[Age-dependent characteristics of the regulation of cytoplasmic NADP+-dehydrogenases in the liver of rats on different diets].
It is established that the activity of malate dehydrogenase and glucose-6-phosphate dehydrogenase in rats aged 1,3 and 12 months lowers under fasting and on high-fatty diet and in old animals (24 months) on a high-fatty diet only the glucose-6-phosphate dehydrogenase activity decreases. When rats after fasting are put on high-carbohydrate diet the activity value of the mentioned enzymes has already returned to the initial level after 12 hours in rats aged 1 and 12 months and in rats aged 3 months it exceeds that activity in intact rats. The rise in the activity of the determined enzymes is completely blocked by the preliminary administration of actinomycin D. The isocitrate dehydrogenase activity remains unchanged under conditions of maintaining animals on different diets.


Quantitative and functional analyses of spastin in the nervous system: implications for hereditary spastic paraplegia.
Spastin and P60-katanin are two distinct microtubule-severing proteins. Autosomal dominant mutations in the SPG4 locus corresponding to spastin are the most common cause of hereditary spastic paraplegia (HSP), a neurodegenerative disease that afflicts the adult corticospinal tracts. Here we sought to evaluate whether SPG4-based HSP is best understood as a "loss-of-function" disease. Using various rat tissues, we found that P60-katanin levels are much higher than spastin levels during development. In the adult, P60-katanin levels plunge dramatically but spastin levels decline only slightly. Quantitative data of spastin expression in specific regions of the nervous system failed to reveal any obvious explanation for the selective sensitivity of adult corticospinal tracts to loss of spastin activity. An alternative explanation relates to the fact that the mammalian spastin gene has two start codons, resulting in a 616 amino acid protein called M1 and a slightly shorter protein called M85. We found that M1 is almost absent from developing neurons and most adult neurons but comprises 20-25% of the spastin in the adult spinal cord, the location of the axons that degenerate during HSP. Experimental expression in cultured neurons of a short dysfunctional M1 polypeptide (but not a short dysfunctional M85 peptide) is deleterious to normal axonal growth. In squid axoplasm, the M1 peptide dramatically inhibits fast axonal transport, whereas the M85 peptide does not. These results are consistent with a "gain-of-function" mechanism underlying HSP wherein spastin mutations produce a cytotoxic protein in the case of M1 but not M85.


The control of copy number of IS6110 in Mycobacterium tuberculosis.
Insertion sequence (IS) elements are bacterial genes that are able to transpose to different locations in the genome. These elements are often used in molecular epidemiology as genetic markers that track the spread of pathogens. Transposable elements have frequently been described as "selfish DNA" because they facilitate their own transposition, causing damage when they insert into coding regions, while contributing little if anything to the bacterial host. According to this hypothesis, the expansion of copy number of insertion sequences is opposed by negative selection against high copy numbers. From an alternative point of view, we might expect IS elements to intrinsically regulate transposition within cells, thereby limiting damage to their bacterial host. Here, we report evidence that the copy number of IS6110 in Mycobacterium tuberculosis is controlled by selection against the element. We first construct 12 different models of marker change resulting from a combination of possible transposition functions and selective regimes. We then compute the Akaike Information Criterion for each model to identify the models that best explain data consisting of serial isolates of M. tuberculosis genotyped with IS6110. We find that the best performing models all include selection against the accumulation of copies. Specifically, our analysis points to the interaction of separate copies of the element causing lethal effects. We discuss the implications of these findings for genome evolution and molecular epidemiology.


Vigilance for pain-related faces in a primary task paradigm: an ERP study.
Pain-related stimuli are supposed to be automatically prioritized over other stimuli. This prioritization has often been tested using primary task paradigms in which pain information is irrelevant for completing the explicitly posed task. Task-irrelevant stimuli are only processed if they are very salient, and pain-related stimuli are assumed to be salient enough. We wanted to further investigate this assumption by assessing event-related brain potentials (ERPs) - a very sensitive method for studying attention and reaction times in response to pictures of people in pain and other emotional faces - using a primary task paradigm. In addition, we assumed that individuals describing themselves as vigilant to pain are especially responsive to pain cues. One hundred pain-free subjects were tested in a primary task paradigm using pictures of facial expressions of pain, anger, happiness, and neutral mood. ERPs were assessed at midline electrodes. Vigilance to pain was assessed by the pain vigilance and awareness questionnaire. In contrast to previous studies (which have used pain words), effects of facial expressions of pain and other emotions on the ERPs and reaction times were surprisingly weak throughout and did not give evidence for a distinct processing of pain-related stimuli. However, hypervigilant subjects appeared to be strongly and cognitively absorbed by all emotional stimuli. Accordingly, it appears that pain-related stimuli are not always of outstanding salience, but that certain characteristics (eg, type of material, emotional richness) have to be present, for pain-related stimuli to be prioritized over stimuli of other emotional content. Hypervigilance to pain may generally predispose individuals to process emotional stimuli in greater depth.


Exercise, weight loss, and changes in body composition in mice: phenotypic relationships and genetic architecture.
The regulation of body weight and composition is complex, simultaneously affected by genetic architecture, the environment, and their interactions. We sought to analyze the complex phenotypic relationships between voluntary exercise, food consumption, and changes in body weight and composition and simultaneously localize quantitative trait loci (QTL) controlling these traits. A large (n = 815) murine advanced intercross line (G(4)) was created from a reciprocal cross between a high-running line and the inbred strain C57BL/6J. Body weight and composition (% fat, % lean) were measured at 4, 6, and 8 wk of age. After measurements at 8 wk of age, mice were given access to running wheels, during which food consumption was quantified and after which body weight and composition were assessed to evaluate exercise-induced changes. Phenotypic correlations indicated that the relationship between exercise and overall change in weight and adiposity depended on body composition before the initiation of exercise. Interval mapping revealed QTL for body weight, % fat, and % lean at 4, 6, and 8 wk of age. Furthermore, QTL were observed for food consumption and changes in weight, % fat, and % lean in response to short-term exercise. Here we provide some clarity for the relationship between weight loss, reduction in adiposity, food consumption, and exercise. Simultaneously, we reinforce the genetic basis for body weight and composition with some independent loci controlling growth at different ages. Finally, we present unique QTL providing insight regarding variation in weight loss and reduction in adiposity in response to exercise.


Roles of connexins and pannexins in (neuro)endocrine physiology.
To ensure appropriate secretion in response to demand, (neuro)endocrine tissues liberate massive quantities of hormones, which act to coordinate and synchronize biological signals in distant secretory and nonsecretory cell populations. Intercellular communication plays a central role in this control. With regard to molecular identity, junctional cell-cell communication is supported by connexin-based gap junctions. In addition, connexin hemichannels, the structural precursors of gap junctions, as well as pannexin channels have recently emerged as possible modulators of the secretory process. This review focuses on the expression of connexins and pannexins in various (neuro)endocrine tissues, including the adrenal cortex and medulla, the anterior pituitary, the endocrine hypothalamus and the pineal, thyroid and parathyroid glands. Upon a physiological or pathological stimulus, junctional intercellular coupling can be acutely modulated or persistently remodeled, thus offering multiple regulatory possibilities. The functional roles of gap junction-mediated intercellular communication in endocrine physiology as well as the involvement of connexin/pannexin-related hemichannels are also discussed.


Development of an improved online comprehensive hydrophilic interaction chromatography × reversed-phase ultra-high-pressure liquid chromatography platform for complex multiclass polyphenolic sample analysis.
In this study, an improved online comprehensive two-dimensional liquid chromatography platform coupled to tandem mass spectrometry was developed for the analysis of complex polyphenolic samples. A narrowbore hydrophilic interaction chromatography column (150 × 2.0 mm, 3.0 μm, cross-linked diol) was employed in the first dimension, while a reversed-phase column based on monodisperse sub-2 μm fully porous particles (50 × 3.0 mm, 1.9 μm d.p.) with high surface area (410 m2 /g) was employed in the second dimension. The combination of a trapping column modulation interface with the high retentive fully porous monodisperse reversed-phase column in the second dimension resulted in higher peak capacity values (1146 versus 867), increased sensitivity, sharper and more symmetrical peaks in comparison with a conventional loop-based method, with the same analysis time (70 min). The system was challenged against a complex polyphenolic extract of a typical Italian apple cultivar, enabling the simultaneous separation of multiple polyphenolic classes, including oligomeric procyanidins, up to degree of polymerization of 10. Hyphenation with an ion trap time-of-flight mass spectrometer led to the tentative identification of 121 analytes, showing how this platform could be a powerful analytical tool for the accurate profiling of complex polyphenolic samples.


Knowledge sources for evidence-based practice in rheumatology nursing.
As rheumatology nursing develops and extends, knowledge about current use of knowledge in rheumatology nursing practice may guide discussions about future knowledge needs. To explore what perceptions rheumatology nurses have about their knowledge sources and about what knowledge they use in their practice, 12 nurses working in specialist rheumatology were interviewed using a semi-structured interview guide. The data were analyzed using conventional qualitative content analysis. The analysis yielded four types of knowledge sources in clinical practice: interaction with others in the workplace, contacts outside the workplace, written materials, and previous knowledge and experience. Colleagues, and physicians in particular, were important for informal learning in daily rheumatology practice. Evidence from the medical arena was accessed through medical specialists, while nursing research was used less. Facilitating informal learning and continuing formal education is proposed as a way toward a more evidence-based practice in extended roles.


Adipose-derived mesenchymal stem cell administration does not improve corneal graft survival outcome.
The effect of local and systemic injections of mesenchymal stem cells derived from adipose tissue (AD-MSC) into rabbit models of corneal allograft rejection with either normal-risk or high-risk vascularized corneal beds was investigated. The models we present in this study are more similar to human corneal transplants than previously reported murine models. Our aim was to prevent transplant rejection and increase the length of graft survival. In the normal-risk transplant model, in contrast to our expectations, the injection of AD-MSC into the graft junction during surgery resulted in the induction of increased signs of inflammation such as corneal edema with increased thickness, and a higher level of infiltration of leukocytes. This process led to a lower survival of the graft compared with the sham-treated corneal transplants. In the high-risk transplant model, in which immune ocular privilege was undermined by the induction of neovascularization prior to graft surgery, we found the use of systemic rabbit AD-MSCs prior to surgery, during surgery, and at various time points after surgery resulted in a shorter survival of the graft compared with the non-treated corneal grafts. Based on our results, local or systemic treatment with AD-MSCs to prevent corneal rejection in rabbit corneal models at normal or high risk of rejection does not increase survival but rather can increase inflammation and neovascularization and break the innate ocular immune privilege. This result can be partially explained by the immunomarkers, lack of immunosuppressive ability and immunophenotypical secretion molecules characterization of AD-MSC used in this study. Parameters including the risk of rejection, the inflammatory/vascularization environment, the cell source, the time of injection, the immunosuppression, the number of cells, and the mode of delivery must be established before translating the possible benefits of the use of MSCs in corneal transplants to clinical practice.


Surgical Management of Endometriosis Involving the Bladder and Ureter.
Endometriosis is a chronic, debilitating condition affecting up to 10% of reproductive-age women. Urinary tract endometriosis is found in 1-6% of women diagnosed with pelvic endometriosis, with the most common sites being the bladder (70-85%), ureter (9-23%), and kidney (4%). Patients typically present with symptoms such as lower abdominal pain, dysuria, and urgency. Unfortunately, urinary tract endometriosis is often asymptomatic, potentially leading to silent obstructive uropathy and kidney failure. To demonstrate a step-by-step approach for the surgical management of urinary tract endometriosis using conventional laparoscopy for partial cystectomy and robotic-assisted laparoscopy for ureteroneocystostomy. Surgical video of two cases managed in an academic tertiary referral center for endometriosis. All authors have no disclosures or conflicts of interest. The first case was a 38-year-old G3P3 with a history of hysterectomy who had an MRI which revealed a T2 hypointense bladder nodule consistent with endometriosis. Patient had significant urinary urgency, dysuria and suprapubpic pain that improved but did not disappear after starting oral progestin therapy (5mg of norethindrone). A cystoscopy was first performed to confirm MRI findings of bladder lesion and to delineate borders and depth of invasion. The second case was a 35-year-old nulliparous woman with chronic pelvic pain and primary infertility. The patient had a history of stage IV endometriosis with deep endometriosis into the bowel and extrinsic encasement of the ureters causing subsequent hydronephrosis requiring bilateral ureteral stents. She had continued daily pelvic pain despite of being on oral contraceptibes for medical management of endometriosis. She subsequently underwent bilateral percutaneous nephrostomy tube placement to allow for ureteral rest prior to surgery. In the first case, conventional laparoscopy was utilized to perform bilateral ureterolysis, bladder mobilization, partial cystectomy for complete excision of the lesion, and two-layered bladder closure. Use of indigo carmine assisted with ureteral orifice identification. In the second case, a cystoscopy was performed with injection of ICG to assist with ureteral identification. After ureterolysis, distal ureteric obstruction due to extensive disease was confirmed on laparoscopy and ureteroscopy. Bilateral ureteroneocystostomy with placement of Double-J ureteral stents was performed using a robotic-assisted approach. Each patient had indwelling foley catheter for bladder decompression during recovery. Pathology in both cases revealed endometriosis. Both patients had an uneventful postoperative course. A postoperative retrograde cystogram confirmed adequate repair prior to removal of each foley catheter. Patient two had uncomplicated office stent removal 6 weeks postoperatively and had a normal renal ultrasound with no hydronephrosis 6 months postoperatively. Endometriosis is an increasingly common condition. It is important for gynecological surgeons to have the proper understanding of anatomy, surgical technique, and multidisciplinary care needed with urology for safe and complete excision of bladder and ureter endometriosis.


An Updated Review on Monkeypox Viral Disease: Emphasis on Genomic Diversity.
Monkeypox virus has remained the most virulent poxvirus since the elimination of smallpox approximately 41 years ago, with distribution mostly in Central and West Africa. Monkeypox (Mpox) in humans is a zoonotically transferred disease that results in a smallpox-like disease. It was first diagnosed in 1970 in the Democratic Republic of the Congo (DRC), and the disease has spread over West and Central Africa. The purpose of this review was to give an up-to-date, thorough, and timely overview on the genomic diversity and evolution of a re-emerging infectious disease. The genetic profile of Mpox may also be helpful in targeting new therapeutic options based on genes, mutations, and phylogeny. Mpox has become a major threat to global health security, necessitating a quick response by virologists, veterinarians, public health professionals, doctors, and researchers to create high-efficiency diagnostic tests, vaccinations, antivirals, and other infection control techniques. The emergence of epidemics outside of Africa emphasizes the disease's global significance. Increased monitoring and identification of Mpox cases are critical tools for obtaining a better knowledge of the ever-changing epidemiology of this disease.


Relationships between renal parameters and serum and urine markers of inflammation in those with and without HIV infection.
We sought to determine the relationships among intrarenal and systemic inflammation and renal disease in HIV. We compared paired serum and urinary levels (normalized to urine creatinine) of monocyte chemotactic protein-1 (MCP-1), regulated on activation normal T cell expressed and secreted (RANTES), interferon-γ-induced protein-10 (IP-10), interleukin-8 (IL-8), and β2-microglobulin (B2M) between two groups of HIV-infected subjects not receiving antiretroviral therapy (ART) [A: not expecting to initiate ART immediately due to having CD4 cell counts ≥350/μl, N=26; B: about to initiate ART, N=19], a group of HIV-infected subjects receiving virologically suppressive antiretroviral therapy [C, N=30], and a group of HIV-uninfected, healthy volunteers [D, N=45]. We then correlated these inflammatory biomarker levels with urine protein/creatinine ratios (uPCR), urine albumin/creatinine ratios (uACR), and estimated glomerular filtration rates (eGFR). Urine inflammatory biomarker levels were highest in Group B. When combining all four study groups, statistically significant positive correlations included uPCR with urine IL-8, urine MCP-1, urine IP-10, and serum IP-10 and uACR with urine IL-8, urine B2M, serum IP-10, and serum B2M. eGFR was statistically significantly negatively correlated with serum MCP-1 and serum B2M. Paired serum and urine levels of IP-10 and B2M (but not IL-8, RANTES, or MCP-1) were significantly correlated with each other in the overall group. The levels of urine inflammatory markers tested differed by HIV status and use of virologically suppressive ART. These urine and serum inflammatory markers were differentially correlated with uPCR, uACR, and eGFR, suggesting that different intrarenal and systemic inflammatory pathways may contribute to different measures of nephropathy.


Stacking resistance to crown gall and nematodes in walnut rootstocks.
Crown gall (CG) (Agrobacterium tumefaciens) and the root lesion nematodes (RLNs) (Pratylenchus vulnus) are major challenges faced by the California walnut industry, reducing productivity and increasing the cost of establishing and maintaining orchards. Current nematode control strategies include nematicides, crop rotation, and tolerant cultivars, but these methods have limits. Developing genetic resistance through novel approaches like RNA interference (RNAi) can address these problems. RNAi-mediated silencing of CG disease in walnut (Juglans regia L.) has been achieved previously. We sought to place both CG and nematode resistance into a single walnut rootstock genotype using co-transformation to stack the resistance genes. A. tumefaciens, carrying self-complimentary iaaM and ipt transgenes, and Agrobacterium rhizogenes, carrying a self-complimentary Pv010 gene from P. vulnus, were used as co-transformation vectors. RolABC genes were introduced by the resident T-DNA in the A. rhizogenes Ri-plasmid used as a vector for plant transformation. Pv010 and Pv194 (transgenic control) genes were also transferred separately using A. tumefaciens. To test for resistance, transformed walnut roots were challenged with P. vulnus and microshoots were challenged with a virulent strain of A. tumefaciens. Combining the two bacterial strains at a 1:1 rather than 1:3 ratio increased the co-transformation efficiency. Although complete immunity to nematode infection was not observed, transgenic lines yielded up to 79% fewer nematodes per root following in vitro co-culture than untransformed controls. Transgenic line 33-3-1 exhibited complete crown gall control and 32% fewer nematodes. The transgenic plants had thicker, longer roots than untransformed controls possibly due to insertion of rolABC genes. When the Pv010 gene was present in roots with or without rolABC genes there was partial or complete control of RLNs. Transformation using only one vector showed 100% control in some lines. CG and nematode resistance gene stacking controlled CG and RLNs simultaneously in walnuts. Silencing genes encoding iaaM, ipt, and Pv010 decrease CG formation and RLNs populations in walnut. Beneficial plant genotype and phenotype changes are caused by co-transformation using A. tumefaciens and A. rhizogenes strains. Viable resistance against root lesion nematodes in walnut plants may be accomplished in the future using this gene stacking technology.


Regulation of Id3 cell cycle function by Cdk-2-dependent phosphorylation.
The functions of basic helix-loop-helix (bHLH) transcription factors in activating differentiation-linked gene expression and in inducing G1 cell cycle arrest are negatively regulated by members of the Id family of HLH proteins. These bHLH antagonists are induced during a mitogenic signalling response, and they function by sequestering their bHLH targets in inactive heterodimers that are unable to bind to specific gene regulatory (E box) sequences. Recently, cyclin E-Cdk2- and cyclin A-Cdk2-dependent phosphorylation of a single conserved serine residue (Ser5) in Id2 has been shown to occur during late G1-to-S phase transition of the cell cycle, and this neutralizes the function of Id2 in abrogating E-box-dependent bHLH homo- or heterodimer complex formation in vitro (E. Hara, M. Hall, and G. Peters, EMBO J. 16:332-342, 1997). We now show that an analogous cell-cycle-regulated phosphorylation of Id3 alters the specificity of Id3 for abrogating both E-box-dependent bHLH homo- or heterodimer complex formation in vitro and E-box-dependent reporter gene function in vivo. Furthermore, compared with wild-type Id3, an Id3 Asp5 mutant (mimicking phosphorylation) is unable to promote cell cycle S phase entry in transfected fibroblasts, whereas an Id3 Ala5 mutant (ablating phosphorylation) displays an activity significantly greater than that of wild-type Id3 protein. Cdk2-dependent phosphorylation therefore provides a switch during late G1-to-S phase that both nullifies an early G1 cell cycle regulatory function of Id3 and modulates its target bHLH specificity. These data also demonstrate that the ability of Id3 to promote cell cycle S phase entry is not simply a function of its ability to modulate bHLH heterodimer-dependent gene expression and establish a biologically important mechanism through which Cdk2 and Id-bHLH functions are integrated in the coordination of cell proliferation and differentiation.


[Pulse therapy in the treatment of optic neuritis in multiple sclerosis].
Corticosteroids are the preparations in the treatment of optic neuritis at multiple sclerosis. The doses and methods of introduction of corticosteroids are chosen individually for the adults and children. A maximum effect is registered in a majority of cases of optic neuritis, when methylprednisolone is prescribed with the pulse-dose introduction, as compared to parabulbar injections or administration of corticosteroids per os. The recovery of vision at relapsing optic neuritis sets on faster and is of a more prolonged nature as compared to other variants of the introduction of corticosteroids. Therefore, the prescription of methylprednisolone as pulse-doses is more effective in both adults and children.


Broad nasal bone reduction: an algorithm for osteotomies.
A persistent problem with nasal bone osteotomies is inadequate reduction of the width of the nasal dorsum. In addition, an algorithm as to which osteotomy to use has not been fully explored. Nine cadavers received a medial oblique osteotomy (15 to 30 degrees off midline) following a humpectomy in six. On one side, the osteotomy was performed on the medial side of the apex of the open roof. On the contralateral side, it was performed on the lateral side of the apex. The osteotome was then pried posteriorly. The resultant hemidorsal widths were compared. Clinically, 53 patients were classified into the following: type I, broad nasal base (lateral osteotomy only); type II, broad nasal base and broad dorsum (lateral and medial oblique osteotomy); and type III, broad dorsum only (medial oblique osteotomy only). The reduction in hemidorsal width was greatest when the osteotome was placed on the lateral side of the apex (t test, p < 0.008). The improved width reduction was attributable to the slippage of the lateral nasal bone under the dorsal hood of the nasal bone. A lateral osteotomy did not have to be performed to reduce the dorsal width alone. After 15 to 32 months, nasal bone width was satisfactory in all but three cases, one of which required a revision. Reduction of the nasal dorsal width is facilitated by a medial oblique osteotomy alone if it is placed at the lateral aspect of the apex of the open roof. A classification of broad nasal bones is given that emphasizes the distinction between dorsal width and nasal base width and suggests which osteotomy to use.


Rh disease: intrauterine intravascular fetal blood transfusion by cordocentesis.
A total of 49 cordocenteses, including 40 intrauterine intravascular fetal blood tranasfusions, were performed in 30 pregnancies complicated by red cell isoimmunization. Transfusions were started at 19-33 weeks' gestation and repeated up to five times, at one-to-four week intervals. The volumes of transfused blood were 20-110 ml, hematocrits were 58-82 percent and the rate of transfusions was 1-15 ml/min. The pretransfusion fetal hemoglobins were 3.5-11.6 g/dl and the posttransfusion fetal hemoglobins were 7.5-15.6 g/dl. There were three intrauterine deaths and two neonatal deaths. The overall survival rate was 83.3 percent including all cordocenteses. The survival rate for the intrauterine transfusions was 81 percent.


Ageritin from poplar mushrooms: scale-up purification and cytotoxicity towards undifferentiated and differentiated SH-SY5Y cells.
Ageritin is the first reported ribotoxin-like protein from basidiomycetes fungi. It can induce ribosomal integrity damage and translation block, and interferes with mitochondrial redox activity of some glioma and neuroblastoma cell lines. Herein, Ageritin has been investigated as a valuable neurotoxin towards either undifferentiated or retinoic acid (RA)-differentiated SH-SY5Y neuroblastoma cells showing a selective cell toxicity against undifferentiated cells. MTT and sulforhodamine B (SRB) assays highlighted that Ageritin markedly decreases the mitochondrial redox activity and viability of undifferentiated cells, meanwhile inducing evident morphological changes eliciting neuronal-like appearance in these cells. Data from lactate dehydrogenase release assay, cytofluorimetric analysis and caspase-3 enzymatic activity measurement suggest that Ageritin promotes cell death through a caspase-dependent apoptotic pathway. The Z-VAD-FMK caspase inhibitor was able to prevent this apoptotic pathway activation. Based on the interesting behaviour of Ageritin vs. SH-SY5Y cells, the development of a scale-up procedure to obtain the purified protein in larger amounts (yield 2.5 mg per 100 g) has been optimized.


Assays of different aspects of haemostasis - what do they measure?
Haemostasis is a complex process affected by many factors including both cellular and plasma components. It is a multistep process starting with platelet adhesion to damaged endothelium and ending in clot fibrinolysis. There are several methods available to study different aspects of haemostasis including adhesion, aggregation, coagulation and fibrinolysis. This review describes the different methods, what aspects of haemostasis they measure and their limitations. Methods discussed include methods to study adhesion (e.g. PFA-100, cone and platelet(let) analyzer and perfusion chambers) and aggregation (e.g. Multiplate, VerifyNow and Plateletworks). Furthermore the principles behind viscoelastic haemostatic assays are presented as well as methods that can analyse aspects of haemostasis in plasma or platelet-rich-plasma samples (thrombin generation, overall haemostasis potential and Thrombodynamics Analyzer).


Childhood high-risk acute lymphoblastic leukemia in first remission: results after chemotherapy or transplant from the AIEOP ALL 2000 study.
The outcome of high-risk (HR) acute lymphoblastic leukemia patients enrolled in the AIEOP-BFM ALL 2000 study in Italy is described. HR criteria were minimal residual disease (MRD) levels ≥10(-3) at day 78 (MRD-HR), no complete remission (CR) at day 33, t(4;11) translocation, and prednisone poor response (PPR). Treatment (2 years) included protocol I, 3 polychemotherapy blocks, delayed intensification (protocol IIx2 or IIIx3), cranial radiotherapy, and maintenance. A total of 312 HR patients had a 5-year event-free survival (EFS) of 58.9% (standard error [SE] = 2.8) and an overall survival of 68.9% (SE = 2.6). In hierarchical order, EFS was 45.9% (4.4) in 132 MRD-HR patients, 41.2% (11.9) in 17 patients with no CR at day 33, 36.4% (14.5) in 11 patients with t(4;11), and 74.0% (3.6) in 152 HR patients only for PPR. No statistically significant difference was found for disease-free survival in patients with very HR features [MRD-HR, no CR at day 33, t(4;11) translocation], given hematopoietic stem cell transplantation (HSCT) (n = 66) or chemotherapy only (n = 88), after adjusting for waiting time to HSCT (5.7 months). Patients at HR only for PPR have a favorable outcome. MRD-HR is associated with poor outcome despite intensive treatment and/or HSCT and may qualify for innovative therapies. The study was registered at www.clinicaltrials.gov as #NCT00613457.


Aggregates in monoclonal antibody manufacturing processes.
Monoclonal antibodies have proved to be a highly successful class of therapeutic products. Large-scale manufacturing of pharmaceutical antibodies is a complex activity that requires considerable effort in both process and analytical development. If a therapeutic protein cannot be stabilized adequately, it will lose partially or totally its therapeutic properties or even cause immunogenic reactions thus potentially further endangering the patients' health. The phenomenon of protein aggregation is a common issue that compromises the quality, safety, and efficacy of antibodies and can happen at different steps of the manufacturing process, including fermentation, purification, final formulation, and storage. Aggregate levels in drug substance and final drug product are a key factor when assessing quality attributes of the molecule, since aggregation might impact biological activity of the biopharmaceutical. In this review it is analyzed how aggregates are formed during monoclonal antibody industrial production, why they have to be removed and the manufacturing process steps that are designed to either minimize or remove aggregates in the final product.


Misconceptions in wetting phenomena.
In a recent paper ('t Mannetje, D.; Banpurkar, A.; Koppelman, H.; Duits, M. H. G.; van den Ende, D.; Mugele, F. Electrically Tunable Wetting Defects Characterized by a Simple Capillary Force Sensor. Langmuir 2013, 29, 9944-9949), there are a few misconceptions regarding the interpretations of theories emanating from Shanahan and de Gennes in describing centrifugal adhesion balance (CAB) experiments, making their results seemingly contradictory to the theory. These are clarified here. We show that their results, if interpreted correctly, do not contradict the theories mentioned above.


The use of cellular matrix in symptomatic knee osteoarthritis.
Knee osteoarthritis is a degenerative "wear and tear" disorder affecting mainly population over 50 years old. It can also present in younger people, especially after an injury or as a part of other diseases. While many therapeutic options exist for knee osteoarthritis, none of them has the potential to cure this condition. Cellular Matrix represents a combination of natural non-crosslinked hyaluronic acid (HA), thixotropic cell separation gel, and sodium citrate anticoagulant solution. A combination of Cellular Matrix with autologous platelet-rich plasma (A-PRP) is a novel therapeutic approach to the management of knee osteoarthritis. It is assumed that the active components HA and PRP have a synergistic effect contributing to a better therapeutic outcome in patients with knee osteoarthritis. Physiotherapy could provide an additional benefit. This is a retrospective pilot study assessing the potential benefit of Cellular Matrix and A-PRP combined with physiotherapy in the management of chronic knee osteoarthritis. Twenty-five patients were enrolled in the study and injected with three doses of Cellular Matrix combined with A-PRP with a time span of 2 weeks between each injection. All patients received standardized physiotherapy. The results showed that 68% of patients achieved more than 50% improvement in pain, stiffness, and function of the knee joints. There were no adverse reactions. This retrospective pilot study confirmed the positive effect of PRP and HA combination in the management of mild and moderate knee osteoarthritis. These preliminary results need to be verified in randomized control trials.


Patients at high risk of tuberculosis recurrence.
Recurrent tuberculosis (TB) continues to be a significant problem and is an important indicator of the effectiveness of TB control. Recurrence can occur by relapse or exogenous reinfection. Recurrence of TB is still a major problem in high-burden countries, where there is lack of resources and no special attention is being given to this issue. The rate of recurrence is highly variable and has been estimated to range from 4.9% to 47%. This variability is related to differences in regional epidemiology of recurrence and differences in the definitions used by the TB control programs. In addition to treatment failure from noncompliance, there are several key host factors that are associated with high rates of recurrence. The widely recognized host factors independent of treatment program that predispose to TB recurrence include gender differences, malnutrition; comorbidities such as diabetes, renal failure, and systemic diseases, especially immunosuppressive states such as human immunodeficiency virus; substance abuse; and environmental exposures such as silicosis. With improved understanding of the human genome, proteome, and metabolome, additional host-specific factors that predispose to recurrence are being identified. Information on temporal and geographical trends of TB cases as well as studies with whole-genome sequencing might provide further information to enable us to fully understand TB recurrence and discriminate between reactivation and new infection. The recently launched World Health Organization End TB Strategy emphasizes the importance of integrated, patient-centered TB care. Continued improvement in diagnosis, treatment approaches, and an understanding of host-specific factors are needed to fully understand the clinical epidemiological and social determinants of TB recurrence.


Consensus statement and recommendations on the treatment of COVID-19: 2021 update.
Many treatments including antiviral and non-antiviral drugs, and critical care are considered for the management of coronavirus disease 2019 (COVID-19). Practice recommendations need to be updated and graded according to the critical evaluation of rapidly emerging literature. In June 2020, Research Center for Epidemic Prevention-National Yang Ming Chiao Tung University formed a task group comprising infectious disease clinicians, pulmonologists, and intensivists with varied areas of expertise. The steering committee prioritized questions and outcomes. The keywords for the searches were COVID-19 and prone position, extracorporeal membrane oxygenation (ECMO), noninvasive positive pressure ventilation (NIPPV), remdesivir, lopinavir, hydroxychloroquine/chloroquine (HCQ/CQ), azithromycin, corticosteroid, tocilizumab, convalescent plasma therapy, and intravenous immunoglobin (IVIG). A systematic review of peer-reviewed literature was performed by the consensus panel. The Grading of Recommendations Assessment, Development and Evaluation (GRADE) approach was used in assessing the certainty of evidence and making recommendations. The effects of COVID-19 treatments on mortality and clinical improvement were summarized in 11 tables, and GRADE was presented to define the strength and quality of evidence for recommendation. The consensus recommended that prone position implanted in COVID-19 patients with hypoxic respiratory failure (IIC), careful selection for the support of ECMO (IIB), NIPPV being feasible but a risk of staff contamination (IIC), remdesivir generally administered in mild-to-moderate COVID-19 patients (IA), the use of dexamethasone in critically ill COVID-19 patients (IA), and the use of tociliziumab in hospitalized severe/critical COVID-19 patient with elevated markers of systemic inflammation (IA). The consensus recommended against the use of lopinavir/ritonavir (IB), HCQ/CQ (IA), azithromycin (IA), convalescent plasma therapy (IA), and IVIG (IA). The inception of the consensus and task group has provided much-needed evidence of the efficacy and safety of various therapies for the management of COVID-19 patients, and make a description about the benefits and harms for most treatments.


Elective cryopreservation of all pronuclear oocytes after GnRH agonist triggering of final oocyte maturation in patients at risk of developing OHSS: a prospective, observational proof-of-concept study.
A bolus dose of GnRH agonist can substitute for hCG as a trigger for the resumption of meiosis in ovarian stimulation with GnRH antagonists, which has been suggested to reduce the risk of ovarian hyperstimulation syndrome (OHSS). As the efficacy of this measure in fresh embryo transfer (ET) cycles is unclear, we evaluated a new clinical concept of GnRH-agonist triggering. In this prospective, observational proof-of-concept study, 20 patients considered at increased risk of developing OHSS (> or = 20 follicles > or = 10 mm or estradiol > or = 4000 pg/ml, or a history of cycle cancellation due to OHSS risk or the development of severe OHSS in a previous cycle) after ovarian stimulation and concomitant GnRH-antagonist administration had final oocyte maturation triggered with 0.2 mg triptorelin s.c. All two pronucleate (2 PN) oocytes were cryopreserved by vitrification, and frozen-thawed ETs (FT-ETs) were performed in an artificial cycle. Main outcome measures were the cumulative ongoing pregnancy rate per patient and the ongoing pregnancy rate per first ET. Secondary outcomes included the incidence of moderate-to-severe OHSS. Of the 20 patients triggered with GnRH agonist, 19 patients underwent 24 FT-ETs in the observational period. The cumulative ongoing pregnancy rate was 36.8% (95% confidence interval: 19.1-59.0%). The ongoing pregnancy rate per first FT-ET was 31.6% (15.4-54.0%). No cases of moderate or severe OHSS were observed. The present study is the proof of the concept that GnRH-agonist triggering of final oocyte maturation in combination with elective cryopreservation of 2 PN oocytes offers OHSS risk patients a good chance of pregnancy achievement, while reducing the risk of moderate and severe OHSS.


ANS binding reveals common features of cytotoxic amyloid species.
Oligomeric assemblies formed from a variety of disease-associated peptides and proteins have been strongly associated with toxicity in many neurodegenerative conditions, such as Alzheimer's disease. The precise nature of the toxic agents, however, remains still to be established. We show that prefibrillar aggregates of E22G (arctic) variant of the Abeta(1-42) peptide bind strongly to 1-anilinonaphthalene 8-sulfonate and that changes in this property correlate significantly with changes in its cytotoxicity. Moreover, we show that this phenomenon is common to other amyloid systems, such as wild-type Abeta(1-42), the I59T variant of human lysozyme and an SH3 domain. These findings are consistent with a model in which the exposure of hydrophobic surfaces as a result of the aggregation of misfolded species is a crucial and common feature of these pathogenic species.


Do mobile units contribute to spatial accessibility to mammography for uninsured women?
Limited spatial accessibility to mammography, and socioeconomic barriers (e.g., being uninsured), may contribute to rural disparities in breast cancer screening. Although mobile mammography may contribute to population-level access, few studies have investigated this relationship. We measured mammography access for uninsured women using the variable two-step floating catchment area (V2SFCA) method, which estimates access at the local level using estimated potential supply and demand. Specifically, we measured supply with mammography machine certifications in 2014 from FDA and brick-and-mortar and mobile facility data from the community-based Breast Screening and Patient Navigation (BSPAN) program. We measured potential demand using Census tract-level estimates of female residents aged 45-74 from 5-year 2012-2016 American Community Survey data. Using the sign test, we compared mammography access estimates based on 3 facility groupings: FDA-certified, program brick-and-mortar only, and brick-and-mortar plus mobile. Using all mammography facilities, accessibility was high in urban Dallas-Ft. Worth, low for the ring of adjacent counties, and high for rural counties outlying this ring. Brick-and-mortar-based estimates were lower for the outlying ring, and mobile-unit contribution to access was observed more in urban tracts. Weak mobile-unit contribution across the study area may indicate suboptimal dispatch of mobile units to locations. Geospatial methods could identify the optimal locations for mobile units, given existing brick-and-mortar facilities, to increase access for underserved areas.


'Getting our voices out there': acceptability of a mental health participation programme for young people with out of home care experience in Australia.
Youth with experience of out-of-home-care (OoHC) typically have poorer mental health than their peers in the general population, and lack opportunities to contribute to service planning. Promoting mental health through leadership training may improve young people's mental health and facilitate system change. The Bounce Project is a pilot youth-leadership mental health training programme co-designed with young people who have experienced OoHC. In this study, we evaluated the Bounce Project from the young people's perspectives to explore the acceptability, successes and limitations of the training to promote the participant's mental health and their contribution to system level change. Thirteen young people aged 18-26 years old who had experienced OoHC and participated in the Bounce Project were interviewed. Semi-structured interviews were conducted and thematically analysed. Four major themes were thereby identified: making their mark; opportunities for growth; redefining roles and pitfalls of research participation. Participants valued the opportunity to have their voices heard, participate in research and learn about mental health. Perceived negative aspects included infrequent participation opportunities, interpersonal difficulties and frustration about the limitations of research including pressure to recruit and restrictive deadlines. Participating in the Bounce Project was a mostly positive experience, but young people also encountered barriers to meaningful participation. Youth with lived experience need more avenues to participate in research and leadership, but research programmes require specific designs that take into consideration the needs of participants and create opportunities for effective and meaningful participation.


Injury pattern analysis of helicopter wire strike accidents (-Gz load).
Injury patterns in rotary wing aircraft wire strike accidents were reviewed to determine mechanisms of injury. It was found that U.S. Army Safety Center data showed that between 1 January 1974 and 31 August 1981 there were 167 wire strikes involving Army helicopters which resulted in 60 injuries and 34 fatalities at a cost of $12,809,100. Updated data on all military rotary wing aircraft accidents investigated between 1978 and 1982 were screened by the Division of Aerospace Pathology to determine the mechanisms of injury to flight deck personnel. From 13 December 1978 to 23 June 1982, three types of rotary wing aircraft were in eight fatal accidents. These mishaps accounted for 28 casualties: 14 fatalities and 14 injuries. Aviators comprised 64.4% of the fatalities. Injury pattern analysis showed 100% had major head and neck injuries with 66% having basilar skull fractures. Two-thirds had associated mandibular fractures or evidence of impact forces transmitted through the mandible to the skull. The same number had wedge-shaped chin lacerations from impact with the cyclic control stick. We postulate transmission of lethal impact forces primarily in the +Gz direction through the mandible to the skull. This suggests either improper use and/or failure of the seat and restraint systems.


Lithium efficacy and adverse effects.
The primary use of lithium is as maintenance treatment of bipolar disorders to prevent relapse. Although it effectively treats acute mania, lithium must often be used in conjunction with neuroleptics. Once-daily lithium therapy may help avoid some side effects and long-term renal damage. Neurotoxicity can be a problem for a small number of sensitive patients when neuroleptics and lithium are combined. Whenever possible, lithium should be avoided in the first trimester of pregnancy because of its potential for inducing cardiovascular abnormalities in the fetus.


Avatars for Teleconsultation: Effects of Avatar Embodiment Techniques on User Perception in 3D Asymmetric Telepresence.
A 3D Telepresence system allows users to interact with each other in a virtual, mixed, or augmented reality (VR, MR, AR) environment, creating a shared space for collaboration and communication. There are two main methods for representing users within these 3D environments. Users can be represented either as point cloud reconstruction-based avatars that resemble a physical user or as virtual character-based avatars controlled by tracking the users' body motion. This work compares both techniques to identify the differences between user representations and their fit in the reconstructed environments regarding the perceived presence, uncanny valley factors, and behavior impression. Our study uses an asymmetric VR/AR teleconsultation system that allows a remote user to join a local scene using VR. The local user observes the remote user with an AR head-mounted display, leading to facial occlusions in the 3D reconstruction. Participants perform a warm-up interaction task followed by a goal-directed collaborative puzzle task, pursuing a common goal. The local user was represented either as a point cloud reconstruction or as a virtual character-based avatar, in which case the point cloud reconstruction of the local user was masked. Our results show that the point cloud reconstruction-based avatar was superior to the virtual character avatar regarding perceived co-presence, social presence, behavioral impression, and humanness. Further, we found that the task type partly affected the perception. The point cloud reconstruction-based approach led to higher usability ratings, while objective performance measures showed no significant difference. We conclude that despite partly missing facial information, the point cloud-based reconstruction resulted in better conveyance of the user behavior and a more coherent fit into the simulation context.


At Least 10-fold Higher Lubricity of Molecularly Thin D2O vs H2O Films at Single-Layer Graphene-Mica Interfaces.
Interfacial water is a widespread lubricant down to the nanometer scale. We investigate the lubricities of molecularly thin H2O and D2O films confined between mica and graphene, via the relaxation of initially applied strain in graphene employing Raman spectroscopy. Surprisingly, the D2O films are at least 1 order of magnitude more lubricant than H2O films, despite the similar bulk viscosities of the two liquids. We propose a mechanism based on the known selective permeation of protons vs deuterons through graphene. Permeated protons and left behind hydroxides may form ion pairs clamping across the graphene sheet and thereby hindering the graphene from sliding on the water layer. This explains the lower lubricity but also the hindering diffusivity of the water layer, which yields a high effective viscosity in accordance with findings in dewetting experiments. Our work elucidates an unexpected effect and provides clues to the behavior of graphene on hydrous surfaces.


The pleuromutilin antibiotics: a new class for human use.
Pleuromutilins were discovered as natural-product antibiotics in 1950. Tiamulin was the first pleuromutilin compound to be approved for veterinary use in 1979, followed by valnemulin in 1999. It was not until 2007 that retapamulin became the first pleuromutilin approved for use in humans. However, retapamulin is limited to topical application. Recent advances in lead optimization have led to the synthesis of pleuromutilins that combine potent antibacterial activity with favorable pharmaceutical properties, making these compounds suitable for oral and intravenous delivery. Most pleuromutilins have an antibacterial spectrum that spans the common pathogens involved in both skin and respiratory tract infections. Two new pleuromutilins, BC-3205 and BC-7013 (both Nabriva Therapeutics AG), have entered clinical trials. In this review, the key properties of pleuromutilin derivatives, designed primarily through modifications at the C(14) side chain, are presented, and the potential of these compounds in systemic therapy in humans is discussed.


The polarity of the proximal tubule cell in rat kidney. Different surface charges for the brush-border microvilli and plasma membranes from the basal infoldings.
Two different membrane fractions were obtained from a brush-border fraction of rat kidney cortex by using their different electrical surface charges in preparative free-flow electrophoresis. One membrane fraction contained only morphologically intact microvilli and was characterized by a high specific activity of alkaline phosphatase. The other fraction morphologically resembled classical plasma membranes by possessing junctional complexes and a high Na-K-ATPase activity The contamination of the isolated membrane fractions by other cell organelles was extremely low These two fractions represent the apical (luminal) and the basal (interstitial) area of the renal proximal tubule cell membrane and clearly demonstrate the polarity of this cell.


Incidence of venous thromboembolism following the neoadjuvant chemotherapy regimen for epithelial type of ovarian cancer.
This study aims to analyze the risk of venous thromboembolism (VTE) in patients receiving neoadjuvant chemotherapy (NACT) for epithelial ovarian cancer (EOC).A retrospective audit was conducted examining 147 patients treated for EOC. Surgical treatment with curative intent, with or without NACT and adjuvant chemotherapy, is the treatment approach, which was modified according to the patient's condition. The incidence of VTE with the most commonly used chemotherapy regimen, carboplatin, cisplatin, paclitaxel, docetaxel, and others were evaluated.This study found a 13.6% incidence of VTE in patients undergoing therapy with curative intent for EOC. No association was seen between NACT and VTE compared to VTE after standard treatment: 2/16 (12.5%) vs 5/131 (3.8%) (P = .16). Univariate and multivariate analyses also demonstrated that NACT has no risk for VTE with odds ratio (OR) = 0.89 (95% CI = 0.18-4.28) and P = 1. Results did not vary significantly with the type of chemotherapy used. Furthermore, increased incidence of VTE as an incidental finding supports the well-established role of malignancy in VTE occurrence. Univariate and multivariate analyses demonstrated that VTE occurred more frequently in menopausal women than nonmenopausal women (17.9% vs 5.8%) with OR = 3.55 (95% CI = 0.99-12.78) and P = .04 in patients aged ≥60 (19.3% vs 10%) with OR = 2.15 (95% CI = 0.83-5.57) and P = .13 but is not statistically significant.We conclude that NACT has no association with VTE and the currently used common chemotherapeutic drug combinations for ovarian cancer carry the minimal risk of thromboembolic events.


[Comments on a report by the BQS on obstetric quality indicators].
When re-evaluating an epidemiologically oriented BQS expertise concerning the quality indicators in obstetrics, we examined three essential methods in obstetrics to verify their scientific evidence and their clinical relevance. In doing so we ascertained that recording the fetal heart rate sub part, analysing fetal blood and determining the blood gas of the umbilical blood are entirely appropriate quality indicators during delivery from a clinical and medical point of view -- although to varying extents. Above all, these three indicators prove to be essentially better evidence-based than described in the BQS expertise. The search for an optimum of evidence-based data in medicine ends in certain fields such as, for example, in obstetrics there where no further randomized studies can be carried out for methodological or ethical reasons. This lack of evidence for the optimum grade I which can clearly be understood from an epidemiological point of view, however, should not lead to an evidence nihilism that fails to accept or admit existing evidence of the weaker grades II and III.


Property-structure relationship of nanoscale ionic materials based on multiwalled carbon nanotubes.
Two categories of nanoscale ionic materials (NIMs) based on multiwalled carbon nanotubes (MWCNTs) were obtained. According to the nature of organic modifier and the type of interaction, these NIMs can be divided into two categories referred to as A-MWCNTs-Fs and I-MWCNTs-Fs, respectively. These MWCNT derivatives were virtually solvent-free and showed good flowability even at room temperature. The liquidlike manner of these MWCNT derivatives was from their relatively high organic content and continual departing-recombining motion of the large organic ions as suggested by the comparison of rheological response of the two categories of MWCNT-based NIMs. Thermal property and temperature-dependent and strain-dependent viscoelasticity of MWCNT derivatives were related to the microscopic structure of their coating layer and subsequently related to the configuration, chemistry, and molecular dimension of modifying molecules to establish the property-structure relationship of MWCNT-based NIMs, which could guide our future work on NIMs to appropriate and promising applications based on their tunable and controllable physical properties.


An iterative solver in the presence and absence of multiplicity for nonlinear equations.
We develop a high-order fixed point type method to approximate a multiple root. By using three functional evaluations per full cycle, a new class of fourth-order methods for this purpose is suggested and established. The methods from the class require the knowledge of the multiplicity. We also present a method in the absence of multiplicity for nonlinear equations. In order to attest the efficiency of the obtained methods, we employ numerical comparisons alongside obtaining basins of attraction to compare them in the complex plane according to their convergence speed and chaotic behavior.


Chemotherapy-induced nausea and vomiting in Italian cancer centers: results of CINVDAY, a prospective, multicenter study.
Guideline consistency in the prevention of chemotherapy-induced nausea and vomiting (CINV) remains low (29% in the Pan European Emesis Registry study) and very low (11%) in regimens with a high emetogenic risk. The aim of this study was to evaluate the guideline consistency of CINV prophylaxis for acute emesis in daily clinical practice in Italy. This was a prospective, observational, multicenter study. Patients scheduled to receive antitumor treatment on a single prespecified day were included. Data on patient characteristics (demographic and clinical), type of anticancer therapy, and type of antiemetic therapy prescribed for acute emesis were collected on electronic data capture forms. Chemotherapy regimens and antiemetic prophylaxis were categorized according to the MASCC 2011 guidelines. The study was approved by the local ethics committees. From July 2013 to February 2014, a total of 502 patients were enrolled at 26 study sites. Median age was 62 years (range 27-87 years). Colorectal cancer and breast cancer were the most common malignancies. The emetogenic potential of the chemotherapy regimens used was high (HEC) (23.7%), moderate (MEC) (40.6%), low (31.3%) or minimal (4.4%). Overall, guideline consistency was 19.3%. Consistency reached 45% when the various 5HT3 receptor antagonists were considered equivalent and interchangeable in MEC regimens. Adherence to guidelines was lowest for MEC and Minimal risk groups. Ten percent of patients in HEC and MEC regimens did not receive any 5HT3 receptor antagonists. NK1 receptor antagonists were used in 8% of all regimens. Our study indicates that antiemetic guideline inconsistency remains an issue in daily clinical oncology practice in Italy.


Efficacy of a diarylheptanoid derivative against Leishmania amazonensis.
The activity of several diarylheptanoid derivatives (curcuminoids) was previously evaluated against Leishmania amazonensis promastigotes and among them the most active compound was the [1-(4-methoxy-phenyl)-7-(3,4-methoxy-4-hydroxy-phenyl)-1,6-heptadien-3, 5-dione]. This derivative was chosen to be assayed in vivo in a treatment trial. For these experiments, the curcuminoid compound was used in a concentration equivalent to the IC50/24 h, obtained from the previous study. Balb/c mice were inoculated subcutaneously in the footpad with L. amazonensis infective promastigotes and 4 weeks after the inoculation, the animals were treated with different schemes, varying from 1 to 3 doses. In all the experiments, Pentamidine Isethionate was used as reference drug under the same experimental conditions. The results showed that one dose was not enough to heal the lesion, however, with 2 and 3 doses the efficiency of the assayed compound was clear. On the other hand, treatment with Pentamidine Isethionate using the three different schemes was not satisfactory when compared to the curcuminoid derivative.


Origin and evolution of the Ig-like domains present in mammalian leukocyte receptors: insights from chicken, frog, and fish homologues.
In mammals many natural killer (NK) cell receptors, encoded by the leukocyte receptor complex (LRC), regulate the cytotoxic activity of NK cells and provide protection against virus-infected and tumor cells. To investigate the origin of the Ig-like domains encoded by the LRC genes, a subset of C2-type Ig-like domain sequences was compiled from mammals, birds, amphibians, and fish. Phylogenetic analysis of these sequences generated seven monophyletic groups in mammals (MI, MII, and FcI, FcIIa, FcIIb, FcIII, FcIV), two in chicken (CI, CII), four in frog (FI-FIV), and five in zebrafish (ZI-ZV). The analysis of the major groups supported the following order of divergence: ZI [or a common ancestor of ZI and F (a cluster composed of the FcIII and FIII groups)], F, CII (or a common ancestor of CII and MII), MII, and MI-CI. The relationships of the remaining groups were unclear, since the phylogenetic positions of these groups were not supported by high bootstrap values. Two main conclusions can be drawn from this analysis. First, the two groups of mammalian LRC sequences must diverged before the separation of the avian and mammalian lineages. Second, the mammalian LRC sequences are most closely related to the Fc receptor sequences and these two groups diverged before the separation of birds and mammals.


Ultrasensitive confocal fluorescence microscopy of C-reactive protein interacting with FcgammaRIIa.
C-Reactive protein (CRP) is an acute phase protein with a suggested pathogenic role in cardiovascular disease. Previous reports proposed that the low-affinity IgG receptor FcgammaRIIa is the major receptor for CRP. However, these reports were met with criticism because the use of anti-CRP antibodies in the detection of CRP binding to FcgammaRIIa may have caused false-positive results. To resolve this controversy, we used ultrasensitive fluorescence microscopy to study the association, dissociation, and equilibrium of CRP binding to FcgammaRIIa. CRP indeed binds to FcgammaRIIa, with low association rates and dissociation rates. Anti-CRP antibodies markedly enhance binding, as is evident from the decrease of the equilibrium dissociation coefficient by 2 orders of magnitude. Our study demonstrates the virtues of single fluorophore labeling and highlights the pitfalls of immunolabeling in investigating CRP/Fc receptor interactions. Importantly, this article provides the first quantitative characterization of CRP binding to FcgammaRIIa and explains and reconciles the diverse and conflicting data presented in the literature.


Detection of nucleosome particles in serum and plasma from patients with systemic lupus erythematosus using monoclonal antibody 4H7.
To develop a monoclonal antibody reagent that would react with nucleosomes but not directly with constituent double stranded DNA (dsDNA) or with histones. Mice were immunized with highly purified chicken mononucleosomes and hybridomas employed to produce Mab that did not react with dsDNA or histones but still showed reactivity with nucleosomes. Murine monoclonal IgG antibody 4H7, generated from a mouse immunized with highly purified chicken erythrocyte nucleosomes, showed no direct ELISA reactivity with either dsDNA or isolated histones or with Sm and RNP antigens or combinations of any of these components. Mab 4H7 did show strong ELISA reactivity for chicken erythrocyte and calf thymus nucleosomes as well as for human leukocyte nucleosomes. The Mab did show strong ELISA reactions with peptides 1-25 of histone H2B and 1-21 of H3, which correspond to sequences known to be located at the surface of nucleosomes. We then measured relative serum levels of 4H7 reactive nucleosome antigen in 140 patients with systemic lupus erythematosus (SLE) in parallel with 50 non-SLE patients with other types of connective tissue disease and 92 healthy subjects. Occasional low levels of serum nucleosomal antigen were seen in 4 of 92 controls, but many patients with SLE (66/140) showed marked elevations of serum nucleosomal antigen. No difference was observed when serum or plasma samples were studied. A marked correlation (R = 0.401, p < 0.0001) was noted when disease activity score (SLEDAI) was plotted against optical density value measured with 4H7 in ELISA. Further, the levels of circulating nucleosomes were raised in SLE patients with very active central nervous system and renal involvement. Presence of nucleosome related antigen in sera from patients with SLE may provide insight into the sequence of disease related antigenic stimuli in active SLE.


Molecular detection of feline calicivirus in clinical samples: A study comparing its detection by RT-qPCR directly from swabs and after virus isolation.
Feline caliciviruses (FCVs) are non-enveloped RNA viruses that exhibit high genetic variation. Two reverse transcription quantitative polymerase chain reaction (RT-qPCR) FCV assays (S1 and S2) were evaluated using samples from 300 field cats. The direct detection of FCV in swabs and after propagation in cell culture, as well as the influence of storage conditions, was assessed. FCV-RNA detectability on dry swabs was similar after storage at either 4°C or -20°C, but viral burdens were maintained for a longer time period when viral transport medium was used. A total of 97 (32%) samples was considered FCV PCR-positive. Of these, 81% and 77% tested positive directly from swabs using S1 and S2, respectively; 84% and 81% tested positive after enrichment in cell culture, respectively. Combined detection by RT-PCR directly from swabs and after VI was most sensitive (up to 96%). Neither of the methods alone were able to detect all FCV-positive samples. In conclusion, clinical samples should be collected in viral transport medium, stored at ≤4°C and processed as soon as possible. The combination of cell culture with RT-qPCR or detection directly from swabs using a combination of different RT-qPCR assays is recommended to reach a high sensitivity of FCV detection.


Anti-Mi-2 antibodies characterize a distinct clinical subset of dermatomyositis with favourable prognosis.
Anti-Mi-2 antibody is a type of myositis-specific autoantibody found in idiopathic inflammatory myopathy patients. To investigate the clinical features and long-term outcomes in anti-Mi-2-positive dermatomyositis (DM) patients. Serum anti-Mi-2β antibodies were detected in 357 DM patients by enzyme-linked immunosorbent assays, and possible associated clinical features were investigated based on cross-sectional and longitudinal studies. Of the DM patients, 40/357 (11.2%) were positive for anti-Mi-2β antibodies and found to have a significantly higher frequency of V sign (72.5% vs 45.7%; p = 0.001), shawl sign (60.0% vs 35.6%; p = 0.003), and muscle weakness (77.5% vs 57.1%; p = 0.013), but a lower incidence of interstitial lung disease (ILD) (37.5% vs 60.9%; p = 0.005) and malignancy (0% vs 12.0%; p = 0.041) than anti-Mi-2β-negative patients. Anti-Mi-2β antibody levels positively correlated with disease activity. After a median follow-up period of 44 months, 97.0% of patients showed clinical remission. Twenty-six anti-Mi-2β-positive patients had a disease course longer than two years, and 16/26 (61.5%) were monocyclic without relapse. Moreover, five patients (15.1%) were drug-free with complete remission for more than three months. Kaplan-Meier survival curves showed that DM patients with positive anti-Mi-2β had a significantly lower mortality rate compared to anti-Mi-2β-negative patients (log-rank; p = 0.035). Interestingly, anti-Mi-2β antibodies did not disappear in all patients over time. Anti-Mi-2β antibodies were associated with a subgroup of DM with a low frequency of ILD and malignancy, good treatment response, and favourable outcome. Moreover, anti-Mi-2β levels correlated with disease activity.


[Sensitivity of phagocytes in the blood and bronchoalveolar tract to hydrocortisone in acute injury of the respiratory organs].
Pneumonia was modelled in experiments on female hybrid mice (CBA X C57Bl)F1 by intratracheal administration of 0.1 ml of 0.4% AgNO3 solution. The functional activity of alveolar macrophages and neutrophils of bronchoalveolar lavage fluid and blood after administration of hydrocortisone (125 mg/kg) was studied in the early stage of pneumonia. The bactericidal and absorptive capacity of neutrophils of the bronchoalveolar lavage fluid and blood diminished considerably under the effect of the hormone. The inhibiting effect of hydrocortisone on the bactericidal activity of alveolar macrophages from the focus of inflammation was more marked than the effect on the cells of the bronchoalveolar lavage fluid from the lungs of intact animals.


Gastroprotective effect and mechanism of patchouli alcohol against ethanol, indomethacin and stress-induced ulcer in rats.
Pogostemonis Herba is an important Chinese medicine widely used in the treatment of gastrointestinal dysfunction. Patchouli alcohol (PA), a tricyclic sesquiterpene, is the major active constituent of Pogostemonis Herba. This study aimed to investigate the possible anti-ulcerogenic potential of PA and the underlying mechanism against ethanol, indomethacin and water immersion restraint-induced gastric ulcers in rats. Gross and histological gastric lesions, biochemical and immunological parameters were taken into consideration. The gastric mucus content and the antisecretory activity were analyzed through pylorus ligature model in rats. Results indicated that oral administration with PA significantly reduced the ulcer areas induced by ethanol, indomethacin and water immersion restraint. PA pretreatment significantly promoted gastric prostaglandin E2 (PGE2) and non-protein sulfhydryl group (NP-SH) levels, upregulated the cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) mRNA expression, and considerably boosted the gastric blood flow (GBF) and gastric mucus production in comparison with vehicle. In addition, PA modulated the levels of interleukin-6 (IL-6), interleukin-10 (IL-10) and tumor necrosis factor-α (TNF-α). The levels of glutathione (GSH), catalase (CAT) and malonaldehyde (MDA) were also restored by PA. However, the gastric secretion parameters (pH, volume of gastric juice and pepsin) did not show any significant alteration. These findings suggest that PA exhibited significant gastroprotective effects against gastric ulceration. The underlying mechanisms might involve the stimulation of COX-mediated PGE2, improvement of antioxidant and anti-inflammatory status, preservation of GBF and NP-SH, as well as boost of gastric mucus production.


TTF-1 is useful for primary site determination in duodenal metastasis.
We report here on a case of duodenal metastasis from primary lung adenocarcinoma. A 69-year old man was diagnosed with primary lung adenocarcinoma. Four courses of combined chemotherapy with carboplatin and paclitaxel associated with irradiation of 60 Gy shrunk the lung tumor. However, soon after,the para-aortic lymph node became swollen. Esophagogastroduodenoscopy revealed three duodenal tumors. Differential diagnosis between malignant lymphoma and metastatic duodenal cancer was endoscopically difficult. The histology of biopsied specimens was poorly differentiated adenocarcinoma. Immunohistochemical analysis revealed a positive reaction for thyroid transcription factor-1 (TTF-1). Thus, we concluded that these were metastatic duodenal tumors from lung adenocarcinoma. Two courses of gemcitabine led to a complete remission in this duodenal metastasis and para-aortic lymph node swelling with only scarring remaining in computed tomography. He is now on the continuous generalized chemotherapy. In conclusion, duodenal metastasis from primary lung adenocarcinoma is rare and hard to diagnose. In such an instance, TTF-1 immunostaining is crucial to obtain the correct diagnosis.


Identification of Dehalobacter reductive dehalogenases that catalyse dechlorination of chloroform, 1,1,1-trichloroethane and 1,1-dichloroethane.
Two novel reductive dehalogenases (RDases) that are highly similar to each other but catalyse distinct dechlorination reactions were identified from Dehalobacter-containing mixed cultures. These two RDases were partially purified from crude protein extracts of anaerobic dechlorinating enrichment cultures using blue native polyacrylamide gel electrophoresis. Gel slices were assayed for dechlorinating activity, and associated proteins were identified using liquid chromatography tandem mass spectrometry with the metagenome of the parent culture as the reference database. The two RDases identified, annotated as CfrA and DcrA, share an amino acid identity of 95.2 per cent, but use different substrates: CfrA dechlorinates chloroform (CF) and 1,1,1-trichloroethane (1,1,1-TCA), but not 1,1-dichloroethane; DcrA dechlorinates 1,1-dichloroethane, but not CF or 1,1,1-TCA. These two novel RDases share no more than 40 per cent amino acid identity to any other known or putative RDases, but both have a twin-arginine motif and two iron-sulfur binding motifs conserved in most RDases. Peptides specific to two putative membrane anchor proteins, annotated as CfrB and DcrB, were also detected in gel slices.


Glucocorticoids inhibit transcription and expression of the UT-A urea transporter gene.
Dexamethasone treatment increases urea excretion and decreases urea permeability and urea transporter UT-A1 protein abundance in the inner medullary collecting duct (IMCD) of adrenalectomized rats. We examined the effect of dexamethasone treatment for 3 days on the abundance of several UT-A mRNA transcripts in rat renal medulla. By Northern blot analysis, a significant decrease in mRNA expression was observed in the inner medulla of dexamethasone-treated rats compared with controls for UT-A1 (71%), UT-A3 (75%), and UT-A3b (75%), but not for UT-A2. We then tested the effect of 100 nM dexamethasone on the activity of promoter I in the UT-A gene, using LLC-PK(1)-GR101 cells that express the glucocorticoid receptor. Dexamethasone significantly decreased the activity of rat UT-A promoter I (72%) but did not affect UT-A promoter II. Deletion analysis and site-directed mutagenesis demonstrated that sequences between -423 and -244 are important for this inhibition and that a 10-bp sequence at -363, which binds a nuclear protein in a gel shift assay, is necessary for basal promoter activity. The specific factors involved in repression of UT-A promoter I activity by glucocorticoids remain to be determined.


Brain-derived neurotrophic factor but not neurotrophin-3 enhances differentiation of somatostatin neurons in hypothalamic cultures.
The present work investigated whether neurotrophins could differentially affect in vitro growth and maturation of two related subsets of hypothalamic neurons, hypophysiotropic somatostatin (SRIH) neurons projecting from the periventricular area and arcuate SRIH interneurons. For this purpose, the hypothalamus of 17-day-old rat fetuses was sampled and separated into a ventral and a dorsal fragment containing respectively periventricular and arcuate regions. Each fragment was dissociated and seeded separately in defined medium. Brain-derived neurotrophic factor (BDNF) or neurotrophin-3 (NT-3), two important members of the neurotrophin family involved in neuronal differentiation and plasticity, were added to the cultures at seeding time. After 6 or 11 days in vitro, neurons were labeled with an anti-SRIH antiserum and submitted to morphometric analysis. In parallel, SRIH mRNA was estimated by semiquantitative reverse-transcriptase-polymerase chain reaction, and neuronal SRIH content, basal and depolarisation-stimulated releases measured by radioimmunoassay. The response of control, non-labeled neurons was estimated by neuronal counts and by assaying glutamic acid decarboxylase, a marker of a large majority of hypothalamic neurons. BDNF markedly increased the size and the branching number of SRIH periventricular cell bodies. Expression of SRIH mRNA, as well as SRIH content and release into the culture medium, were also stimulated by the neurotrophin. Non-SRIH neurons were not affected by the treatment. Under the same conditions, arcuate neurons exhibited a weak, mostly transient response to BDNF. NT-3 was ineffective on either neuronal subset. Immunoneutralization of Trk receptors provided further evidence for BDNF effect specificity. The results indicate that BDNF is a selective activator of the differentiation of hypophysiotropic SRIH neurons in the periventricular area of the hypothalamus.


[Genetic carrier screening for hemoglobinopathies in the Netherlands is not opportune].
To investigate whether screening for carriership of sickle-cell anaemia and thalassaemia is desirable in the Netherlands, given the prevalence of the diseases, the possibilities of a screening test and the knowledge of the population at risk. Descriptive. Wilhelmina Children's Hospital, Clinical Genetic Centre, Utrecht, the Netherlands. In September 1992 Dutch paediatricians were treating 128 children with sickle-cell anaemia and 31 with thalassaemia major. Between October 1992 and December 1994, sickle-cell anaemia was diagnosed in 40 children and thalassaemia major in 14; of these, 16 and 4, respectively, had been born in the Netherlands. A protocol for the screening for carriership adjusted to the Dutch situation was drawn up in the past, with an estimate of the corresponding costs. Patients with sickle-cell anaemia or thalassaemia major, their families, care givers and persons without disease but originating from Surinam, the Netherlands Antilles, Turkey or Morocco were interviewed about their knowledge of these two diseases. The knowledge of the diseases was low among the Turkish and Moroccan populations, and very global among the Surinamese and Antillian populations. There was more belief in supernatural causes than in a genetic predisposition. Given the low number of newly diagnosed haemoglobinopathies in children born in the Netherlands, the poor knowledge of the disease of the population at risk, the difficulty in identifying the best moment in life for screening and the rather complicated screening method, a screening programme for carriership of haemoglobinopathies in the Netherlands is not opportune. Where sickle-cell anaemia is concerned, screening for this disease during the neonatal period might be considered.


[Antinociceptive components of general anesthesia and postoperative analgesia].
The philosophy of full-value antinociceptive protection of patients during and after surgical interventions is discussed. The author presents her viewpoint on the problem with due consideration for recent published reports. She advocates a prophylactic approach to attaining the antinociceptive protection (the so-called "preemptive analgesia"). Analgesics of peripheral action are the main means for protection at the peripheral level. Peripheral antialgogens contrycal (trasilol), acelysin (aspisol), baralgine, diclofenac, and ketorolac were effectively used during the pre-, intra-, and postoperative periods: the course of total anesthesia and postoperative period improved, intensity of the painful syndrome and the need in opioids was decreased, and wound healing improved. For preventing secondary (central) hyperalgesia and decreasing the intensity of postoperative painful syndrome, NMDA-receptor antagonist ketamine in microdoses is recommended before and during surgery. Afferent nociceptive flow can be decreased by local and regional anesthesia. Practical application of the preemptive analgesia principle improves the efficacy and safety of analgesia at different stages of surgical treatment, decreases total and local (at the level of operated tissues) consequences of surgical trauma, and decreases the probability of complications.


Cultural consonance, constructions of science and co-existence: a review of the integration of traditional, complementary and alternative medicine in low- and middle-income countries.
This review examined the determinants, patterns and imports of official recognition, and incorporation of different traditional, complementary and alternative systems of medicine (TCAM) in the public health establishment of low- and middle-income countries, with a particular focus on India. Public health systems in most countries have tended to establish health facilities centred on allopathy, and then to recognize or derecognize different TCAM based on evidence or judgement, to arrive at health-care configurations that include several systems of medicine with disparate levels of authority, jurisdiction and government support. The rationale for the inclusion of TCAM providers in the public health workforce ranges from the need for personnel to address the disease burden borne by the public health system, to the desirability of providing patients with a choice of therapeutic modalities, and the nurturing of local culture. Integration, mostly described as a juxtaposition of different systems of medical practice, is often implemented as a system of establishing personnel with certification in different medical systems, in predominantly allopathic health-care facilities, to practise allopathic medicine. A hierarchy of systems of medicine, often unacknowledged, is exercised in most societies, with allopathy at the top, certain TCAM systems next and local healing traditions last. The tools employed by TCAM practitioners in diagnosis, research, pharmacy, marketing and education and training, which are seen to increasingly emulate those of allopathy, are sometimes inappropriate for use in therapeutic systems with widely divergent epistemologies, which call for distinct research paradigms. The coexistence of numerous systems of medicine, while offering the population greater choice, and presumably enhancing geographical access to health care as well, is often fraught with tensions related to the coexistence of philosophically disparate, even opposed, disciplines, with distinct and unaligned notions of evidence and efficacy, and ethical and operational challenges of the administration of a plural workforce.


Lyme Carditis: A Reversible Cause of Acquired Third-Degree AV Block.
BACKGROUND Lyme borreliosis, caused by spirochetes of the Borrelia burgdorferi genospecies complex, is the most commonly reported tickborne infection in North America and those infected may present with cutaneous, cardiac, articular, and neuropsychiatric abnormalities. The protean nature of many of its clinical manifestations presents a diagnostic conundrum. Lyme disease can affect the heart, albeit rarely, with cardiac abnormalities usually manifesting as varying degrees of heart block or arrhythmias. CASE REPORT We present a case of complete heart block in a young man who participated in outdoor activities in a Lyme-endemic area and developed fatigue and palpitations weeks after a flu-like illness. He noticed that his heart rate was low; he had an intermediate suspicious index in Lyme carditis (SILC) score with positive Lyme serologies. His initial electrocardiogram when he presented to the emergency department showed a complete heart block. In this case, he was successfully managed with intravenous ceftriaxone, amoxicillin, and a transcutaneous pacemaker, obviating the need for a permanent pacemaker. CONCLUSIONS Electrocardiographic changes such as heart block and arrhythmias with or without symptoms may be the initial manifestation of Lyme carditis in a patient who may or may not remember a tick bite or have a typical skin rash. The SILC score may assist in recognizing these cases and prompt initiation of antibiotics usually leads to the resolution of these electrocardiographic abnormalities and symptoms that may be present.


Effects of beta-blockers on the concentration and oxidizability of plasma lipids.
1. beta-blockers improve morbidity and mortality after myocardial infarction, probably by several mechanisms. We investigated potentially relevant effects of beta-blockers in vivo and in vitro on plasma lipid oxidizability. Forty-two healthy men were randomized to receive placebo (13), metoprolol (14) or propranolol (15). 2. At 4 weeks, the effects on heart rate, blood pressure and lipids appeared similar and subjects taking a beta-blocker were combined. Compared with placebo, those on a beta-blocker gained 0.5 kg in weight (P = 0.04), heart rate fell from 63 to 52 beats/min (P < 0.0001) and blood pressure fell from 116/74 to 113/69 mmHg (P < 0.005); high-density lipoprotein (HDL)-cholesterol fell from 1.26 to 1.11 mmol/l (P = 0.005), there being no change in the ratio of free to esterified cholesterol in HDL, and there was an apparent rise in serum triacylglycerols from 1.18 to 1.43 mmol/l (P = 0.15 when adjusted for weight gain). Low-density lipoprotein (LDL)-cholesterol and lipoprotein (a) did not change. In this study, the oxidizability of LDL was unaffected by beta-blocker therapy. beta-blockade was not associated with any change in LDL fatty acid profile, or beta-carotene or alpha-tocopherol content which might account for the reduced LDL oxidizability previously reported in patients treated with beta-blockers. Furthermore, neither atenolol nor propranolol, at concentrations up to 100 mumol/l, had any effect on in vitro oxidizability of LDL obtained from healthy volunteers. 3. In contrast to the favourable haemodynamic effects conferred by beta-blockers, the effects on weight and serum triacylglycerols and HDL-cholesterol appear to be adverse and we did not demonstrate any changes in lipid oxidizability which might be relevant to the protective effects of beta-blockers in patients with coronary disease.


The influence of thermal processing on emulsion properties of defatted hazelnut flour.
In this study, the influences of roasting and the amount of hazelnut flour on the formation and stabilization of emulsions containing different amounts of oil were investigated. After hazelnuts were roasted in an oven at 140°C for 40 min, the oil content was removed. The emulsions with defatted hazelnut flour containing corn oil at 3%, 10% and 50% were prepared. Roasting process significantly decreased the interfacial tension values of samples down to 1.9 mN/m due to protein denaturation. There was no significant difference between the particle sizes of oil droplets in emulsions with roasted and raw hazelnut flour at the same concentration. However, diffusion coefficients of oil droplets increased for emulsions containing roasted defatted flour samples. The zeta (ζ) potential values of all emulsions increased when roasted hazelnut flour was used, indicating the stabilization of suspensions and the solution resistance against aggregation. Storage modulus (G'), loss modulus (G″) and complex viscosity (η(∗)) of emulsions were measured. G' value was found to be greater than the G″ value, which fits into weak gel model. The roasting process resulted with lower transition temperatures but with increased transition enthalpies of the flour samples based on differential scanning (DSC) measurements. Lower transition temperatures may be attributed to the partial gelatinization of starch in the flour and partial denaturation of proteins. These results may help to tailor the properties of defatted hazelnut flour when it is used in food products containing emulsions such as sauces, dressings and creams for stabilizing purposes.


In-Field Evaluation of Drone-Released Lacewings for Aphid Control in California Organic Lettuce.
Aphids are one of the most economically important pests to California's Central Coast lettuce industry. Aphids vector the lettuce mosaic virus and are crop contaminants in the packaging of the product. Lettuce aphid, Nasonovia ribisnigri (Mosley) (Hemiptera: Aphididae), is one of the predominant aphid species in lettuce, and it poses unique management challenges forming colonies inside the lettuce head. Current management practices rely on repeated foliar insecticide applications to reduce aphid densities per plant. Some organic growers have explored the release of laboratory-reared beneficial insects to manage aphids in their commercial fields. This project sought to document the effects of drone-released lacewing eggs on lettuce aphid densities in organic romaine lettuce fields. Commercially reared lacewing eggs were released at a rate of 74,131 eggs/ha and organic-certified insecticides were sprayed following their respective label recommendations. Our results could be interpreted as preliminary evidence that drone release of lacewings could reduce aphid densities (15.6-150.0 aphids/lettuce head) when compared to the untreated plots (32.1-257.9 aphids/lettuce head). Aphid densities were also decreased after the application of foliar organic-certified insecticides (11.77-143.5). Traditionally, the cost of labor has limited the use of beneficials in the lettuce production system, but the use of drones for these releases may make this strategy more attractive. Based on our calculations, spraying an organic-certified insecticide is less expensive (direct operational costs of U.S. $ 116.36/ha) than releasing beneficial insects by hand ($ 185.40) or by drone ($ 176.00) when mimicking the conditions of this study.


Molecular-Scale Dynamics of Long Range Retrograde Brain-Derived Neurotrophic Factor Transport Shaped by Cellular Spatial Context.
Retrograde neurotrophin (NT) transport is a specialized form of signal transduction used to conduct information from axons to the cell bodies of central and peripheral nervous system neurons. It is activated upon NT-Trk receptor binding, NT-Trk internalization into signaling endosomes, and their motion along the axon toward the cell body. Brain-derived neurotrophic factor (BDNF) is an abundant NT that modulates key brain and spinal cord functions, and defects in BDNF trafficking are associated with neuronal death, neurodegenerative diseases and in nerve injury. Decades of study have yielded impressive progress in elucidating NT retrograde transport; however, much information remains unclear. For example, while it is known that NT function is dependent on tight control of NT-receptor intracellular trafficking, data describing the precise spatiotemporal molecular dynamics of their axonal to somatic transport are lacking. In past work, we showed the use of discrete, photo-bleaching-resistant quantum dot (QD)-BNDF probes to activate and track BDNF-TrkB receptor internalization; this revealed a rich diversity of molecular motions that intracellular BDNF signaling endosomes undergo within the soma of nodose ganglia sensory neurons. Here, we used combined techniques of discrete QD-BDNF tracking with compartmented microfluidic chambers to characterize retrograde BDNF-TrkB transport over long-ranging distances of primary dorsal root ganglion sensory neuronal axons. Our new findings show that axonal retrograde motion is comprised of heterogeneous mixtures of diffusive behaviors, pauses, and variations in net molecular-motor-dependent transport speeds. Notably, specific molecular dynamic features such as NT speed were dependent on spatial context that could be categorized in distance from distal axons and proximity to the soma and were not entirely dictated by active motor transport speed. The important implication is recognition that NT-receptor retrograde transport is comprised of molecular dynamics, which change over the course of long-range trafficking to shape overall transport and possibly signaling.


[Ejaculatory duct dilation combined with seminal vesicle clysis for refractory hematospermia: A report of 32 cases].
To evaluate the effect of ejaculatory duct dilation combined with seminal vesicle clysis in the treatment of refractory hematospermia. Using ureteroscopy, we treated 32 patients with refractory hematospermia by transurethral dilation of the ejaculatory duct combined with clysis of the seminal vesicle with diluent gentamicin. The operation was successfully accomplished in 31 cases, with the mean operation time of 32 (26-47) minutes. The patients were followed up for 6-39 (mean 23.6) months. No complications, such as urinary incontinence and retrograde ejaculation, were found after operation. Hematospermia completely disappeared in 27 cases, was relieved in 1, and recurred in 3 after 3 months postoperatively. Those with erectile dysfunction or mental anxiety symptoms showed significantly decreased scores of IIEF-Erectile Function (IIEF-EF) and Self-Rating Anxiety Scale (SAS). Ejaculatory duct dilation combined with seminal vesicle clysis under the ureteroscope, with its the advantages of high effectiveness and safety, minimal invasiveness, few complications, and easy operation, deserves general clinical application in the treatment of refractory hematospermia.


[Multiple sclerosis--a disease with complex genetics].
Multiple sclerosis (MS) is an inflammatory disease of the central nervous system, characterised by demyelinisation, gliosis and various degrees of axonal damage. The cause of the disease is unknown though there is evidence that MS is an immune mediated disease in which genetic, environmental and stochastic factors are involved. Evidence of genetic influence in MS is reviewed. About 20% of patients have other family members with MS. First-degree relatives of MS patients have increased risk (2-5%) of developing MS compared to the risk in the general population (0.1%). The concordance rate in monozygotic twins (25-30%) is higher than in dizygotic twins (2-5%). Genetic analysis has shown association and linkage of MS to the HLA-DR2, DQ6 haplotype. Genome-wide screens have shown chromosome regions of potential importance. MS is a complex disease in which several genes are involved. Despite extensive candidate gene studies and genome-wide linkage screens, the HLA-DR2, DQ6 haplotype is the only genetic region that has shown unequivocal association and linkage to the disease. Further studies aimed at defining other MS susceptibility and disease-modifying genes are in progress.


Paternal determinants of early embryo development.
Existing research has primarily focused on investigating the impacts of the maternal environment, female fertility phenotype, and genetics on pregnancy loss in dairy cattle. Recently, attention has been directed toward understanding the role the sire has on embryo quality and viability. Studies have shown there is a paternal influence on early pregnancy loss, but the specific mechanisms impacting pregnancy establishment and maintenance remain unclear. Despite clear differences that sires have on pregnancy outcomes, there is a lack of evidence regarding specifically how sires influence pregnancy. Sperm characteristics, such as motility, concentration, and morphology, have been extensively studied, but further research is needed to understand what makes one sire more or less fertile than another sire and how this affects pregnancy. To effectively address pregnancy loss, a deeper understanding of the processes involved from fertilisation to blastocyst formation is essential, particularly for understanding early pregnancy loss.


Preparation of molecularly imprinted polymer nanobeads for selective sensing of carboxylic acid vapors.
The detection and discrimination of volatile carboxylic acid components, which are the main contributors to human body odor, have a wide range of potential applications. Here, a quartz crystal microbalance (QCM) sensor array based on molecularly imprinted polymer (MIP) nanobeads is developed for highly sensitive and selective sensing of typical carboxylic acid vapors, namely: propionic acid (PA), hexanoic acid (HA) and octanoic acid (OA). The MIP nanobeads were prepared by precipitation polymerization with methacrylic acid (MAA) as a functional monomer, trimethylolproane trimethacrylate (TRIM) as a crosslinker, and carboxylic acids (PA, HA and OA) as the template molecules. The precipitation polymerization resulted in nano-sized (150-200 nm) polymer beads with a regular shape. The polymerization conditions were optimized to give a functional monomer, crosslinker, and template ratio of 1:1:2. We investigated the imprinting effect using both QCM and GC/MS measurements comparing vapor absorption characteristics between the imprinted and non-imprinted (NIP) nanobeads. A four-channel QCM sensory array based on the NIP and the three types of MIP nanobeads was fabricated for sensing the three types of carboxylic acid vapor at concentrations on the ppm level. The output of the sensor array was analyzed by both a non-supervised method (principle component analysis: PCA) and supervised method (linear discrimination analysis: LDA). LDA showed a better discrimination ability than PCA. A 96%-classification rate was achieved by applying leave-one-out cross-validation to the LDA model. The high sensitivity and selectivity of the sensor array was attributed to the imprinting effect of the nano-sized polymer beads. The developed MIP nanobeads, together with other types of MIPs, show promise as materials for artificial receptors in vapor and odorant sensing.