Time course of retinal degeneration associated with the absence of 1, 4, 5-inositol trisphosphate receptor in Drosophila melanogaster.
The absence of the inositol trisphosphate receptor is associated with a gradual retinal degeneration in Drosophila melanogaster. To characterize the time-course profile of this process, mosaic flies expressing a null allele of the itp gene in the eye were studied by electroretinograms and electronic microscopy. Membrane contour alterations, disrupted mitochondria, altered morphology and even loss of photoreceptors were increased progressively starting 5 d after hatching, were more evident during days 10-15 and promoted highly disorganized structures thereafter. Comparison between electroretinograms recorded in wild type and mutant tissues showed progressive differences in the on and off transients as well as in the magnitude of the summed receptor potentials of photoreceptor cells from day 5 of hatching, [corrected] and the functional defects became progressively more severe. Unexpectedly, these alterations were detected not only in the non-pigmented mutant ommatidia, but also in the pigmented ommatidia, including heterozygous and twin clones expressing 1, 4, 5-inositol trisphosphate receptor (IP(3)R). To explore the mechanism underlying this degenerative process, the progression of pro-oxidant and apoptotic reactions was characterized by immunohistochemical techniques. Mutant ommatidia showed intermittent episodes of increased pro-oxidant reactions (detected as adducts of 4-hydroxy-nonenal) throughout the fly's life. Similarly, several episodes of active caspase 3, an apoptotic effector, were evident with the same time pattern. Episodes of enhanced lipid peroxidation and apoptosis were also observed in the pigmented ommatidia of the mosaic eyes. The results indicate that photoreceptors lacking IP(3)R suffer episodes of increased lipid peroxidation, which eventually perturb the retinal subcellular organization and disrupt the phototransduction process and cell viability. Pigmented ommatidia also showed a similar pattern of damage, indicating that the degenerative process is non-autonomous and is so intense that it propagated to the non-mutant retinal cells in the mosaic eyes. In conclusion, ommatidia with a null mutation of IP(3)R degenerate by a process associated with intermittent lipid peroxidation and apoptotic activities.


Multidrug resistant pathogenic Escherichia coli status in water sources and Yamuna River in and around Mathura, India.
The present study was taken to understand the status of water resources in the holy city of Mathura, India. As it is a tourist place and pilgrims come from all around the world, there are more chances of spread of pathogens through them. The pathogens which are particularly excreted through urine and feces are most commonly excreted out. E. coli is one of them which causes many severe diseases particularly in neonatal calves or neonates of human. As the city has its limitation of accommodation and river Yamuna is also shrinking, the pressure of millions of pilgrims has enormous, drastic and stipulated effect on water resources particularly river Yamuna. The samples collected showed the presence of pathogenic E. coli. Out of total 100 samples 40 were found positive for E. coli. The 60 samples of different water resources showed the presence of E. coli in 26 samples while 14 were found positive out of 40 samples of Yamuna water. The total coliforms count ranged between 1.08 log10 CFU m L(-1) to 6.34 log10 CFU m L(-1) in drinking water sources and waste water, respectively. The high percentage of positive samples and coliforms count may be because of the method and place of sample collection. When these isolates were analyzed for antibiotic sensitivity pattern against some commonly used antibiotics. They showed a variable effectiveness against them. The number of resistant isolate is significant enough to make an alert at the earliest to protect the water resources and also to secure better and brighter future of human population.


Tonsillectomy and the value of peritonsillar infiltrations.
In this prospective double-blind trial, the effect of peritonsillar infiltrations was assessed with regard to intra-operative bleeding and postoperative pain in 100 young adults who underwent elective tonsillectomy under balanced general anaesthesia. Patients were randomly assigned to 1 of 5 groups (20 patients in each group) and received infiltrations as follows: group 1--bupivacaine (0.5%) with adrenaline (1:200,000); group II--bupivacaine (0.5%); group III--normal saline with adrenaline (1:200,000); group IV--normal saline; group V--no infiltration (control group). With regard to blood loss, groups I-IV lost a mean of 47 ml (95% CI = 69.78), while group V lost a mean of 121 ml (95% CI = 78.10) (P = 0.0002). Group V had the highest pain score (average 4.62) measured in recovery, 4 and 24 hours after tonsillectomy (P = 0.0051) and required more narcotic analgesia. Groups II and III had the lowest score (average 1.72) after 24 hours. The bupivacaine group (II) had the highest incidence (71.4%) of nausea and vomiting. Peritonsillar infiltrations decrease intra-operative bleeding and pain, independent of the type of solution infiltrated, by providing a better defined plane of dissection which minimises trauma to the surrounding tissue. Normal saline infiltrations with or without adrenaline should be used since they have no side-effects and are inexpensive and easily available.


Switch from stress response to homeobox transcription factors in adipose tissue after profound fat loss.
In obesity, impaired adipose tissue function may promote secondary disease through ectopic lipid accumulation and excess release of adipokines, resulting in systemic low-grade inflammation, insulin resistance and organ dysfunction. However, several of the genes regulating adipose tissue function in obesity are yet to be identified. In order to identify novel candidate genes that may regulate adipose tissue function, we analyzed global gene expression in abdominal subcutaneous adipose tissue before and one year after bariatric surgery (biliopancreatic diversion with duodenal switch, BPD/DS) (n = 16). Adipose tissue from lean healthy individuals was also analyzed (n = 13). Two different microarray platforms (AB 1700 and Illumina) were used to measure the differential gene expression, and the results were further validated by qPCR. Surgery reduced BMI from 53.3 to 33.1 kg/m(2). The majority of differentially expressed genes were down-regulated after profound fat loss, including transcription factors involved in stress response, inflammation, and immune cell function (e.g., FOS, JUN, ETS, C/EBPB, C/EBPD). Interestingly, a distinct set of genes was up-regulated after fat loss, including homeobox transcription factors (IRX3, IRX5, HOXA5, HOXA9, HOXB5, HOXC6, EMX2, PRRX1) and extracellular matrix structural proteins (COL1A1, COL1A2, COL3A1, COL5A1, COL6A3). The data demonstrate a marked switch of transcription factors in adipose tissue after profound fat loss, providing new molecular insight into a dichotomy between stress response and metabolically favorable tissue development. Our findings implicate homeobox transcription factors as important regulators of adipose tissue function.


Apigenin inhibits TGF-β1-induced proliferation and migration of airway smooth muscle cells.
It is well known that the proliferation and migration of ASM cells (ASMCs) plays an important role in the pathogenesis of airway remodeling in asthma. Previous studies reported that apigenin can inhibit airway remodeling in a mouse asthma model. However, its effects on the proliferation and migration of ASMCs in asthma remain unknown. Therefore, the aim of our present study was to investigate the effects of apigenin on ASMC proliferation and migration, and explore the possible molecular mechanism. We found that apigenin inhibited transforming growth factor-β1 (TGF-β1)-induced ASMC proliferation. The cell cycle was blocked at G1/S-interphase by apigenin. It also suppressed TGF-β1-induced ASMCs migration. Furthermore, apigenin inhibited TGF-β1-induced Smad 2 and Smad 3 phosphorylation in ASMCs. Taken together, these results suggested that apigenin inhibited the proliferation and migration of TGF-β1-stimulated ASMCs by inhibiting Smad signaling pathway. These data might provide useful information for treating asthma and show that apigenin has potential for attenuating airway remodeling.


Rare Case of Multiple Perirenal, Extra-Adrenal Myelolipoma: Case Report, Current Management Options, and Literature Review.
Extra-adrenal myelolipomas are rare, asymptomatic entities, although large tumors may cause local symptoms or hemorrhage. When these lesions occur outside the adrenals in the retroperitoneum, they are radiographically easily confused with both primary and secondary retroperitoneal tumors, which tend to be aggressive. Although myelolipomas are benign and can be managed conservatively, if malignancy is suspected, a surgical procedure is an option. We report a case of a 68-year-old patient with multiple perirenal extra-adrenal myelolipomas. Initial abdominal ultrasound reviled an inhomogeneous mass surrounding the left kidney. Subsequent CT examination of the abdomen showed four separate, extrarenal, well-circumscribed, round-shaped, fat-containing retroperitoneal tumors. Given the significant size of the masses, that compressed major abdominal vessels and the suspicion of liposarcoma, a surgical excision of the lesions was performed. The tumors were easily separated, all surrounding structures were spared, and they were removed completely. Histologically, all masses consisted of hematopoietic and mature fat tissue and the final diagnosis was extra-adrenal myelolipoma. The patient was released from the hospital 7th day after surgery in good condition and at his baseline. Since myelolipomas are, by definition, nonfunctional benign tumors, there was no need for further follow-up. The radiological evaluation and fine needle biopsy are usually sufficient to establish the diagnosis, but in some cases of well-differentiated liposarcoma, the differentiation between myelolipoma and liposarcoma can be challenging. Therefore, considering that myelolipomas and liposarcomas have opposite prognoses, which affects the surgeon's decision on the extent of surgical procedure and further treatment, we also emphasize the importance of intraoperative assessment of the tumor, both by the surgeon and by intraoperative pathology consultation.


Experimental and numerical predictions of the ultimate strength of a low-cost composite transtibial prosthesis.
Saito et al. (Modern Plastics 1997;74:175-7) have developed a low-cost transtibial prosthesis made of fiber-reinforced plastic (FRP). The prosthesis is comprised of an aluminum pylon, a cosmetic cover, and a constant cross-section composite foot into which aluminum supports are screwed to increase load-bearing capacity. Replacing these supports with a single integrated FRP stiffener significantly reduced manufacturing cost while providing high strength, great durability, and smooth walking. The optimal location and orientation of the proposed FRP stiffener were determined by finite element (FE) analysis. When a replica of this component was tested according to ISO standard 10328, the experimental prosthesis catastrophically failed under 6,600 N of force. Maximum percent error of strain between experimental and numerical results was 18.6%, showing good correlation between the two data sets. The optimized design provides sufficient strength and reduces the cost of manufacturing and thus can be used to replace the original design.


Lupus vasculopathy: Diagnostic, pathogenetic and therapeutic considerations.
A rare form of vascular disease in systemic lupus erythematosus (SLE), lupus vasculopathy is characterized by necrosis and accumulation of immunoglobulins (IGs) and complements in the wall of arterioles and small arteries resulting in luminal narrowing. Lupus vasculopathy often accompanies lupus nephritis and portends a poor prognosis. Although there is general agreement on the treatment of lupus nephritis, effective treatment strategies for lupus vasculopathy remain to be defined. We report a 20-year-old woman with SLE who presented with generalized tonic-clonic seizure. Her immunosuppressive regimen consisted of mycophenolate mofetil, prednisone and hydroxychloroquine. On physical examination, she was Cushingoid in appearance and hypertensive. Laboratory tests indicated renal disease. Coagulation studies disclosed de novo lupus anticoagulant. Magnetic resonance imaging of the brain demonstrated acute focal cerebral hemorrhage. Echocardiography revealed reduced ejection fraction and severe mitral regurgitation. Despite high-dose glucocorticoids and mycophenolate mofetil, renal function remained poor. Kidney biopsy demonstrated lupus vasculopathy and glomerulonephritis. Plasma exchange therapy and intravenous cyclophosphamide were administered. Over the ensuing four weeks, renal function improved, complement levels increased, autoantibody titers decreased and lupus anticoagulant disappeared. In conclusion, lupus vasculopathy can occur in SLE despite a heavy immunosuppressive regimen. Antiphospholipid antibodies might be involved in the pathogenesis of lupus vasculopathy. Plasma exchange therapy in conjunction with intravenous cyclophosphamide may represent an effective treatment strategy for lupus vasculopathy.


Unfolding dynamics of proteins under applied force.
Understanding the mechanisms of protein folding is a major challenge that is being addressed effectively by collaboration between researchers in the physical and life sciences. Recently, it has become possible to mechanically unfold proteins by pulling on their two termini using local force probes such as the atomic force microscope. Here, we present data from experiments in which synthetic protein polymers designed to mimic naturally occurring polyproteins have been mechanically unfolded. For many years protein folding dynamics have been studied using chemical denaturation, and we therefore firstly discuss our mechanical unfolding data in the context of such experiments and show that the two unfolding mechanisms are not the same, at least for the proteins studied here. We also report unexpected observations that indicate a history effect in the observed unfolding forces of polymeric proteins and explain this in terms of the changing number of domains remaining to unfold and the increasing compliance of the lengthening unstructured polypeptide chain produced each time a domain unfolds.


Resolution enhancement method for lensless in-line holographic microscope with spatially-extended light source.
We propose a resolution enhancement method for lensless in-line holographic microscope (LIHM) with spatially-extended light source, where the resolution is normally deteriorated by the insufficient spatial coherence of the illumination. In our LIHM setup, a light-emitting diode (LED), which was a spatially-extended light source, directly illuminated the sample, and the in-line hologram were recorded by a CMOS imaging sensor located behind the sample. In our holographic reconstruction process, the in-line hologram was first deconvoled with a properly resized image of the LED illumination area, and then back-propagated with scalar diffraction formula to reconstruct the sample image. We studied the hologram forming process and showed that the additional deconvolution process besides normal scalar diffraction reconstruction in LIHM can effectively enhance the imaging resolution. The resolution enhancements capability was calibrated by numerical simulations and imaging experiments with the U.S. air force target as the sample. We also used our LIHM to image the wing of a green lacewing to further demonstrate the capability of our methods for practical imaging applications. Our methods provide a way for LIHM to achieve satisfactory resolution with less stringent requirement for spatial coherence of the source and could reduce the cost for compact imaging system.


Redrawing the Iceland-Scotland Overflow Water pathways in the North Atlantic.
Iceland-Scotland Overflow Water (ISOW) is a primary deep water mass exported from the Norwegian Sea into the North Atlantic as part of the global Meridional Overturning Circulation. ISOW has historically been depicted as flowing counter-clockwise in a deep boundary current around the subpolar North Atlantic, but this single-boundary-following pathway is being challenged by new Lagrangian observations and model simulations. We show here that ISOW leaves the boundary and spreads into the interior towards the central Labrador and Irminger basins after flowing through the Charlie-Gibbs Fracture Zone. We also describe a newly observed southward pathway of ISOW along the western flank of the Mid-Atlantic Ridge. The partitioning of these pathways is shown to be influenced by deep-reaching eddies and meanders of the North Atlantic Current. Our results, in tandem with previous studies, call for a revision in the historical depiction of ISOW pathways throughout the North Atlantic.


Angiogenesis in chronic obstructive pulmonary disease: a translational appraisal.
Angiogenesis is a crucial component of lung pathophysiology, not only in cancer but also in other disorders, such as chronic obstructive pulmonary disease (COPD). In COPD angiogenesis is definitely able to control and orchestrate the progression of airway remodeling. Herein, we provide several remarkable translational aspects of angiogenesis in COPD, exploring both basic and clinical research in this field. Indeed, we present a number of pro- and anti-angiogenic factors, which can be also used as potential biomarkers to monitor disease progression.


Adherence to HIV antiretroviral therapy among pregnant and postpartum women during the Option B+ era: 12-month cohort study in urban South Africa and rural Uganda.
We conducted a cohort study to understand patterns of anti-retroviral therapy (ART) adherence during pregnancy, postpartum and non-pregnancy follow-up among women initiating ART in public clinics offering Option B+ in rural Uganda and urban South Africa. We collected survey data, continuously monitored ART adherence (Wisepill), HIV-RNA and pregnancy tests at zero, six and twelve months from women initiating ART in Uganda and South Africa, 2015 to 2017. The primary predictor of interest was follow-up time categorized as pregnant (pregnancy diagnosis to pregnancy end), postpartum (pregnancy end to study exit) or non-pregnancy-related (neither pregnant nor postpartum). Fractional regression models included demographics and socio-behavioural factors informed by the Behavioral Model for Vulnerable Populations. We evaluated HIV-RNA at 12 months by ever- versus never-pregnant status. In Uganda, 247 women contributed 676, 900 and 1274 months of pregnancy, postpartum and non-pregnancy-related follow-up. Median ART adherence was consistently ≥90%: pregnancy, 94% (interquartile range [IQR] 78,98); postpartum, 90% (IQR 70,97) and non-pregnancy, 90% (IQR 80,98). Poorer adherence was associated with younger age (0.98% [95% CI 0.33%, 1.62%] average increase per year of age) and higher CD4 cell count (1.01% [0.08%, 1.94%] average decrease per 50 cells/mm3 ). HIV-RNA was suppressed among 91% (N = 135) ever-pregnant and 86% (N = 85) never-pregnant women. In South Africa, 190 women contributed 259, 624 and 1247 months of pregnancy, postpartum and non-pregnancy-related follow-up. Median adherence was low during pregnancy, 74% (IQR 31,96); postpartum, 40% (IQR 4,65) and non-pregnancy, 77% (IQR 47,92). Poorer adherence was associated with postpartum status (22.3% [95%CI 8.6%, 35.4%] average decrease compared to non-pregnancy-related follow-up) and less emotional support (1.4% [0.22%, 2.58%] average increase per unit increase). HIV-RNA was suppressed among 57% (N = 47) ever-pregnant and 86% (N = 93) never-pregnant women. Women in rural Uganda maintained high adherence with 91% of ever-pregnant and 86% of never-pregnant women suppressing HIV-RNA at 12 months. Women in urban South Africa struggled with adherence, particularly during postpartum follow-up with median adherence of 40% and 57% of women with HIV-RNA suppression at one year, suggesting a crisis for postpartum women with HIV in South Africa. Findings suggest that effective interventions should promote emotional support.


[Problems of hemotransfusion from the evidence-based medicine point of view].
The operational analysis of hospital is conducted on blood transfusion for 5 years. For this time the main parameters of activity were improved and the quantity of blood transfusions has decreased The parameters of activity have not worsened. The number of AIDS--carriers was considerably increased, high there is still the level of a morbidity by hepatitis B and hepatitis C. Presently blood transfusion is equated to transplantation of an organs. From the attitude of evidential medicine it is necessary to prune the number of blood transfusions. The complications of a blood transfusion have been analyzed. The alternate methods of treatment are induced.


[Self-organization of physiological, pathological and ambivalent functional systems].
Development of P.K. Anokhin's theory of functional systems rejects the idea of adaptation result as a universal system-forming factor. This integrates the theory with conception on pathological and ambivalent systems. The existence of several system-forming factors is validated as well as their individual ability to create physiological, pathological and ambivalent systems. Perspectives of further transformation of the idea on the mechanism of system formation is discussed.


Elevated sweat chloride concentration in children without cystic fibrosis who are receiving topiramate therapy.
Topiramate, which is used as an anticonvulsant and for migraine prophylaxis in children, causes oligohydrosis as a side-effect, but its effect on sweat chloride concentrations has not been studied systematically. Twenty-one children receiving topiramate and 20 healthy controls with no signs or symptoms of pulmonary or gastrointestinal disease and a negative family history for cystic fibrosis (CF) underwent bilateral pilocarpine iontophoresis and sweat collection via Macroduct® system. Adequate samples (>15 µl volume) were obtained from 17/19 topiramate subjects (89%), and 19/20 (95%) controls. The mean sweat chloride concentration was 37.7 ± 18.8 mmol/L for patients receiving topiramate, and 15.9 ± 6.9 mmol/L for controls (p = 0.0001). The mean sweat volume was 29.1 ± 17.4 µl for patients receiving topiramate, and 41.2 ± 17.5 µl for controls (p = 0.037). Overall 8/17 (47%) of patients on topiramate with a measurable sweat chloride had either an intermediate (>40 mmol/L but <60 mmol/L) or elevated (>60 mmol/L) sweat chloride test result, while 0/19 control subjects had elevated sweat chloride (p = 0.0008). Further analysis of the in vitro activity of topiramate on cultured human bronchial epithelial cells in modified Ussing chambers showed no differences in chloride conductance measured in cells exposed to 10 or 50 µg/ml of topiramate when compared to non-exposed cells. This is the first report of a medication affecting sweat chloride values and shows that topiramate therapy can cause elevated sweat chloride concentrations in the absence of clinical manifestations of CF.


Evaluation of a New All-in-One Optical Biometer and Comparison With a Validated Swept-source OCT Biometer.
To assess agreement between a new all-in-one non-contact optical biometer based on optical low coherence reflectometry (SW-9000 µm Plus; Suoer) and a swept-source optical coherence tomography biometer (OA-2000; Tomey). Each eye was scanned three times in a row by each device at random. The measured ocular parameters included central corneal thickness (CCT), anterior chamber depth (ACD), lens thickness (LT), axial length (AL), flat keratometry (Kf), steep keratometry (Ks), mean keratometry (Km), astigmatism, corneal diameter (CD), and pupil diameter (PD). The paired t test was used to show the differences between the SW-9000 and OA-2000. Bland-Altman plots and the 95% limits of agreement (LoA) were applied to assess the consistency of the measurements. Sixty eyes from 60 healthy participants were examined, with a mean spherical equivalent refraction of -5.58 ± 2.31 diopters and a mean age of 30.40 ± 6.07 years. The Bland-Altman plots showed high agreement for AL, ACD, LT, Kf, Ks, Km, astigmatism, and CD measurements (95% LoA: -0.06 to 0.04 mm, -0.10 to 0.06 mm, -0.12 to 0.11 mm, -0.30 to 0.29 D, -0.35 to 0.38 D, -0.29 to 0.30 D, -0.30 to 0.34 D, and -0.50 to 0.06 mm, respectively), whereas the agreement for CCT and PD were moderate (95% LoA: 7.12 to 20.43 µm, -0.75 to 1.19 mm, respectively). The new all-in-one non-contact biometer had high agreement with the OA-2000 biometer on the AL, ACD, LT, Kf, Ks, Km, astigmatism, and CD measurements. For most of the ocular parameters assessed, they were clinically interchangeable. [J Refract Surg. 2023;39(12):825-830.].


Bridging the gap between emotion and joint action.
Our daily human life is filled with a myriad of joint action moments, be it children playing, adults working together (i.e., team sports), or strangers navigating through a crowd. Joint action brings individuals (and embodiment of their emotions) together, in space and in time. Yet little is known about how individual emotions propagate through embodied presence in a group, and how joint action changes individual emotion. In fact, the multi-agent component is largely missing from neuroscience-based approaches to emotion, and reversely joint action research has not found a way yet to include emotion as one of the key parameters to model socio-motor interaction. In this review, we first identify the gap and then stockpile evidence showing strong entanglement between emotion and acting together from various branches of sciences. We propose an integrative approach to bridge the gap, highlight five research avenues to do so in behavioral neuroscience and digital sciences, and address some of the key challenges in the area faced by modern societies.


Pudilan Keyanning mouthwash inhibits dextran-dependent aggregation and biofilm organization of Streptococcus mutans.
This research aimed to investigate the inhibitory effects of Pudilan mouthwash (PDL) on Streptococcus mutans (S. mutans) biofilms and identify its chemical components. The impacts of 100% concentrated PDL on S. mutans biofilm were detected by colony-forming unit (CFU) assays, crystal violet staining, confocal laser scanning microscopy (CLSM), scanning electron microscopy (SEM) and quantitative real-time PCR (qRT‒PCR). The biocompatibility with human gingival fibroblasts (HGFs) was evaluated by Cell-Counting-Kit-8 (CCK-8) assay. And chemical components were identified by UPLC-HRMS. PBS and 0.12% chlorhexidine were used as negative control and positive control, respectively. Results indicate early 8 h S. mutans biofilms are sensitive to PDL. Additionally, it leads to a decrease in bacterial activities and dextran-dependent aggregation in 24 h S. mutans biofilms. PDL significantly downregulates the gene expression of gtfB/C/D and smc. And 114 components are identified. PDL has an inhibitory effect on S. mutans and favorable biocompatibility. It has potential to be exploited as a novel anti-biofilm agent.


Interactions with soil biota shift from negative to positive when a tree species is moved outside its native range.
Studies evaluating plant-soil biota interactions in both native and introduced plant ranges are rare, and thus far have lacked robust experimental designs to account for several potential confounding factors. Here, we investigated the effects of soil biota on growth of Pinus contorta, which has been introduced from Canada to Sweden. Using Swedish and Canadian soils, we conducted two glasshouse experiments. The first experiment utilized unsterilized soil from each country, with a full-factorial cross of soil origin, tree provenance, and fertilizer addition. The second experiment utilized gamma-irradiated sterile soil from each country, with a full-factorial cross of soil origin, soil biota inoculation treatments, tree provenance, and fertilizer addition. The first experiment showed higher seedling growth on Swedish soil relative to Canadian soil. The second experiment showed this effect was due to differences in soil biotic communities between the two countries, and occurred independently of all other experimental factors. Our results provide strong evidence that plant interactions with soil biota can shift from negative to positive following introduction to a new region, and are relevant for understanding the success of some exotic forest plantations, and invasive and range-expanding native species.


Disruption and therapeutic rescue of autophagy in a human neuronal model of Niemann Pick type C1.
An unresolved issue about many neurodegenerative diseases is why neurons are particularly sensitive to defects in ubiquitous cellular processes. One example is Niemann Pick type C1, caused by defects in cholesterol trafficking in all cells, but where neurons are preferentially damaged. Understanding this selective failure is limited by the difficulty in obtaining live human neurons from affected patients. To solve this problem, we generated neurons with decreased function of NPC1 from human embryonic stem cells and used them to test the hypothesis that defective cholesterol handling leads to enhanced pathological phenotypes in neurons. We found that human NPC1 neurons have strong spontaneous activation of autophagy, and, contrary to previous reports in patient fibroblasts, a block of autophagic progression leading to defective mitochondrial clearance. Mitochondrial fragmentation is an exceptionally severe phenotype in NPC1 neurons compared with fibroblasts, causing abnormal accumulation of mitochondrial proteins. Contrary to expectation, these abnormal phenotypes were rescued by treatment with the autophagy inhibitor 3-methyladenine and by treatment with the potential therapeutic cyclodextrin, which mobilizes cholesterol from the lysosomal compartment. Our findings suggest that neurons are especially sensitive to lysosomal cholesterol accumulation because of autophagy disruption and accumulation of fragmented mitochondria, thus defining a new route to effective drug development for NPC1 disease.


Myocardial extravascular extracellular volume fraction measurement by gadolinium cardiovascular magnetic resonance in humans: slow infusion versus bolus.
Myocardial extravascular extracellular volume fraction (Ve) measures quantify diffuse fibrosis not readily detectable by conventional late gadolinium (Gd) enhancement (LGE). Ve measurement requires steady state equilibrium between plasma and interstitial Gd contrast. While a constant infusion produces steady state, it is unclear whether a simple bolus can do the same. Given the relatively slow clearance of Gd, we hypothesized that a bolus technique accurately measures Ve, thus facilitating integration of myocardial fibrosis quantification into cardiovascular magnetic resonance (CMR) workflow routines. Assuming equivalence between techniques, we further hypothesized that Ve measures would be reproducible across scans. In 10 volunteers (ages 20-81, median 33 yr, 3 females), we compared serial Ve measures from a single short axis slice from two scans: first, during a constant infusion, and second, 12-50 min after a bolus (0.2 mmol/kg gadoteridol) on another day. Steady state during infusion was defined when serial blood and myocardial T1 data varied <5%. We measured T1 on a 1.5 T Siemens scanner using a single-shot modified Look Locker inversion recovery sequence (MOLLI) with balanced SSFP. To shorten breath hold times, T1 values were measured with a shorter sampling scheme that was validated with spin echo relaxometry (TR = 15 sec) in CuSO4-Agar phantoms. Serial infusion vs. bolus Ve measures (n = 205) from the 10 subjects were compared with generalized estimating equations (GEE) with exchangeable correlation matrices. LGE images were also acquired 12-30 minutes after the bolus. No subject exhibited LGE near the short axis slices where Ve was measured. The Ve range was 19.3-29.2% and 18.4-29.1% by constant infusion and bolus, respectively. In GEE models, serial Ve measures by constant infusion and bolus did not differ significantly (difference = 0.1%, p = 0.38). For both techniques, Ve was strongly related to age (p < 0.01 for both) in GEE models, even after adjusting for heart rate. Both techniques identically sorted older individuals with higher mean Ve values. Myocardial Ve can be measured reliably and accurately 12-50 minutes after a simple bolus. Ve measures are also reproducible across CMR scans. Ve estimation can be integrated into CMR workflow easily, which may simplify research applications involving the quantification of myocardial fibrosis.


Soluble fusion expression and characterization of human beta-defensin 3 using a novel approach.
Human β-defensin 3 (DEFB103) is a recently identified small cysteine-rich cationic peptide expressed ubiquitously upon local microbial invasion. A number of accumulating evidences indicate that this peptide is involved in many biological processes, including microbicidal activities, chemoattraction, and immunomodulation. In this article, we describe a novel approach through which we performed the expression and purification of the recombinant DEFB103 peptide in Escherichia coli (E. coli) based on the pTWIN1 expression system. This approach does not introduce any extra residues to the peptide product, and also eliminates the requirement of removing the fusion tag by exogenous proteases. A high yield of 112 mg of soluble fusion DEFB103 was obtained in 1 liter of Luria-Bertani (LB) medium. By one-step affinity chromatography and on-column, auto-cleavage of the fusion tag, the mature DEFB103 peptide was produced with a yield of 30 mg/L LB. The purified DEFB103 peptide demonstrated strong antimicrobial activities against E. coli, S. aureus and C. albicans, which were representatives of Gram-negative and Gram-positive bacteria and fungi, respectively. Using this novel approach, we have successfully expressed and purified several human defensins with significant bioactivities. Our work may be helpful for structural and functional studies of other human defensins, and also for the production of human defensins.


Optimizing Enrollment of Patients into Nephrology Research Studies.
Advances in medical care and biomedical research depend on the participation of human subjects. Poor patient enrollment in research has limited past clinical and translational research endeavors in nephrology. Simultaneously, patients and their caregivers are seeking better diagnostic, monitoring, and therapeutic approaches to improve or restore kidney and overall health. This manuscript will discuss a framework and strategies to optimize patient enrollment within nephrology research and provide examples of success from existing nephrology research programs.


[Detection of tick-borne encephalitis virus in Ixodes ticks collected in a natural focus of Gornyi Altai].
Enzyme immunoassay of tick-borne encephalitis virus (TBE) in the samples of Ixodes ticks collected in the outskirts of the settlement of Manzherok, Maiminsk District, Republic of Altai, revealed TBE antigen in 16.9 +/- 1.9% of the talga ticks. Real-time reverse transcription-polymerase chain reaction (RT-PCR) with specific fluorescent probes and phylogenetic analysis of the nucleotide sequences of RT-RCR products corresponding to 5'-terminal fragment of the E gene of TBE, all the virus strains isolated from the ticks collected in Gomyi Altai were referred to as the Siberian genetic type that was dominant in virus-carrying ticks in the majority of endemic areas of Russia and near abroad. Viral load assays using the real-time RT-PCR with the probes indicated the threshold cycles Ct = 25.34-28.98, which, with regard to the efficiency of RNA identification and reverse transcription, was equal to about 10(4)-10(5) viral RNA copies per tick.


Mass Customization of Polylactic Acid (PLA) Parts via a Hybrid Manufacturing Process.
Mass customization is the development of items tailored to specific customers, but produced at low unit cost in high-volume. In this context, hybrid manufacturing (HM) combines fused deposition modeling (FDM) and injection molding (IM) to fabricate a single personalized part with minimum manufacturing cost. In this technique, inserts with different physical features are first FDM-fabricated and then IM-overmolded. This study investigated the effect of hybrid FDM-IM production technology, FDM insert geometry on mechanical properties, and micro-structural evolution of Polylactic Acid (PLA) samples. The findings indicated a comparable tensile properties of FDM-IM samples (68.38 MPa) to IM batch (68.95 MPa), emphasizing the potential of HM in the manufacturing industry. Maximum tensile stress of FDM-IM specimens shows an upward trend due to the increased infill density of preforms. In addition, overmolding interface direction results in a big gap for the maximum tensile strengths between half-length series specimens (12.99 MPa to 19.09 MPa) and half-thickness series specimens (53.83 MPa to 59.92 MPa). Furthermore, four joint configurations resulted in different mechanical performances of finished specimens, in which the female cube sample exhibits the highest tensile stress (68.38 MPa), while the batch with male T joint shows a lower value in maximum tensile strength (59.51 MPa), exhibiting a similar tensile performance with the half-thickness 75% batch without joint configuration. This study lays the groundwork for using HM to produce bespoke and mechanically improved parts over FDM alone.


Serum Galectin-3 Level Is Positively Associated with Endothelial Dysfunction in Patients with Chronic Kidney Disease Stage 3 to 5.
Galectin-3, which is a novel biomarker of cardiovascular stress and related to inflammation, could predict adverse cardiovascular events. However, its relationship with endothelial function in patients with chronic kidney disease (CKD) remains inconclusive. This study aimed to investigate the association between serum galectin-3 levels and endothelial function in patients with stages 3-5 CKD. Fasting blood samples were obtained from 130 patients. Serum galectin-3 levels were determined using the enzyme-linked immunosorbent assay. The endothelial function, demonstrated as a vascular reactivity index (VRI), was measured noninvasively through digital thermal monitoring test. Then, we sorted the patients into poor, intermediate, and good vascular reactivity (VRI < 1.0, 1.0 ≤ VRI < 2.0, and VRI ≥ 2.0), accounting for 24 (18.5%), 44 (33.8%), and 62 (47.7%) patients, respectively. As the VRI decreased, the serum galectin-3 and C-reactive protein (CRP) levels significantly increased. The galectin-3 value positively correlated with the CRP value but negatively correlated with estimated glomerular filtration rate. In multivariable stepwise linear regression analysis, serum log-transformed galectin-3 level and log-transformed CRP were significantly negatively associated with VRI values. Therefore, galectin-3 together with CRP is associated with VRI values and is a potential endothelial function modulator and a valuable biomarker of endothelial dysfunction in patients with CKD.


Regulation of the catechol 1,2-dioxygenase- and phenol monooxygenase-encoding pheBA operon in Pseudomonas putida PaW85.
In Pseudomonas putida PaW85, the ortho-cleavage pathway is used for catechol degradation. The 11.4-kb XhoI fragment cloned from phenol degradation plasmid pEST1226 into pKT240 (recombinant plasmid pAT1140) contains the inducible pheBA operon that encodes catechol 1,2-dioxygenase (gene pheB) and phenol monooxygenase (gene pheA), the first two enzymes for the phenol degradation pathway. The promoter of the pheBA operon is mapped 1.5 kb upstream of the pheB gene. The plasmid pAT1140, when introduced into P. putida PaW85, enables the bacteria to use the hybrid plasmid-chromosome-encoded pathway for phenol degradation. The synthesis of the plasmid-encoded phenol monooxygenase and catechol 1,2-dioxygenase is induced by cis,cis-muconate. The expression studies of the deletion subclones derived from pAT1140 revealed that the transcription of the pheBA operon is positively controlled by a regulatory protein that is chromosomally encoded in P. putida. cis,cis-Muconate in cooperation with positive transcription factor CatR activates the transcription of the chromosomal ortho-pathway genes catA and catBC in P. putida (R. K. Rothmel, T. L. Aldrich, J. E. Houghton, W. M. Coco, L. N. Ornston, and A. M. Chakrabarty, J. Bacteriol. 172:922-931, 1990). The inability to express the pheBA operon in a P. putida CatR- background and activation of transcription of the pheBA operon in Escherichia coli in the presence of the catR-expressing plasmid demonstrated that the transcription of the pheBA operon in P. putida PaW85 carrying pEST1226 is controlled by the chromosomally encoded CatR.


Immunoregulatory Effect of Calcitriol on Experimental Autoimmune Encephalomyelitis (EAE) Mice.
Previous studies noted an imbalance in T helper (Th) 17 and regulatory T cells (Tregs) in experimental autoimmune encephalomyelitis (EAE), a multiple sclerosis animal model. calcitriol, vitamin D's active form, was found to ameliorate EAE symptoms by favoring Tregss over Th17 cells, suggesting immunomodulatory effects. This study aimed to assess calcitriol's impact on EAE manifestations and cytokine profile in mice. In this study, we recruited twenty-eight C57BL/6 mice and divided them into 4 groups: healthy controls, EAE, EAE with calcitriol treatment, and healthy mice with calcitriol treatment. CD4+ T cells were isolated from splenocytes using magnetic-activated cell sorting. Real-time polymerase chain reaction was employed to quantify the genes associated with Th9 cells (i.e., SPI1 encoding PU.1 and IL9 encoding interleukin [IL]-9). Moreover, the levels of IL-17 and transforming growth factor beta (TGF-β) were evaluated through enzyme-linked immunosorbent assay in the supernatant of CD4+ T cell culture stimulated by anti-CD3 and anti-CD28 antibodies for 72 hours. In the supernatant of CD4+ T cell cultures, the levels of interleukin-17 (IL-17) were significantly increased, while the levels of transforming growth factor beta (TGF-β) were decreased in the EAE Group compared to the healthy control group. Calcitriol treatment reversed these changes and attenuated EAE symptoms, as confirmed in hematoxylin and eosin, and luxol fast blue stains. Notably, calcitriol increased IL9 gene expression in both EAE and healthy mice. This study provides further evidence of the anti-inflammatory effects of calcitriol and its role in attenuating EAE.


Picturing pathogen infection in plants.
Several imaging techniques have provided valuable tools to evaluate the impact of biotic stress on host plants. The use of these techniques enables the study of plant-pathogen interactions by analysing the spatial and temporal heterogeneity of foliar metabolism during pathogenesis. In this work we review the use of imaging techniques based on chlorophyll fluorescence, multicolour fluorescence and thermography for the study of virus, bacteria and fungi-infected plants. These studies have revealed the impact of pathogen challenge on photosynthetic performance, secondary metabolism, as well as leaf transpiration as a promising tool for field and greenhouse management of diseases. Images of standard chlorophyll fluorescence (Chl-F) parameters obtained during Chl-F induction kinetics related to photochemical processes and those involved in energy dissipation, could be good stress indicators to monitor pathogenesis. Changes on UV-induced blue (F440) and green fluorescence (F520) measured by multicolour fluorescence imaging in pathogen-challenged plants seem to be related with the up-regulation of the plant secondary metabolism and with an increase in phenolic compounds involved in plant defence, such as scopoletin, chlorogenic or ferulic acids. Thermal imaging visualizes the leaf transpiration map during pathogenesis and emphasizes the key role of stomata on innate plant immunity. Using several imaging techniques in parallel could allow obtaining disease signatures for a specific pathogen. These techniques have also turned out to be very useful for presymptomatic pathogen detection, and powerful non-destructive tools for precision agriculture. Their applicability at lab-scale, in the field by remote sensing, and in high-throughput plant phenotyping, makes them particularly useful. Thermal sensors are widely used in crop fields to detect early changes in leaf transpiration induced by both air-borne and soil-borne pathogens. The limitations of measuring photosynthesis by Chl-F at the canopy level are being solved, while the use of multispectral fluorescence imaging is very challenging due to the type of light excitation that is used.


Digital localisation in an illicit market space: interactional creation of a psychedelic assemblage in a darknet community of exchange.
Sociology of drugs and digital sociology-albeit for different reasons-need the analysis of interactions, an approach underdeveloped in current scholarship. We address this gap by providing a specific analytical framework for the analysis of digital interactions which enables an ethnomethodological account of micro-interactional dynamics within a cryptomarket: an anonymous darknet market of illicit drugs. As a case study we chose the 'PsychForumMarket' which is unusual in that it operates as a forum based market space and explicitly rejects centralised technical market solutions such as escrow and encryption systems. Instead, it emphasises personal relationships between buyers and vendors as the basis of trust. Hence it forms a community of exchange, both material and cultural. The data were collected through a process of manual scraping from the forum from 2017 to 2020. The data was purposefully sampled to construct a 'thick data' set, and analysed thematically to examine the micro interactional turn taking, sanctioning and norming processes by which the market culture is normalized and embedded. This market is a laboratory to investigate the constitutive nature of digital group interactions. Due to the very nature of this market the disciplining process cannot lie with external authorities. Interactions between community members are permeated with mutual monitoring and policing. We find that in and through digital communication a particular culture emerges to which individuals who wish to join this community have to ascribe. We refer to this particular culture as a 'psychedelic assemblage,' i.e., a local constellation of cultural constructs which frames the experience of drug using and trading. Our investigation reveals the constitutive methods which enable the norming of members' practices and underpin the emergence of a shared lifeworld which in turn ensures the operability of this cryptomarket.


Delivery of eupenifeldin via polymer-coated surgical buttresses prevents local lung cancer recurrence.
Lung cancer is the leading cause of cancer deaths worldwide. Unfortunately, high recurrence rates and poor survival remain despite surgical resection and conventional chemotherapy. Local drug delivery systems are a promising intervention for lung cancer treatment with the potential for improved efficacy with reduced systemic toxicity. Here, we describe the development of a chemotherapy-loaded polymer buttress, to be implanted along the surgical margin at the time of tumor resection, for achieving local and prolonged release of a new anticancer agent, eupenifeldin. We prepared five different formulations of buttresses with varying amounts of eupenifeldin, and additional external empty polymer coating layers (or thicknesses) to modulate drug release. The in vitro eupenifeldin release profile depends on the number of external coating layers with the formulation of the greatest thickness demonstrating a prolonged release approaching 90 days. Similarly, the long-term cytotoxicity of eupenifeldin-loaded buttress formulations against murine Lewis lung carcinoma (LLC) and human lung carcinoma (A549) cell lines mirrors the eupenifeldin release profiles and shows a prolonged cytotoxic effect. Eupenifeldin-loaded buttresses significantly decrease local tumor recurrence in vivo and increase disease-free survival in a lung cancer resection model.


Effect of vasopressin on hemodynamic variables, organ blood flow, and acid-base status in a pig model of cardiopulmonary resuscitation.
Based upon the hypothesis that vasopressin (antidiuretic hormone) may increase vascular resistance during ventricular fibrillation, the effects of this potent vasoconstrictor were studied in a porcine model of ventricular fibrillation. Vasopressin therapy was compared to epinephrine by randomly allocating 14 pigs to receive either 0.045 mg/kg of epinephrine (n = 7) or 0.8 U/kg of vasopressin (n = 7) after 4 min of ventricular fibrillation and 3 min of open-chest cardiopulmonary resuscitation. During cardiopulmonary resuscitation, myocardial blood flow before and 90 s and 5 min after drug administration was 57 +/- 11, 84 +/- 11, and 59 +/- 9 mL.min-1 x 100 g-1 (mean +/- SEM) in the epinephrine group, and 61 +/- 5, 148 +/- 26, and 122 +/- 22 mL.min-1 x 100 g-1 in the vasopressin group (P < 0.05 at 90 s and 5 min). At the same times, mean cardiac index was not significantly different between the groups. After drug administration, coronary venous PCO2 was significantly higher and coronary venous pH was significantly lower in the epinephrine as compared to the vasopressin group. All pigs in both groups were resuscitated and survived the 2-h observation period. We conclude that vasopressin improves vital organ perfusion during ventricular fibrillation and cardiopulmonary resuscitation. Vasopressin seems to be at least as effective as epinephrine in this pig model of ventricular fibrillation.


HIV prevention services in correctional drug treatment programs: do they change risk behaviors?
This study evaluated receipt of HIV prevention services in correctional substance abuse treatment programs and examined their impact on short-term risk behaviors. The authors performed a secondary analysis of the National Treatment Improvement Evaluation Study (NTIES), a prospective cohort study conducted during 1993 to 1995. The sample included 1,223 adult non-HIV-positive inmates, enrolled in nine correctional substance abuse treatment programs. A composite index modeled after the validated Risk Assessment Battery measured HIV risk behavior at treatment entry and at 12-month follow-up. Overall, most inmates received HIV prevention services while in treatment. Controlling for potential confounders, HIV prevention services were significantly associated with reduced risk behavior among inmates who were out of custody at follow-up, but no such association was observed among those still in custody. This analysis suggests that HIV prevention services are beneficial in reducing risk behavior among incarcerated individuals whose discharge is expected in the near future.


Effect of ankle joint fixation on tibialis anterior muscle activity during split-belt treadmill walking in healthy subjects: A pilot study.
This study aims to examine the characteristics of muscle activity change of the tibialis anterior (TA) muscle in healthy adults while they walked on a split-belt treadmill with one fixed ankle. This randomized controlled trial was conducted between November 2017 and July 2018. Fourteen healthy male individuals (mean age 31.4 years; range, 23 to 50 years) were divided into two groups: right ankle joint fixed by ankle-foot orthosis (fixation group) and no orthosis (control group). Both groups were asked to walk on a treadmill with the same belt speed. After familiarizing with walking on both belts at 5.0 km/h, they walked for 6 min with the right belt slower (2.5 km/h) and the left faster (5.0 km/h). For analysis, the 6 min were divided equally among three time periods. The TA muscle activity was calculated at first and last time periods. We compared muscle activities in time periods (early and late phase) and in groups (fixation and control) using two-way mixed analysis of variance. The TA muscle activity decreased in the late phase regardless of ankle joint fixation, and also decreased in the fixation group regardless of the time periods. There was an interaction between these factors. These data show that changes in the TA muscle activity were smaller in the fixation group, suggesting that the ankle joint fixation reduces the adaptation.


Long thoracic nerve injury due to an electric burn.
A 19-year-old white man was burned over 7.5% of his body when he sustained an electric injury from a transformer. There was no associated fall or loss of consciousness. Debridement and grafting were required. The patient had some transient weakness of the muscles of his right arm associated with lower cervical nerve-root injury. This subsequently improved. He also was found to have paralysis of the serratus anterior muscle, with winging of the scapula due to long thoracic nerve injury. This has not improved. A surgical procedure suggested to improve function of the shoulder was rejected by the patient. This is only the second case reported of long thoracic nerve injury due to an electric burn of which we are aware.


Embryonic loss of human females with partial trisomy 19 identifies region critical for the single active X.
To compensate for the sex difference in the number of X chromosomes, human females, like human males have only one active X. The other X chromosomes in cells of both sexes are silenced in utero by XIST, the Inactive X Specific Transcript gene, that is present on all X chromosomes. To investigate the means by which the human active X is protected from silencing by XIST, we updated the search for a key dosage sensitive XIST repressor using new cytogenetic data with more precise resolution. Here, based on a previously unknown sex bias in copy number variations, we identify a unique region in our genome, and propose candidate genes that lie within, as they could inactivate XIST. Unlike males, the females who duplicate this region of chromosome 19 (partial 19 trisomy) do not survive embryogenesis; this preimplantation loss of females may be one reason that more human males are born than females.


Conjunctival Microbiome Changes Associated With Soft Contact Lens and Orthokeratology Lens Wearing.
Usage of different types of contact lenses is associated with increased risk of sight-threatening complications. Changes in the ocular microbiome caused by contact lens wear are suggested to affect infection development in those individuals. To address this question, this study compares conjunctival microbial communities in contact lens wearers with those in noncontact lens wearers. Paired-end sequencing of the V3 region of the 16S rRNA gene was used to characterize the bacterial communities on the conjunctival surfaces of contact lens wearers and nonwearers. No differences in microbial diversity were detected between contact lens wearers and nonwearers. Nevertheless, some slight microbe variability was evident between these two different groups. Bacillus, Tatumella and Lactobacillus abundance was less in orthokeratology lens (OKL) wearers than in nonwearers. In soft contact lenses (SCL) wearers, Delftia abundance decreased whereas Elizabethkingia levels increased. The difference in the SCL and nonwearer group was smaller than that in the OKL group. Variations in the conjunctival taxonomic composition between SCL wearers were larger than those in other groups. Sex differences in the conjunctival microbiota makeup were only evident among nonwearers. Even though there were slight percentage changes between contact lens wearers and nonwearers in some microbes, there were no differences in their diversity. On the other hand, contact lens usage might cause relative abundance of some taxa to change. Our results will help assess whether or not conjunctival microbiome changes caused by contact lens wear affect infection risk.


Effects of axotomy and testosterone on androgen receptor mRNA expression in hamster facial motoneurons.
We have previously demonstrated that testosterone propionate (TP) treatment accelerates the rate of regeneration following facial nerve crush axotomy in adult male hamsters. These effects are mediated by androgen receptor (AR) activation and are blocked by pretreatment with the AR antagonist, flutamide. In addition to its beneficial effects on regeneration, TP regulates AR mRNA levels in facial motor neurons (FMN). Gonadectomized (gdx) male hamsters have been shown to have approximately 50% of the AR mRNA levels found in gonadally intact males. Administration of TP to gdx males results in an upregulation in AR mRNA levels after 1 day of treatment. Recent reports in the literature suggest that axotomy also may regulate the expression of AR in motor neurons. In this study, we examined the effects of axotomy and exogenous steroid treatment on the regulation of AR mRNA in hamster FMN. Five days after castration, adult male hamsters were subjected to a right facial nerve axotomy. Half the animals received one 10-mm Silastic capsule filled with 100% crystalline TP, and the remainder were sham implanted. Postoperative survival times were 6 h or 1, 2, 4, 7, or 14 days. In situ hybridization in conjunction with an AR riboprobe and computerized image analysis were used to quantify AR mRNA levels. The contralateral FMN served as internal controls for these experiments, and FMN of gonadally intact males served as additional nonaxotomized controls. As predicted, AR mRNA levels were upregulated in contralateral control FMN after TP treatment. However, this TP-induced upregulation of AR mRNA levels did not occur in the axotomized FMN. These results indicate that axonal injury can disrupt the normal regulatory pattern of AR mRNA expression by exogenous steroids in motoneurons. We conclude that the potentiation of regenerative events by TP does not require augmented synthesis of AR, but, instead, enhanced stabilization of existing receptors.


Evaluating children's rehabilitation services: an application of a programme logic model.
To apply a programme logic model to evaluate the effectiveness of a new therapy service for children with special needs who were in transition from pre-school to kindergarten. A children's outpatient rehabilitation centre in Ontario, Canada. The short-term outcomes included parents' perceptions of the transition process itself and the information they required, the children's skill development for the transition to kindergarten, and parents' perceptions of services and satisfaction with resources. A combination of quantitative methods [Goal Attainment Scaling (GAS), Measure of Processes of Care (MPOC), Client Satisfaction Questionnaire (CSQ)] and qualitative interviews were used to evaluate both the process ('Outputs') and outcomes ('Short-term objectives') of the new therapy service. The children involved in the evaluation met or exceeded goals that were set by therapists and parents. Parents' perceptions of, and satisfaction with, the new service were higher than the provincial average. Qualitative data from interviews with parents and service providers supported the findings from standardized measures, and provided suggestions for future service delivery. The programme logic model provided researchers and service providers a collaborative and systematic approach to conducting programme evaluation in a relatively short-time frame. It appears to be a useful option for evaluation of other children's services.


Robust Self-Ensembling Network for Hyperspectral Image Classification.
Recent research has shown the great potential of deep learning algorithms in the hyperspectral image (HSI) classification task. Nevertheless, training these models usually requires a large amount of labeled data. Since the collection of pixel-level annotations for HSI is laborious and time-consuming, developing algorithms that can yield good performance in the small sample size situation is of great significance. In this study, we propose a robust self-ensembling network (RSEN) to address this problem. The proposed RSEN consists of two subnetworks including a base network and an ensemble network. With the constraint of both the supervised loss from the labeled data and the unsupervised loss from the unlabeled data, the base network and the ensemble network can learn from each other, achieving the self-ensembling mechanism. To the best of our knowledge, the proposed method is the first attempt to introduce the self-ensembling technique into the HSI classification task, which provides a different view on how to utilize the unlabeled data in HSI to assist the network training. We further propose a novel consistency filter to increase the robustness of self-ensembling learning. Extensive experiments on three benchmark HSI datasets demonstrate that the proposed algorithm can yield competitive performance compared with the state-of-the-art methods.


An epidemiological study of dental caries and associated risk factors among primary school children in the Aileu Municipality, Timor-Leste.
Recent evidence indicates that the oral health for children in Timor-Leste is deteriorating, with 40% of school children experiencing toothache during 2014. Timorese have easy access to sugar, poor food security and lack of water fluoridation, all of which exacerbate the risk of dental caries. A lack of quality epidemiological data is available to confirm anecdotal information of high caries rates in rural and remote Timor-Leste. Such data are required to inform oral health issues and health policy at both the local and national levels. This study investigated the caries status and potential risk factors among primary school children in the rural Aileu municipality of Timor-Leste. The objectives of this study were to determine caries prevalence and experience, the status (active/arrested) of existing caries lesions and associations between dental caries and potential risk factors, among primary school children in the Aileu municipality, Timor-Leste. This study analysed secondary data. De-identified data for this analysis were obtained from North Richmond Community Health (NRCH), Melbourne, Australia. North Richmond Community Health (NRCH) has been working with the Friends of Aileu (a government-to-government partnership between an Australian local government area and the municipality of Aileu) to improve the oral health of school children in the municipality of Aileu. NRCH conducts an outreach school-based oral health promotion program, called Kose Nehan, at six primary schools in the Aileu municipality. Caries was diagnosed using the International Caries Detection and Assessment System (ICDAS) and reported using the decayed, missing and filled teeth (DMFT/dmft) index. Examiners were trained and calibrated. A brief interviewer-administered questionnaire was used to capture information on child oral hygiene and diet behaviours. For the analysis, dental caries was defined as 'any caries lesions' (ICDAS caries codes 1-6). Descriptive and inferential analyses were conducted using STATA 14. Multivariable logistic regression analysis predicting the odds of dental caries (yes/no) was used to determine independent associations between the exposures and the outcome. Data were analysed for 685 children. In the primary dentition, the overall prevalence of caries was 64% and the mean dmft score was 2.74 (standard deviation (SD) 3.08). In the permanent dentition, the overall prevalence was 53% and the mean DMFT score was 1.74 (SD 2.46). Overall, approximately 84% of caries lesions were identified as being active. The multivariable regression analysis did not identify independent predictors of caries. Dental caries was highly prevalent among this population and urgent action is required to reduce the population burden of this disease. Malnutrition, which was not measured for this study, is highly prevalent among children in Timor-Leste and could explain the high caries rates in this population. The effect of malnutrition on dental caries and vice-versa needs further investigation. Programs and policies are urgently needed for oral health promotion and also the prevention and management of dental caries in Timorese children. These strategies should also address the urgent need for emergency dental services aimed at pain relief, first aid for oral infections and restorations, given the high prevalence of advanced disease in this child population.


Cytometric assessment of mitochondria using fluorescent probes.
Mitochondria are most important organelles in the survival of eukaryotic aerobic cells because they are the primary producers of ATP, regulators of ion homeostasis or redox state, and producers of free radicals. The key role of mitochondria in the generation of primordial ATP for the survival and proliferation of eukaryotic cells has been proven by extensive biochemical studies. In this context, it is crucial to understand the complexity of the mitochondrial compartment and its functionality and to develop experimental tools allowing the assessment of its nature and its function and metabolism. This review covers the role of the mitochondria in the cell, focusing on its structure, the mechanism of the mitochondrial respiratory chain, the maintenance of the transmembrane potential and the production of reactive oxygen species. The main probes used for mitochondrial compartment monitoring are described. In addition, various applications using mitochondrial-specific probes are detailed to illustrate the potential of flow and image cytometry in the study of the mitochondrial compartment. This review contains a panel of tools to explore mitochondria and to help researchers design experiments, determine the approach to be employed, and interpret their results.


Ceftriaxone-induced hemolytic anemia and hepatitis in an adolescent with hemoglobin SC disease.
To describe a case of a ceftriaxone-induced hemolytic anemia and hepatitis leading to multiple organ failure and death in an adolescent with hemoglobin SC disease and to review the previous cases of this rare and potentially fatal disorder in children. Case report and literature review. Intensive care unit. Adolescent with hemoglobin SC. Emergency treatment. After 4 days of ceftriaxone therapy, the adolescent experienced an acute hemolytic reaction (hemoglobin decreased to 5 g/dL with hemoglobinuria) and severe hepatitis (all enzymes increasing dramatically including aminoaspartate transferase >20,000 IU/L). Renal failure and ultimately multiple organ failure ensued, and the patient died on hospital day 19. Direct antiglobulin tests on red cells obtained from the patient on hospital day 2 showed microscopic agglutination with polyspecific and anticomplement (C3) antiglobulin reagents. Plasma samples showed macroscopic agglutination reactions when incubated in the presence of ceftriaxone, many days after cessation of ceftriaxone, indicating the continued presence of ceftriaxone-dependent antibodies. Drug reactions leading to hemolysis are relatively uncommon, and a total of ten cases of ceftriaxone-induced hemolytic anemia have been reported in children. The present case describes an adolescent who ultimately died on hospital day 19 from multiple organ failure, although the presentation of this case seems atypical in several respects. Children with clinical syndromes that place them at risk for hemolysis and children who frequently require broad spectrum antibiotics present unique diagnostic challenges, and the possibility that hemolytic syndromes may be due to ceftriaxone must be considered.


Outcomes of allogeneic hematopoietic cell transplantation under letermovir prophylaxis for cytomegalovirus infection.
Cytomegalovirus (CMV) infection is a major infectious complication following allogeneic hematopoietic cell transplantation (allo-HCT). Although letermovir (LMV) prophylaxis dramatically reduces the incidence of early clinically significant CMV (csCMV) infection, it remains unclear whether it has a beneficial effect on nonrelapse mortality (NRM) and overall survival (OS). Herein, we evaluated the impact of LMV prophylaxis on posttransplant outcomes using the registry database of the Japanese Society for Transplantation and Cellular Therapy. Adult patients who underwent allo-HCT between 2017 and 2019 were analyzed (n = 6004). LMV prophylaxis was administered to 1640 patients (LMV group) and it significantly reduced the incidence of csCMV infection compared with those not administered LMV prophylaxis (15.4% vs 54.1%; p < 0.01). However, it did not improve the 1-year NRM (hazard ratio [HR], 0.93; p = 0.40) and OS (HR, 0.96; p = 0.49). In the LMV group, 74 patients had breakthrough csCMV infection and showed inferior NRM (HR, 3.44; p < 0.01) and OS (HR, 1.93; p = 0.02) compared with those without infection. After completing LMV prophylaxis, 252 patients had late csCMV infection and showed inferior NRM (HR, 1.83; p < 0.01) and OS (HR, 1.58; p < 0.01). Our findings suggest that managing breakthrough and late csCMV infections is important for improving long-term outcomes.


Meningococcal outer membrane protein NhhA is essential for colonization and disease by preventing phagocytosis and complement attack.
Neisseria meningitidis is a leading cause of meningitis and septicemia worldwide, with a rapid onset of disease and a high morbidity and mortality. NhhA is a meningococcal outer membrane protein included in the family of trimeric autotransporter adhesins. The protein binds to the extracellular matrix proteins heparan sulfate and laminin and facilitates attachment to host epithelial cells. In this study, we show that NhhA is essential for bacterial colonization of the nasopharyngeal mucosa in a murine model of meningococcal disease. Successful colonization depends on bacterial attachment but also to the capacity to overcome innate host immune responses. We found that NhhA protected bacteria from phagocytosis, which is important for the mucosal survival of bacteria. In addition, NhhA mediated extensive serum resistance that increased bacterial survival in blood and promoted lethal sepsis. The presence of NhhA protected bacteria from complement-mediated killing by preventing the deposition of the membrane attack complex. Taken together, the results of this work reveal that NhhA inhibits phagocytosis and protects bacteria against complement-mediated killing, which enhances both nasal colonization and the development of sepsis in vivo.


Does the Degree of Septal Deviation Affect Cribriform Plate Dimensions and Middle Turbinate Length?
To investigate the effect of septal deviation severity on the dimensions of the lateral lamina of the cribriform plate (LLCP), middle turbinate length (MTL), and the angle of the lateral lamella of the cribriform plate (ALLCP). Paranasal computed tomography (CT) scans of 148 patients with septum deviation (102 males, 46 females; age range 18-63 years) were retrospectively evaluated. The patients were divided into 3 groups according to the measured angle of nasal septal deviation as mild (<9°), moderate (9-15°), or severe (>15°). Height and width of LLCP, MTL, and ALLCP on both sides were measured and these parameters were compared between the groups according to the septal deviation side. The nasal septum was deviated to the right in 73 patients (49.3%) and to the left in 75 patients (50.7%). The LLCP dimensions, MTL, and ALLCP between the groups at the contralateral side were not statistically different (P > 0.05). The LLCP height, ALLCP between the groups at the ipsilateral side were not statistically different (P > 0.05). There was a significant difference in the LLCP width at the ipsilateral side between the groups (P = 0.039). The MTL at the ipsilateral side was significantly different between the groups (P = 0.003). The severity of nasal septum deviation in patients with nasal septum deviation affects the ipsilateral LLCP width and the ipsilateral MTL. These findings suggest that the increase in the severity of septum deviation in patients undergoing endoscopic sinus surgery does not increase the risk of possible damages that may occur in LLCP.


Procalcitonin performance in detecting serious and invasive bacterial infections in children with fever without apparent source: a systematic review and meta-analysis.
The management of children with fever without apparent sources (FWAS) is difficult. This study aimed to evaluate the diagnostic accuracy of procalcitonin (PCT) in detecting serious or invasive bacterial infection (SBI or IBI) in children with FWAS. Areas covered: A systematic review of literature from 2007 to 2017 was performed on Medline. Twelve studies were included, involving 7,260 children with FWAS and analyzing the diagnostic performance of PCT. Four meta-analyses were performed to calculate pooled sensitivity, specificity, diagnostic odds ratio, and summary receiver operating characteristic curve for PCT in detecting SBI and IBI, at two different thresholds. Considering IBI, PCT sensitivity and specificity at a threshold of 0.5 ng/mL were 0.82 and 0.86, respectively; at a threshold of 2 ng/mL sensitivity and specificity were 0.61 and 0.94, respectively. In detecting SBI, PCT performance was lower, with 55% sensitivity and 85% specificity at a threshold of 0.5 ng/mL, and 30% sensitivity and 95% specificity at a threshold of 2 ng/mL. Expert commentary: Considering IBI, results showed high diagnostic accuracy for PCT. Conversely, PCT performance in diagnosis of SBI was poor. These findings suggest that PCT level determination could be helpful in detecting meningitis, bacteremia and sepsis in children with FWAS.


Characterization, antioxidant activity and potential application fractionalized Szechuan pepper on fresh beef meat as natural preservative.
The pericarp of Szechuan pepper is rich in phenols and alkylamides, making it a potential source of antioxidant compounds. Despite being recognized as the primary antioxidants in Szechuan pepper, there is still limited knowledge about their application in real food systems. This study aims to identify, separate, and apply polyphenol and alkylamide fractions derived from Szechuan extracts to beef meat. Using HPLC-MS2, we identified 5 phenols and 11 alkylamides in Szechuan extracts. The quality of the minced meat was evaluated based on color, thiobarbituric acid reactive substances (TBARS), conjugated dienes, carbonyl content, Sulfhydryl content, microbiological content, and total volatile basic nitrogen content (TVB-N). Compared to the polyphenol fraction (1.25 mg/mL), alkylamide fraction (25 mg/mL), and control samples, beef samples incorporated with the polyphenol fraction (6.25 mg/mL) significantly reduced carbonyl content, TBARS, and TVB-N values at the end of storage. Furthermore, they exhibited a significant slowdown in microbial development, improved meat color stability, and preserved pH. Therefore, the use of Szechuan pepper fractions as natural preservatives in meat and meat products is an important area of research and has the potential to enhance the safety and quality of meat products.


The value of endoscopic study of the bile ducts and the pancreas in the elderly.
Endoscopic retrograde cholangiopancreatography was performed in 46 patients age 60 or older with various suspected pancreaticobiliary or duodenal diseases. Thirty-one patients (67.4%) had diagnostically satisfactory opacification of a duct system. Pathologic conditions obstruction injection of the contrast medium prevented opacification in eight patients (17.4%). Seven cannulations (15.2%) were technical failures. There was one complication, a Gram-negative septicemia that was successfully treated. The procedure is of value in the geriatric patient because it may provide preoperative evidence of pancreatic or biliary disease that cannot be diagnosed by any other technique. In jaundiced patients, identification of the site of obstruction facilites the surgical approach and thus reduces operating time. If the common bile duct is shown to be patent, the patient is spared exploratory surgery or a prolonged hospital stay for observation. Although a normal pancreatic duct does not exclude cancer of the pancreas, it should influence the physician to search for other explanations for abdomianl pain or weight loss befror proceeding with abdominal surgery.


[Appendicular metastatic involvement in breast cancer].
The paper describes a case of appendicular metastatic involvement in breast cancer. The 60-year-old female patient developed the signs of acute appendicitis 18 years after detection of primary tumor; after appendectomy the intraoperative appendular specimens exhibited trains of tumor cells that infiltrated the mid- and lower muscle layer third. Immunohistochemical study revealed a moderate expression of Er and Pgr receptors (150 H scores), a positive reaction with EMA, K7, K6, and lactoalbumin The authors review the data available in the literature on the specific features of metastatic spread into the appendix and on management tactics in these patients.


Molecularly imprinted polymeric micro- and nano-particles for the targeted delivery of active molecules.
Molecular imprinting (MI) represents a strategy to introduce a 'molecular memory' in a polymeric system obtaining materials with specific recognition properties. MI particles can be used as drug delivery systems providing a targeted release and thus reducing the side effects. The introduction of molecular recognition properties on a polymeric drug carrier represents a challenge in the development of targeted delivery systems to increase their efficiency. This review will summarize the limited number of drug delivery MI particles described in the literature along with an overview of potential solutions for a larger exploitation of MI particles as targeted drug delivery carriers. Molecularly imprinted drug carriers can be considered interesting candidates to significantly improve the efficiency of a controlled drug treatment.


Large-area broadband saturable Bragg reflectors by use of oxidized AlAs.
Broadband saturable Bragg reflectors (SBRs) are designed and fabricated by monolithic integration of semiconductor saturable absorbers with broadband Bragg mirrors. The wet oxidation of AlAs creates low-index AlxOy layers for broadband, high-index-contrast AlGaAs/AlxOy or InGaAlP/AlxOy mirrors. SBR mirror designs indicate greater than 99% reflectivity over bandwidths of 294, 466, and 563 nm for center wavelengths of 800, 1300, and 1550 nm, respectively. Highly strained and unstrained absorbers are stably integrated with the oxidized mirrors. Large-scale lateral oxidation techniques permit the fabrication of SBRs with diameters of 500 microm. Large-area, broadband SBRs are used to self-start and mode lock a variety of laser systems at wavelengths from 800 to 1550 nm.


Early evidence of royal purple dyed textile from Timna Valley (Israel).
In the context of a broad study aimed at examining dyeing technologies in the Timna textiles collection, three samples of prestigious fibers dyed with murex sea snail were identified. Our identification is based on the presence of 6-monobromoindigotin and 6,6-dibromoindigotin components (detected using HPLC analysis), which is considered unequivocal evidence for the use of murex-derived purple dyestuff. Furthermore, by comparing the analytical results with those obtained in a series of controlled dyeing experiments we were able to shed more light on the specific species used in the dyeing process and glean insights into the ancient dyeing technology. The samples originated from excavations at the extensive Iron Age copper smelting site of "Slaves' Hill" (Site 34), which is tightly dated by radiocarbon to the late 11th-early 10th centuries BCE. While evidence for the important role of purple dyes in the ancient Mediterranean goes back to the Middle Bronze Age (early 2nd millennium BCE), finds of dyed textiles are extremely rare, and those from Timna are the oldest currently known in the Southern Levant. In conjunction with other observations of the very high quality of the Timna textiles, this provides an exceptional opportunity to address questions related to social stratification and organization of the nomadic society operating the mines (early Edom), the "fashion" of elite in the region during the early Iron Age, trade connections, technological capabilities, and more.


Use of letrozole in assisted reproduction: a systematic review and meta-analysis.
Letrozole is the third-generation aromatase inhibitor (AI) most widely used in assisted reproduction. AIs induce ovulation by inhibiting estrogen production; the consequent hypoestrogenic state increases GnRH release and pituitary follicle-stimulating hormone (FSH) synthesis. A systematic search of the literature was performed for both prospective and retrospective studies. Meta-analyses of randomized clinical trials (RCTs) were performed for three comparisons: letrozole versus clomiphene citrate (CC), letrozole + FSH versus FSH in intrauterine insemination (IUI) and letrozole + FSH versus FSH in IVF. In the absence of RCTs, non-randomized studies were pooled. Nine studies were included in the meta-analysis. Four RCTs compared the overall effect of letrozole with CC in patients with polycystic ovary syndrome. The pooled result was not significant for ovulatory cycles (OR = 1.17; 95% CI 0.66-2.09), or for pregnancy rate per cycle (OR = 1.47; 95% CI 0.73-2.96) or for pregnancy rate per patient (OR = 1.37; 95% CI 0.70-2.71). In three retrospective studies which compared L + FSH with FSH in ovarian stimulation for IUI, the pooled OR was 1.15 (95% CI 0.78-1.71). A final meta-analysis included one RCT and one cohort study that compared letrozole + gonadotrophin versus gonadotrophin alone: the pooled pregnancy rate per patient was not significantly different (OR = 1.40; 95% CI 0.67-2.91). Letrozole is as effective as other methods of ovulation induction. Further randomized-controlled studies are warranted to define more clearly the efficacy and safety of letrozole in human reproduction.


Lipid droplets, potential biomarker and metabolic target in glioblastoma.
Lipid droplets (LDs) are subcellular organelles that store large amounts of the neutral lipids, triglycerides (TG) and/or cholesteryl esters (CE). LDs are commonly formed in adipocytes, liver cells and macrophages, and their formation has been shown to be associated with the progression of metabolic diseases, i.e., obesity, fatty liver and atherosclerosis. Interestingly, LDs are also found in some tumor tissues. We recently showed that LDs are prevalent in glioblastoma (GBM), the most deadly brain tumor, but are not detectable in low-grade gliomas and normal brain tissues, suggesting that LDs may serve as a novel diagnostic biomarker for GBM. This short review will briefly introduce LD biology, summarize recent observations about LDs in several types of cancer tissues, and discuss LD formation in GBM. Moreover, we will highlight the role of SOAT1 (sterol-O transferase 1), a key enzyme regulating CE synthesis and LD formation in GBM, in the regulation of SREBP (sterol regulatory-element binding protein) activation. The therapeutic potential of LDs and SOAT1 will be discussed.


The Rac-GEF Tiam1 controls integrin-dependent neutrophil responses.
Rac GTPases are required for neutrophil adhesion and migration, and for the neutrophil effector responses that kill pathogens. These Rac-dependent functions are impaired when neutrophils lack the activators of Rac, Rac-GEFs from the Prex, Vav, and Dock families. In this study, we demonstrate that Tiam1 is also expressed in neutrophils, governing focal complexes, actin cytoskeletal dynamics, polarisation, and migration, in a manner depending on the integrin ligand to which the cells adhere. Tiam1 is dispensable for the generation of reactive oxygen species but mediates degranulation and NETs release in adherent neutrophils, as well as the killing of bacteria. In vivo, Tiam1 is required for neutrophil recruitment during aseptic peritonitis and for the clearance of Streptococcus pneumoniae during pulmonary infection. However, Tiam1 functions differently to other Rac-GEFs. Instead of promoting neutrophil adhesion to ICAM1 and stimulating β2 integrin activity as could be expected, Tiam1 restricts these processes. In accordance with these paradoxical inhibitory roles, Tiam1 limits the fMLP-stimulated activation of Rac1 and Rac2 in adherent neutrophils, rather than activating Rac as expected. Tiam1 promotes the expression of several regulators of small GTPases and cytoskeletal dynamics, including αPix, Psd4, Rasa3, and Tiam2. It also controls the association of Rasa3, and potentially αPix, Git2, Psd4, and 14-3-3ζ/δ, with Rac. We propose these latter roles of Tiam1 underlie its effects on Rac and β2 integrin activity and on cell responses. Hence, Tiam1 is a novel regulator of Rac-dependent neutrophil responses that functions differently to other known neutrophil Rac-GEFs.


What has changed with riluzole?
Riluzole, after two significant trials, was introduced as the first standard treatment of amyotrophic lateral clerosis (ALS) in the early 95'. After 5 years what has changed in the field of ALS? In the field of basic science, riluzole as an active drug has largely contributed to stimulate the research of the possible role of glutamate in the genesis of ALS. However, the apparent simplicity of the relation between the drug and its mechanisms has to modulated in the light of the negativity of other trials (gabapentin) and the display of other mechanisms of the disease and of the compound. Possible relation with other putative mechanisms of ALS, as oxydative stress or growth factors, could be (and probably are) also involved. In the field of its activity, riluzole has an impact on the survival rate which has been largely debated. Comparison with historical databases are supporting the results of the two initial trials. Other information have been published supporting the probable activity of the drug on the muscle strength decline, a controversial matter. They strengthen the initial data and give additional reasons to use riluzole as a standard treatment of patients. In the field of the daily care, riluzole provided a real and unique hope for ALS sufferers. Even if its activity is not as complete as patients would have expected, it provides a hope for slowing down the rate of evolution and abolishes the myth of "no hope, no cure" which was the leitmotiv of patients care until recently. We have to better define the mode of administration with regard to the clinical status of the patients (respiratory disorders, fatigue, stiffness). In the field of care givers, riluzole was one major factor which provided the basis for national and international collaborations either for therapeutic trials or for standard of care. It made possible large collaborative programs in and among many countries. We do hope that this impulse will continue and be stimulated by additional results both in the field of basic science and clinical research.


Developing a system for documenting teaching effectiveness.
Faculty evaluation in nursing education is becoming increasingly important as economic realities and accountability efforts hit higher education. Efforts to document teaching effectiveness in nursing are essential to demonstrate nursing education's accountability to the profession and the public it serves. The purposes of faculty evaluation and its interface with the institutional mission and program goals are discussed. Characteristics of effective teaching, evaluation models, and the effects of teaching styles and student learning styles are presented. Also, variables in the instructional setting and evaluation participants re discussed. Components of teaching effectiveness evaluation including student, peer, self, and administrative evaluation are described. Finally, input, outcome and process measures are presented as a model for teaching effectiveness evaluation and guidelines are provided to assist with the development of a system for documenting effective teaching in nursing.


An amphiphilic dendrimer for effective delivery of small interfering RNA and gene silencing in vitro and in vivo.
An amphiphilic dendrimer bearing a hydrophobic alkyl chain and hydrophilic poly(amidoamine) dendrons is able to combine the advantageous features of lipid and dendrimer vectors to deliver a heat shock protein 27 siRNA and produce potent gene silencing and anticancer activity in vitro and in vivo in a prostate cancer model. This dendrimer can be used alternatively for treating various diseases.


The Yin and Yang of Cannabis: A Systematic Review of Human Neuroimaging Evidence of the Differential Effects of Δ9-Tetrahydrocannabinol and Cannabidiol.
Cannabidiol (CBD) and Δ9-tetrahydrocannabinol (THC) have been the most investigated cannabinoids at the human and preclinical levels, although the neurobiological mechanisms underlying their effects remain unclear. Human experimental evidence complemented by observational studies suggests that THC may have psychotogenic effects while CBD may have antipsychotic effects. However, whether their effects on brain function are consistent with their opposing behavioral effects remains unclear. To address this, here we synthesize neuroimaging evidence investigating the acute effects of THC and CBD on human brain function using a range of neuroimaging techniques, with an aim to identify the key brain substrates where THC and CBD have opposing effects. After a systematic search, a review of the available studies indicated marked heterogeneity. However, an overall pattern of opposite effect profiles of the two cannabinoids was evident with some degree of consistency, primarily attributed to the head-to-head challenge studies of THC and CBD. While head-to-head comparisons are relatively few, collectively the evidence suggests that opposite effects of THC and CBD may be present in the striatum, parahippocampus, anterior cingulate/medial prefrontal cortex, and amygdala, with opposite effects less consistently identified in other regions. Broadly, THC seems to increase brain activation and blood flow, whereas CBD seems to decrease brain activation and blood flow. Given the sparse evidence, there is a particular need to understand the mechanisms underlying their opposite behavioral effects because it may not only offer insights into the underlying pathophysiological mechanisms of psychotic disorders but also suggest potentially novel targets and biomarkers for drug discovery.


Diversity-driven chemical probe development for biomolecules: beyond hypothesis-driven approach.
Bioprobes are indispensable tools for biological study and clinical diagnosis. A conventional strategy for probe development is hypothesis-driven approach based on known molecular mechanisms of recognition for individual analytes. However, even the most sophisticated rational design does not always guarantee the applicability of probes in complex biological systems, therefore the efficiency and scope of probe development has been intrinsically limited. Diversity-driven approach is a rapidly emerging alternative and has been employed for the development of new probes even in the absence of the knowledge about target recognition mechanism. This tutorial review summarizes the recent advances in probe development along with conceptual advantages and perspectives of the diversity-driven approach.


Potential worldwide distribution of Fusarium dry root rot in common beans based on the optimal environment for disease occurrence.
Root rots are a constraint for staple food crops and a long-lasting food security problem worldwide. In common beans, yield losses originating from root damage are frequently attributed to dry root rot, a disease caused by the Fusarium solani species complex. The aim of this study was to model the current potential distribution of common bean dry root rot on a global scale and to project changes based on future expectations of climate change. Our approach used a spatial proxy of the field disease occurrence, instead of solely the pathogen distribution. We modeled the pathogen environmental requirements in locations where in-situ inoculum density seems ideal for disease manifestation. A dataset of 2,311 soil samples from commercial farms assessed from 2002 to 2015 allowed us to evaluate the environmental conditions associated with the pathogen's optimum inoculum density for disease occurrence, using a lower threshold as a spatial proxy. We encompassed not only the optimal conditions for disease occurrence but also the optimal pathogen's density required for host infection. An intermediate inoculum density of the pathogen was the best disease proxy, suggesting density-dependent mechanisms on host infection. We found a strong convergence on the environmental requirements of both the host and the disease development in tropical areas, mostly in Brazil, Central America, and African countries. Precipitation and temperature variables were important for explaining the disease occurrence (from 17.63% to 43.84%). Climate change will probably move the disease toward cooler regions, which in Brazil are more representative of small-scale farming, although an overall shrink in total area (from 48% to 49% in 2050 and 26% to 41% in 2070) was also predicted. Understanding pathogen distribution and disease risks in an evolutionary context will therefore support breeding for resistance programs and strategies for dry root rot management in common beans.


The effects of contrast on visual orientation and spatial frequency discrimination: a comparison of single cells and behavior.
We have compared the effects of contrast on human psychophysical orientation and spatial frequency discrimination thresholds and on the responses of individual neurons in the cat's striate cortex. Contrast has similar effects on orientation and spatial frequency discrimination: as contrast is increased above detection threshold, orientation and spatial frequency discrimination performance improves but reaches maximum levels at quite low contrasts. Further increases in contrast produce no further improvements in discrimination. We measured the effects of contrast on response amplitude, orientation and spatial frequency selectivity, and response variance of neurons in the cat's striate cortex. Orientation and spatial frequency selectivity vary little with contrast. Also, the ratio of response variance to response mean is unaffected by contrast. Although, in many cells, response amplitude increases approximately linearly with log contrast over most of the visible range, some cells show complete or partial saturation of response amplitude at medium contrasts. Therefore, some cells show a clear increase in slope of the orientation and spatial frequency tuning functions with increasing contrast, whereas in others the slopes reach maximum values at medium contrasts. Using receiver operating characteristic analysis, we estimated the minimum orientation and spatial frequency differences that can be signaled reliably as a response change by an individual cell. This analysis shows that, on average, the discrimination of orientation or spatial frequency improves with contrast at low contrasts more than at higher contrasts. Using the optimal stimulus for each cell, we estimated the contrast threshold of 48 neurons. Most cells had contrast thresholds below 5%. Thresholds were only slightly higher for nonoptimal stimuli. Therefore, increasing the contrast of sinusoidal gratings above approximately 10% will not produce large increases in the number of responding cells. The observed effects of contrast on the response characteristics of nonsaturating cortical cells do not appear consistent with the psychophysical results. Cells that reach their maximum response at low-to-medium contrasts may account for the contrast independence of psychophysical orientation and spatial frequency discrimination thresholds at medium and high contrasts.


Regulation of IRE1 RNase activity by the Ribonuclease inhibitor 1 (RNH1).
Adaptation to endoplasmic reticulum (ER) stress depends on the activation of the sensor inositol-requiring enzyme 1α (IRE1), an endoribonuclease that splices the mRNA of the transcription factor XBP1 (X-box-binding protein 1). To better understand the protein network that regulates the activity of the IRE1 pathway, we systematically screened the proteins that interact with IRE1 and identified a ribonuclease inhibitor called ribonuclease/angiogenin inhibitor 1 (RNH1). RNH1 is a leucine-rich repeat domains-containing protein that binds to and inhibits ribonucleases. Immunoprecipitation experiments confirmed this interaction. Docking experiments indicated that RNH1 physically interacts with IRE1 through its cytosolic RNase domain. Upon ER stress, the interaction of RNH1 with IRE1 in the ER increased at the expense of the nuclear pool of RNH1. Inhibition of RNH1 expression using siRNA mediated RNA interference upon ER stress led to an increased splicing activity of XBP1. Modulation of IRE1 RNase activity by RNH1 was recapitulated in a cell-free system, suggesting direct regulation of IRE1 by RNH. We conclude that RNH1 attenuates the activity of IRE1 by interacting with its ribonuclease domain. These findings have implications for understanding the molecular mechanism by which IRE1 signaling is attenuated upon ER stress.


RNA-mediated gene duplication: the rat preproinsulin I gene is a functional retroposon.
Rats and mice have two, equally expressed, nonallelic genes encoding preproinsulin (genes I and II). Cytological hybridization with metaphase chromosomes indicated that both genes reside on rat chromosome I but are approximately 100,000 kilobases apart. In mice the two genes reside on two different chromosomes. DNA sequence comparisons of the gene-flanking regions in rats and mice indicated that the preproinsulin gene I has lost one of the two introns present in gene II, is flanked by a long (41-base) direct repeat, and has a remnant of a polydeoxyadenylate acid tract preceding the downstream direct repeat. These structural features indicated that gene I was generated by an RNA-mediated duplication-transposition event involving a transcript of gene II which was initiated upstream from the normal capping site. Sequence divergence analysis indicated that the pair of the original gene and its retroposed, but functional, counterpart (which appeared about 35 million years ago) is maintained by strong negative selection operating primarily on the segments encoding the chains of the mature hormone, whereas the segments encoding the parts of the polypeptide that are eliminated during processing and also the introns and the flanking regions are evolving neutrally.