-hemolysin was maximal against human erythrocytes at pH 6.5. Hemolytic activity was characterized in time by a distinct lag-phase and the phase of the greatest rate of reaction. The duration of the lag-phase and also the rate of hemolysis depended on the concentration of alpha-hemolysin (with the increase of the hemolysin concentration lag-phase was shortened and the reaction was accelerated). There proved to be a linear relationship between the amount of erythrocytes taken into the reaction and the rate of hemoglobin release, and also there was noted a temperature activation of the hemolytic reaction.


Cloning and characterization of the ponA gene encoding penicillin-binding protein 1 from Neisseria gonorrhoeae and Neisseria meningitidis.
The ponA gene encoding penicillin-binding protein 1 (PBP 1) from Neisseria gonorrhoeae was cloned by a reverse genetic approach. PBP 1 was purified from solubilized membranes of penicillin-susceptible strain FA19 by covalent ampicillin affinity chromatography and used to obtain an NH2-terminal amino acid sequence. A degenerate oligonucleotide based on this protein sequence and a highly degenerate oligonucleotide based on a conserved amino acid motif found in all class A high-molecular-mass PBPs were used to isolate the PBP 1 gene (ponA). The ponA gene encodes a protein containing all of the conserved sequence motifs found in class A PBPs, and expression of the gene in Escherichia coli resulted in the appearance of a new PBP that comigrated with PBP 1 purified from N. gonorrhoeae. A comparison of the gonococcal ponA gene to its homolog isolated from Neisseria meningitidis revealed a high degree of identity between the two gene products, with the greatest variability found at the carboxy terminus of the two deduced PBP 1 protein sequences.


Alterations in rabbit alveolar macrophages as a result of traumatic shock.
Biochemical and electron microscope studies were conducted to determine the effects of traumatic shock on rabbit alveolar macrophages. Both resting and phagocytosing macrophages from the shocked animals, in comparison to comparable control macrophages, showed increased release of acid phosphatase from the cells into medium upon incubation in vitro, but decreases in the total content of acid phosphatase and beta-glucuronidase. Studies by electron microscopy showed ultrastructural alterations in macrophages from shocked animals consisting of a reduction in the number or a complete absence of lysosomes and, in some cases, increased amounts of rough endoplasmic reticulum and free ribosomes. In vitro incubation of macrophages from shocked animals with Pseudomonas aeruginosa showed that the process of bacterial ingestion was not impaired nor were the numbers of bacteria ingested decreased as compared to control macrophages. However, the ability of macrophages from shocked animals to destroy ingested bacteria appeared to be significantly altered. Extensive degradation of Pseudomonas was observed within phagocytic vacuoles of control macrophages after 15 minutes of incubation. In contrast, the majority of ingested organisms in macrophages from shocked animals showed no evidence of degradative changes.


Anticancer Drugs for Intra-Arterial Treatment of Colorectal Cancer Liver Metastases: In-Vitro Screening after Short Exposure Time.
To treat colorectal liver metastases, intra-arterial chemotherapies may complete therapeutic arsenal. Drugs using intra-arterially are very heterogeneous. The aim of this study was to select the most efficient drug on a panel of colorectal cancer (CRC) cell lines (Caco-2, HCT 116, HT 29, SW 48, SW 480, SW 620) exposed for 30 min to 12 cytotoxic agents (doxorubicin, epirubicin, idarubicin, 5-FU, raltitrexed, gemcitabine, cisplatin, oxaliplatin, mitomycin C, irinotecan, streptozocin, paclitaxel) at different concentrations. The effect on cell viability was measured using the WST-1 cell viability assay. For each drug and cell line, the IC50 and IC90 were calculated, which respectively correspond to the drug concentration (mg/mL) required to obtain 50% and 90% of cell death. We also quantified the cytotoxic index (CyI90 = C Max/IC90) to compare drug efficacy. The main findings of this study are that idarubicin emerged as the most cytotoxic agent to most of the tested CRC cell lines (Caco-2, HT29, HCT116, SW620 and SW480). Gemcitabine seemed to be the most efficient chemotherapy for SW48. Interestingly, the most commonly used cytotoxic agents in the systemic and intra-arterial treatment of colorectal liver metastasis (CRLM) (oxaliplatin, 5-FU, irinotecan) showed very limited cytotoxicity to all the cell lines.


Plasminogen binding inhibitors demonstrate unwanted activities on GABAA and glycine receptors in human iPSC derived neurons.
Plasminogen binding inhibitors (PBIs) reduce the risk of bleeding in hemorrhagic conditions. However, generic PBIs are also associated with an increased risk of seizures, an adverse effect linked to unwanted activities towards inhibitory neuronal receptors. Development of novel PBIs serve to remove compounds with such properties, but progress is limited by a lack of higher throughput methods with human translatability. Herein we apply human induced pluripotent stem cell (hiPSC) derived neurons in combination with dynamic mass redistribution (DMR) technology to demonstrate robust and reproducible modulation of both GABAA and glycine receptors. These cells respond to GABA (EC50 0.33 ± 0.18 μM), glycine (EC50 11.0 ± 3.7 μM) and additional ligands in line with previous reports from patch clamp technologies. Additionally, we identify and characterize a competitive antagonistic behavior of the prototype inhibitor and drug tranexamic acid (TXA). Finally, we demonstrate proof of concept for effective counter-screening of lead series compounds towards unwanted GABAA receptor activities. No activity was observed for a previously identified PBI candidate drug, AZD6564, whereas a discontinued analog, AZ13267257, could be characterized as a potent GABAA receptor agonist.


Paclitaxel plus carboplatin in the treatment of ovarian cancer.
Second-line treatment with paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) may achieve remissions in patients suffering from ovarian cancer who have failed primary chemotherapy with cisplatin- or carboplatin-based regimens. Introduction of paclitaxel in combination with cisplatin into the first-line treatment strategy was therefore the next logical step in the development of chemotherapy against ovarian cancer. Data already have shown that this may result in better survival. Since carboplatin may replace cisplatin, the combination of paclitaxel with carboplatin seemed a further necessary step. We therefore embarked on a dose-finding study of paclitaxel and carboplatin. Fourteen patients with International Federal of Gynecology and Obstetrics stage III and IV ovarian cancer with a median age of 55.5 years entered this study of escalating doses of either carboplatin or paclitaxel. Doses of carboplatin could be escalated from 300 to 450 mg/m2 and paclitaxel could be escalated from 125 to 175 mg/m2 without dose-limiting myelosuppression. At the highest dose level reported here, only transient short-lived leukopenia was observed. Other toxicities consisted of nausea and vomiting, peripheral neurotoxicity, and arthralgia, all mild. In the first 14 patients, 10 of whom are evaluable, complete remissions were seen in two patients and partial remissions in six. This study will escalate the doses of paclitaxel and carboplatin further. This treatment is well tolerated and yields satisfactory antitumor results.


Nonmedical Prescription Drug Use Among Female Adolescents: The Relative Influence of Maternal Factors, Social Norms, and Perceptions of Risk and Availability.
Increasing understanding of the risk and protective factors for adolescent nonmedical use of prescription drugs (NMUPD) could inform prevention efforts. Several correlates have been identified, including parental factors, perceptions about use and accessibility, social norms, and age. However, these constructs have rarely been simultaneously examined using paired data from parents and adolescents. We aimed to examine the relative influence of these correlates among dyads (N=349) of mothers and adolescent daughters. Using multiple logistic regression, daughters' past NMUPD and inclination for future NMUPD were regressed onto descriptive norms for friend use, perceived drug accessibility and risk of harm from use, daughter age, mothers' disapproval about use, mothers' past NMUPD and inclination for future NMUPD, and the mother-daughter relationship quality. Akaike weights and lasso regressions were also estimated to evaluate the relative importance of each correlate. Higher descriptive norms for friend use, older age, and mothers' inclination for NMUPD were risk factors for daughters' NMUPD, while a closer mother-daughter relationship and mothers' disapproving attitudes towards NMUPD were protective factors. The three analysis approaches were corroborative. Results suggest friend descriptive norms, mother-daughter relationship quality, and mothers' attitudes about NMUPD are important prevention targets.


Proposed revision of the Wauters et al. antigenic scheme for serotyping of Yersinia enterocolitica.
The antigenic scheme of Wauters and co-workers was applied to serotype a total of 585 strains of Yersinia enterocolitica, 43 strains of Yersinia kristensenii, 40 strains of Yersinia frederiksenii, and 25 strains of Yersinia intermedia. The O antigens 11,23; 11,24; 12,25; 12,26; and 28 were exclusively associated with Y. kristensenii. O antigen 16 was associated with Y. kristensenii and Y. frederiksenii. The serogroups O:4,33 and O:17 were only found in Y. intermedia, whereas the remaining serogroups were associated with Y. enterocolitica. The O antigen 34 proved to be identical with O:10; O factor 29 was poorly expressed and is suggested to be excluded from the list of recognized antigens. As sera of O factors 1, 2a, and 2b of Y. enterocolitica are difficult to prepare, we propose to combine the serogroups O:1,2a,3 and O:2a,2b,3 with O group 3. The H antigens l, r, s, and t were characteristic of Y. kristensenii, whereas H:p was exclusively found in Y. frederiksenii. H:q belonged to Y. intermedia, whereas H:o occurred in Y. kristensenii, Y. frederiksenii, and Y. intermedia. The remaining H factors a to k, m, and n occurred in Y. enterocolitica in various combinations. Up to 15 different combinations of H antigen factors were identified in a single serogroup of Y. enterocolitica. We propose the definition of a new H antigen, u. We suggest the exclusion from the typing scheme of those O and H antigens which are not associated with Y. enterocolitica. For O typing, we present a revised antigenic scheme of Y. enterocolitica with 18 serogroups containing 20 O factors. We emphasize the necessity of its continuous usage, taking into account the international situation.


Detection of Pyrethroid Resistance Mutations in the Major Leishmaniasis Vector Phlebotomus papatasi.
Phlebotomine sand flies (Diptera: Psychodidae) are primary vectors of leishmaniasis. Greece and Turkey are both endemic for visceral and cutaneous leishmaniasis and are widely affected by the disease. Measures commonly applied for controlling sand flies rely on the use of insecticides, predominantly pyrethroids. A worldwide problem associated with the intensive use of insecticides is the development of resistance. Scarce information is available regarding the resistance status in sand fly populations. Sand flies were collected from Greece (Thessaloniki, Peloponnese, Chios island) and Turkey (Sanliurfa) and analyzed for the presence and frequency of target-site knockdown resistance mutations on the voltage-gated sodium channel (Vgsc) gene. Five sand fly species were included in the analysis: Phlebotomus perfiliewi Parrot, Phlebotomus neglectus Tonnoir, Phlebotomus simici Nitzulescu, Phlebotomus tobbi Adler and Theodor, and Phlebotomus papatasi Scopoli. Their Vgsc gene-domain II was analyzed for the presence of known pyrethroid resistance mutations. The mutation 1014F, associated with pyrethroid-resistant phenotypes, was detected in P. papatasi sand flies from Sanliurfa at an allele frequency of 48%. Homozygotes for the wild type allele 1014L (Leu/Leu) represented 36% of the population, while homozygotes for the resistant allele 1014F (Phe/Phe) and heterozygotes encompassing both alleles (Leu/Phe) each had a frequency of 32%. In all other sand fly species, only the wild type allele 1014L was detected. This is the first report for the detection of resistance mutations in the major leishmaniasis vector P. papatasi and is of major concern regarding leishmaniasis control.


Effect of smoking habit on the frequency of micronuclei in human lymphocytes: results from the Human MicroNucleus project.
The effect of tobacco smoking on the frequency of micronuclei (MN) in human lymphocytes has been the object of many population studies. In most reports, the results were unexpectedly negative, and in many instances smokers had lower frequencies of MN than non-smokers. A pooled re-analysis of 24 databases from the HUMN international collaborative project has been performed with the aim of understanding the impact of smoking habits on MN frequency. The complete database included 5710 subjects, with 3501 non-smokers, 1409 current smokers, and 800 former smokers, among subjects in occupational and environmental surveys. The overall result of the re-analysis confirmed the small decrease of MN frequencies in current smokers (frequency ratio (FR) = 0.97, 95% confidence interval (CI) = 0.93-1.01) and in former smokers (FR = 0.96, 95% CI = 0.91-1.01), when compared to non-smokers. MN frequency was not influenced by the number of cigarettes smoked per day among subjects occupationally exposed to genotoxic agents, whereas a typical U-shaped curve is observed for non-exposed smokers, showing a significant increase of MN frequency in individuals smoking 30 cigarettes or more per day (FR = 1.59, 95% CI = 1.35-1.88). This analysis confirmed that smokers do not experience an overall increase in MN frequency, although when the interaction with occupational exposure is taken into account, heavy smokers were the only group showing a significant increase in genotoxic damage as measured by the micronucleus assay in lymphocytes. From these results some general recommendations for the design of biomonitoring studies involving smokers can be formulated. Quantitative data about smoking habit should always be collected because, in the absence of such data, the simple comparison of smokers versus non-smokers could be misleading. The sub-group of heavy smokers (> or =30 cigarettes per day) should be specifically evaluated whenever it is large enough to satisfy statistical requirements. The presence of an interaction between smoking habit and occupational exposure to genotoxic agents should be always tested.


Evidence that Brain-Reactive Autoantibodies Contribute to Chronic Neuronal Internalization of Exogenous Amyloid-β1-42 and Key Cell Surface Proteins During Alzheimer's Disease Pathogenesis.
Blood-brain barrier (BBB) permeability is a recognized early feature of Alzheimer's disease (AD). In the present study, we examined consequences of increased BBB permeability on the development of AD-related pathology by tracking selected leaked plasma components and their interactions with neurons in vivo and in vitro. Histological sections of cortical regions of postmortem AD brains were immunostained to determine the distribution of amyloid-β1-42 (Aβ42), cathepsin D, IgG, GluR2/3, and alpha7 nicotinic acetylcholine receptor (α7nAChR). Results revealed that chronic IgG binding to pyramidal neurons coincided with internalization of Aβ42, IgG, GluR2/3, and α7nAChR as well as lysosomal compartment expansion in these cells in regions of AD pathology. To test possible mechanistic interrelationships of these phenomena, we exposed differentiated SH-SY5Y neuroblastoma cells to exogenous, soluble Aβ42 peptide and serum from AD and control subjects. The rate and extent of Aβ42 internalization in these cells was enhanced by serum containing neuron-binding IgG autoantibodies. This was confirmed by treating cells with individual antibodies specific for α7nAChR, purified IgG from AD or non-AD sera, and sera devoid of IgG, in the presence of 100 nM Aβ42. Initial co-localization of IgG, α7nAChR, and Aβ42 was temporally and spatially linked to early endosomes (Rab11) and later to lysosomes (LAMP-1). Aβ42 internalization was attenuated by treatment with monovalent F(ab) antibody fragments generated from purified IgG from AD serum and then rescued by coupling F(ab) fragments with divalent human anti-Fab. Overall, results suggest that cross-linking of neuron-binding autoantibodies targeting cell surface proteins can accelerate intraneuronal Aβ42 deposition in AD.


Physiological responses of Pseudomonas putida KT2442 to phosphate starvation.
The physiological responses of Pseudomonas putida KT2442 to phosphate starvation were examined with respect to cell morphology, qualitative demonstration of the accumulation of the intracellular storage component poly-3-hydroxyalkanoate (PHA), cellular ATP and ribosome content, and the rate of total protein synthesis. Upon prolonged incubation under phosphate-limiting conditions, the number of viable cells decreased by two to three orders of magnitude during the first 3 weeks. However, after this decline, viability of the cultures remained remarkably constant for many weeks. The cells remained rod-shaped under phosphate starvation conditions with a tendency to swell in parallel with the accumulation of PHA. Protein synthesis and ribosome concentration were gradually reduced, and ATP levels dropped to very low values after the onset of starvation; later, however, there was a return to near-normal ATP concentrations. Evidence was obtained that the strong selective pressure imposed by phosphate deprivation forces the selection of mutants with a competitive advantage. These mutants are able to grow, possibly utilizing nutrients derived from dead cells, and eventually take over the cultures. One frequently encountered mutant formed smaller colonies on rich solidified medium and displayed an altered cell morphology. This mutant was isolated and further characterized. By employing a bioluminescence-based marker system, we demonstrated that this mutant is able to replace wild-type cells in mixed culture experiments. Thus, long-term phosphate-deprived cultures represent dynamic regimes that can undergo population shifts.


Impaired Synthesis of Stromal Components in Response to Minnelide Improves Vascular Function, Drug Delivery, and Survival in Pancreatic Cancer.
Pancreatic cancer stromal microenvironment is considered to be the major reason for failure of conventional and targeted therapy for this disease. The desmoplastic stroma, comprising mainly collagen and glycosaminoglycans like hyaluronan (HA), is responsible for compression of vasculature in the tumor resulting in impaired drug delivery and poor prognosis. Minnelide, a water-soluble prodrug of triptolide currently in phase I clinical trial, has been very effective in multiple animal models of pancreatic cancer. However, whether Minnelide will have efficacious delivery into the tumor despite the desmoplastic stroma has not been evaluated before. Patient tumor-derived xenografts (PDX) and spontaneous pancreatic cancer mice were treated with 0.42 and 0.21 mg/kg body weight for 30 days. Stromal components were determined by IHC and ELISA-based assays. Vascular functionality and drug delivery to the tumor were assessed following treatment with Minnelide. Our current study shows that treatment with Minnelide resulted in reduction of ECM components like HA and collagen in the pancreatic cancer stroma of both the spontaneous KPC mice as well as in patient tumor xenografts. Furthermore, treatment with Minnelide improved functional vasculature in the tumors resulting in four times more functional vessels in the treated animals compared with untreated animals. Consistent with this observation, Minnelide also resulted in increased drug delivery into the tumor compared with untreated animals. Along with this, Minnelide also decreased viability of the stromal cells along with the tumor cells in pancreatic adenocarcinoma. In conclusion, these results are extremely promising as they indicate that Minnelide, along with having anticancer effects is also able to deplete stroma in pancreatic tumors, which makes it an effective therapy for pancreatic cancer.


Methotrexate Aspasomes Against Rheumatoid Arthritis: Optimized Hydrogel Loaded Liposomal Formulation with In Vivo Evaluation in Wistar Rats.
Aspasomes of methotrexate with antioxidant, ascorbyl palmitate, were developed and optimized using factorial design by varying parameters such as lipid molar ratio, drug to lipid molar ratio, and type of hydration buffer for transdermal delivery for disease modifying activity in rheumatoid arthritis (RA). Aspasomes were characterized by drug-excipients interaction, particle size analysis, determination of zeta potential, entrapment efficiency, and surface properties. The best formulation was loaded into hydrogel for evaluation of in vitro drug release and tested in vivo against adjuvant induced arthritis model in wistar rats, by assessing various physiological, biochemical, hematological, and histopathological parameters. Optimized aspasome formulation exhibited smooth surface with particle size 386.8 nm, high drug loading (19.41%), negative surface potential, and controlled drug release in vitro over 24 h with a steady permeation rate. Transdermal application of methotrexate-loaded aspasome hydrogel for 12 days reduced rat paw diameter (21.25%), SGOT (40.43%), SGPT (54.75%), TNFα (33.99%), IL β (34.79%), cartilage damage (84.41%), inflammation (82.37%), panus formation (84.38%), and bone resorption (80.52%) as compared to arthritic control rats. Free methotrexate-treated group showed intermediate effects. However, drug-free aspasome treatment did not show any effect. The experimental results indicate a positive outcome in development of drug-loaded therapeutically active carrier system which presents a non-invasive controlled release transdermal formulation with good drug loading, drug permeation rate, and having better disease modifications against RA than the free drug, thereby providing a more attractive therapeutic strategy for rheumatoid disease management.


The impact of leadership training programs on physicians in academic medical centers: a systematic review.
To identify the impact of leadership training programs at academic medical centers (AMCs) on physicians' knowledge, skills, attitudes, behaviors, and outcomes. In 2011, the authors conducted a systematic review of the literature, identifying relevant studies by searching electronic databases (MEDLINE, EMBASE, CINAHL, Cochrane Central Register), scanning reference lists, and consulting experts. They deemed eligible any qualitative or quantitative study reporting on the implementation and evaluation of a leadership program for physicians in AMCs. Two independent reviewers conducted the review, screening studies, abstracting data, and assessing quality. The authors initially identified 2,310 citations. After the screening process, they had 11 articles describing 10 studies. Three were controlled before-and-after studies, four were before-and-after case series, and three were cross-sectional surveys. The authors did not conduct a meta-analysis because of the methodological heterogeneity across studies. Although all studies were at substantial risk of bias, the highest-quality ones showed that leadership training programs affected participants' advancement in academic rank (48% versus 21%, P=.005) and hospital leadership position (30% versus 9%, P=.008) and that participants were more successful in publishing papers (3.5 per year versus 2.1 per year, P<.001) compared with nonparticipants. The authors concluded that leadership programs have modest effects on outcomes important to AMCs. Given AMCs' substantial investment in these programs, rigorous evaluation of their impact is essential. High-quality studies, including qualitative research, will allow the community to identify which programs are most effective.


Phase I Study of Alvocidib Followed by 7+3 (Cytarabine + Daunorubicin) in Newly Diagnosed Acute Myeloid Leukemia.
Alvocidib is a cyclin-dependent kinase 9 inhibitor leading to downregulation of the antiapoptotic BCL-2 family member, MCL-1. Alvocidib has shown clinical activity in a timed sequential regimen with cytarabine and mitoxantrone in relapsed/refractory and newly diagnosed acute myeloid leukemia (AML) but has not been studied in combination with traditional 7+3 induction therapy. A multiinstitutional phase I dose-escalation study of alvocidib on days 1-3 followed by 7+3 (cytarabine 100 mg/m2/day i.v. infusion days 5-12 and daunorubicin 60 mg/m2 i.v. days 5-7) was performed in newly diagnosed AML ≤65 years. Core-binding factor AML was excluded. There was no MTD on this study; the recommended phase II dose of alvocidib was 30 mg/m2 i.v. over 30 minutes followed by 60 mg/m2 i.v. infusion over 4 hours. There was one dose-limiting toxicity of cytokine release syndrome. The most common grade ≥3 nonhematologic toxicities were diarrhea (44%) and tumor lysis syndrome (34%). Overall, 69% (22/32) of patients achieved complete remission (CR). In an exploratory cohort, eight of nine (89%) patients in complete remission had no measurable residual disease, as determined by a centralized flow cytometric assay. Clinical activity was seen in patients with secondary AML, AML with myelodysplastic syndrome-related changes, and a genomic signature of secondary AML (50%, 50%, and 92% CR rates, respectively). Alvocidib can be safely administered prior to 7+3 induction with encouraging clinical activity. These findings warrant further investigation of alvocidib combinations in newly diagnosed AML. This study was registered at clinicaltrials.gov identifier NCT03298984.


"Kind of empowered": Perceptions of socio-emotional development in children driving ride-on cars.
Early powered mobility (PM) experiences can be essential facilitators of self-initiated mobility, socialization, and exploration for young children with disabilities. Cerebral palsy (CP) and developmental delay are two of the most common diagnoses associated with motor disability in young children with 1 in 345 children diagnosed with CP and 1 in 6 with developmental delay in the US. The purpose of this study was to explore the longitudinal experiences and caregiver perceptions of socio-emotional development in particular, in young children with disabilities during modified ride-on car (ROC) use. A qualitative, grounded theory approach was used. Semi-structured interviews were conducted with 15 families (children ages 1-4 with CP or developmental delay) at baseline, 6 months (as able due to COVID), and 1 year following ROC introduction. Data were coded independently by three researchers using constant comparison until data saturation occurred and themes emerged. Four themes emerged from the data: "Leveling the Playing Field," "Breaking Down Barriers," "Fun and Work: ROC as Toy and Therapy Device," and "Mobility is a Pathway to Autonomy." Conclusions/Implication: Children and caregivers viewed ROCs as both fun and therapeutic, consistently identifying perceived benefits for children's socio-emotional development. This qualitative study provides a better understanding of the complexities and impact of ROCs on children and their families in the socio-emotional domain and may help facilitate clinical decision-making when introducing PM to young children with disabilities as part of a multimodal approach to early intervention. (PsycInfo Database Record (c) 2023 APA, all rights reserved).