Study of thermal pre-treatment on anaerobic digestion of slaughterhouse waste by TGA-MS and FTIR spectroscopy.
Thermogravimetric analysis coupled to mass spectrometry (TGA-MS) and Fourier-transform infrared spectroscopy (FTIR) were used to describe the effect of pasteurization as a hygienic pre-treatment of animal by-products over biogas production. Piggery and poultry meat wastes were used as substrates for assessing the anaerobic digestion under batch conditions at mesophilic range. Poultry waste was characterized by high protein and carbohydrate content, while piggery waste presented a major fraction of fat and lower carbohydrate content. Results from anaerobic digestion tests showed a lower methane yield for the pre-treated poultry sample. TGA-MS and FTIR spectroscopy allowed the qualitative identification of recalcitrant nitrogen-containing compounds in the pre-treated poultry sample, produced by Maillard reactions. In the case of piggery waste, the recalcitrant compounds were not detected and its biodegradability test reported higher methane yield and production rates. TGA-MS and FTIR spectroscopy were demonstrated to be useful tools for explaining results obtained by anaerobic biodegradability test and in describing the presence of inhibitory problems.


Copper(II) complexes of N-terminal protected tri- and tetra-peptides containing histidine residues.
Copper(II) complexes of peptides containing two or three histidyl residues (Ac-HisGlyHis-OH, Ac-HisGlyHis-NHMe, Ac-HisHisGlyHis-OH and Ac-HisHisGlyHis-NHMe) have been studied by potentiometric, UV-Vis, EPR and CD spectroscopic measurements. The imidazole nitrogen atoms are described as the primary metal binding sites of all ligands resulting in the formation of various macrochelates in the pH range 4 to 7. The (Nim, N-, Nim)-co-ordinated [CuH-1L]0+ complexes were mainly detected in samples containing free carboxylates at the C-termini, whilst the [CuH-2L]-(0) complexes were the predominant species in slightly alkaline solution and their binding modes were described via 4N-co-ordination (Nim, N-, N-, Nim) in (7,5,6)-membered fused chelate rings. Deprotonation and co-ordination of the third amide nitrogens were detected above pH approximately 9 in all cases.


The sugars in chromomycin A3 stabilize the Mg(2+)-dimer complex.
Chromomycin A3 (CRA3) is a glycosylated antitumor antibiotic that binds as a dimer to the minor groove of DNA, with a Mg2+ cation (or another divalent cation with a radius less than 0.85 A) forming the center of the dimer. It has been shown that the chromose sugars are necessary for DNA binding [Kaziro & Kamiyama (1967) J. Biochem. (Tokyo) 62, 424-429; Kamiyama (1968) J. Biochem. (Tokyo) 63, 566-572], although the reason for this has not been addressed. We have investigated the role that the chromose sugars play in metal complexation in solution (methanol) by comparing the optical behavior of CRA3 and its aglycon, CRN, in the presence of various divalent metals (Mg2+, Ni2+, and Ca2+). The results show that CRA3 forms a dimeric complex [i.e., (CRA3)2M, where M is a metal ion] in the presence of 1 mol equiv of either Ni2+ or Mg2+ but a 1:1 complex in the presence of the much larger Ca2+. In contrast, CRN forms a 1:1 complex (CRN.M)+ with all three metals under identical conditions (1:1 mole ratio of drug to metal). Thus, for the smaller metal ions the sugars stabilize the 2:1 CRA3-metal complex in solution. NMR data on the 2:1 CRA3-Mg2+ complex show that the trisaccharide of one CRA3 molecule lies in close proximity to the chromophore of the other CRA3 molecule. This interaction, which is also present in the Mg(2+)-CRA3-DNA complex [Gao & Patel (1989) Biochemistry 28, 751-762], appears to be related to the stability of the dimer in solution.(ABSTRACT TRUNCATED AT 250 WORDS)


Critical limb ischaemia in patients over 80 years of age: options in a district general hospital.
A total of 114 reconstructions were performed in 82 octogenarian patients and the results compared with those of 33 patients who had primary amputation. The operative mortality rate was higher after amputation (45 versus 11 per cent) and the mean survival less (25 versus 34 months). Quality of life assessment using a Rosser scale suggested that, although there was no improvement in the 38 per cent with a failed reconstruction, there was a significant improvement in the 62 per cent whose reconstruction remained patent. There was minimal improvement in quality of life after primary amputation and this was due to relief of pain. Costs (including the costs of revisions and community costs) were assessed in detail. Although the mean total operative costs of reconstruction were higher than those of amputation (10,222 pounds versus 6475 pounds) this was more than offset by the high community costs of amputation. The total cost of reconstruction was 13,546 pounds, compared with 33,095 pounds for amputation. Following reconstruction 66 per cent of those patients independent before critical limb ischaemia occurred were able to return to their own home; only 33 per cent of amputees were able to do so.


Systemic Response to Infection Induces Long-Term Cognitive Decline: Neuroinflammation and Oxidative Stress as Therapeutical Targets.
In response to pathogens or damage signs, the immune system is activated in order to eliminate the noxious stimuli. The inflammatory response to infectious diseases induces systemic events, including cytokine storm phenomenon, vascular dysfunction, and coagulopathy, that can lead to multiple-organ dysfunction. The central nervous system (CNS) is one of the major organs affected, and symptoms such as sickness behavior (depression and fever, among others), or even delirium, can be observed due to activation of endothelial and glial cells, leading to neuroinflammation. Several reports have been shown that, due to CNS alterations caused by neuroinflammation, some sequels can be developed in special cognitive decline. There is still no any treatment to avoid cognitive impairment, especially those developed due to systemic infectious diseases, but preclinical and clinical trials have pointed out controlling neuroinflammatory events to avoid the development of this sequel. In this minireview, we point to the possible mechanisms that triggers long-term cognitive decline, proposing the acute neuroinflammatory events as a potential therapeutical target to treat this sequel that has been associated to several infectious diseases, such as malaria, sepsis, and, more recently, the new SARS-Cov2 infection.


The dimensionality of alcohol abuse and dependence: factor analysis of DSM-III-R and proposed DSM-IV criteria in the 1988 National Health Interview Survey.
Decisions on the final version of the DSM-IV alcohol abuse and dependence criteria will be determined largely by the APA's substance abuse field trials, conducted primarily in treated, clinical samples. Among the major objectives of the field trials are to study the boundaries between abuse and dependence, and to identify specific criteria that define the abuse category. The decisions on revisions of the abuse and dependence criteria in DSM-IV should, however, be informed by data from non-treated or general population samples as well. The present study addresses the field trial objectives using recent data from a large general population survey, the 1988 National Health Interview Survey (NHIS88). The paper reports on factor analyses to assess the dimensions underlying the DSM-III-R and DSM-IV dependence and abuse criteria as operationalized in the NHIS88. The focus of the analyses is on whether models with more than one dimension are needed and if so, the correspondence of the dimensions to criteria sets defined in the DSM-III-R and DSM-IV. The analyses show that a two-dimensional model is required. The dimensions are interpreted as abuse and dependence, but the sets of criteria that define each of the dimensions show important deviations from the criteria sets used in the DSM definitions.


Effective depth controls the nitrate removal rates in a water supply reservoir with a high nitrate load.
The Occoquan Reservoir is part of an indirect potable reuse system where a water reclamation plant (WRP) discharges a nitrified product water to prevent the onset of anaerobic conditions in the bottom sediments during the summer months. The elongated narrow shape of the reservoir combined with water temperature gradients in the inlet results in density currents that enhance the transport of nitrate from the surface to the bottom waters. The morphology of the reservoir also causes a longitudinal change in the ratio of water volume to sediment area, herein defined as the effective depth (ZED). Field observations revealed that first-order nitrate removal rate coefficients (k) varied inversely with ZED, suggesting that the upper reaches of the reservoir have a higher potential for nitrate removal compared to the areas closer to the dam. A similar relationship between k (d-1) and ZED was confirmed during laboratory experiments. Differences in k values were attributed mainly to the change in the nitrate supply rate as a result of the increase in water volume flowing over a specific sediment area, which limited nitrate transport to the sediments. The low variability found between the mass transfer coefficients for nitrate (Coefficient of Variation = 0.25) suggested a nearly constant biotic nitrogen removal and confirmed that k values were mainly affected by changes in ZED. Finally, similarities in k values between field and laboratory samples with similar ZED values suggested that different segments of natural systems may be properly downscaled to laboratory-sized configurations for analytical purposes by means of the ZED concept.


Conceptualizing the digitalization of healthcare work: A metaphor-based Critical Interpretive Synthesis.
The digitalization of healthcare work has gained center stage in academic debates spanning disciplines as diverse as medicine, sociology and STS. The different analytical interests and methodological traditions of these three strains of scholarship have, however, resulted in quite diverging approaches to this issue. Points of interest have ranged from the (disattended) promise of increased efficiency of healthcare work, to dynamics of task delegation, (re-)professionalization and (re-)distribution of invisible work, to the disruption of informal organization. Instead of studying these dynamics in practice, in this paper we foreground the potentiality for theory-making inherent in the systematic cross-contamination of different theoretical and disciplinary perspectives. We perform a Critical Interpretive Synthesis (CIS) centering the ways the digitalization of healthcare work has been investigated in recent STS, sociological and medical literature. To open up assumptions and insights intrinsic to each body of literature for scholars and practitioners in other fields, we propose here a metaphor-based variation on CIS approaches. We probe, in turn, what slime molds can teach us about STS's focus on interconnections and materiality, how we can better understand sociological analyses of invisible work exploring them through theatrical performances, and which lessons river engineering offers concerning medical scholarship's discussion of efficiency and proper healthcare work. Thinking through these metaphors, we conceptualize the digitalization of healthcare work as a phenomenon spanning, at once, the directionality of technological innovation trajectories and the open-endedness of situated changes in work practices. Based on our analysis, we propose focusing on technological scripts, and various forms of invisible work and informal organization as entry points into the study of the tension between directionality and open-endedness in the context of the digitalization of healthcare work.


The vertebrate alcohol dehydrogenase system: variable class II type form elucidates separate stages of enzymogenesis.
A mixed-class alcohol dehydrogenase has been characterized from avian liver. Its functional properties resemble the classical class I type enzyme in livers of humans and animals by exhibiting low Km and kcat values with alcohols (Km = 0.7 mM with ethanol) and low Ki values with 4-methylpyrazole (4 microM). These values are markedly different from corresponding parameters of class II and III enzymes. In contrast, the primary structure of this avian liver alcohol dehydrogenase reveals an overall relationship closer to class II and to some extent class III (69 and 65% residue identities, respectively) than to class I or the other classes of the human alcohol dehydrogenases (52-61%), the presence of an insertion (four positions in a segment close to position 120) as in class II but in no other class of the human enzymes, and the presence of several active site residues considered typical of the class II enzyme. Hence, the avian enzyme has mixed-class properties, being functionally similar to class I, yet structurally similar to class II, with which it also clusters in phylogenetic trees of characterized vertebrate alcohol dehydrogenases. Comparisons reveal that the class II enzyme is approximately 25% more variable than the "variable" class I enzyme, which itself is more variable than the "constant" class III enzyme. The overall extreme, and the unusual chromatographic behavior may explain why the class II enzyme has previously not been found outside mammals. The properties define a consistent pattern with apparently repeated generation of novel enzyme activities after separate gene duplications.


[Effect of intensive insulin therapy on high mobility group box-1/nuclear factor-ΚB pathway in severe traumatic brain injury patient with stress hyperglycemia].
To explore the effect of intensive insulin therapy (IIT) on high mobility group box-1/nuclear factor-ΚB (HMGB1/NF-ΚB) signaling pathway in severe traumatic brain injury (sTBI) patient with stress hyperglycemia. Sixty-one sTBI patients with stress hyperglycemia [Glasgow coma scale (GCS) ≤ 8, three times of random blood glucose levels > 11.1 mmoL/L, glycosylated hemoglobin (HbA1c) < 0.065] admitted to the Affiliated Huaian No.1 People's Hospital of Nanjing Medical University from July 2015 to October 2017 were enrolled. Patients were divided into IIT group (29 cases, keeping blood glucose at 4.4-7.8 mmol/L) and conventional glycemic therapy (CGT) group (32 cases, keeping blood glucose at 7.8-12.2 mmo/L) according to the random number table method. Before treatment and 1, 7 and 14 days after treatment, the levels of plasma HMGB1 and tumor necrosis factor-α (TNF-α) were measured by enzyme linked immunosorbent assay (ELISA); C-reactive protein (CRP) was determined by automatic biochemical analyzer, and NF-ΚB p65 gene expression in peripheral blood mononuclear cells was detected by real-time quantitative polymerase chain reaction (PCR). Nine patients were withdrawn from the observation because the 4 consecutive blood glucose monitoring did not reach the target value, combined with severe infection, or abandoned the treatment with serious brain damage. Finally, 52 patients were enrolled in the analysis, including 28 in CGT group and 24 in IIT group. The levels of plasma HMGB1, TNF-α, CRP and the expression of NF-ΚB gene in monocytes of the two groups at 1 day after treatment were significantly higher than those before treatment, and reached the peak value, then gradually decreased. After 7 days of treatment, they were significantly lower than 1 day. The levels of plasma CRP and TNF-α in the IIT group were significantly lower than those in the CGT group [CRP (mg/L): 36.7±4.4 vs. 45.1±6.1, TNF-α (ng/L): 42.4±9.7 vs. 53.2±9.1, both P < 0.05], the level of HMGB1 in plasma and the expression of NF-ΚB p65 in monocytes were significantly lower than those in the CGT group after 14 days of treatment [HMGB1 (μg/L): 60.1±8.7 vs. 80.5±9.1, NF-ΚB p65 (ΔCt): 35.8±8.5 vs. 53.5±7.3, both P < 0.05]. IIT inhibits the inflammatory response in sTBI patients with stress hyperglycemia through HMGB1/NF-ΚB pathway.


A Fuzzy Robust Optimization Model for Waste Allocation Planning Under Uncertainty.
In this study, a fuzzy robust optimization (FRO) model was developed for supporting municipal solid waste management under uncertainty. The Development Zone of the City of Dalian, China, was used as a study case for demonstration. Comparing with traditional fuzzy models, the FRO model made improvement by considering the minimization of the weighted summation among the expected objective values, the differences between two extreme possible objective values, and the penalty of the constraints violation as the objective function, instead of relying purely on the minimization of expected value. Such an improvement leads to enhanced system reliability and the model becomes especially useful when multiple types of uncertainties and complexities are involved in the management system. Through a case study, the applicability of the FRO model was successfully demonstrated. Solutions under three future planning scenarios were provided by the FRO model, including (1) priority on economic development, (2) priority on environmental protection, and (3) balanced consideration for both. The balanced scenario solution was recommended for decision makers, since it respected both system economy and reliability. The model proved valuable in providing a comprehensive profile about the studied system and helping decision makers gain an in-depth insight into system complexity and select cost-effective management strategies.


Management and prognosis of liposarcomas of the spermatic cord: an experience with six cases.
Liposarcoma of the spermatic cord (LSC) is a rare tumor with no consensus on therapeutic management. This study reports six new cases of LSC. The patients' age ranged from 56 to 80 years. All patients presented with a scrotal mass, and it was ultrasound that oriented the diagnosis. The initial treatment consisted of an inguinal orchiectomy. Anatomopathological study coupled with immunohistochemistry using the anti-MDM2 antibody confirmed that the tumors were well-differentiated LSC in four cases and dedifferentiated LSC in the other two cases. Adjuvant radiotherapy was performed in two patients. No recurrence was noted in these two patients at 14 and 34 months of follow-up. The only recurrence we had was local and occurred at 44 months of follow-up in a patient who had a dedifferentiated form ofLSC.


Damage Control Laparotomy in the Cirrhotic Trauma Patient is Highly Lethal: A Matched Cohort Study.
Damage control laparotomy (DCL) has revolutionized trauma care and is considered the standard of care for severely injured patients requiring laparotomy. The role of DCL in cirrhotic patients has not been investigated. A matched cohort study using American College of Surgeons Trauma Quality Improvement Program database including patients undergoing DCL within 24 hours of admission. A 1:2 cohort matching of cirrhotic vs. non-cirrhotic patients was matched for the following criteria: age (>55, ≤55 years), gender, mechanism of injury (blunt and penetrating), injury severity score (ISS) (≤25, >25), head/face/neck Abbreviated Injury Scale (AIS) (<3, ≥3), chest AIS (<3, ≥3), abdominal AIS (<3, ≥3), and overall comorbidities. Outcomes between the 2 cohorts were subsequently compared with univariable analysis. Overall, 1151 patients with DCL within 24 hours were identified, 29 (2.5%) with liver cirrhosis. Six cirrhotic patients were excluded because there were no suitable matching controls. The remaining 23 cirrhotic patients were matched with 46 non-cirrhotic patients. Overall mortality in the cirrhotic group was 65% vs. 26% in the non-cirrhotic group (P = .002). The higher mortality rate in cirrhotic vs. non-cirrhotic patients was accentuated in the group with ISS >25 (83% vs. 33%; P = .005). 40% of the deaths in cirrhotic patients occurred after 10 days of admission, compared to only 8% in non-cirrhotic patients (P = .091). The total blood product use within 24 hours was significantly higher in cirrhotic than non-cirrhotic patients [33 (14-46) units vs. 19.9 (4-32) units; P = .044]. Cirrhotic trauma patients undergoing DCL have a very high mortality. A significant number of deaths occur late and alternative methods of physiological support should be considered.


Arteriographic view of treatment to achieve regression of coronary atherosclerosis and to prevent plaque disruption and clinical cardiovascular events.
Lipid-lowering therapy, as assessed by angiography, clearly benefits the arterial disease process. For example, among intensively treated patients in FATS the frequency of definite progression per lesion at risk was reduced by 75% among mild and moderate lesions, which form the great preponderance of the lesion population. Regression frequency per lesion was more than doubled by intensive therapy in mild and moderate subgroups and quadrupled in the subgroup with severe lesions. Clinical events were reduced by 73%. This was clearly due to a 15-fold reduction in the likelihood that a mildly or moderately diseased arterial segment would undergo abrupt and substantial progression to a severe lesion at the time of the clinical event. It has been shown that the process of plaque fissuring, leading to plaque disruption, thrombosis, and clinical coronary events, is predicted by the size of the plaque core lipid pool and the abundance of lipid-laden macrophages in its fibrous cap. Experimentally, lipid lowering therapy decreases the number of lipid-laden intimal macrophages and more slowly depletes core cholesteryl ester deposits. Thus the composite of new and previously published data presented here supports the idea that lipid-lowering therapy selectively lipid-depletes (causes regression of) those fatty lesions containing a large lipid core and abundant intimal foam cells. By doing so, these lesions, which are most vulnerable to fissuring, are rendered much more stable and the clinical event rate is accordingly decreased.


Specificities of β1 integrin signaling in the control of cell adhesion and adhesive strength.
Cells exert actomyosin contractility and cytoskeleton-dependent force in response to matrix stiffness cues. Cells dynamically adapt to force by modifying their behavior and remodeling their microenvironment. This adaptation is favored by integrin activation switch and their ability to modulate their clustering and the assembly of an intracellular hub in response to force. Indeed integrins are mechanoreceptors and mediate mechanotransduction by transferring forces to specific adhesion proteins into focal adhesions which are sensitive to tension and activate intracellular signals. α(5)β(1) integrin is considered of major importance for the formation of an elaborate meshwork of fibronectin fibrils and for the extracellular matrix deposition and remodeling. Here we summarize recent progress in the study of mechanisms regulating the activation cycle of β(1) integrin and the specificity of α(5)β(1) integrin in mechanotransduction.


[Cerebrovascular imaging to facilitate stroke reperfusion therapy in Japan].
Imaging diagnosis is essential to perform appropriate reperfusion therapy for acute ischemic stroke. To equally perform reperfusion therapy all over in Japan, it is important to properly facilitate acute imaging evaluation for stroke suspected patients by medical staff not only in stroke-specialized hospitals but also in non-stroke-specialized hospitals. It is unique that CT and MRI are available in most of Japanese hospitals. Even in non-stroke-specialized hospitals, inpatients may suffer from in-hospital stroke. We review statements and recommendation items for a diagnostic imaging to appropriately perform reperfusion therapy based on major clinical trials, stroke guidelines and the current status of acute stroke imaging in Japan.


Psychometric properties of the Separation-Individuation Test of Adolescence within a clinical population.
Completed forms of the Separation Individuation Test of Adolescence (SITA) and the Millon Adolescent Personality Inventory were obtained from 117 clinical subjects. In addition, clinicians who were working with the subjects rated them on the SITA's subscale dimensions. The results were compared with those previously found for a nonclinical population. Moderately strong internal-structural properties were attained for six of the nine SITA subscales. Personality external criteria correlations showed a great deal of convergence with findings previously obtained from a nonclinical sample and were consistent with the theoretical foundations of the SITA subscales. Clinician ratings did not tend to correlate with the complementary SITA subscales, although they did with SITA subscales that were conceptually similar.


[Psychoanalysis, adolescence and subsequent psychological effects].
Theories of the avant-garde in art aim at discovering new meanings in what appear to be securely established traditions. This approach also plays an important role in psychoanalysis, inquiring as it does into the human capacity for subsequently relating to the past in such a way as to create new traditions with a view to restructuring the present. Erdheim's contention is that this historical capacity is a product of the two-phase nature of human sexual development and comes to fruition in adolescence: "Adolescence is the avant-garde of the individual." With reference to three specific factors--male-female polarity, the meaning of aggression and omnipotence, antagonism between family and culture--the author traces the way in which this past-oriented and future-related structure takes shape in adolescence.


Dominance of Replica Off-Diagonal Configurations and Phase Transitions in a PT Symmetric Sachdev-Ye-Kitaev Model.
We show that, after ensemble averaging, the low temperature phase of a conjugate pair of uncoupled, quantum chaotic, non-Hermitian systems such as the Sachdev-Ye-Kitaev (SYK) model or the Ginibre ensemble of random matrices is dominated by saddle points that couple replicas and conjugate replicas. This results in a nearly flat free energy that terminates in a first-order phase transition. In the case of the SYK model, we show explicitly that the spectrum of the effective replica theory has a gap. These features are strikingly similar to those induced by wormholes in the gravity path integral which suggests a close relation between both configurations. For a nonchaotic SYK, the results are qualitatively different: the spectrum is gapless in the low temperature phase and there is an infinite number of second order phase transitions unrelated to the restoration of replica symmetry.


Development of a Novel Friction Model for Machining Simulations in Unidirectional Composite Materials.
Constant coefficients of friction (COFs) are currently used in the literature to describe the contact mechanics between tool and workpiece for finite element (FE) machining simulation of carbon fibre-reinforced polymers (CFRPs). However, these are solely based on closed-loop tribology experimentation, which insufficiently represent machining conditions. To overcome this gap in the knowledge, this work proposes a novel experimental open-loop tribological testing method to produce a dynamic FE friction model for CFRP machining simulations. The newly proposed dynamic friction model is based on a function of fibre angle, contact pressure and slip rate, and it has been validated to both experimental results and constant COF FE simulations. The main aim of this article is to create a link between machining, tribology and FE simulation, by implementing cutting-edge tribological testing that results in highly accurate FE simulations. This dynamic model has been shown to improve the accuracy of open-loop tribological simulations, giving confidence in future implantation in CFRP machining simulations.


Emerald ash borer (Coleoptera: Buprestidae) attraction to stressed or baited ash trees.
Emerald ash borer, Agrilus planipennis Fairmaire (Coleoptera: Buprestidae), has killed millions of ash (Fraxinus sp.) trees in North America since its discovery in Michigan in 2002. Efficient methods to detect low-density A. planipennis populations remain a critical priority for regulatory and resource management agencies. We compared the density of adult A. planipennis captured on sticky bands and larval density among ash trees that were girdled for 1 or 2 yr, wounded, exposed to the stress-elicitor methyl jasmonate, baited with Manuka oil lures, or left untreated. Studies were conducted at four sites in 2006 and 2007, where A. planipennis densities on untreated trees ranged from very low to moderate. In 2006, 1-yr girdled trees captured significantly more adult A. planipennis and had higher larval densities than untreated control trees or trees treated with methyl jasmonate or Manuka oil. Open-grown trees captured significantly more A. planipennis beetles than partially or fully shaded trees. In 2007, A. planipennis population levels and captures of adult A. planipennis were substantially higher than in 2006. The density of adults captured on sticky bands did not differ significantly among canopy exposure classes or treatments in 2007. Larval density was significantly higher in untreated, wounded, and 1-yr girdled trees (girdled in 2007) than in 2-yr girdled trees (girdled in 2006), where most phloem was consumed by A. planipennis larvae the previous year. A total of 36 trees (32 in 2006, 4 in 2007) caught no beetles, but 16 of those trees (13 in 2006, 3 in 2007) had A. planipennis larvae. In 2006, there was a positive linear relationship between the density of adults captured on sticky bands and larval density in trees. Our results show that freshly girdled and open grown trees were most attractive to A. planipennis, especially at low-density sites. If girdled trees are used for A. planipennis detection or survey, debarking trees to locate larval galleries is crucial.


Enhanced cytotoxicity of natural killer cells following the acquisition of chimeric antigen receptors through trogocytosis.
Natural killer (NK) cells have the capacity to target tumors and are ideal candidates for immunotherapy. Viral vectors have been used to genetically modify in vitro expanded NK cells to express chimeric antigen receptors (CARs), which confer cytotoxicity against tumors. However, use of viral transduction methods raises the safety concern of viral integration into the NK cell genome. In this study, we used trogocytosis as a non-viral method to modify NK cells for immunotherapy. A K562 cell line expressing high levels of anti-CD19 CARs was generated as a donor cell to transfer the anti-CD19 CARs onto NK cells via trogocytosis. Anti-CD19 CAR expression was observed in expanded NK cells after these cells were co-cultured for one hour with freeze/thaw-treated donor cells expressing anti-CD19 CARs. Immunofluorescence analysis confirmed the localization of the anti-CD19 CARs on the NK cell surface. Acquisition of anti-CD19 CARs via trogocytosis enhanced NK cell-mediated cytotoxicity against the B-cell acute lymphoblastic leukemia (B-ALL) cell lines and primary B-ALL cells derived from patients. To our knowledge, this is the first report that describes the increased cytotoxicity of NK cells following the acquisition of CARs via trogocytosis. This novel strategy could be a potential valuable therapeutic approach for the treatment of B-cell tumors.


Impact of seizure duration in maintenance electroconvulsive therapy.
Acute electroconvulsive therapy has anticonvulsive effects. The aim of the present study was to investigate the anticonvulsive efficiency of maintenance electroconvulsive therapy. Records of patients treated with maintenance electroconvulsive therapy were screened retrospectively, and the changes in seizure duration were measured. The patients were subdivided into responders and non-responders. Responders had no significant seizure duration changes within the first week of maintenance electroconvulsive therapy. In contrast, the seizure duration of the non-responder group increased significantly within the first week. It was concluded that the early increase in seizure duration at constant energy could be predictive of relapse.


[The impact of human umbilical cord-derived mesenchymal stem cells on the pancreatic function of type 2 diabetic mice and their regulatory role on NLRP3 inflammasomes].
Objective: To investigate the effect and regulation of umbilical cord-derived mesenchymal stem cells (UC-MSCs) on islets function and NOD-like receptor family, pyrin domain containing 3 (NLRP3) and autophagy in type 2 diabetic mellitus (T2DM) mice. Methods: Experimental study. Twenty, 8-week-old, male C57BL/6J mice were selected and divided into a normal control group (n=5) and a high-fat feeding modeling group (n=15). The model of T2DM was established by high-fat feeding combined with intraperitoneal injection of low-dose streptozotocin. After successful modeling, those mice were divided into a diabetes group (n=7) and a UC-MSCs treatment group (n=7). The UC-MSCs treatment group was given UC-MSCs (1×106/0.2 ml phosphate buffer solution) by tail vein infusion once a week for a total of 4 weeks; the diabetes group was injected with the same amount of normal saline, and the normal control group was not treated. One week after the treatment, mice underwent intraperitoneal glucose tolerance tests and intraperitoneal insulin tolerance tests, and then the mice were sacrificed to obtain pancreatic tissue to detect the expressions of interleukin-1β (IL-1β) and pancreatic and duodenal homeobox 1 (PDX-1) by immunofluorescence. The bone marrow-derived macrophages were stimulated with lipopolysaccharide and adenosine triphosphate (experimental group) in vitro, then co-cultured with UC-MSCs for 24 h (treatment group). After the culture, enzyme-linked immunosorbent assay was used to detect the secretion level of IL-1β in the supernatant, and immunofluorescence staining was used to detect the expression of NLRP3 inflammasome, and related autophagy proteins. Statistical analysis was performed using unpaired one-way analysis of variance, repeated measure analysis of variance. Results: In vivo experiments showed that compared with the diabetes group, the UC-MSCs treatment group partially repaired islet structure, improved glucose tolerance and insulin sensitivity (all P<0.05), and the expression of PDX-1 increased and IL-1β decreased in islets under confocal microscopy. In vitro experiments showed that compared with the experimental group, the level of IL-1β secreted by macrophages in the treatment group was decreased [(85.9±74.6) pg/ml vs. (883.4±446.2) pg/ml, P=0.001], the expression of NLRP3 inflammasome and autophagy-related protein P62 was decreased, and the expressions of microtubule-associated protein 1 light chain 3β (LC3) and autophagy effector Beclin-1 were increased under confocal microscopy. Conclusions: UC-MSCs can reduce the level of pancreatic inflammation in T2DM mice, preserving pancreatic function. This might be associated with the ability of UC-MSCs to inhibit the activity of NLRP3 inflammasomes in macrophages and enhance autophagy levels.


Risk of pneumonia in patients with gastroesophageal reflux disease: A population-based cohort study.
The prevalence of gastroesophagel reflux disease (GERD) has steadily increased. However, the association between GERD itself and the risk of pneumonia remains unclear. This study aimed to investigate the association between GERD and long-term risk of pneumonia and to identify the major risk factors for pneumonia in GERD patients. Using the Taiwan National Health Insurance Research Database, we identified patients who were newly diagnosed with GERD and treated with proton pump inhibitors (PPIs) from January 1, 2004 through December 31, 2010. Two groups comprising 15,715 GERD cases and 15,715 non-GERD matched controls were generated using propensity score matching, thereby making the differences in basic demographics, concomitant medication use, and comorbidities between the two groups inconsiderable. Cumulative incidence of pneumonia was significantly higher in the patients with GERD than that in the non-GERD matched controls, with an adjusted HR of 1.48 (95% confidence interval [CI] = 1.31-1.67; P < 0.001) within 6-year follow-ups. Multivariate stratified analyses revealed similar results in many subgroups, with a highest risk in individuals younger than 40 years of age (HR = 2.17, 95% CI = 1.48-3.19). Crucially, patients with GERD using PPIs longer than 4 months were at a significantly increased risk of pneumonia than those who did not use PPIs or took PPIs less than 4 months. GERD was significantly associated with long-term risk of pneumonia, especially in GERD with PPI use longer than 4 months or in the young population. Further prospective longitudinal studies should be conducted for validation and implementing clinical practice guidelines.


Cortical structure predicts the pattern of corticocortical connections.
Cortical areas are linked through pathways which originate and terminate in specific layers. The factors underlying which layers are involved in specific connections are not well understood. Here we tested whether cortical structure can predict the pattern as well as the relative distribution of projection neurons and axonal terminals in cortical layers, studied with retrograde and anterograde tracers. We used the prefrontal cortices in the rhesus monkey as a model system because their laminar organization varies systematically, ranging from areas that have only three identifiable layers, to those that have six layers. We rated each prefrontal area based on the number and definition of its cortical layers (level 1, lowest; level 5, highest). The structural model accurately predicted the laminar pattern of connections in approximately 80% of the cases. Thus, projection neurons from a higher-level cortex originated mostly in the upper layers and their axons terminated predominantly in the deep layers (4-6) of a lower-level cortex. Conversely, most projection neurons from a lower-level area originated in the deep layers and their axons terminated predominantly in the upper layers (1-3) of a higher-level area. In addition, the structural model accurately predicted that the proportion of projection neurons or axonal terminals in the upper to the deep layers would vary as a function of the number of levels between the connected cortices. The power of this structural model lies in its potential to predict patterns of connections in the human cortex, where invasive procedures are precluded.


Diagnosis of a bleeding Dieulafoy lesion on computed tomography and its subsequent embolization.
Dieulafoy lesions are a potentially serious cause of gastrointestinal bleeding. Because they may bleed intermittently, and only be endoscopically evident during hemorrhage, their diagnosis can be challenging. This is the first case to be reported in the English literature of a patient with a Dieulafoy lesion diagnosed during computed tomography (CT) examination. Reduced acquisition times required for multislice helical CT allow the application of CT angiography in the diagnosis of gastrointestinal bleeding. CT scans are now widely used in the diagnostic algorithm for acute gastrointestinal hemorrhage, and the present case illustrates that with fortuitous timing, they can provide critical information and an opportunity for selected angiography and coil embolization.


Acquired hypertrichosis lanuginosa associated with malignancy.
A patient is described with acquired hypertrichosis lanuginosa associated with malignancy; the case history and pathologic findings are described. The literature, consisting of 35 cases, is analyzed in respect to the hormonal and pathologic findings, and the age and sex of the patients. No definite hormonal cause is as yet assignable to this syndrome.


Identification of bZIP interaction partners of viral proteins HBZ, MEQ, BZLF1, and K-bZIP using coiled-coil arrays.
Basic-region leucine-zipper transcription factors (bZIPs) contain a segment rich in basic amino acids that can bind DNA, followed by a leucine zipper that can interact with other leucine zippers to form coiled-coil homo- or heterodimers. Several viruses encode proteins containing bZIP domains, including four that encode bZIPs lacking significant homology to any human protein. We investigated the interaction specificity of these four viral bZIPs by using coiled-coil arrays to assess self-associations as well as heterointeractions with 33 representative human bZIPs. The arrays recapitulated reported viral-human interactions and also uncovered new associations. MEQ and HBZ interacted with multiple human partners and had unique interaction profiles compared to any human bZIPs, whereas K-bZIP and BZLF1 displayed homospecificity. New interactions detected included HBZ with MAFB, MAFG, ATF2, CEBPG, and CREBZF and MEQ with NFIL3. These were confirmed in solution using circular dichroism. HBZ can heteroassociate with MAFB and MAFG in the presence of MARE-site DNA, and this interaction is dependent on the basic region of HBZ. NFIL3 and MEQ have different yet overlapping DNA-binding specificities and can form a heterocomplex with DNA. Computational design considering both affinity for MEQ and specificity with respect to other undesired bZIP-type interactions was used to generate a MEQ dimerization inhibitor. This peptide, anti-MEQ, bound MEQ both stably and specifically, as assayed using coiled-coil arrays and circular dichroism in solution. Anti-MEQ also inhibited MEQ binding to DNA. These studies can guide further investigation of the function of viral and human bZIP complexes.


The Role of Neurotransmitters in the Protection of Caenorhabditis Elegans for Salmonella Infection by Lactobacillus.
Salmonellosis is a common foodborne disease. We previously reported the protection of Caenorhabditis elegans from Salmonella Typhimurium DT104 infection by Lactobacillus zeae LB1. However, the mechanism is not fully understood. C. elegans exhibits behavior plasticity when presented with diverse pathogenic or commensal bacteria. Whether it can exert approach avoidance to S. Typhimurium through altering its neurological activity remains to be determined. In the current study, both the wild type and mutants defective in serotonin or dopamine production of C. elegans were used to investigate olfactory preference of the nematode to L. zeae LB1, DT104, and Escherichia coli OP50 by choice assays, and its resistance to DT104 infection and the protection offered by L. zeae LB1 using a life-span assay. The expression of target genes in C. elegans was also examined by real-time quantitative PCR. Results showed that pre-exposure to L. zeae LB1 did not elicit aversive olfactory behavior of the nematode toward DT104. Both mutants tph-1 and cat-2 succumbed faster than the wild type when infected with DT104. While pre-exposure to L. zeae LB1 significantly increased the survival of both the wild type and mutant tph-1, it provided no protection to mutant cat-2. Supplementation of dopamine resulted in both the resistance of mutant cat-2 to S. Typhimurium infection and the protection from L. zeae LB1 to the same mutant. Gene expression data also supported the observations in the life-span assay. These results suggest that both serotonin and dopamine play a positive role in the host defense of C. elegans to S. Typhimurium infection and that the L. zeae LB1 protection is not dependent on modifying olfactory preference of the nematode but mediated by dopamine that may have involved the regulation of p38-mitogen-activated protein kinase and insulin/insulin-like growth factor signaling pathways.


A Retrospective Study of Clinical and Histopathological Features of 81 Cases of Canine Apocrine Gland Adenocarcinoma of the Anal Sac: Independent Clinical and Histopathological Risk Factors Associated with Outcome.
Canine apocrine gland anal sac adenocarcinoma (AGASAC) is a malignant tumour with variable clinical progression. The objective of this study was to use robust multivariate models, based on models employed in human medical oncology, to establish clinical and histopathological risk factors of poor survival. Clinical data and imaging of 81 cases with AGASAC were reviewed. Tissue was available for histological review and immunohistochemistry in 49 cases. Tumour and lymph node size were determined using the response evaluation criteria in the solid tumours system (RECIST). Modelling revealed tumour size over 2 cm, lymph node size grouped in three tiers by the two thresholds 1.6 cm and 5 cm, surgical management, and radiotherapy were independent clinical variables associated with survival, irrespective of tumour stage. Tumour size over 1.3 cm and presence of distant metastasis were independent clinical variables associated with the first progression-free interval. The presence of the histopathological variables of tumour necrosis, a solid histological pattern, and vascular invasion in the primary tumour were independent risk factors of poor survival. Based upon these independent risk factors, scoring algorithms to predict survival in AGASAC patients are presented.


Prenatal Exposure to Endocrine Disruptors and Reprogramming of Adipogenesis: An Early-Life Risk Factor for Childhood Obesity.
Obesity is a global health problem. It is characterized by excess adipose tissue that results from either increase in the number of adipocytes or increase in adipocytes size. Adipocyte differentiation is a highly regulated process that involves the activation of several transcription factors culminating in the removal of adipocytes from the cell cycle and induction of highly specific proteins. Several other factors, including hormones, genes, and epigenetics, are among the most important triggers of the differentiation process. Although the main contributing factors to obesity are high caloric intake, a sedentary lifestyle, and genetic predisposition, strong evidence supports a role for life exposure to environmental pollutants. Endocrine-disrupting chemicals are exogenous, both natural and man-made, chemicals that disrupt the body signaling processes, thus interfering with the endocrine system. Several studies have shown that prenatal exposure to endocrine disruptors modulates the mechanisms, by which multipotent mesenchymal stem cells differentiate into adipocytes. This review discusses adipocytes differentiation and highlights the possible mechanisms of prenatal exposure to endocrine disruptors in reprogramming of adipogenesis and induction of obesity later in life. Therefore, this review provides knowledge that reduction of early life exposure to these chemicals could open the door for new strategies in the prevention of obesity, especially during childhood.


The "Brain Stress Timing" phenomenon and other misinterpretations of randomized clinical trial on aneurysmal subarachnoid hemorrhage.
Clipping and coiling are currently the two alternatives in treatment of ruptured cerebral aneurysms. In spite of some meritorious analysis, further discussion is helpful to understand the actual state of art. Retreatment and rebleeding rates clearly favors clipping, although short-term functional outcome seems to be beneficial for clipping, while this different is not such if we perform the comparison at a longer follow up. Long-term follow ups and cost analysis are mandatory to have a clear view of the current picture in treatment of subarachnoid hemorrhage. Treatment strategy should be made by a multi-disciplinary team in accredited centers with proficient experience in both techniques.


Mycobacterium tuberculosis cAMP receptor protein (Rv3676) differs from the Escherichia coli paradigm in its cAMP binding and DNA binding properties and transcription activation properties.
The pathogen Mycobacterium tuberculosis produces a burst of cAMP upon infection of macrophages. Bacterial cyclic AMP receptor proteins (CRP) are transcription factors that respond to cAMP by binding at target promoters when cAMP concentrations increase. Rv3676 (CRP(Mt)) is a CRP family protein that regulates expression of genes (rpfA and whiB1) that are potentially involved in M. tuberculosis persistence and/or emergence from the dormant state. Here, the CRP(Mt) homodimer is shown to bind two molecules of cAMP (one per protomer) at noninteracting sites. Furthermore, cAMP binding by CRP(Mt) was relatively weak, entropy driven, and resulted in a relatively small enhancement in DNA binding. Tandem CRP(Mt)-binding sites (CRP1 at -58.5 and CRP2 at -37.5) were identified at the whiB1 promoter (PwhiB1). In vitro transcription reactions showed that CRP1 is an activating site and that CRP2, which was only occupied in the presence of cAMP or at high CRP(Mt) concentrations in the absence of cAMP, is a repressing site. Binding of CRP(Mt) to CRP1 was not essential for open complex formation but was required for transcription activation. Thus, these data suggest that binding of CRP(Mt) to the PwhiB1 CRP1 site activates transcription at a step after open complex formation. In contrast, high cAMP concentrations allowed occupation of both CRP1 and CRP2 sites, resulting in inhibition of open complex formation. Thus, M. tuberculosis CRP has evolved several distinct characteristics, compared with the Escherichia coli CRP paradigm, to allow it to regulate gene expression against a background of high concentrations of cAMP.


Improving the diastereoselectivity of penicillin G acylase for ampicillin synthesis from racemic substrates.
Semi-synthetic β-lactam antibiotics are synthesized enzymatically with the use of penicillin G acylase (PGA). Currently, PGA only exhibits weak diastereoselectivity with respect to the alpha amino group of rac-phenylglycine methyl ester (rac-PGME) when it is coupled with 6-aminopenicillanic acid to synthesize ampicillin. Therefore, we sought to improve the diastereoselectivity of PGA by targeting residues for site-saturation based on the proximity to the substrate's chiral center. Four variants with improved selectivity for (R)-ampicillin synthesis were identified, all resulting from a mutation at the β24 position. βPhe24Ala, while not identified from our library screening, was obtained with site-directed mutagenesis because it has been previously shown to be selective for (R)-enantiomers with substituents other than an amino group. The diastereomeric excess (d.e.(R)) value of 37% for the wild-type enzyme was improved to a d.e.(R) value of 98% for our most selective mutant, βPhe24Ala. Also, four mutations at the α146 position that resulted in (S)-selective PGA variants were identified. βPhe24 and αPhe146 are on opposite sides of the alpha carbon of the substrate, and we have shown that altering these residues results in enhanced selectivity in opposite directions. All variants that showed selectivity for (S)-ampicillin synthesis showed decreased synthetic activity for pure substrates and a decreased synthesis-to-hydrolysis ratio. In contrast, the mutants that were selective for (R)-ampicillin showed significantly decreased primary and secondary hydrolysis when synthesizing ampicillin from pure (R)-PGME, resulting in up to 4-fold decrease in the synthesis to hydrolysis ratio and up to 2-fold increase in the yield achieved. Finally, it was discovered that the selective PGA variants have racemase or epimerase activity, a fascinating phenomenon that has never been reported.


Reaction to Koppad SN et al. Bilateral spermatocytic seminoma: an update on "synchronous" and "sequential" presentation of this rare variant of testicular tumour.
We read with interest the report of Koppad and colleagues in the Pan African Medical Journal describing a case of bilateral synchronous presentation of spermatocytic seminoma in an elderly Indian male. While we appreciate their efforts in documenting this rare presentation, we disagree with the reported figures as outlined in the report and wish to draw attention of the authors as well as the readers of the journal to the gross inaccuracies in the reported statistics. We present our data, following a comprehensive literature review, to unveil the magnitude of bilateral presentation (synchronous and sequential) of this unique variant of testicular tumor as reported in medical literature to facilitate dissemination of precise information on the topic.


Universal predictive models on octanol-air partition coefficients at different temperatures for persistent organic pollutants.
Owing to the importance of octanol-air partition coefficients (KOA) in describing the partition of organic pollutants from air to environmental organic phases, the paucity of KOA data at different environmental temperatures, and the difficulty or high expenditures involved in experimental determination, the development of predictive models for KOA is necessary. Approaches such as this are greatly needed to evaluate the environmental fate of the ever-increasing list of production chemicals. Partial least squares (PLS) regression with 18 molecular structural descriptors was used to develop predictive models based on directly measured KOA values of selected chlorobenzenes, polychlorinated biphenyls (PCBs), polychlorinated naphthalenes, polychlorinated dibenzo-p-dioxins/dibenzofurans, polybrominated diphenyl ethers, polycyclic aromatic hydrocarbons, and organochlorine pesticides (OPs). An optimization procedure resulted in two temperature-dependent universal predictive models that explained at least 91 % of the variance of log KOA. Model 1 was the more general of the two models that could be used for all the persistent organic pollutant (POP) classes investigated. Although model 1 performed poorly for select OPs, this was attributed to wide variability in structural types within this subset of POPs and their diversity compared to the other POP classes that were investigated. The exclusion of the structurally complex OP subset resulted in a more precise model, model 5. Intermolecular dispersive interactions (induced dipole-induced dipole forces) between octanol and solute molecules play a decisive role in governing KOA and its temperature dependence. Further investigations are needed to better characterize the steric structures of the POPs under study, especially of OPs.


Control of Adipogenic Differentiation in Mesenchymal Stem Cells via Endogenous Gene Activation Using CRISPR-Cas9.
Mesenchymal stem cells (MSCs) are of interest in regenerative medicine owing to their multilineage differentiation and self-renewal properties. Understanding the in vivo differentiation process is necessary for clinical applications including cell therapy and transplantation. This remains challenging owing to the lack of induction methods that imitate the natural programming process. Endogenous gene regulation of tissue-specific transcription factors is therefore desirable. In the present study, we demonstrated endogenous activation of adipogenic genes through the dCas9-based transcription system and achieved efficient induction of different types of adipocyte-like cells from MSCs. Interestingly, the MSCs converted via single-gene activation exhibited morphological and molecular properties of white adipocytes, while beige adipocyte-like cells were induced via multiplex gene activation of three specific transcription factors. These results reveal that the fate of MSCs can be effectively manipulated by direct activation of specific endogenous gene expression using a dCas9-based activator with reduced exogenous additives.


Conservation of lipid functions in cytochrome bc complexes.
Lipid binding sites and properties are compared in two sub-families of hetero-oligomeric membrane protein complexes known to have similar functions in order to gain further understanding of the role of lipid in the function, dynamics, and assembly of these complexes. Using the crystal structure information for both complexes, we compared the lipid binding properties of the cytochrome b(6)f and bc(1) complexes that function in photosynthetic and respiratory membrane energy transduction. Comparison of lipid and detergent binding sites in the b(6)f complex with those in bc(1) shows significant conservation of lipid positions. Seven lipid binding sites in the cyanobacterial b(6)f complex overlap three natural sites in the Chlamydomonas reinhardtii algal complex and four sites in the yeast mitochondrial bc(1) complex. The specific identity of lipids is different in b(6)f and bc(1) complexes: b(6)f contains sulfoquinovosyldiacylglycerol, phosphatidylglycerol, phosphatidylcholine, monogalactosyldiacylglycerol, and digalactosyldiacylglycerol, whereas cardiolipin, phosphatidylethanolamine, and phosphatidic acid are present in the yeast bc(1) complex. The lipidic chlorophyll a and β-carotene (β-car) in cyanobacterial b(6)f, as well as eicosane in C. reinhardtii, are unique to the b(6)f complex. Inferences of lipid binding sites and functions were supported by sequence, interatomic distance, and B-factor information on interacting lipid groups and coordinating amino acid residues. The lipid functions inferred in the b(6)f complex are as follows: (i) substitution of a transmembrane helix by a lipid and chlorin ring, (ii) lipid and β-car connection of peripheral and core domains, (iii) stabilization of the iron-sulfur protein transmembrane helix, (iv) n-side charge and polarity compensation, and (v) β-car-mediated super-complex with the photosystem I complex.


Th17 cells and neutrophils: Close collaborators in chronic Leishmania mexicana infections leading to disease severity.
Cutaneous leishmaniasis caused by Leishmania mexicana is associated with an important inflammatory response. We here analysed the kinetics of Th17 cells and neutrophils in ear lobe lesions caused by Leishmania mexicana throughout 90 days of disease progression in susceptible BALB/c and semi-resistant C57BL/6 mice infected with 1 × 105 Leishmania mexicana promastigotes. Cells in the lesions were extracted and quantified by flow cytometry, whereas their distribution in the tissues in relation to the parasites was analysed by immunohistochemistry. Our results show that in BALB/c mice, both Th17 cells and neutrophils increase concomitantly and to significantly higher levels on day 90 post-infection, as compared to C57BL/6 mice. Our results provide novel evidence on the cells causing chronic inflammation throughout Leishmania mexicana infections, resulting as a consequence of neutrophil recruitment together with Th17 cell differentiation and recruitment, both of which remain in the infection site throughout the late phase of the infection. We conclude that the more enhanced levels of Th17 cells and neutrophils during chronic inflammatory lesions in BALB/c mice participate in their enhanced susceptibility towards a progressive disease evolution, whereas the more controlled response of these cells in C57BL/6 mice possibly relates to the more resistant profile of this mouse strain.


Mycobacterium tuberculosis malate synthase is a laminin-binding adhesin.
Mycobacterium tuberculosis (M. tb) uses the glyoxalate bypass for intracellular survival in vivo. These studies provide evidence that the M. tb malate synthase (MS) has adapted to function as an adhesin which binds to laminin and fibronectin. This binding is achieved via the unique C-terminal region of the M. tb MS. The ability to function as an adhesin necessitates extracellular localization. We provide evidence that despite the absence of a Sec-translocation signal sequence the M. tb MS is secreted/excreted, and is anchored on the cell wall by an undefined mechanism. The MS of Mycobacterium smegmatis is cytoplasmic but the M. tb MS expressed in M. smegmatis localizes to the cell wall and enhances the adherence of the bacteria to lung epithelial A549 cells. Antibodies to the C-terminal laminin/fibronectin-binding domain interfere with the binding of the M. tb MS to laminin and fibronectin and reduce the adherence of M. tb to A549 cells. Coupled to the earlier evidence of in vivo expression of M. tb MS during active but not latent infection in humans, these studies show that a housekeeping enzyme of M. tb contributes to its armamentarium of virulence promoting factors.


Ridge Augmentation in a Site of a Previous Implant Failure Using Tenting Screws With Allograft and Collagen Membrane.
Rehabilitation of a severely resorbed mandibular posterior ridge with implants poses a challenge to the clinician. Several techniques to address this challenge have been described in the literature. This case report describes the successful vertical and horizontal bone augmentation of a site of a previous implant failure with severe vertical and horizontal ridge deficiencies using tenting screws, cortico-cancellous particulate bone allograft, and a resorbable collagen membrane. A bone core was obtained at the time of implant placement, 8 months postoperatively, and histological findings showed the highest concentration of lamellar bone at the apical third; a 50-50% graft and lamellar bone proportion in the middle third; and a higher concentration of bone allograft at the coronal third of the bone core. Successful implant placement was achieved at the site. This case report demonstrates the effective use of tenting screws for vertical and horizontal bone augmentation and consequent implant placement in a severely resorbed ridge in the posterior mandible.


Out-of-home placement of children with intellectual disability: Israeli-born parents vs. new immigrants from the ex-USSR.
Although the policy of deinstitutionalization encourages parents to raise a child with development disability at home, professionals are becoming increasingly aware of these parents' care-giving roles. Immigrant parents from the ex-USSR who have children with developmental disabilities are potential clients for placement, but to date, the tendency in Israel to out-of-home placement by immigrant parents, compared with Israeli-born parents, has not been studied. The placement variables were examined as a function of place of birth, emotional stress and social support. The sample was 100 parents who have children aged 6-21 years with mild to moderate mental retardation (according to the American Association on Mental Retardation definition) who live at home. Fifty parents were immigrants (arrived in Israel after 1990) and 50 were Israeli-born. They live in the region of Haifa and the north of Israel. The following measurements were carried out: (1) Demographic Questionnaire for data such as place of birth, age, socioeconomic status, etc., (2) Questionnaire of Resources and Stress, (3) Family Support Scale and (4) Out-of-Home Placement Questionnaire. The results showed that parents' readiness to apply for out-of-home placement proved to be related to their stress level and child's age, regardless of their origin (immigrants or Israeli-born) and regardless of social support resources. The older the children with mental retardation and the higher the stress levels felt by their parents, the higher was these parents' potential to apply for out-of-home placement. Immigrant parents tended to report a significantly higher level of stress than Israeli-born parents. No difference was found in the social support resources available to both research groups. Immigrant parents' stress level was higher as the child's mental retardation level was more moderate and their time in Israel was longer. Stress level among Israeli-born parents was higher among those who were religious, were of lower socioeconomic status and/or when, in addition to the mental retardation, the child suffered from other medical problems. In conclusion, families with children with mental retardation should be referred to services in an integrative way, regardless of the origin of the children's parents. Professionals must increase their own awareness of parents' level of stress and its impact on their readiness to apply for out-of-home placement. Recommendations for practical work and for future researches are made.


Dual Wavelength Differential Detection of Fiber Bragg Grating Sensors with a Pulsed DFB Laser.
We show how dual wavelength differential detection can be used to measure fiber Bragg grating sensors using nanosecond pulses from a single DFB laser diode, by taking advantage of its dynamic chirp. This can be performed in two ways: by measuring the reflected power from two separate pulses driven by two different currents, or by taking two delayed digitized samples within a single pulse. A prototype instrument using fast digitizing and processing with an FPGA is used to characterize the chirp, from which the performance can be optimized for both measurement schemes.


Amniotic membrane transplantation in corneal melting after anterior lamellar keratoplasty assisted by femtosecond laser in children.
To evaluate the efficacy of amniotic membrane transplantation in lamella melting after anterior lamellar keratoplasty (ALK) assisted by femtosecond laser in a pediatric patient. An 11-year-old girl with progressive keratoconus underwent ALK assisted by femtosecond laser on the right eye. The surgical procedure was performed under general anesthesia. The 60-KHz IntraLase femtosecond laser (Abbott Medical Optics) created both the donor (thickness 350 µm; diameter 8.1 mm; side cut 70°) and recipient (thickness 260 µm; diameter 8.0 mm; side cut 70°) lamellae. The recipient lamella was then gently removed and donor was fitted into place and sutured using 4 interrupted sutures added to a running suture in nylon 10-0. The surgery was uneventful. Two months after surgery, a lamella melting was observed. One month after topical steroid treatment, amniotic membrane transplantation was performed. When lamella melting was observed, the best-corrected visual acuity (BCVA) was 0.1 and 0.5 with foramen and no inflammatory reaction in the anterior chamber was reported. Eleven months after amniotic membrane transplantation, BCVA was 0.9 and no visual acuity increase was recorded with foramen. Our findings show that amniotic membrane transplantation could be considered in lamella melting after ALK assisted by femtosecond laser in children.


The construction and implementation of a psychosocial interventions care pathway within a low secure environment: a pilot study.
This paper builds on previous articles that explore the use of psychosocial interventions within inpatient settings. There are obvious difficulties in the implementation of psychosocial interventions in practice. The authors suggest that one method of developing psychosocial interventions within services is to provide organizational support to clinicians. This paper explores the development and initial implementation of a psychosocial care pathway within an inpatient setting to provide such support. It concludes with a summary of the immediate effect of the pathway on care.


Statins are not associated with a reduced risk of pancreatic cancer at the population level, when taken at low doses for managing hypercholesterolemia: evidence from a meta-analysis of 12 studies.
Recent experimental research on a class of pharmacological agents that reduce plasma cholesterol, 3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors (statins), has shown promise in pancreatic cancer chemoprevention. While the mechanism remains unclear, several epidemiological studies have also evaluated the relationship between statin use and pancreatic cancer. Our aim was to examine the strength of this association through a detailed meta-analysis of the studies published on the subject in peer-reviewed literature. A comprehensive search for articles published up to December 2007 was performed, reviews of each study were conducted, and data were abstracted. Prior to meta-analysis, the studies were evaluated for publication bias and heterogeneity. Pooled relative risk (RR) estimates and 95% confidence intervals (CIs) were calculated using the random-effects model. Twelve studies (3 randomized placebo-controlled trials [RCTs], 4 cohort, and 5 case-control studies) contributed to the analysis. The studies were grouped on the basis of study design, and separate meta-analyses were conducted. There was no evidence of an association between statin use and pancreatic cancer among either the RCTs (RR 0.99, 95% CI 0.44-2.21) or the observational studies (RR 0.86, 95% CI 0.60-1.24). Similarly, we found no evidence of publication bias. However, a high heterogeneity was detected among the observational studies. Despite the chemopreventive potential of statins demonstrated in experimental studies, our results do not support the hypothesis that these agents reduce the risk of pancreatic cancer at the population level, when taken at low doses for managing hypercholesterolemia.


Plant RanGAPs are localized at the nuclear envelope in interphase and associated with microtubules in mitotic cells.
In animals and yeast, the small GTP-binding protein Ran has multiple functions - it is involved in mediating (i) the directional passage of proteins and RNA through the nuclear pores in interphase cells; and (ii) the formation of spindle asters, the polymerization of microtubules, and the re-assembly of the nuclear envelope in mitotic cells. Nucleotide binding of Ran is modulated by a series of accessory proteins. For instance, the hydrolysis of RanGTP requires stimulation by the RanGTPase protein RanGAP. Here we report the complementation of the yeast RanGAP mutant rna1 with Medicago sativa and Arabidopsis thaliana cDNAs encoding RanGAP-like proteins. Confocal laser microscopy of Arabidopsis plants overexpressing chimeric constructs of GFP with AtRanGAP1 and 2 demonstrated that the fusion protein is localized to patchy areas at the nuclear envelope of interphase cells. In contrast, the cellular distribution of RanGAPs in synchronized tobacco cells undergoing mitosis is characteristically different. Double-immunofluorescence shows that RanGAPs are co-localized with spindle microtubules during anaphase, with the microtubular phragmoplast and the surface of the daughter nuclei during telophase. Co-assembly of RanGAPs with tubulin correlates with these in vivo observations. The detected localization pattern is consistent with the postulated function of plant RanGAPs in the regulation of nuclear transport during interphase, and suggests a role for these proteins in the organization of the microtubular mitotic structures.


Macrophage and lymphocyte potentiation of syngeneic tumor cell and host fibroblast collagenolytic activity in rats.
The collagenolytic responses of normal rat skin fibroblasts (NRS-F) and rat mammary MTLn3 tumor-derived fibroblasts (Ln3-F) were examined following exposure to rat macrophage (M phi-CM)- and lymphocyte (LYM-CM)-conditioned culture medium and/or tumor cell-conditioned medium. Alveolar, intratumoral, and peritoneal macrophages were prepared from mammary adenocarcinoma-bearing rats, as were the peritoneal lymphocytes. Incubation of the two fibroblast populations with LYM-CM produced a 10- and 7-fold stimulation of collagenolytic activity by NRS-F and Ln3-F cells, respectively. Similarly, exposure of NRS-F and Ln3-F fibroblasts to peritoneal M phi-CM produced a 7- and 4-fold increase in the expression of collagenolytic activity, respectively. Conditioned medium from MTLn2 tumor cells also stimulated the collagenolytic expression of both fibroblast populations. Incubation of tumor-associated Ln3-F or NRS-F fibroblasts with MTLn2 tumor cell-conditioned medium enhanced fibroblast collagenolytic activity approximately 20 and 17 times, respectively. When M phi-CM and LYM-CM were further "conditioned" by a subsequent incubation with MTLn2 tumor cells, each stimulated the expression of collagenolytic activity by both fibroblast populations and this was especially pronounced (120-fold increase) in the response of Ln3-F to LYM-CM further conditioned by MTLn2 tumor cells. The conditioned media derived from M phi, LYM, and MTLn2 tumor cells with or without trypsin activation contained low levels of interstitial-type collagenolytic activity which made no significant contribution to the collagenolytic activity of the stimulated fibroblasts. Some collagenase inhibitory activity, however, was detected in the M phi-CM, suggesting that the actual stimulation of collagenolysis by host fibroblasts is underestimated. We conclude that macrophages, lymphocytes, and tumor cells all have the potential to produce stimulatory factor(s) which enhance the collagenolytic activity of normal fibroblast populations. This study provides further evidence of the multifactorial control of collagenase production and supports the concept that host cell-tumor cell interactions can enhance the expression of collagenolytic enzymes.


[Reactive SH-groups in clinical practice. Methodological notes].
Experimental investigations have been performed in order to point out the extrinsic factors influencing the results of -SH group determination. It has been observed that the anticoagulants normally used to obtain plasma samples, influence the serum -SH groups; in particular, heparin increases and citrate depresses the levels; the most reproducible data were obtained in serum samples. In addition it has been shown that the exposure to light for 6-8 hours lowered the serum -SH groups, whereas darkness prevented these changes. Serum-SH groups were not influenced by the administration of non steroidal anti-inflammatory agents, whereas D-Penicillamine and Tiopronin induced an increase of the -SH levels, ranging between 15-20%. The levels returned to the basal values within two hours from the drug intake. Following the methodological criteria here described it is possible to obtain a good information about the -SH group reserve, which free radicals generated in several diseases could consume. The practical usefulness of the serum -SH group determination in oxidative-peroxidative processes in discussed.


Prediction driven functional annotation of hypothetical proteins in the major facilitator superfamily of S. aureus NCTC 8325.
Antibiotic resistance Staphylococcus aureus strains cause several life threatening infections. New drug treatment options are needed, but are slow to develop because 50% of the S. aureus genome is hypothetical. The goal of this is to aid in the annotation of the S. aureus NCTC 8325 genome by identifying hypothetical proteins related to the Major Facilitator Superfamily (MFS). The MFS is a broad protein group with members involved in drug efflux mechanisms causing resistance. To do this, sequences for three MFS proteins with x-ray crystal structures in E. coli were PSI-BLASTed against the S. aureus NCTC 8325 genome to identify homologs. Eleven identified hypothetical protein homologs underwent BLASTP against the non-redundant NCBI database to fit homologs specific to each hypothetical protein. ExPASy characterized the physiochemical features, CDD-BLAST and Pfam identified domains, and the SOSUI server defined transmembrane helices of each hypothetical protein. Based on size (300 - 700 amino acids), number of transmembrane helices (>7), CD06174 and MFS domains in CDD-BLAST and Pfam, respectively, and close relation to well-defined homologs, SAOUHSC_00058, SAOUHSC_00078, SAOUHSC_00952, SAOUHSC_02435, SAOUHSC_02752, and ABD31642.1 are members of the MFS. Further multiple-alignment and phylogeny analyses show SAOUHSC_00058 to be a quinolone resistance protein (NorB), SAOUHSC_00058 a siderophore biosynthesis protein (SbnD), SAOUHSC_00952 a glycolipid permease (LtaA), SAOUHSC_02435 a macrolide MFS transporter, SAOUHSC_02752 a chloramphenicol resistance (DHA1), and ABD31642.1 is a Bcr/CflA family drug resistance efflux transporter. These findings provide better annotation for the existing genome, and identify proteins related to antibiotic resistance in S. aureus NCTC 8325.


Orphanin FQ: a neuropeptide that activates an opioidlike G protein-coupled receptor.
A heptadecapeptide was identified and purified from porcine brain tissue as a ligand for an orphan heterotrimeric GTP-binding protein (G protein)-coupled receptor (LC132) that is similar in sequence to opioid receptors. This peptide, orphanin FQ, has a primary structure reminiscent of that of opioid peptides. Nanomolar concentrations of orphanin FQ inhibited forskolin-stimulated adenylyl cyclase activity in cells transfected with LC132. This inhibitory activity was not affected by the addition of opioid ligands, nor did the peptide activate opioid receptors. Orphanin FQ bound to its receptor in a saturable manner and with high affinity. When injected intracerebroventricularly into mice, orphanin FQ caused a decrease in locomotor activity but did not induce analgesia in the hot-plate test. However, the peptide produced hyperalgesia in the tail-flick assay. Thus, orphanin FQ may act as a transmitter in the brain by modulating nociceptive and locomotor behavior.


[The role of hematopoietic growth factors in the regulation of normal and pathological hematopoiesis].
Normal hematopoiesis is regulated by a family of hematopoietic growth factors that mediate the proliferation and maturation of blood cell precursors and pluripotential stem cells. Several of these factors have been isolated, biochemically characterized and are now being produced by recombinant DNA techniques in quantities sufficient for preclinical studies and clinical use. Three factors in particular have recently received considerable attention for their potential clinical use: granulocyte-macrophage colony-stimulating factor (GM-CSF), granulocyte colony-stimulating factor (G-CSF) and Multi-CSF or Interleukin 3 (IL-3). G-CSF is apparently lineage-specific for the proliferation of and differentiation of neutrophil granulocyte precursors. GM-CSF and IL-3 are less restricted in their action, affecting various lineages, such as eosinophil granulocytes, neutrophil granulocytes, monocytes, erythrocytes and megakaryocytes. The CSFs have many potential clinical applications including enhancing granulocyte production and function in neutropenic patients and rendering myeloid leukemic cells more susceptible to killing by cycle-specific agents.


p53 controls CDC7 levels to reinforce G1 cell cycle arrest upon genotoxic stress.
DNA replication initiation is a key event in the cell cycle, which is dependent on 2 kinases - CDK2 and CDC7. Here we report a novel mechanism in which p53 induces G1 checkpoint and cell cycle arrest by downregulating CDC7 kinase in response to genotoxic stress. We demonstrate that p53 controls CDC7 stability post-transcriptionally via miR-192/215 and post-translationally via Fbxw7β E3 ubiquitin ligase. The p53-dependent pathway of CDC7 downregulation is interlinked with the p53-p21-CDK2 pathway, as p21-mediated inhibition of CDK2-dependent phosphorylation of CDC7 on Thr376 is required for GSK3ß-phosphorylation and Fbxw7ß-dependent degradation of CDC7. Notably, sustained oncogenic high levels of active CDC7 exert a negative feedback onto p53, leading to unrestrained S-phase progression and accumulation of DNA damage. Thus, p53-dependent control of CDC7 levels is essential for blocking G1/S cell-cycle transition upon genotoxic stress, thereby safeguarding the genome from instability and thus representing a novel general stress response.


Kinin B1 receptors: key G-protein-coupled receptors and their role in inflammatory and painful processes.
Kinins are a family of peptides implicated in several pathophysiological events. Most of their effects are likely mediated by the activation of two G-protein-coupled receptors: B(1) and B(2). Whereas B(2) receptors are constitutive entities, B(1) receptors behave as key inducible molecules that may be upregulated under some special circumstances. In this context, several recent reports have investigated the importance of B(1) receptor activation in certain disease models. Furthermore, research on B(1) receptors in the last years has been mainly focused in determining the mechanisms and pathways involved in the process of induction. This was essentially favoured by the advances obtained in molecular biology studies, as well as in the design of selective and stable peptide and nonpeptide kinin B(1) receptor antagonists. Likewise, development of kinin B(1) receptor knockout mice greatly helped to extend the evidence about the relevance of B(1) receptors during pathological states. In the present review, we attempted to remark the main advances achieved in the last 5 years about the participation of kinin B(1) receptors in painful and inflammatory disorders. We have also aimed to point out some groups of chronic diseases, such as diabetes, arthritis, cancer or neuropathic pain, in which the strategic development of nonpeptidic oral-available and selective B(1) receptor antagonists could have a potential relevant therapeutic interest.


Conformational transition in DNA on a cold surface.
The contour length of DNA fragments, deposited and imaged on mica under buffer, was measured as a function of deposition temperature. Extended DNA molecules (on Ni- and silane-treated surfaces) contract rapidly with falling temperature, approaching the contour length of A-DNA at 2 degrees C. The contraction is not unique to a specific sequence and does not occur in solution at 2 degrees C or on a surface at 25 degrees C, indicating that it arises from a combination of low temperature and surface contact. It is probably a consequence of reduced water activity at a cold surface.


Efficacy and safety of olanzapine for treatment of patients with bipolar depression: Chinese subpopulation analysis of a double-blind, randomized, placebo-controlled study.
Depression in bipolar I disorder responds to the atypical antipsychotic olanzapine. This subpopulation analysis assessed whether olanzapine is superior to placebo specifically in the treatment of Chinese patients with bipolar I depression. This was a subpopulation analysis of a 6-week, multicenter, double-blind, parallel, randomized, placebo-controlled trial among 12 Chinese study centers. Eligible inpatients and outpatients were randomized to olanzapine (5 to 20 mg/day) or placebo. Patients were primarily assessed by the Montgomery-Åsberg Depression Rating Scale total score. Secondary assessments used a range of other efficacy and safety measures. This subpopulation analysis was underpowered to show statistically significant differences between treatment groups. In total, 210 patients (mean age 32.9 years at baseline, 54.3% females) were random-ized. Similar proportions of patients treated with olanzapine (75.0%) and placebo (72.9%) completed the double-blind phase. Baseline-to-endpoint least-squares mean ± standard error decrease in the Montgomery-Åsberg Depression Rating Scale total score in the olanzapine group (-13.55±0.80) was similar to that noted in the parent trial (-13.82±0.65). However, the difference between olanzapine and placebo groups was not statistically significant (P=0.44); this finding was also true for the secondary efficacy measures. A post hoc analysis showed a greater emergence of mania in the placebo group, which likely reduced the treatment difference between olanzapine and placebo in the primary efficacy measure. Safety data were consistent with the known safety profile of olanzapine, including a higher incidence of weight gain (≥7%) in the olanzapine group (24.1% vs 1.4%, P<0.001). Olanzapine provides similar improvement in depression among Chinese and non-Chinese bipolar I patients. The lack of a statistically significant difference between the olanzapine and placebo groups in this Chinese subpopulation analysis may relate to an a priori lack of study power, and underestimation of the effect of olanzapine because of a greater emergence of mania in placebo-treated patients and missing data associated with a high early discontinuation rate.


Auditory symptoms in whiplash patients - could earwax occlusion be a benign cause?
Excess cerumen (earwax) in the external auditory canal is associated with symptoms of earache, fullness in the ears, and diminished hearing. These symptoms, and tinnitus, are commonly associated with whiplash injury. Eighty-six whiplash patients were examined to determine if there was a correlation between symptoms of earache, fullness in the ear, diminished hearing, and tinnitus, and the degree of cerumen occlusion. Cerumen occlusion was measured by visualisation of the tympanic membrane and graded according to a 4 point scale. Of 71 subjects reporting no acute onset (within 7 days of the collision that caused their whiplash) earache, fullness in the ears, hearing loss, or tinnitus, 62 had little or no cerenum occlusion. Of seven subjects reporting tinnitus but no other auditory symptoms, none had greater than moderate cerenum occlusion. Of eight subjects reporting one or more of acute onset earache, fullness in the ears, diminished hearing, and tinnitus, seven had complete cerenum occlusion in the affected ear. The findings suggest high grade cerumen occlusion frequently occurs in the ear affected by acute auditory symptoms. However, tinnitus alone has no apparent association with cerumen occlusion. It is possible that a significant number of acute onset auditory symptoms reported in whiplash patients have a benign cause.


AHA to fight HCFA's proposed uniform reporting rules.
Events at the AHA's 1979 annual meeting included approval of a strategy for mobilizing response to the uniform reporting requirement of the 1977 Medicare-Medicaid Antifraud and Abuse Amendments, reaffirmation of AHA's commitment to the Voluntary Effort, an AHA pledge to investigate whether Health Systems Agencies can dictate hospital board composition, and talks by government and association leaders concerning regulation, national health insurance, and the VE.


Comparing the Utility of Mitochondrial and Nuclear DNA to Adjust for Genetic Ancestry in Association Studies.
Mitochondrial genome-wide association studies identify mitochondrial single nucleotide polymorphisms (mtSNPs) that associate with disease or disease-related phenotypes. Most mitochondrial and nuclear genome-wide association studies adjust for genetic ancestry by including principal components derived from nuclear DNA, but not from mitochondrial DNA, as covariates in statistical regression analyses. Furthermore, there is no standard when controlling for genetic ancestry during mitochondrial and nuclear genetic interaction association scans, especially across ethnicities with substantial mitochondrial genetic heterogeneity. The purpose of this study is to (1) compare the degree of ethnic variation captured by principal components calculated from microarray-defined nuclear and mitochondrial DNA and (2) assess the utility of mitochondrial principal components for association studies. Analytic techniques used in this study include a principal component analysis for genetic ancestry, decision-tree classification for self-reported ethnicity, and linear regression for association tests. Data from the Health and Retirement Study, which includes self-reported White, Black, and Hispanic Americans, was used for all analyses. We report that (1) mitochondrial principal component analysis (PCA) captures ethnic variation to a similar or slightly greater degree than nuclear PCA in Blacks and Hispanics, (2) nuclear and mitochondrial DNA classify self-reported ethnicity to a high degree but with a similar level of error, and 3) mitochondrial principal components can be used as covariates to adjust for population stratification in association studies with complex traits, as demonstrated by our analysis of height-a phenotype with a high heritability. Overall, genetic association studies might reveal true and robust mtSNP associations when including mitochondrial principal components as regression covariates.


Laparoscopic left hemi-nephroureterectomy for a patient with suspected urothelial carcinoma in a horseshoe kidney.
A 76-year-old gentleman with a background of benign prostatic hyperplasia, hiatus hernia and anxiety was referred to the rapid access haematuria service following an episode of painless visible haematuria. Flexible cystoscopy did not reveal any concerning bladder lesions. CT Urogram demonstrated a horseshoe kidney with a filling defect in the left upper pole moiety suspicious for an urothelial carcinoma. The patient was subsequently referred to the urology services in a tertiary centre. Flexible ureterorenoscopy was performed, with findings of a likely urothelial carcinoma corresponding to the suspicious area on imaging. Biopsy of this lesion revealed a low grade urothelial cancer. The patient proceeded to have a laparoscopic left heminephroureterectomy with an open bladder cuff. The patient recovered well and urinary catheter was removed Day 12 post procedure after the performance of a cystogram. Histology revealed a favourable pTa low grade malignancy, and the patient will require ongoing follow-up moving forward. This case report highlights the operative intricacies in managing patients with horseshoe kidney due to anatomic variations associated with this condition.


Mutations in the small subunit of cyanobacterial ribulose-bisphosphate carboxylase/oxygenase that modulate interactions with large subunits.
In the cyanobacterium Anacystis nidulans (Synechococcus PCC6301), ribulose 1,5-bisphosphate carboxylase/oxygenase (Rbu-P2 carboxylase) is composed of eight large subunits and eight small subunits. There are three regions of the small subunit that contain amino acids that are conserved throughout evolution, from bacteria to higher plants. Since the function of the small subunit is not fully understood, site-directed mutagenesis was performed on highly conserved residues in the first and second conserved regions. Ser-16, Pro-19, Leu-21, and Tyr-54 were replaced by Asp-16, His-19, Glu-21, and Ser-54, respectively. Crude extracts containing the recombinant His-19 mutant enzyme indicated that there was little effect on either Rbu-P2 carboxylase activity or interactions between large and small subunits. However, the Asp-16, Glu-21, and Ser-54 mutations showed effects on Rbu-P2 carboxylase activity and the interaction between large and small subunits. The large and small subunits of the Asp-16, Glu-21, and Ser-54 enzymes were found to dissociate during nondenaturing gel electrophoresis or sucrose density gradient centrifugation. However, the dissociated small subunits remained functional and were capable of reconstituting Rbu-P2 carboxylase activity when added to large subunits. These results indicated that Ser-16, Leu-21, and Tyr-54 might play an important role in interactions between large and small subunits of the A. nidulans enzyme.


CXCL11-armed oncolytic adenoviruses enhance CAR-T cell therapeutic efficacy and reprogram tumor microenvironment in glioblastoma.
Glioblastoma (GBM) is the most aggressive primary malignant brain cancer and urgently requires effective treatments. Chimeric antigen receptor T (CAR-T) cell therapy offers a potential treatment method, but it is often hindered by poor infiltration of CAR-T cells in tumors and highly immunosuppressive tumor microenvironment (TME). Here, we armed an oncolytic adenovirus (oAds) with a chemokine CXCL11 to increase the infiltration of CAR-T cells and reprogram the immunosuppressive TME, thus improving its therapeutic efficacy. In both immunodeficient and immunocompetent orthotopic GBM mice models, we showed that B7H3-targeted CAR-T cells alone failed to inhibit GBM growth but, when combined with the intratumoral administration of CXCL11-armed oAd, it achieved a durable antitumor response. Besides, oAd-CXCL11 had a potent antitumor effect and reprogramed the immunosuppressive TME in GL261 GBM models, in which increased infiltration of CD8+ T lymphocytes, natural killer (NK) cells, and M1-polarized macrophages, while decreased proportions of myeloid-derived suppressor cells (MDSCs), regulatory T cells (Tregs) and M2-polarized macrophages were observed. Furthermore, the antitumor effect of the oAd-CXCL11 was CD8+ T cell dependent. Our findings thus revealed that CXCL11-armed oAd can improve immune-virotherapy and can be a promising adjuvant of CAR-T therapy for GBM.


Effects of hardwood smoke exposure on allergic airway inflammation in mice.
Hardwood smoke (HWS) from wood burning stoves and fireplaces can be a significant contributor to the composition of ambient air pollution. We hypothesize that the inhalation of HWS by ovalbumin (OVA)-sensitized mice with preexisting lung inflammation leads to the exacerbation of allergic airway responses. Two different models were employed to characterize the effects of inhaled wood smoke on allergic airway inflammation. In both models, male BALB/c mice were sensitized by injection with OVA and alum. In one model, mice were challenged by inhalation with OVA 1 day prior to exposure to HWS (30, 100, 300, or 1000 microg particulate matter [PM]/m(3)) for 6 h/day on 3 consecutive days. In the other model, mice were exposed by inhalation to OVA, rested for 11 days, were exposed to HWS for 3 consecutive days, and then were exposed to OVA immediately after the final HWS exposure. Bronchoalveolar lavage (BAL), and blood collection were performed approximately 18 h after the last HWS or OVA exposure. HWS exposure after the final allergen challenge (first model) led to a significant increase in BAL eosinophils only at the 300 microg/m(3) level. In contrast, changes in BAL cells did not reach statistical significance in the second model. There were no HWS-induced changes in BAL interleukin (IL)-2, IL-4, IL-13, and interferon (IFN)gamma levels in either model following OVA challenge. These results suggest that acute HWS exposure can minimally exacerbate some indices of allergic airway inflammation when a final OVA challenge precedes HWS exposure, but does not alter Th1/Th2 cytokine levels.


Isolation and biochemical composition of the cuticle of Onchocerca volvulus.
A preparation of the cuticle of Onchocerca volvulus was obtained by extracting worm fragments in an series of buffers with 1.5% Triton-X-100 and 3% Sodium dodecyl sulfate (SDS). Electron micrographs of worm fragments, treated with the detergents or collagenase showed that our methods had been effective in isolating the cuticle from the other organs of the parasite. The cuticular preparation was found to contain 19 different amino acids with glycine (23.4%); proline (11.23%); hydroxyproline (10%); and glutamic acid (9.4%) being the most abundant. Hydroxylysine was present in small amounts (0.04%). Total reducing sugar was determined to be 5.3 mg per gram dry weight of the preparation. The cuticular preparation was solubilized by boiling in 2-mercaptoethanol and shown by SDS-PAGE to contain at least 10 different polypeptides in the Mr range 17,000-163,000. Five of these polypeptides with apparent Mr respectively of 33,000; 67,000; 74,000, 88,500 and 114,000 were isolated by preparative gel electrophoresis and their amino acid compositions shown to be similar to that of invertebrate collagens. We conclude that the cuticle of O. volvulus contains collagen-like proteins held together by disulfide bridges.


Photocontrol of Anthocyanin Synthesis: IV. Dose Dependence and Reciprocity Relationships in Anthocyanin Synthesis.
Under continuous far red light, anthocyanin synthesis in young, dark-grown cabbage seedlings (Brassica oleracea cv. Red Acre) is irradiance-dependent and fails to follow the reciprocity (irradiance x time = constant) relationships. Under intermittent far red treatments extended over a prolonged period of time, anthocyanin synthesis becomes dose dependent, and reciprocity relationships are valid. Intermittent far red treatments with short dark intervals between successive irradiations are as effective as continuous treatments, if the total radiation doses applied with the two types of treatments are equal and are applied over equally long periods of time. The high effectiveness of inter-mittent treatments, the dose dependence, and the validity of the reciprocity relationships suggest that cycling between red-absorbing form of phytochrome and far red-absorbing form of phytochrome and the formation of electronically excited far red-absorbing form of phytochrome, or the involvement of a second photoreactive system, besides phytochrome, may play only a minor role in high irradiance reaction anthocyanin synthesis brought about by prolonged exposures to far red irradiation.


Flexible adaptation to changing task demands in severe closed head injury patients: a driving simulator study.
The assessment of fitness to drive in patients with closed head injury (CHI) does not usually include executive functioning. Executive functions may be particularly important for the tactical (e.g. anticipatory adaptation of speed) and strategical (e.g. choice of route and time) aspects of driving. The literature lacks evidence on the tactical aspects, which are critical in the assessment of actual driving. Executive functioning was therefore assessed in a task context relevant to tactical decision making in driving. Twenty very severe chronic CHI patients and 20 healthy control subjects were tested with a driving simulator test and an extended neuropsychological test battery. In the driving simulator test, executive function was operationally defined as the flexible adaptation to changing task demands. A specific feature of the test was the experimental control for slow information processing, a persistent sequel of severe CHI. Given this control for slow information processing, it was found that patients were unimpaired in their flexible adaptations. Similar findings were obtained with a neuropsychological test battery. Patients were consistently slower, but planning and flexibility were found to be unimpaired.