In vivo magnetic resonance imaging of the effects of photodynamic therapy.
Nuclear magnetic resonance (NMR) proton imaging and measurements of the parameters T1 and T2, have been carried out in vivo on the murine mammary tumour T50/80. Tumours had been treated 24 h previously by photodynamic therapy (PDT, using haematoporphyrin derivative and 630 nm laser light). Proton images clearly demarcated a high signal-intensity region on the side of the tumour closest to the incident light beam, while the parts of the tumour more remote from the beam resembled the images from untreated controls. Both T1 and T2 values were raised in the high-intensity region. This high-intensity region was shown to correspond to PDT-induced histological necrosis, the low-intensity region to histologically intact tumour. Linear regression analysis of the relationship of depth of necrosis measured histologically and 'depth of necrosis' measured from the NMR images, yielded a slope of 0.93 (r2 = 0.95).


Use of postmortem radiographs for the investigation of underwater and hyperbaric deaths.
The use of postmortem radiographs as a useful adjunct in the investigation of hyperbaric and immersion deaths is discussed. This technique enables accurate identification of gas within cavities that otherwise could not be detected at routine autopsy, which may also be artifact. In addition it is possible to define bone lesions that need a histologic diagnosis.


[Study of the interaction between the organophosphorus insecticide GA-41 and model membranes using NMR-spectroscopy].
Interaction between organophosphorous insecticide GA-41 and liposomes formed from lecithin and multilamellar dispersion from lecithin and cardiolipin mixture (molar ratio is 7:3) has been studied by 1H- and 31P-NMR spectroscopy. The results obtained showed that while interacting with model membranes the GA-41 molecules intercalate into the range of hydrocarbon chains of phospholipids. At certain concentration of GA-41 its molecules induce the formation of some defects on bilayer structure, leading to permeability for Pr3+ ions.


Synergy of antithrombin III concentrate and antivenom in preventing coagulopathy in a rat model of Malayan pit viper envenoming.
The effects of unrefined equine antivenom and antithrombin III (AT-III) concentrate on the coagulopathy induced by systemic envenomation by Malayan pit viper (Calloselasma rhodostoma; MPV) venom were investigated in a rat model. 37 rats received an intramuscular injection of MPV venom and serial blood samples were taken from the femoral vein for simple whole blood clotting tests and measurement of AT-III activity. 30 min after venom injection, treatment (antivenom, AT-III or both) was given intravenously. 6 rats were untreated and all developed uncoagulable blood and AT-III depletion 90-210 (median 180) min after venom injection. A combination of high dose AT-III concentrate (0.5 units/g) and antivenom (20 micrograms/g) prevented abnormal clotting (P less than 0.001), whereas AT-III alone, antivenom alone, or a combination of low dose AT-III (0.25 units/g) and antivenom did not (P less than 0.05). These results suggest that the coagulation abnormality in MPV envenomation is secondary to activation of the coagulation cascade at several levels, and that treatment with antivenom alone may not be sufficient to reverse or prevent this phenomenon.


The hippocampal formation as a source of the slow endogenous potentials.
Magnetic fields were detected with a SQUID sensor at the temporal and occipital areas of the head in response to a frequent and an infrequent attended visual stimulus. The time-course of the magnetic field for the infrequent stimulus correlated highly with the simultaneously measured electrical potential that showed the commonly observed N2-P3 complex. Analysis of the pattern of the magnetic field showed that the sources of N2 and P3 lay deep in the brain within the hippocampal formation.


Near-infrared co-illumination of fluorescent proteins reduces photobleaching and phototoxicity.
Here we present a method to reduce the photobleaching of fluorescent proteins and the associated phototoxicity. It exploits a photophysical process known as reverse intersystem crossing, which we induce by near-infrared co-illumination during fluorophore excitation. This dual illumination method reduces photobleaching effects 1.5-9.2-fold, can be easily implemented on commercial microscopes and is effective in eukaryotic and prokaryotic cells with a wide range of fluorescent proteins.


Expectant management of viable cesarean scar pregnancies: a systematic review.
Cesarean scar pregnancy (CSP) is a potentially life-threatening disease that has been steadily increasing in prevalence. Pregnancy termination is usually recommended given the risk of life-threatening complications. In some cases, patients refuse to terminate viable CSPs, even after counseling. Recent studies report that, even with a high burden of possible complications and maternal morbidity, many CSPs progress to live, close to term births. The aim of this study is to further demonstrate the natural history of viable cesarean scar pregnancies. We conducted a systematic review of original studies reporting cases of expectant management of CSPs with positive fetal heartbeats. After selection, 28 studies were included in the review, with a total of 398 cases of CSP, 136 managed expectantly and 117 with positive fetal heartbeat managed expectantly. This study confirmed that the majority of patients experience live births, as 78% of patients selected for expectant management experienced live births at or close to term, with 79% developing morbidly adherent placenta, 55% requiring hysterectomy, and 40% having severe bleeding. The optimal management protocol for CSP is still to be defined and more studies are needed to further elucidate this rare but rising disease. Our study provides information on the natural history of untreated CSPs and suggests that termination may not be the only option offered to the patient.


Wrist and forearm range of motion commencement time following primary triangular fibrocartilage complex foveal repair surgery: A scoping review.
Scoping review. Rehabilitation guidelines following triangular fibrocartilage complex (TFCC) foveal repair surgery have been inconsistently reported in the published literature, with no consensus regarding wrist or forearm range of motion (ROM) commencement time. To scope the available literature to identify the extent and strength of the evidence supporting the clinical guidelines for wrist and forearm ROM commencement time following primary TFCC foveal repair surgery. A systematic search produced 26 studies (3 retrospective cohort studies, 1 prospective cohort study, 1 retrospective comparative study, and 21 retrospective case series) that described specific rehabilitation protocols following TFCC foveal repair surgery. No supporting evidence was identified regarding rehabilitation protocol recommendations across all the included studies. Postsurgery wrist ROM commencement ranged from 2 to 8 weeks; forearm ROM commencement ranged from 2 to 12 weeks. ROM commencement times did not appear to systematically influence the rate of adverse events, although adverse events were poorly reported. TFCC rehabilitation protocols were poorly reported and varied widely between the included studies. Additional research is recommended to comprehensively evaluate the association between wrist and/or forearm ROM and the rate of adverse events for this complex and multifaceted condition.


Highly selective glycosylated prodrugs of cytostatic CC-1065 analogues for antibody-directed enzyme tumor therapy.
Novel prodrugs of the cytotoxic antibiotic CC-1065 for an antibody-directed enzyme prodrug therapy (ADEPT) were prepared that show an excellent selectivity with a high toxicity of the corresponding drug. In particular, the seco-CBI analogue of CC-1065, 1-chloromethyl-5-hydroxy-1,2-dihydro-3H-benz[e]indole, as well as the novel methyl-seco-CBI analogue 1-(1'-chloroethyl)-5-hydroxy-1,2-dihydro-3H-benz[e]indole, were synthesized and transformed into their galactosides 10 a and 10 b, respectively. These galactosides can be cleaved with beta-D-galactosidase to give the free cytotoxic compounds. They were tested in in vitro cytotoxicity assays by using human bronchial carcinoma cells of line A549 in the presence and in the absence of beta-D-galactosidase. While the seco-CBI prodrugs revealed only modest selectivity, prodrugs of the methyl-seco-CBI analogue bearing an anti orientation of the substituents at the two stereogenic centers of the N-heterocycle displayed an excellent selectivity with an ED(50) quotient of about 750. The cytotoxicity of the corresponding phenol was rather high, with an ED(50) of 1.3 nM. The diastereomer with a syn orientation at the stereogenic centers was much less toxic.


Rapid frequency-domain FLIM spinning disk confocal microscope: lifetime resolution, image improvement and wavelet analysis.
A spinning disk confocal attachment is added to a full-field real-time frequency-domain fluorescence lifetime-resolved imaging microscope (FLIM). This provides confocal 3-D imaging while retaining all the characteristics of the normal 2-D FLIM. The spinning disk arrangement allows us to retain the speed of the normal 2-D full field FLIM while gaining true 3-D resolution. We also introduce the use of wavelet image transformations into the FLIM analysis. Wavelets prove useful for selecting objects according to their morphology, denoising and background subtraction. The performance of the instrument and the analysis routines are tested with quantitative physical samples and examples are presented with complex biological samples.


Stroke among cancer patients.
We identify cancer patients at highest risk of fatal stroke. This is a population-based study using nationally representative data from the Surveillance, Epidemiology, and End Results program, 1992-2015. Among 7,529,481 cancer patients, 80,513 died of fatal stroke (with 262,461 person-years at risk); the rate of fatal stroke was 21.64 per 100,000-person years, and the standardized mortality ratio (SMR) of fatal stroke was 2.17 (95% CI, 2.15, 2.19). Patients with cancer of the prostate, breast, and colorectum contribute to the plurality of cancer patients dying of fatal stroke. Brain and gastrointestinal cancer patients had the highest SMRs (>2-5) through the follow up period. Among those diagnosed at <40 years of age, the plurality of strokes occurs in patients treated for brain tumors and lymphomas; if >40, from cancers of the prostate, breast, and colorectum. For almost all cancers survivors, the risk of stroke increases with time.


Multiplex real-time SYBR Green I PCR assays for simultaneous detection of 15 common enteric pathogens in stool samples.
Diarrheal diseases account for more than 50% of foodborne diseases worldwide, the majority of which occur in infants and young children. The traditional bacterial detection method is complex and time-consuming; therefore, it is necessary to establish a rapid and convenient detection method that can detect multiple pathogens simultaneously. In this study, we developed a set of five multiplex real-time SYBR Green I PCR assays to simultaneously detect 15 common enteric pathogens based on the Homo-Tag Assisted Non-Dimer system. These assays effectively reduced primer-dimer formation and improved the stability, uniformity, and amplification efficiency of multiplex PCR. The detection limit of the multiplex SYBR Green I PCR system was approximately 104-106 CFU/mL for stool specimens. Furthermore, we vitrified heat-unstable components on the cap of a reaction tube, showing that Taq DNA polymerase, dNTPs, primers, and SYBR Green I remained stable at 25 °C. In summary, we developed multiplex SYBR Green I PCR assays that can simultaneously detect 15 enteric pathogens. This method is comprehensive, rapid, inexpensive, accurate, and simple and displays high specificity.


Extracellular matrix stiffness mediates radiosensitivity in a 3D nasopharyngeal carcinoma model.
Radiotherapy is one of the essential treatment modalities for nasopharyngeal carcinoma (NPC), however, radioresistance still poses challenges. Three-dimensional (3D) tumor culture models mimic the in vivo growth conditions of cells more accurately than 2D models. This study is to compare the tumor biological behaviors of NPC cells in 2D, On-Surface 3D and Embedded 3D systems, and to investigate the correlation between radioresistance and extracellular matrix (ECM) stiffness. The morphology and radioresistance of the human NPC cell line CNE-1 were observed in 2D and 3D systems. The CCK-8 assay, wounding healing assays, flow cytometry, soft agar assays, and western blot analysis were used to evaluate differences in biological behaviors such as proliferation, migration, cell cycle distribution, and stem cell activity. Different ECM stiffness systems were established by co-blending collagen and alginate in varying proportions. ECM stiffness was evaluated by compressive elastic moduli measurement and colony formation assay was used to assess radioresistance of NPC cells in systems with different ECM stiffness after irradiation. Compared to 2D models, the morphology of NPC cells in 3D culture microenvironments has more in common with in vivo tumor cells and 3D cultured NPC cells exhibit stronger radioresistance. Integrin β1 but not the epithelial-to-mesenchymal transition pathway in 3D models boost migration ability. Cell proliferation was enhanced, the proportion of tumor stem cells was increased, and G1/S phase arrest occurred in 3D models. NPC cells cultured in softer ECM systems (with low alginate proportions) exhibit striking resistance to ionizing radiation. The tumor biological behaviors of NPC cells in 3D groups were obviously different from that of 2D. Radioresistance of NPC cells increased with the stiffness of ECM decreasing.


Use of an inner preputial free graft to extend the indications of Snodgrass hypospadias repair (Snodgraft).
Tubularized incised plate (tip) urethroplasty or Snodgrass repair has become the most commonly used procedure for distal hypospadias repair. However, where the urethral plate is narrow or shallow, the good results may be compromised. We present our experience of using a small inner preputial graft to deepen and widen the urethral plate (Snodgraft). This procedure has certainly reduced the number of two-stage hypospadias repairs in patients who lack a urethral groove or have a small glans. The cosmetic and functional results are excellent.


Histone demethylase KDM5D upregulation drives sex differences in colon cancer.
Sex exerts a profound impact on cancer incidence, spectrum and outcomes, yet the molecular and genetic bases of such sex differences are ill-defined and presumptively ascribed to X-chromosome genes and sex hormones1. Such sex differences are particularly prominent in colorectal cancer (CRC) in which men experience higher metastases and mortality. A murine CRC model, engineered with an inducible transgene encoding oncogenic mutant KRASG12D and conditional null alleles of Apc and Trp53 tumour suppressors (designated iKAP)2, revealed higher metastases and worse outcomes specifically in males with oncogenic mutant KRAS (KRAS*) CRC. Integrated cross-species molecular and transcriptomic analyses identified Y-chromosome gene histone demethylase KDM5D as a transcriptionally upregulated gene driven by KRAS*-mediated activation of the STAT4 transcription factor. KDM5D-dependent chromatin mark and transcriptome changes showed repression of regulators of the epithelial cell tight junction and major histocompatibility complex class I complex components. Deletion of Kdm5d in iKAP cancer cells increased tight junction integrity, decreased cell invasiveness and enhanced cancer cell killing by CD8+ T cells. Conversely, iAP mice engineered with a Kdm5d transgene to provide constitutive Kdm5d expression specifically in iAP cancer cells showed an increased propensity for more invasive tumours in vivo. Thus, KRAS*-STAT4-mediated upregulation of Y chromosome KDM5D contributes substantially to the sex differences in KRAS* CRC by means of its disruption of cancer cell adhesion properties and tumour immunity, providing an actionable therapeutic strategy for metastasis risk reduction for men afflicted with KRAS* CRC.


My path in the company of chemistry.
Experiencing the honor of this international recognition in chemistry, I wonder how this came to be. I reflect on my imperfect but rewarding path to where I am now, and on those who have helped me along the way.


Neurophysiologic and quantitative sensory testing in the diagnosis of trigeminal neuropathy and neuropathic pain.
This study investigated the utility of neurophysiologic examination and thermal quantitative sensory testing (QST) in the diagnosis of trigeminal neuropathy and neuropathic pain. Fifty-eight patients (14 men), 34 with sensory deficit within the inferior alveolar nerve (IAN) and 24 within the lingual nerve (LN) distribution, were included. Twenty-six patients (45%) reported neuropathic pain. Patients underwent blink reflex (BR) test and thermal QST; sensory neurography was done to the IAN patients. Results of clinical sensory testing were available from the charts of 48 patients revealing abnormal findings in 77% of the IAN and in 94% of the LN patients. The BR test was abnormal in 41%, neurography in 96%, and QST in 91% of the IAN patients. In the LN group, BR was abnormal in 33%, and QST in 100% of the patients tested. Neurophysiologic tests and QST verified the subjective sensory alteration in all but 2 IAN patients, both with old injuries, and 4 LN patients who did not undergo QST. When abnormal, thermal QST showed elevation of warm and cold detection thresholds (hypo/anesthesia), hypoalgesia was less marked, and heat allodynia was only occasionally present. Contralateral thermal hypoesthesia after unilateral injury was found in 14 patients. It was associated with the occurrence of neuropathic pain (P=0.016). Axonal Abeta afferent damage was less severe in the IAN patients with pain than in those without pain (P=0.012). Neurophysiologic tests and thermal QST provide sensitive tools for accurate diagnosis of trigeminal neuropathy and study of pathophysiological features characteristic to human neuropathic pain.


Construction of quality-assured infant feeding process of care data repositories: Construction of the perinatal repository (Part 2).
This is the second in a series of two papers regarding the construction of data quality (DQ) assured repositories, based on population data from Electronic Health Records (EHR), for the reuse of information on infant feeding from birth until the age of two. This second paper describes the application of the computational process of constructing the first quality-assured repository for the reuse of information on infant feeding in the perinatal period, with the aim of studying relevant questions from the Baby Friendly Hospital Initiative (BFHI) and monitoring its deployment in our hospital. The construction of the repository was carried out using 13 semi-automated procedures to assess, recover or discard clinical data. The initial information consisted of perinatal forms from EHR related to 2048 births (Facts of Study, FoS) between 2009 and 2011, with a total of 433,308 observations of 223 variables. DQ was measured before and after the procedures using metrics related to eight quality dimensions: predictive value, correctness, duplication, consistency, completeness, contextualization, temporal-stability, and spatial-stability. Once the predictive variables were selected and DQ was assured, the final repository consisted of 1925 births, 107,529 observations and 73 quality-assured variables. The amount of discarded observations mainly corresponds to observations of non-predictive variables (52.90%) and the impact of the de-duplication process (20.58%) with respect to the total input data. Seven out of thirteen procedures achieved 100% of valid births, observations and variables. Moreover, 89% of births and ~98% of observations were consistent according to the experts׳ criteria. A multidisciplinary approach along with the quantification of DQ has allowed us to construct the first repository about infant feeding in the perinatal period based on EHR population data.


Latissimus dorsi dynamic cardiomyoplasty.
Cardiomyoplasty is a new surgical technique that uses an electrostimulated skeletal muscle to reinforce or partially replace the heart muscle. Since the first clinical case performed in 1985, long-term beneficial cardiac effects of latissimus dorsi cardiomyoplasty have been documented in patients with various myocardial diseases (ischemic, neoplastic, dilated cardiomyopathy). In this article we describe the surgical technique that we have developed to biomechanically assist severe and irreversible ventricular failure.


Sterile inflammation and the NLRP3 inflammasome in cardiometabolic disease.
Inflammation plays an important role in the pathophysiology of cardiometabolic diseases. Sterile inflammation, a non-infectious and damage-associated molecular pattern (DAMP)-induced innate response, is now well-established to be closely associated with development and progression of cardiometabolic diseases. The NOD-like receptor (NLR) family pyrin domain-containing 3 (NLRP3) inflammasome is well-established as a major player in sterile inflammatory responses. It is a multimeric cytosolic protein complex which regulates the activation of caspase-1 and subsequently promotes cleavage and release of interleukin (IL)-1 family cytokines, which have a deleterious impact on the development of cardiometabolic diseases. Therefore, targeting NLRP3 itself or the downstream consequences of NLRP3 activation represent excellent potential therapeutic targets in inflammatory cardiometabolic diseases. Here, we review our current understanding of the role which NLRP3 inflammasome regulation plays in cardiometabolic diseases such as obesity, diabetes, non-alcoholic steatohepatitis (NASH), atherosclerosis, ischemic heart disease and cardiomyopathy. Finally, we highlight the potential of targeting NLPR3 or related signaling molecules as a therapeutic approach.


Application of carbon fiber composite minielectrodes for measurement of kinetic constants of nitric oxide decay in solution.
Carbon fiber microelectrodes and carbon fiber composite minielectrodes (CFM/CFCM) have been generally used for measurements of nitric oxide (NO) concentration in chemical and biological systems. The response time of a CFM/CFCM is usually from milliseconds to seconds depending on the electrode size, the thickness of coating layers on the electrode, and NO diffusion coefficients of the coating layers. As a result, the time course of recoded current changes (I-t curves) by the CFM/CFCM may be different from the actual time course of NO concentration changes (c-t curves) if the half-life of NO decay is close to or shorter than the response time of the electrode used. This adds complexity to the process for determining rate constants of NO decay kinetics from the recorded current curves (I-t curves). By computer simulations based on a mathematical model, an approximation method was developed for determining rate constants of NO decay from the recorded current curves. This method was first tested and valuated using a commercial CFCM in several simple reaction systems with known rate constants. The response time of the CFCM was measured as 4.7±0.7 s (n=5). The determined rate constants of NO volatilization and NO autoxidation in our measurement system at 37 °C are (1.9±0.1)×10(-3) s(-1) (n=4) and (2.0±0.3)×10(3) M(-1) s(-1) (n=7), which are close to the reported rate constants. The method was then applied to determine the rate of NO decay in blood samples from control and smoking exposed mice. It was observed that the NO decay rate in the smoking group is >20% higher than that in control group, and the increased NO decay rate in the smoking group was reversed by 10 μM diphenyleneiodonium chloride (DPI), an inhibitor of flavin enzymes such as leukocyte NADPH oxidase.


A case series of flow-through free anterolateral thigh flap to augment the vascularity of ischaemic limbs with soft tissue defect.
In a world of fast moving vehicles, heavy machinery and industries crush injury to limbs with vascular compromise and soft tissue defect is common. The traditional treatment is a 2 step one dealing with vascular repair and soft tissue cover separately, in the same operation. We report a series of single step vascular repair and soft tissue cover with flow through anterolateral thigh flap (ALT) flap for limb salvage. Ten patients with soft tissue defect and vascular injury were included in this study. A two team approach was used to minimise operative time, team one prepared the vessels and team 2 harvested the flap. Of the ten patients operated (8 males), eight flaps were done for upper limb and two for lower limb salvage. Six anastomosis were done with ulnar vessels, two with radial and two with posterior tibial vessels. Nine extremities could be salvaged while one patient developed progressive thrombosis leading to amputation. The ALT flow-through flap is a versatile single step procedure that can be used to salvage an ischemic limb with soft tissue loss avoiding the need for interpositional vein graft.


Comparative Safety and Effectiveness of Biologic Therapy for Crohn's Disease: A CA-IBD Cohort Study.
We compared the safety and effectiveness of tumor necrosis factor α (TNF-α) antagonists vs vedolizumab vs ustekinumab in patients with Crohn's disease (CD) in a multicenter cohort (CA-IBD). We created an electronic health record-based cohort of adult patients with CD who were initiating a new biologic agent (TNF-α antagonists, ustekinumab, vedolizumab) from 5 health systems in California between 2010 and 2017. We compared the risk of serious infections (safety) and all-cause hospitalization and inflammatory bowel disease-related surgery (effectiveness) between different biologic classes using propensity score (PS) matching. As compared with TNF-α antagonists (n = 1030), 2:1 PS-matched, ustekinumab-treated patients with CD (n = 515) experienced a lower risk of serious infections (hazard ratio [HR], 0.36; 95% CI, 0.20-0.64), without any difference in the risk of hospitalization (HR, 0.99; 95% CI, 0.89-1.21) or surgery (HR, 1.08; 95% CI, 0.69-1.70). Compared with vedolizumab (n = 221), 1:1 PS-matched, ustekinumab-treated patients with CD (n = 221) experienced a lower risk of serious infections (HR, 0.20; 95% CI, 0.07-0.60), without significant differences in risk of hospitalization (HR, 0.76; 95% CI, 0.54-1.07) or surgery (HR, 1.42; 95% CI, 0.54-3.72). Compared with TNF-α antagonists (n = 442), 2:1 PS-matched, vedolizumab-treated patients with CD (n = 221) had a similar risk of serious infections (HR, 1.53; 95% CI, 0.84-2.78), hospitalization (HR, 1.32; 95% CI, 0.98-1.77), and surgery (HR, 0.63; 95% CI, 0.27-1.47). High comorbidity burden, concomitant opiate use, and prior hospitalization were associated with serious infections and hospitalization in biologic-treated patients with CD. In a multicenter cohort of biologic-treated patients with CD, ustekinumab was associated with a lower risk of serious infections compared with TNF-α antagonists and vedolizumab, without any differences in risk of hospitalization or surgery. The risk of serious infections was similar for TNF-α antagonists vs vedolizumab.


Organic-inorganic hybrid gels for the selective absorption of oils from water.
Organic-inorganic hybrid gels were synthesized by the condensation of a linear aliphatic diol (1,8-octanediol) and altering the chain length of the alkyltriethoxysilanes (from ethyltriethoxysilane to hexadecyltrimethoxysilane) through a bulk polymerization process without using any initiator, activator, catalyst, or solvent for the selective removal of oils from water. Fourier transform infrared spectroscopy (FTIR) and solid-state (13)C and (29)Si cross-polarization magic-angle spinning nuclear magnetic resonance (CPMAS NMR) were used for the structural analysis of hybrid gels. Thermal properties of the hybrid gels were determined by thermogravimetric analysis (TGA). Oil absorbency of organic-inorganic hybrid gels was determined by oil absorption tests. The results showed that hybrid gels have high and fast absorption capacities and excellent reusability. Good selectivity, high thermal stability, low density, and excellent recyclability for the oil removal give the material potential applications.


[Three-step column chromatographic method for separation and purification of thrombin-like enzyme from venom of Agkistrodon halys pallas].
A three-step column chromatographic method, utilizing CM Sepharose CL-6B, DEAE Sepharose Fast Flow and Sephadex G-75 in sequence, for the separation and purification of thrombin-like enzyme from crude venom of Agkistrodon halys pallas was developed. Based on the separation results on lab-scale chromatographic column, this isolation and purification process was amplified according to the diameter of the chromatographic columns used in labscale. It was found that when the amount of the sample loaded on the large-scale columns was 25 times as that loaded on the lab-scale columns, the quality level of the thrombin-like enzyme obtained with the large-scale columns was almost the same as that obtained with lab-scale columns. The relative molecular mass of thrombin-like enzyme obtained was about 33500, and its purity was about 96%. When using this method to isolate thrombin-like enzyme from crude venom of Agkistrodon halys pallas, the total protein yield and total activity yield of thrombin-like enzyme were about 0.3% and 64% respectively, and specific activity was more than 2000 U/mg.


The amygdalo-brainstem pathway: selective innervation of dopaminergic, noradrenergic and adrenergic cells in the rat.
The present study investigated the organization and distribution of amygdaloid axons within the various brainstem dopaminergic, noradrenergic and adrenergic cell groups. This was accomplished via Phaseolus vulgaris leucoagglutinin lectin (PHA-L) anterograde tracing technique combined with glucose-oxidase immunocytochemistry to catecholamine markers (i.e. tyrosine hydroxylase, dopamine beta-hydroxylase, and phenylethanolamine N-methyltransferase). Injections of PHA-L within the medial part of the central amygdaloid nucleus resulted in axonal labeling within most catecholamine containing cell groups within the brainstem. The most heavily innervated catecholaminergic groups were the A9 (lateral) cells of the substantia nigra, the A8 dopaminergic cells of the retrorubral field and the C2 adrenergic cells of nucleus of the solitary tract. Amygdaloid terminals frequently contacted cells within these regions. A moderate amount of amygdaloid terminals were located within the rostral A6 (locus coeruleus) and A2 (nucleus of the solitary tract) groups. Amygdaloid terminal contacts were apparent on the majority of the rostral A6 and A2 neurons. Light or no amygdaloid terminal labeling was observed within the other brainstem catecholaminergic cell groups. Thus, the amygdala mainly innervates the A8 and lateral A9 dopaminergic cells of midbrain, rostral locus coeruleus (A6) noradrenergic neurons and the adrenergic (C2) and noradrenergic (A2) cells within the nucleus of the solitary tract. Selective innervation of these brainstem catecholaminergic systems may be important for integration of amygdaloid-mediated defensive and stress-induced behaviors.


Clinical study of autonomic dysfunction in patients with autoimmune encephalitis.
Autoimmune encephalitis (AE) is a collective name, covering an emerging spectrum of autoimmune-mediated neurological diseases related to antibodies and synaptic or intracellular proteins. Anti-NMDAR, anti-LGI1, and anti-GABABR are three types of neuronal cell surface antibodies. Autonomic dysfunction represents a frequently occurring clinical manifestation. This observational study purposes to investigate comparisons between two groups with or without autonomic dysfunction and detect the autonomic dysfunction and other indexes in anti-NMDAR, anti-LGI1, and anti-GABABR cohorts. Patients with anti-NMDAR, anti-LGI1 and anti-GABABR encephalitis were recruited from the May 2017 to the April 2022. The following information was recorded: age, age at onset, tumor presence, gender, prodromal symptoms, clinical manifestations, cranial magnetic resonance imaging, cerebrospinal fluid and blood examinations, and immunotherapy. There were totally 161 patients enrolled in this study. Among these participants, 104 individuals (64.6%) presented autonomic dysfunction and the remaining 57 (35.4%) were free of autonomic dysfunction. Sinus tachycardia was the most common autonomic dysfunction, followed by pollakiuria/uroclepsia, feverscence, central hypoventilation, sinus bradycardia, constipation, uroschesis, hyperhidrosis, hypersalivation, hypotension, and early satiety/emesis. Compared to patients without autonomic dysfunction, those with autonomic dysfunction had a higher incidence of central hypoventilation and ICU admissions. Meanwhile, in both groups with or without autonomic dysfunction, meatal behavior disorder, cognitive impairment, and epileptic seizure were three most common clinical manifestations. There were no significant differences in cranial magnetic resonance imaging (MRI), cerebrospinal fluid (CSF) examination, antibody titers and number of immunotherapy types. Further analysis of AE mediated by distinct neuronal surface antibodies demonstrated that there were 85 anti-NMDAR, 56 anti-LGI1, and 20 anti-GABABR encephalitis patients. The significant differences between these three cohorts appeared in age, tumor presence, fervescence presence and antibody titers. This study demonstrated the comparisons between autonomic dysfunction group and autonomic dysfunction-free group and provided insights into better diagnosis and treatment.


Anthelmintics and control.
Anthelmintic control of Ostertagia ostertagi in cattle presents some special problems because the arrested larval stage (hypobiotic EL4) tolerates all of the older anthelmintics. The only anthelmintics on the North American market that are effective against this stage as well as the adult and developing stages are ivermectin and fenbendazole. In addition to these, the newer broad-spectrum benzimidazoles and probenzimidazoles, albendazole, febantel, netobimin and oxfendazole, are effective and available in other countries. These anthelmintics can be used for prophylaxis of Type II ostertagiasis. The older anthelmintics, levamisole, morantel tartrate, thiabendazole, coumaphos, haloxon and phenothiazine, are effective against the adults and to some extent the developing stages of O. ostertagi so that they can be used to treat Type I ostertagiasis. They can also be used to prevent incoming larvae from establishing if they are administered continuously over a long period of time. This is possible with long term in-feed administration or by the use of the morantel slow release bolus. In cooler temperature regions, where cattle are housed over the winter, this bolus is given at turnout to remove any parasites in the animals and to kill incoming larvae for 60-90 days. This can prevent the build up of significant infective larvae on pasture so that very few arrested L4 larvae accumulate in the summer/autumn, effectively preventing Type II ostertagiasis from occurring later. The use of ivermectin and the newer benzimidazoles in intermittent or slow release devices should prove highly effective in the control of Type I and II ostertagiasis, as well as of subclinical ostertagiasis. To achieve maximum economic benefit, the use of anthelmintics should be based on sound epidemiological considerations, so that stock are not rapidly reinfected after treatment.


Quantifying stretching and rearrangement in epithelial sheet migration.
Although understanding the collective migration of cells, such as that seen in epithelial sheets, is essential for understanding diseases such as metastatic cancer, this motion is not yet as well characterized as individual cell migration. Here we adapt quantitative metrics used to characterize the flow and deformation of soft matter to contrast different types of motion within a migrating sheet of cells. Using a Finite-Time Lyapunov Exponent (FTLE) analysis, we find that - in spite of large fluctuations - the flow field of an epithelial cell sheet is not chaotic. Stretching of a sheet of cells (i.e., positive FTLE) is localized at the leading edge of migration and increases when the cells are more highly stimulated. By decomposing the motion of the cells into affine and non-affine components using the metric D2min , we quantify local plastic rearrangements and describe the motion of a group of cells in a novel way. We find an increase in plastic rearrangements with increasing cell densities, whereas inanimate systems tend to exhibit less non-affine rearrangements with increasing density.


Porous and highly responsive cross-linked β-cyclodextrin based nanomatrices for improvement in drug dissolution and absorption.
Aim of the study was to enhance the solubility of Chlorthalidone by developing beta-cyclodextrin cross-linked hydrophilic nanomatrices. Nine different formulations were fabricated by free radical polymerization technique. All formulations were characterized through different studies. FTIR spectroscopy of unloaded and loaded nanomatrices was processed to determine compatibility of constituents and that of the drug with the system. Surface morphology of the nanomatrices was studied by SEM. The size of the optimized formulation was determined by zeta sizer. Swelling, in-vitro release and solubility studies were carried out in different media and results of in-vitro release profiles of nanomatrices and commercially available tablet of Chlorthalidone were compared. For determination of biocompatibility, toxicity studies were proclaimed in rabbits. Main peaks of corresponding functional groups of individual constituents and that of drug were depicted in FTIR spectra of unloaded and loaded nanomatrices. Porous and fluffy structure was visualized through SEM images. Particle size of the optimized formulation was in the range of 175 ± 5.27 nm. Percent loading of optimized formulation showed the best result. Comparing the in-vitro drug release profiles of nanomatrices and commercially available tablet, the results of the synthesized nanomatrices were quite satisfactory. Solubility profiles were also high as compared to the drug alone. Moreover, toxicity studies confirmed that nanomatrices were biocompatible and no sign of any toxic effect was found. We concluded that our developed nanomatrices had successfully enhanced the solubility of Chlorthalidone and can also be used for other poorly aqueous soluble drugs.


Mirror image atrial dilatation in adult patients with atrial fibrillation and congenital heart disease.
Atrial fibrillation (AF) is largely regarded to be initiated from left atrial (LA) dilatation, with subsequent dilatation of the right atrium (RA) in those who progress to chronic AF. We hypothesized that in adult patients with right-sided congenital heart disease (CHD) and AF, RA dilatation will predominate with subsequent dilatation of the left atrium, as a mirror image. Adult patients with diagnosis of right-sided, ASD or left-sided CHD who had undergone an echocardiographic study and electrocardiographic recording in 2007 were included. RA and LA area were measured from the apical view. AF was diagnosed from a 12-lead electrocardiogram or Holter recording. A multivariate logistic regression model was used to identify predictors of AF and linear regression models were performed to measure relationship between RA and LA area and AF. A total of 291 patients were included in the study. Multivariate analysis showed that age (p=0.0001), RA (p=0.025) and LA area (p=0.0016) were significantly related to AF. In patients with pure left-sided pathologies, there was progressive and predominant LA dilatation that paralleled the development of AF from none to paroxysmal to chronic AF. In patients with pure right-sided pathologies, there was a mirror image of progressive and predominant RA dilatation with the development of AF. We observed a mirror image atrial dilatation in patients with right sided disease and AF. This may provide novel mechanistic insight as to the origin of AF in these patients and deserves further studying in the form of targeted electrophysiological studies.


Riluzole produces distinct anxiolytic-like effects in rats without the adverse effects associated with benzodiazepines.
In this study, we investigated the anxiolytic-like effect of riluzole using three different innate anxiety models in rats. In the elevated plus-maze test, riluzole significantly increased the time spent in, and entries into, the open arm after 60 min administration. This finding was supported by results obtained from light/dark and open-field tests. The magnitude of the anxiolytic-like effects of riluzole in each of the behavioral models was similar to those produced by a benzodiazepine, diazepam, suggesting that riluzole has a robust anxiolytic-like activity in rats. To clarify the involvement of sodium channels in this anxiolytic activity, we examined the effect of a co-administered sodium channel activator, veratrine. The anxiolytic-like action of riluzole was diminished by veratrine in the elevated plus-maze, light/dark and open-field tests. Based on these results, it is suggested that the anxiolytic mechanism of riluzole is clearly distinct from that of diazepam. In addition, to examine whether riluzole directly and non-selectively affected the GABA(A)-benzodiazepine receptor complex, we performed three behavioral tests (footprint analysis, Y-maze test and the ethanol-induced sleeping time test) that are closely related to the GABA(A)-benzodiazepine pathways. In contrast to diazepam, riluzole produced no significant effects in these tests. Here, we provide the first report demonstrating that riluzole produces distinct anxiolytic-like effects in rats without the adverse effects associated with benzodiazepines.


Four-Color STED Super-Resolution RNA Fluorescent In Situ Hybridization and Immunocytochemistry to Visualize Mitochondrial mRNAs in Context with Mitochondrial Ribosomes.
High-resolution imaging has enabled scientists to explore the mitochondrial network at remarkable resolution. This has been exploited to help increase our knowledge of how mitochondrial gene expression is compartmentalized in cultured cells. Here, we provide detailed methodology to simultaneously visualize up to four components including mtDNA-encoded transcripts, submitochondrial marker proteins, mitoribosomal subunits, or core members of the translational apparatus using STED super-resolution nanoscopy.


Avian monocytic leukemia cells release fibroblast growth factor: implications to associated myelofibrosis.
The marrow of chicks with leukemia induced by avian 'myeloblastosis' virus (AMV) exhibited a 5-10-fold increase in the number of fibroblast colony-forming cells (CFU-F). The increased CFU-F correlated with a mild fibrosis which can be seen in the marrow of these animals. Fibroblast proliferation likely was not simply due to the presence of leukemic cells because addition of formaldehyde-fixed peripheral leukemic cells failed to initiate CFU-F growth. Conditioned medium (CM) from day-4 cultures of peripheral leukemic cells was markedly stimulatory to CFU-F growth. The stimulatory activity was not due to virus released from the leukemic cells as, (1) removal of virus by pelleting had no effect on the CM activity and (2) direct inoculation of CFU-F cultures with virus failed to stimulate CFU-F growth. Normal avian marrow macrophage monolayers also released high levels of a fibroblast growth factor and both the macrophage-derived and leukemic cell-derived factors were heat-labile (65 degrees, 30 min).


Mice overexpressing human uncoupling protein-3 in skeletal muscle are hyperphagic and lean.
Uncoupling protein-3 (UCP-3) is a recently identified member of the mitochondrial transporter superfamily that is expressed predominantly in skeletal muscle. However, its close relative UCP-1 is expressed exclusively in brown adipose tissue, a tissue whose main function is fat combustion and thermogenesis. Studies on the expression of UCP-3 in animals and humans in different physiological situations support a role for UCP-3 in energy balance and lipid metabolism. However, direct evidence for these roles is lacking. Here we describe the creation of transgenic mice that overexpress human UCP-3 in skeletal muscle. These mice are hyperphagic but weigh less than their wild-type littermates. Magnetic resonance imaging shows a striking reduction in adipose tissue mass. The mice also exhibit lower fasting plasma glucose and insulin levels and an increased glucose clearance rate. This provides evidence that skeletal muscle UCP-3 has the potential to influence metabolic rate and glucose homeostasis in the whole animal.


Diacylglycerol acyltransferase in maturing safflower seeds: its influences on the fatty acid composition of triacylglycerol and on the rate of triacylglycerol synthesis.
Diacylglycerol acyltransferase in a particulate preparation from maturing safflower seeds showed no strict selectivity for acyl-CoA when acyl-CoA substrates were administered as mixtures. This suggested that the fatty acid composition of position 3 in safflower triacyl-sn-glycerol exclusively depends on the acyl-CoA composition in the cell. The specific activity of the acylation was approximately 3 nmol/min per mg protein of the preparation under the optimum assay conditions. This low activity and other data appeared to indicate that the diacylglycerol acyltransferase reaction may be the rate-limiting step in triacylglycerol synthesis in vivo.


Adiponectin pathway polymorphisms and risk of breast cancer in African Americans and Hispanics in the Women's Health Initiative.
Adiponectin, a protein secreted by the adipose tissue, is an endogenous insulin sensitizer with circulating levels that are decreased in obese and diabetic subjects. Recently, circulating levels of adiponectin have been correlated with breast cancer risk. Our previous work showed that polymorphisms of the adiponectin pathway are associated with breast cancer risk. We conducted the first study of adiponectin pathways in African Americans and Hispanics in the Women's Health Initiative SNP Health Association Resource cohort of 3,642 self-identified Hispanic women and 8,515 self-identified African American women who provided consent for DNA analysis. Single nucleotide polymorphisms (SNPs) from three genes were included in this analysis: ADIPOQ, ADIPOR1, and ADIPOR2. The genome-wide human SNP array 6.0 (909,622 SNPs) ( www.affymetrix.com ) was used. We found that rs1501299, a functional SNP of ADIPOQ that we previously reported was associated with breast cancer risk in a mostly Caucasian population, was also significantly associated with breast cancer incidence (HR for the GG/TG genotype: 1.23; 95 % CI 1.059-1.43) in African American women. We did not find any other SNPs in these genes to be associated with breast cancer incidence. This is the first study assessing the role of adiponectin pathway SNPs in breast cancer risk in African Americans and Hispanics. RS1501299 is significantly associated with breast cancer risk in African American women. As the rates of obesity and diabetes increase in African Americans and Hispanics, adiponectin and its functional SNPs may aid in breast cancer risk assessment.


Gastric fluid volume and pH in elective inpatients. Part I: Coffee or orange juice versus overnight fast.
In order to assess the effect of preoperative oral fluids, with and without ranitidine, on gastric fluid volume and pH, 300 elective surgical inpatients, ASA physical status I or II, were randomly assigned to one of six groups. The three groups that received placebo are discussed in this paper, Part I, and the three that received ranitidine in Part II. Between two and three hours before the scheduled time of surgery patients received either 150 ml coffee (Group 1), or 150 ml orange juice (Group 2), while the control group continued their overnight fast (Group 3). No opiate or belladonna premedication was given. Immediately following induction of anaesthesia the residual gastric fluid was obtained by suction on a nasogastric tube and its volume and pH measured. Residual gastric fluid volumes showed no statistically significant differences among the groups (Group 1: 24.5 +/- 21.6 ml; Group 2: 23.7 +/- 18.4 ml; Group 3: 23.2 +/- 17.3 ml; p greater than 0.1). Values for pH among the groups were also similar (Group 1: 2.18 +/- 1.58; Group 2: 1.95 +/- 1.24; Group 3: 1.95 +/- 1.62; p greater than 0.1).


Complete diastereodivergence in asymmetric 1,6-addition reactions enabled by minimal modification of a chiral catalyst.
Catalytic systems that allow selective generation of any diastereomer of a reaction product bearing multiple stereocentres through minimal modification of a single catalyst scaffold remain elusive, particularly for carbon-carbon bond formations requiring simultaneous control of multiple selectivity factors. Here, we report a catalyst-directed pinpoint inversion of diastereochemical preference in the 1,6-addition of azlactones to δ-aryl dienyl carbonyl compounds with full control over other selectivities preserved. This rigorous diastereodivergence is enabled by the slight structural adjustment of a chiral iminophosphorane catalyst, providing access to all the stereoisomers with high regio-, distereo- and enantioselectivity. The utility of this method is demonstrated in the facile stereodivergent preparation of densely functionalized proline derivatives. The experimental and computational elucidation of the origin of the diastereodivergence is also reported.


Critical differences in HIV-1 and HIV-2 protease specificity for clinical inhibitors.
Clinical inhibitor amprenavir (APV) is less effective on HIV-2 protease (PR₂) than on HIV-1 protease (PR₁). We solved the crystal structure of PR₂ with APV at 1.5 Å resolution to identify structural changes associated with the lowered inhibition. Furthermore, we analyzed the PR₁ mutant (PR(1M) ) with substitutions V32I, I47V, and V82I that mimic the inhibitor binding site of PR₂. PR(1M) more closely resembled PR₂ than PR₁ in catalytic efficiency on four substrate peptides and inhibition by APV, whereas few differences were seen for two other substrates and inhibition by saquinavir (SQV) and darunavir (DRV). High resolution crystal structures of PR(1M) with APV, DRV, and SQV were compared with available PR₁ and PR₂ complexes. Val/Ile32 and Ile/Val47 showed compensating interactions with SQV in PR(1M) and PR₁, however, Ile82 interacted with a second SQV bound in an extension of the active site cavity of PR(1M). Residues 32 and 82 maintained similar interactions with DRV and APV in all the enzymes, whereas Val47 and Ile47 had opposing effects in the two subunits. Significantly diminished interactions were seen for the aniline of APV bound in PR₁ (M) and PR₂ relative to the strong hydrogen bonds observed in PR₁, consistent with 15- and 19-fold weaker inhibition, respectively. Overall, PR(1M) partially replicates the specificity of PR₂ and gives insight into drug resistant mutations at residues 32, 47, and 82. Moreover, this analysis provides a structural explanation for the weaker antiviral effects of APV on HIV-2.


Resistance of maize landraces to the maize weevil Sitophilus zeamais Motsch. (Coleoptera: Curculionidae).
The maize weevil Sitophilus zeamais Motsch. is an important pest of maize that attacks the grain both in the field and during storage. The damage caused by the maize weevil S. zeamais on maize landraces, Amarelo Antigo, Asteca, Caiano, Carioca, and Ferrinho, was evaluated by no-choice tests under laboratory conditions. The commercial varieties Sol da Manhã, BR 106, BR 451, and the synthetics PC 0203 and PC 9903 were evaluated for comparisons with the maize landraces. The parameters evaluated were susceptibility index, number of weevil progeny, development time, weevil progeny dry weight, and grain dry weight loss. The landraces were more susceptible to the maize weevil as compared to the commercial varieties. Based on the cluster analysis, two groups of susceptibility to the maize weevil were observed: one of more susceptible populations formed by local landraces and BR 451, and another less susceptible, with commercial varieties, synthetics, and the landrace Amarelo.


Thermal constraints on foraging in adult european starlings.
The operative temperature of the environment was estimated for starlings using hollow, unheated taxidermic mounts. On average, adults foraging in full sun were characterized by shorter foraging bouts than those adults foraging in full shade. Simultaneous observations of air temperature, operative temperature, and the foraging duration of adults indicated that air temperature was a poor predictor of the maximum length of a foraging bout. The operative temperature of the environment was not correlated to the maximum and mean length of foraging bouts for temperatures below 31.5°C, but was negatively related to maximum and mean foraging duration for values above 31.5°C. I also found that foraging adults experiencing high thermal loads (T e≥31.5°C) were less likely to return to the nest with food. These results raise the possibility that adults may be limited in their utilization of foraging sites due to an inability to cope with heat load, and that much of the loafing behavior observed for breeding birds may be behavior to avoid heat stress. The implication for seasonal variation of clutch size is discussed.


Prevalence of gastroesophageal reflux disease in children with extraesophageal manifestations using combined-video, multichannel intraluminal impedance-pH study.
Gastroesophageal reflux disease (GERD) might be either a cause or comorbidity in children with extraesophageal problems especially as refractory respiratory symptoms, without any best methods or criterion for diagnosing it in children. To evaluate the prevalence of extraesophageal GERD using conventional and combined-video, multichannel intraluminal impedance-pH (MII-pH), and to propose novel diagnostic parameters. The study was conducted among children suspected of extraesophageal GERD at King Chulalongkorn Memorial Hospital between 2019 and 2022. The children underwent conventional and/or combined-video MII-pH. The potential parameters were assessed and receiver operating characteristic was used for the significant parameters. Of 51 patients (52.9% males), aged 2.24 years were recruited. The common problems were cough, recurrent pneumonia, and hypersecretion. Using MII-pH, 35.3% of the children were diagnosed with GERD by reflux index (31.4%), total reflux events (3.9%), and symptom indices (9.8%) with higher symptom recorded in the GERD group (94 vs 171, P = 0.033). In the video monitoring group (n = 17), there were more symptoms recorded (120 vs 220, P = 0.062) and more GERD (11.8% vs 29.4%, P = 0.398) by symptom indices. Longest reflux time and mean nocturnal baseline impedance were significant parameters for diagnosis with receiver operating characteristic areas of 0.907 (P = 0.001) and 0.726 (P = 0.014). The prevalence of extraesophageal GERD in children was not high as expected. The diagnostic yield of symptom indices increased using video monitoring. Long reflux time and mean nocturnal baseline impedance are novel parameters that should be integrated into the GERD diagnostic criteria in children.


[Altered expression of the HSD17B4 gene in esophageal squamous cell carcinoma and loss of heterozygosity analysis].
To investigate the alteration of the gene HSD17B4 in esophageal squamous cell carcinoma and its potential significance. The mRNA expression and loss of heterozygosity (LOH) of HSD17B4 in 40 primary esophageal tumors were detected by reverse transcriptase-polymerase chain reaction (RT-PCR) and microsatellite analysis with the intragenic marker D5S1384 of the gene. The frequencies of allelic loss of D5S1384 and the rate of down-regulation of gene HSD17B4 were 46.2% and 62.5%, respectively. HSD17B4 may be a candidate tumor suppressor gene associated with esophageal squamous cell carcinoma.


Interferon-γ Production by Peripheral Lymphocytes Predicts Survival of Tumor-Bearing Mice Receiving Dual PD-1/CTLA-4 Blockade.
Immune checkpoint inhibitors are transforming the way cancer is treated. However, these therapies do not benefit all patients and frequently cause significant immune-related adverse events. Biomarkers that identify patients with a favorable early response to therapy are essential for guiding treatment decisions and improving patient outcomes. In this report of our study, we present evidence that shortly after administration of dual PD-1/CTLA-4 blockade, the proinflammatory capacity of peripheral lymphocytes is predictive of tumor progression and survival outcomes in multiple murine models. Specifically, we observed that the quantity of interferon-γ (IFNγ) produced by peripheral lymphocytes in response to CD3/CD28 stimulation was robustly correlated with subsequent survival outcomes. In the tumor models and early time points assessed in this study, this relationship was considerably more predictive than a host of other potential biomarkers, several of which have been previously reported. Overall, these findings suggest that measuring the capacity of peripheral lymphocytes to produce IFNγ may help identify which patients are benefitting from combination anti-PD-1/anti-CTLA-4 immunotherapy. Cancer Immunol Res; 4(8); 650-7. ©2016 AACR.


SUNCT syndrome: A cohort of 15 Portuguese patients.
In this paper, we describe a cohort of patients with short-lasting unilateral neuralgiform headache attacks with conjunctival injection and tearing (SUNCT), a rare trigeminal autonomic headache, managed in the outpatient clinic of a tertiary hospital. Patients were identified through review of individual records between January 1, 2008 and June 30, 2014. Fifteen patients were identified (eight males:seven females), with mean age at onset of 49.7 years, mean number of attacks per day of 7.5 and mean attack duration of 54.6 seconds. Pain was mostly orbital, periorbital or temporal. Cranial autonomic signs/symptoms were universally present; one patient reported ipsilateral epistaxis. Two symptomatic cases were identified and treated surgically. Most patients responded to lamotrigine, one to topiramate and another to eslicarbazepine. Our case series is among the largest reported, with findings similar to others already published, but the first to report epistaxis during SUNCT attack and response to eslicarbazepine.


[Endoscopic endonasal surgery for tumors of petroclival region and infratemporal fossa].
Nasal endoscope has been used increasingly during skull base surgery. However, most of endoscopic surgery limited to the repair of cerebrospinal rhinorrhea, decompression of traumatic optic nerve, hypophysectomy, etc. This study was undertaken to determine whether endoscopic endonasal approach was safe and effective for the resection of tumors located in petroclival region and infratemporal fossa. Seventeen patients with tumors of petroclival region and infratemporal fossa treated by endoscopic endonasal surgery between January 2002 and February 2005 were studied prospectively. The operative technique was described in detail. There were 5 chordoma, 1 esthesioneuroblastoma, 1 chondrosarcoma, 1 lymphoma, 1 craniopharyngioma, 1 hemangioblastoma, 4 meningioma, 1 schwannoma, and 2 metastatic carcinoma. 3 patients were selected for neuronavigation-aided endoscopic endonasal surgery. Total tumor removal was obtained in 15 cases, subtotal removal in 2 case. With follow-up of 5 to 43 months, 1 case with chordoma was recurrent 5 months later postoperatively and underwent reoperation subsequently. The other cases with benign tumors were no recurrence. All of 5 cases with malignant tumors followed up for longer than 2 years were no recurrence and death. The complications included subarachnoid hemorrhage in 1 patient, transient cerebrospinal leakage in 2 cases. The endoscopic endonasal surgery provides satisfied treatment for selected tumors of petroclival region and infratemporal fossa. This approach promises a simple and rapid access to petroclival region and infratemporal fossa. It is a safe, minimally invasive and efficient procedure. Using neuronavigation system, it is helpful to determining anatomical landmark and removing the tumor completely and securely.


Optimization of a GC/MS procedure that uses parallel factor analysis for the determination of bisphenols and their diglycidyl ethers after migration from polycarbonate tableware.
Bisphenol A (BPA), bisphenol F (BPF) and their corresponding diglycidyl ethers (BADGE and BFDGE) are simultaneously determined using a programmed-temperature vaporizer-gas chromatography/mass spectrometry (PTV-GC/MS) system. BPA is used in the production of polycarbonate (PC), whereas BADGE and BFDGE are for manufacturing epoxy resins. Several food alerts caused by the migration of this kind of substances from contact food materials have led to the harmonization of the European legislation in Commission Regulation (EU) No. 10/2011, in force from 14 January 2011. In consequence, the use of BPA has been prohibited in the manufacture of plastic infant feeding bottles from 1 May 2011 and from 1 June 2011 regarding the placing on the market and importation into the European Union. Recently, the French Parliament has decreed that the presence of BPA in any food containers will be banned. Similarly, the use and/or presence of BFDGE are not allowed. In this work, a GC/MS method has been developed for the simultaneous determination of BPF, BPA, BFDGE and BADGE. For each one of the I samples that are analyzed, the abundance of J characteristic m/z ratios is recorded at K times around the retention time of each peak, so a data tensor of dimension I×J×K is obtained for every analyte. The decomposition of this tensor by means of parallel factor analysis (PARAFAC) enables to: (a) identify unequivocally each analyte according to the maximum permitted tolerances for relative ion intensities, and (b) quantify each analyte, even in the presence of coeluents. This identification, based on the mass spectrum and the retention time, guarantees the specificity of the analysis. This specificity could fail if the total ion chromatogram (TIC) is considered when there is poor resolution between some peaks or whether interferents coelute. With the aim of studying the effect of shortening the time of the analysis on the quality of the determinations while maintaining the specificity of the identifications, two of the heating ramps in the oven temperature program are changed according to a two-level factorial design. Each analyte is identified by means of a PARAFAC decomposition of a data tensor obtained from several concentration levels, in such a way that five figures of merit are calculated for each experiment of the design. The analysis of these figures of merit for the 16 objects (4 compounds×4 heating ramps) using principal component analysis (PCA) shows that the shortest temperature program should be considered, since this is the one the best figures of merit for BPA and BFDGE (both banned) are achieved with. At these conditions and with probabilities of false positive and false negative fixed at 0.05, values of detection capability (CCβ) between 2.65 and 4.71 μg L(-1) when acetonitrile is the injection solvent, and between 1.97 and 5.53 μg L(-1) when acetone, are obtained. This GC/MS method has been applied to the simultaneous determination of BPF, BPA, BFDGE and BADGE in food simulant D1 (ethanol-H2O, 1:1 v/v), which had been previously in contact with PC tableware for 24h at 70 °C and then pretreated by a solid-phase extraction (SPE) step. The migration of BPA from the new PC containers analyzed is confirmed, and values between 104.67 and 181.46 μg L(-1) (0.73 and 1.27 μg L(-1) after correction) of BPA have been estimated. None of the results obtained exceeds the specific migration limit of 600 μg L(-1) established by law for BPA in plastic food materials different from PC infant feeding bottles. Severe problems of coelution of interferents have been overcome using PARAFAC decompositions in the analysis of these food simulant samples.


Cervical pregnancy successfully treated with chemotherapy.
A case of cervical pregnancy in a 16-year-old woman is reported. Non-surgical management with actinomycin-D, methotrexate and cyclophosphamide chemotherapy resolved the pregnancy without significant morbidity. Subsequent pregnancy occurred with premature birth of viable normal infant.


A new method for fast blood cell counting and partial differentiation by flow cytometry.
A new blood counting method by flow cytometry is described which determines absolute counts and relative proportions of erythrocytes, reticulocytes, thrombocytes, lymphocytes and granulocytes from one sample of saline diluted human or animal blood. Staining time is 2 to 5 min and measuring time between 1 and 2 additional minutes. Measured simultaneously are the electrical cell volume, the green and optionally also the red fluorescence of the transmembrane potential sensitive dye 3,3-dihexyloxacarbocyanine DiOC6(3) and the RNA/DNA stain acridine orange (AO). Work is under way to fully automate staining, measurement and data evaluation. The use of stains by which blood cell counting and biochemical analysis can be combined offers new possibilities for routine blood cell counting without requirement for additional time. The potential of such stains is that pathologic cell conditions which are not, or not yet reflected in the cell count may be earlier detectable by biochemical stains.


Clinical characteristics of open-angle glaucoma progression with peripapillary microvasculature dropout in different locations.
The study attempted to identify clinical characteristics associated with structural progression in open-angle glaucoma (OAG) in the presence of MvD in different locations. A total of 181 consecutive OAG eyes (follow-up 7.3 ± 4.0 years), which demonstrated peripapillary choroidal MvD (defined as a focal capillary loss with no visible microvascular network in choroidal layer) on optical coherence tomography (OCT) angiography (OCTA), were divided based on the location of MvD. Structural progression was determined using trend-based analysis of the Guided Progression Analysis software of Cirrus OCT. MvD was identified in the temporal quadrant in 110 eyes (temporal MvD; 60.5 ± 12.6 years), and in the inferior quadrant in 71 eyes (inferior MvD; 60.3 ± 11.1 years). After adjusting for age, average intraocular pressure (IOP) and baseline retinal nerve fibre layer (RNFL) thickness and visual field mean deviation, inferior MvD eyes showed faster rates of thinning in the inferior RNFL (mean (95% CI); -0.833 (-1.298 to -0.367)) compared to temporal MvD eyes (-0.144 (-0.496 to 0.207)) when long-term IOP fluctuation was larger than the median value (1.7 mmHg; P = 0.022). Long-term IOP fluctuations were independently associated with inferior RNFL thinning in eyes with inferior MvD (P = 0.002) but not in eyes with temporal MvD. In OAG eyes, the rates of RNFL and GCIPL thinning were comparable regardless of MvD locations. However, inferior MvD is associated with faster RNFL and GCIPL thinning in the same quadrant when long-term IOP fluctuation is present. Structural progression in the presence of temporal MvD was less associated with IOP fluctuation.


Stirrups to minimize complications of prolonged dorsal lithotomy positioning.
We evaluated the incidence of postoperative complications associated with positioning in 150 patients who underwent radical retropubic prostatectomy or radical cystectomy with urinary diversion in the modified dorsal lithotomy position. English stirrups were used in the first 75 patients, whereas a new type of stirrup modified from a Krauss arm support was used in the second 75 patients. Use of the new stirrups in the second group significantly decreased the postoperative morbidity associated with the dorsal lithotomy position from 13.3 to 2.6 per cent (p less than 0.05).


Epigenetic regulation of metabotropic glutamate 2/3 receptors: Potential role for ultra-resistant schizophrenia?
Schizophrenia is a severe and debilitating psychiatric disorder characterized by early cognitive deficits, emotional and behavioral abnormalities resulted by a dysfunctional gene x environment interaction. Genetic and epigenetic abnormalities in cortical parvalbumin-positive GABAergic interneurons lead to alterations in glutamate-mediated excitatory neurotransmission, synaptic plasticity, and neuronal development. Epigenetic alterations during pregnancy or early phases of postnatal life are associated with schizophrenia vulnerability as well as inflammatory processes which are at the basis of brain pathology. An epigenetic animal model of schizophrenia showed specific changes in promoter DNA methylation activity of genes related to schizophrenia such as reelin, BDNF and GAD67, and altered expression and function of mGlu2/3 receptors in the frontal cortex. Although antipsychotic medications represent the main treatment for schizophrenia and generally show an optimal efficacy profile for positive symptoms and relatively poor efficacy for negative or cognitive symptoms, a considerable percentage of individuals show poor response, do not achieve a complete remission, and approximately 30 % of patients show treatment-resistance. Here, we explore the potential role of epigenetic abnormalities linked to metabotropic glutamate 2/3 receptors changes in expression and function as key molecular factors underlying the difference in response to antipsychotics.


Population-Based Estimates and Predictors of Child and Adolescent Linkage to HIV Care or Death in Western Kenya.
Population-level estimates of linkage to HIV care among children and adolescents (CAs) can facilitate progress toward 95-95-95 goals. This study was conducted in Bunyala, Chulaimbo, and Teso North subcounties, Western Kenya. Linkage to care was defined among CAs diagnosed with HIV through Academic Model Providing Access to Healthcare (AMPATH)'s home-based counseling and testing initiative (HBCT) by merging HBCT and AMPATH Medical Record System data. Using follow-up data from Bunyala, we examined factors associated with linkage or death, using weighted multinomial logistic regression to account for selection bias from double-sampled visits. Based on the estimated model, we imputed the trajectory for each person in 3 subcounties until a simulated linkage or death occurred or until the end of 8 years when an individual was simulated to be censored. Of 720 CAs in the analytic sample, 68% were between 0 and 9 years and 59% were female. Probability of linkage among CAs in the combined 3 subcounties was 48%-49% at 2 years and 64%-78% at 8 years while probability of death was 13% at 2 years and 19% at 8 years. Single or double orphanhood predicted linkage (adjusted odds ratio [aOR]: 2.66, 95% confidence interval [CI]: 1.33 to 5.32) and death (aOR: 9.85 [95% CI: 2.21 to 44.01]). Having a mother known to be HIV-positive also predicted linkage (aOR = 1.94, 95% CI: 0.97 to 3.86) and death (aOR: 14.49, 95% CI: 3.32 to 63.19). HIV testers/counselors should continue to ensure linkage among orphans and CAs with mothers known to be HIV-positive and also to support other CAs to link to HIV care.


1996 Peritoneal Dialysis Core Indicators Study: report on nutritional indicators.
The 1996 Peritoneal Dialysis Core Indicators Study illustrates the conduct of peritoneal dialysis in the United States during 1996. DESIGN AND PATIENT POPULATION: The survey is a medical records audit of 1317 randomly selected adult U.S.A. Medicare patients using peritoneal dialysis during 1996. Abstracted data included basic demographic characteristics, dialysis prescription, delivered dialysis dose, residual renal function, serum albumin, hematocrit, anemia management, and patient status. The survey included 785 patients using continuous ambulatory peritoneal dialysis (CAPD) and 423 using automated peritoneal dialysis (APD) primarily in the form of continuous cycling peritoneal dialysis (CCPD). Except for the prescription mechanics and a greater likelihood that African-Americans would use CAPD, the groups did not differ substantially from one another. Evaluation of patient weight (W), body mass index (BMI), residual renal function, average serum albumin, protein equivalent of nitrogen appearance (nPNA), and dialysis efficiency as weekly fractional urea nitrogen removal (wKt/Vurea) and weekly creatinine clearance (wCrCl) revealed a picture of reasonable dialysis delivery and marginal protein nutrition. Additionally, there was little evidence that "dialysis efficiency," over the range assessed, had a major influence on nutritional status. Despite a tendency toward obesity (body weight = 76.6+/-20.0 kg and BMI = 27+/-7), 47% of patients had an average serum albumin below"normal" (3.5 g/dL by bromcresol green) and 70% had a nPNA below 1.0 g/kg/day. Peritoneal dialysis patients appear to have marginal protein reserves despite surfeit energy stores.


Toxicity and physiological actions of biflavones on potassium current in insect neuronal cells.
Stellera chamaejasme L. is a Chinese traditional herb. It has a long history and many medicinal usages. Biflavones, one of the main active ingredients in S. chamaejasme's roots, possess excellent insecticidal activities both in vivo and in vitro. However, the mechanism of these compounds and its potential molecular targets on insect cell were still not clear. Here the whole cell patch clamp technique was used to investigate whether biflavones affects voltage-gated potassium channels (Kv) on insect neuronal cells (AW1 and WG2). The results confirmed that both the three biflavones: neochamaejasmin A (NCA), neochamaejasmin B (NCB) and isochamaejasmin A (ICM) can significantly inhibit the A-type potassium current (IA) than delayed rectifier potassium current (IK) expressed on insect cells. Moreover, ICM stood out as the strongest inhibition activity on IA with IC50 value of 106.75 μM. Multiple results suggest that the inhibition of potassium current was related to the gating modification of biflavones. ICM produced concentration dependent hyperpolarizing shifts in the voltage dependence of channel steady-state activation and inactivation. Maximal shifts of the ICM-induced V0.5, were -15.1 mV for activation and -6.93 mV for inactivation. ICM also prolonged recovery from inactivation of current. Moreover, the biflavones could inhibited AW1 cell survival in both dose- and time-dependent manners with well correlation of K+ inhibitory activity. Our study showed that biflavones from S. chamaejasme exhibiting significant blocked effects on Kv of AW1 cells and inhibited cell proliferation. These findings may not only show the toxic mechanisms of biflavones on insect cells, but also suggest that Kv channel play an important role in biflavones' mode of action and may be the new targets for designing novel insecticides.


Application of biomarkers for improving risk assessments of chemicals under the Water Framework Directive: a case study.
To answer the requirement of the European Commission's Water Framework Directive (WFD) for biological-effects endpoints to classify the ecological health of aquatic ecosystems, we propose the biomarker response index (BRI). The BRI, based on a suite of biomarkers at different levels of biological response at the individual level, provides an integrated relative measure of the general health status of coastal invertebrates. Using the BRI, the health of mussels (Mytilus edulis) from 10 estuaries classified by the Environment Agency of England and Wales under the WFD was compared. Eight sites were healthier than predicted and two showed a similar health status to that of the predicted point-source pollution risk classification. Results indicate that the BRI offers a potential measure of organism health that can be used in monitoring under the WFD as an additional aid to reduce uncertainty in defining risk classification and to provide better evidence of existing impact.


Hepatitis C virus core protein expression leads to biphasic regulation of the p21 cdk inhibitor and modulation of hepatocyte cell cycle.
Hepatitis C virus (HCV) Core protein is implicated in viral pathogenesis by the modulation of hepatocyte gene expression and function. To determine the effect of Core protein on the cell-cycle control of hepatocytes, a HepG2 cell line containing a Flag-tagged Core under the control of an inducible promoter was generated. Initial Core protein expression included the presence of unprocessed (191 aa) and processed (173 aa) forms of the Core proteins with the processed form becoming dominant later. Expression of the 191 aa form of Core protein corresponded to an increase in the expression of the p21, a decrease in cdk2-dependent kinase activity, and a decrease in the percentage of cells in S-phase along with an accumulation of cells in the G(0)/G(1) phase of the cell cycle. As the processed form accumulated, the p21 levels started to decline, suggesting that Core protein regulates p21 expression in a biphasic manner. These findings implicate Core protein in potentially modulating hepatocyte cell cycle differentially in the early stages of infection through biphasic regulation of p21 cdk kinase inhibitor.


Treatment of menopausal oestrogen deficiency symptoms in hysterectomised women by means of 17beta-oestradiol pellet implants.
Ninety-four women having undergone hysterectomy, with or without simultaneous ovariectomy, were treated for menopausal symptoms by means of subcutaneous implantation of sterile pellets containing 20 mg 17beta-oestradiol, 589 implantations in all. The subjective effect was good and lasted generally for about 6 months. The Karyopyknotic index, serum FSH and plasma oestradiol were checked in a few of the patients. The method was judged to be valuable in appropriate cases.


A microcosm investigation of fe (iron) removal using macrophytes of ramsar lake: A phytoremediation approach.
The present study deals with the microcosm study of Fe (Iron) phytoremediation using Eichhornia crassipes, Lemna minor, Pistia stratiotes and Salvinia cucullata species collected from the Loktak Lake, a Ramsar Site which exists in north-eastern India (an Indo-Burma hotspot region). Efficiency of these four macrophytes was compared using different Fe concentrations of 1 mg L(-1), 3 mg L(-1) and 5 mg L(-1) for 4 days, 8 days and 12 days, respectively. E. crassipes was the most efficient macrophyte whereas L. minor was the least efficient. E. crassipes removed the highest percentage of Fe, i.e. 89% from 1 mg L(-1), 81.3% from 3 mg L(-1) and 73.2% from 5 mg L(-1) in 12-day experiment.


Mutagenesis of individual pentatricopeptide repeat motifs affects RNA binding activity and reveals functional partitioning of Arabidopsis PROTON gradient regulation3.
Pentatricopeptide repeat (PPR) proteins bind RNA and act in multiple eukaryotic processes, including RNA editing, RNA stability, and translation. Here, we investigated the mechanism underlying the functional versatility of Arabidopsis thaliana proton gradient regulation3 (PGR3), a chloroplast protein harboring 27 PPR motifs. Previous studies suggested that PGR3 acts in (1) stabilization of photosynthetic electron transport L (petL) operon RNA, (2) translation of petL, and (3) translation of ndhA. We showed here that replacement of the 4th amino acid of the 12th PPR with nonpolar or charged amino acids abolished functions (1) and (2) but not (3) of PGR3 by compromising the function of this specific PPR. This discovery enabled us to knock out the RNA binding ability of individual PPR motifs. Consequently, we showed that the 16 N-terminal PPRs were sufficient for function (1) via sequence-specific RNA binding, whereas the 11 C-terminal motifs were essential for functions (2) and (3) by activating translation. We also clarified that the 14th amino acid of the 12th PPR should be positively charged to make the PPR functionally active. Our finding opens up the possibility of selectively manipulating the functions of PPR proteins.


Psychophysical estimate of plantar vibration sensitivity brings additional information to the detection threshold in young and elderly subjects.
Vibration detection threshold of the foot sole was compared to the psychophysical estimate of vibration in a wide range of amplitudes in young (20-34 years old) and elderly subjects (53-67 years old). The vibration detection threshold was determined on the hallux, 5th metatarsal head, and heel at frequencies of 25, 50 and 150 Hz. For vibrations of higher amplitude (reaching 360 μm), the Stevens power function (Ψ = k * Φn ) allowed to obtain regression equations between the vibration estimate (Ψ) and its physical magnitude (Φ), the n coefficient giving the subjective intensity in vibration perception. We searched for age-related changes in the vibration perception by the foot sole. In all participants, higher n values were measured at vibration frequencies of 150 Hz and, compared to the young adults the elderly had lower n values measured at this frequency. Only in the young participants, the vibration detection threshold was lowered at 150 Hz. The psychophysical estimate brings further information than the vibration detection threshold which is less affected by age. The clinical interest of psychophysical vibration estimate was assessed in a patient with a unilateral alteration of foot sensitivity.


Effects of ZnCl2 on membrane interactions in myelin of normal and shiverer mice.
X-ray diffraction was used to record the effects of metal cations on the structure of peripheral nerve myelin. Acidic saline (pH 5.0) either with or without added metal cations caused myelin to swell by 10-20 A from its native period of 178 A. The X-ray patterns usually showed broad reflections, and higher orders were either weak or unobserved. With added ZnCl2, however, the swollen myelin gave diffraction patterns that retained sharp reflections to approx. 15 A spacing. Alkaline saline (pH 9.7) containing ZnCl2 produced a reduction of the myelin period by approx. 5 A which was at least twice as much as that produced by other metals. To examine the underlying chemical basis for these unique interactions of Zn2+ with myelin, we carried out parallel X-ray experiments on sciatic nerve from the shiverer mutant mouse, which lacks the major myelin basic proteins. Shiverer myelin responded like normal myelin to ZnCl2 in acidic saline; however, in alkaline saline shiverer myelin showed broadened X-ray reflections which indicated disordering of the regularity of the membrane arrays, and additional reflections were recorded which indicated lipid phase separation. This breakdown may come about by the binding of Zn2+ to negatively-charged lipids which could be more exposed due to the absence of myelin basic proteins. Electron density profiles were calculated on the assumption that, except for changes in their packing, the myelin membranes were minimally altered in structure. For both normal and shiverer myelins, treatments under acidic conditions resulted in swelling at the extracellular apposition and a slight narrowing of the cytoplasmic space. This swelling is likely due to adsorption of protons and divalent cations. Interaction between Zn2+ and myelin P0 glycoprotein could preserve an ordered arrangement of the apposed membrane surfaces. Alkaline saline containing ZnCl2 produced compaction at the cytoplasmic apposition in both normal and shiverer myelins possibly through interactions with a portion of P0 glycoprotein which extends into the cytoplasmic space between membranes.


Heparin binding to protein C inhibitor.
Protein C inhibitor is a plasma protein whose ability to inhibit activated protein C, thrombin, and other enzymes is stimulated by heparin. These studies were undertaken to further understand how heparin binds to protein C inhibitor and how it accelerates proteinase inhibition. The region of protein C inhibitor from residues 264-283 was identified as the heparin-binding site. This differs from the putative heparin-binding site in the related proteins antithrombin and heparin cofactor. The glycosaminoglycan specificity of protein C inhibitor was relatively broad, including heparin and heparan sulfate, but not dermatan sulfate. Non-sulfated and non-carboxylated polyanions also enhanced proteinase inhibition by protein C inhibitor. Heparin accelerated inhibition of alpha-thrombin, gamma T-thrombin, activated protein C, factor Xa, urokinase, and chymotrypsin, but not plasma kallikrein. The ability of glycosaminoglycans to accelerate proteinase inhibition appeared to depend on the formation of a ternary complex of inhibitor, proteinase, and glycosaminoglycan. The optimum heparin concentration for maximal rate stimulation varied from 10 to 100 micrograms/ml and was related to the apparent affinity of the proteinase for heparin. There was no obvious relationship between heparin affinity and maximum inhibition rate or degree of rate enhancement. The affinity of the resultant protein C inhibitor-proteinase complex was also not related to inhibition rate enhancement, and the results showed that decreased heparin affinity of the complex is not an important part of the catalytic mechanism of heparin. The importance of protein C inhibitor as a regulator of the protein C system may depend on the relatively large increase in heparin-enhanced inhibition rate for activated protein C compared to other proteinases.


Intramolecular Azo Coupling Reaction of Binaphthyl Compounds: Synthesis of Pyrazole-Containing Helicene-Like Molecules.
A method for accessing pyrazole-containing helicene-like molecules from easily accessible NOBIN derivatives was developed. The reaction proceeded efficiently via diazonium salt intermediates, which provided a series of helicene-like molecular products in yields of 77%-89% regardless of their steric and electronic natures. The photophysical properties of the products were investigated. The 3,3'-disubstituted molecules showed a characteristic blue shift in their emission spectra. Product derivatizations were conducted, and interesting reactivities toward nucleophiles were observed.


Assessing data in the informal e-waste sector: The Agbogbloshie Scrapyard.
The Old-Fadama-Scrapyard, better known as Agbogbloshie, is located in Accra, Ghana. Over the last 20 years, the area has developed into a large scrapyard, where the informal sector processes mainly electronic waste (e-waste) and scrap metals. However, unsafe treatment methods, such as the open burning of cables and foams, and the spilling of hazardous liquids onto the ground, cause environmental pollution and create health risks by releasing persistent organic pollutants (POPs) and heavy metals. There is a recognized lack in literature of data on e-waste mass and material flows due to the lack of applicable methods to measure e-waste quantities in an informally managed treatment system. However, to establish sustainable e-waste management, e-waste mass and material flow data are crucial prerequisites. Therefore, the material flow analysis (MFA) methodology is proposed as a means for data collection within a limited time frame in the informal e-waste recycling context. In this case study, mass and material flows of e-waste processed at Agbogbloshie were estimated using two different approaches: Firstly, the kind, measures, constitution of load and number of loaded entering and exiting vehicles was observed and documented, and second, to validate the data collected, the mass and material flow of e-waste treatment processes on site were observed and documented. The resulting annual mass flows range between 13,090 t/a and 17,094 t/a of e-waste. Based on the data for Ghana from the Global E-waste Monitor, an average of 15,092 t/a (approximately 39% of the Ghanaian e-waste generation) is treated in Agbogbloshie.


Toward objective quantification of perfusion-weighted computed tomography in subarachnoid hemorrhage: quantification of symmetry and automated delineation of vascular territories.
Perfusion-weighted computed tomography (CTP) is a relatively recent innovation that estimates a value for cerebral blood flow (CBF) using a series of axial head CT images tracking the time course of a signal from an intravenous contrast bolus. CTP images were obtained using a standard imaging protocol and were analyzed using commercially available software. A novel computer-based method was used for objective quantification of side-to-side asymmetries of CBF values calculated from CTP images. Our method corrects for the inherent variability of the CTP methodology seen in the subarachnoid hemorrhage (SAH) patient population to potentially aid in the diagnosis of cerebral vasospasm (CVS). This method analyzes and quantifies side-to-side asymmetry of CBF and presents relative differences in a construct termed a Relative Difference Map (RDM). To further automate this process, we have developed a unique methodology that enables a computer to delineate vascular territories within a brain image, regardless of the size and shape of the brain. While both the quantification of image symmetry using RDMs and the automated assignment of vascular territories were initially designed for the analysis of CTP images, it is likely that they will be useful in a variety of applications.


Efficacy and safety of ainuovirine versus efavirenz combination therapies with lamivudine/tenofovir disoproxil fumarate for medication of treatment-naïve HIV-1-positive adults: week 48 results of a randomized controlled phase 3 clinical trial followed by an open-label setting until week 96.
Ainuovirine (ANV) is a new non-nucleoside reverse transcriptase inhibitor (NNRTI), which was initially synthesized in Korea and later further developed in both Korea and China. A randomized, double-blind, double-dummy, positive parallel group, non-inferiority, phase 3 trial was conducted in 7 sites across China. Eligible HIV-1-positive antiretroviral therapy (ART)-naïve adults aged 18-65 years were randomly assigned in a 1:1 ratio to receive tenofovir disoproxil fumarate and lamivudine (TDF+3TC) in combination with either ANV (ANV group) or efavirenz (EFV group) for up to 48 weeks. Subsequently, participants in both groups received one of the two drug combinations according to their choice until week 96 in an observational study under an open-label setting. The primary endpoint was the proportion of participants achieving HIV RNA <50 copies/mL at week 48, with non-inferiority pre-specified at a margin of 10%. The secondary efficacy endpoints were logarithmic changes in HIV RNA, percentage of participants with HIV RNA levels ≤400 copies/mL and changes in the CD4 T-cell count after 48 and 96 weeks of treatment, as well as the percentage of participants with HIV RNA levels <50 copies/mL at 96 weeks of treatment. Safety endpoints were the incidence of adverse events and laboratory abnormalities evaluated according to the Division of AIDS criteria. This study was registered with the Chinese Clinical Trial Registry (Registration number: ChiCTR1800019041). Between November 27, 2018 and March 11, 2021, a total of 826 participants were screened, and 630 were finally enrolled and randomly assigned (1:1) to either ANV (n = 315) or EFV (n = 315) groups. The mean age was 30.6 ± 9.4 years and most participants were male (94.6%). At week 48, 274 (87.0%) of 315 participants in the ANV group and 288 (91.7%) of 314 in the EFV group achieved HIV-1 RNA <50 copies/mL and non-inferiority was established (difference: -4.7%, 95% CI: -9.6 to 0.1%). In the period, 293 participants continued to take the ANV regimen and 287 switched from the EFV to the ANV regimen. During the open-label period, 92.5% (271/293) of participants in the continued ANV group and 95.1% (273/287) in the ANV to EFV transfer group remained virologically suppressed (HIV-1 RNA <50 copies/mL) at week 96 (p = 0.189). The incidence of NNRTI treatment-related adverse events (TEAEs) at week 48 was 67.6% in 315 participants in the ANV group, which was significantly lower than in 91.4% of 314 participants in the EFV group (p < 0.001). The most common TEAEs (weeks 0-48) were dizziness (10.5%) and dyslipidemia (22.2%) in the ANV group vs. 51.0% and 34.4% in the EFV group, respectively, followed by transaminase elevation (9.2% vs. 29.0%), γ-glutamyl transferase elevation (8.3% vs. 19.1%), and rash (7.9% vs. 18.8%) (all p < 0.001). After switching from EFV to ANV, TEAEs in the former EFV participants were significantly reduced in the following observational period of 48-96 weeks. The week 48 results indicated that the efficacy of ANV was non-inferior to EFV when combined with two NRTIs. The per-protocol risk difference at week 48 for the primary endpoint also supported non-inferiority. TEAEs in ANV treated participants were less frequent with regard to liver toxicity, dyslipidemia, neuropsychiatric symptoms and rash compared to the EFV group during the first 48 weeks of therapy. The effects were maintained during the 48-96 weeks of therapy. Jiangsu Aidea Pharmaceutical Co., Ltd.


Detection of sucrase-isomaltase in stains on a knife by sandwich enzyme immunoassay to determine penetration of the small intestine in a pig model.
Using 2 kitchen knives, 2 stab wounds were made into the abdomen of a pig that had been sacrificed under sanctioned processing methods of a meat packing plant. One knife wound penetrated the jejunum and the other, the liver. After leaving the knives at room temperature for a week, the stains on the blades were extracted and subjected to a sandwich enzyme immunoassay to detect the presence of sucrase-isomaltase (sucrase-alpha-dextrinase or SI), a major digestive enzyme of the small intestine. Results of this assay revealed that sufficient SI could be detected only on the knife that had penetrated the jejunum. This animal experiment thus suggests the potential usefulness of an SI assay to identify a weapon that has penetrated the small intestine. Further investigation will be pursued to determine whether this detection method is also valid in humans.


Conditions of Confinement in U.S. Carceral Facilities During COVID-19: Individuals Speak-Incarcerated During the COVID-19 Epidemic.
We aimed to describe conditions of confinement among people incarcerated in the United States during the coronavirus disease 2019 (COVID-19) pandemic using a community-science data collection approach. We developed a web-based survey with community partners to collect information on confinement conditions (COVID-19 safety, basic needs, support). Formerly incarcerated adults released after March 1, 2020, or nonincarcerated adults in communication with an incarcerated person (proxy) were recruited through social media from July 25, 2020 to March 27, 2021. Descriptive statistics were estimated in aggregate and separately by proxy or formerly incarcerated status. Responses between proxy and formerly incarcerated respondents were compared using Chi-square or Fisher's exact tests based on α=0.05. Of 378 responses, 94% were by proxy, and 76% reflected state prison conditions. Participants reported inability to physically distance (≥6 ft at all times; 92%), inadequate access to soap (89%), water (46%), toilet paper (49%), and showers (68%) for incarcerated people. Among those receiving prepandemic mental health care, 75% reported reduced care for incarcerated people. Responses were consistent between formerly incarcerated and proxy respondents, although responses by formerly incarcerated people were limited. Our findings suggest that a web-based community-science data collection approach through nonincarcerated community members is feasible; however, recruitment of recently released individuals may require additional resources. Our data obtained primarily through individuals in communication with an incarcerated person suggest COVID-19 safety and basic needs were not sufficiently addressed within some carceral settings in 2020-2021. The perspectives of incarcerated individuals should be leveraged in assessing crisis-response strategies.


The A1 receptor agonist R-Pia reduces the imbalance between cerebral glucose metabolism and blood flow during status epilepticus: could this mechanism be involved with neuroprotection?
It is well known that the uncoupling between local cerebral glucose utilization (LCGU) and local cerebral blood flow (LCBF), i.e. decrease in LCBF rates with high LCGU, is frequently associated with seizure-induced neuronal damage. This study was performed to assess if the neuroprotective effect of the adenosinergic A(1) receptor agonist R-N-phenylisopropyladenosine (R-Pia) injected prior to pilocarpine is able to reduce the uncoupling between LCGU and LCBF during status epilepticus (SE). Four groups of rats were studied: Saline, Pilo, R-Pia+Saline and R-Pia+Pilo. For LCGU and LCBF studies, rats were subjected to autoradiography using [(14)C]-2-deoxyglucose and [(14)C]-iodoantypirine, respectively. Radioligands were injected 4 h after SE onset. Neuronal loss was evaluated by Fluorojade-B (FJB) at two time points after SE onset (24 h and 7 days). The results showed a significant increase in LCGU in almost all brain regions studied in the Pilo and R-Pia+Pilo groups compared to controls. However, in R-Pia pretreated rats, the uncoupling between LCGU and LCBF was moderated in a limited number of structures as substantia nigra pars reticulata and hippocampal formation rather in favor of hyperperfusion. Significant increases in LCBF were observed in the entorhinal cortex, thalamic nuclei, mammillary body, red nucleus, zona incerta, pontine nucleus and visual cortex. The neuroprotective effect of R-Pia assessed by FJB showed a lower density of degenerating cells in the hippocampal formation, piriform cortex and basolateral amygdala. In conclusion our data shows that the neuroprotective effect of R-Pia was accompanied by a compensatory metabolic input in brain areas involved with seizures generation.