Temporary changes in neutrophil-to-lymphocyte, platelet-to-lymphocyte ratios, and mean platelet volume reflecting the inflammatory process after radioiodine therapy.
Data on the effects of radioiodine (RAI) therapy on systemic inflammation are very limited. The aim of this study is to explore alterations of subclinical systemic inflammatory markers, such as neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), and mean platelet volume (MPV), after RAI therapy in patients with differentiated thyroid carcinoma (DTC). We evaluated 57 DTC patients treated with RAI (RAI group), 37 DTC patients not treated with RAI (non-RAI control group), and 37 age-matched healthy individuals (healthy control group). NLR, PLR, and MPV levels were compared among the study groups; these were also examined after RAI in the RAI group. Initially, NLR was significantly higher in the RAI group than in the healthy controls. NLR and PLR increased significantly and MPV decreased significantly 2 months after RAI therapy (P=0.021, 0.001, and 0.008, respectively). Although NLR and PLR levels decreased, they were still high compared with the preoperative values. MPV returned to normal levels at 6 months. These parameters did not change significantly in the non-RAI control group. This is the first study to evaluate changes in NLR, PLR, and MPV after RAI therapy. Our findings suggest that NLR, PLR, and MPV changes indicate systemic inflammation that occurs after RAI therapy because of thyroid remnant tissue ablation.


Erythroleukaemia induction by the Friend spleen focus-forming virus.
The Friend spleen focus-forming virus has been a valuable tool for understanding the molecular events involved in the multiple stages of leukaemia. As summarized in Figure 3, the primary effect of SFFV, which occurs within days, is to cause a polyclonal proliferation of erythroid precursor cells that can proliferate in the absence of their normal regulator erythropoietin. This is the direct result of the unique envelope glycoprotein encoded by SFFV, which is transported to the cell surface and apparently interacts with the EpoR or another component of the multimeric EpoR complex, resulting in the constitutive activation of the Epo signal transduction pathway. Within this proliferating population of erythroid cells is a rare cell that has undergone several genetic changes due to the integration of the viral genome in specific sites in the mouse DNA. This leads to the activation of a gene encoding the PU.1 transcription factor, whose high expression in erythroid cells may be the cause of the block in differentiation that is characteristic of SFFV-transformed erythroid cells. SFFV integration can also lead to the inactivation of the p53 tumour supressor gene, giving these cells a growth advantage in the mouse. The disease induced by SFFV in mice is very similar to polycythaemia vera in humans (Golde et al, 1981). The major clinical feature of polycythaemia vera is the continuous expansion of the number of mature red blood cells in the presence of low serum Epo levels. Also, BFU-E and CFU-E from these patients can form in the absence of Epo like the analogous cells from SFFV-infected mice (Casadevall et al, 1982). It is possible that haematopoietic cells from individuals suffering from this disease express a protein similar to the envelope glycoprotein of SFFV that can interact with the EpoR and lead to its constitutive activation. Alternatively, these patients may contain a mutant EpoR gene that is constitutively activated like the mutant EpoR described earlier. As we understand more fully how the SFFV envelope protein constitutively activates te EpoR complex, we can begin to design therapies to counteract its action that can then be applied to treating patients with polycythaemia vera or other human diseases associated with uncontrolled erythropoiesis.


Transdermal HRT and Doppler findings in normotensive and hypertensive postmenopausal patients.
To evaluate the effects of transdermal hormone replacement therapy (HRT) on plasma viscosity, serum levels of thromboxane B2 (TXB2) and vascular impedance in the uterine, bladder wall, internal carotid and ophthalmic arteries in normotensive and hypertensive postmenopausal patients. Thirty postmenopausal patients underwent continuous estradiol transdermal supplementation at a dose of 50 microg/day and 12-day courses of medroxyprogesterone acetate 10 mg/day every 2 months. The women were divided into two groups according to their blood pressure: normotensive women (Group 1, n=14) and hypertensive subjects (Group 2, n=16). Before starting HRT and after 6 months of therapy, the patients underwent: transvaginal ultrasonographic examination of the pelvic organs; Doppler examination of the blood flow velocities in the uterine, bladder wall, internal carotid and ophthalmic arteries; and analysis of plasma viscosity and plasma TXB2. After 6 months of HRT plasma viscosity had decreased in both groups (mean reduction in Group 1, (14+/-1)%, P=0.005; mean reduction in Group 2, (10+/-1)%, P=0.005) as had the TXB2 levels (mean reduction in Group 1, (93+/-2)%, P<0.001; mean reduction in Group 2, (92+/-3)%, P<0.001). The mean percentage reduction in plasma viscosity was smaller in hypertensive women than in normotensive women (P<0.05). There was also a significant reduction in vascular impedance in the uterine artery (mean reduction in Group 1, (16+/-1)%, P=0.005; mean reduction in Group 2, (19+/-1)%, P=0.005), the bladder wall arteries (mean reduction in Group 1, (23+/-2)%, P=0.005; mean reduction in Group 2, (18+/-1)%, P=0.005), the internal carotid artery (mean reduction in Group 1, (25+/-1)%, P=0.005; mean reduction in Group 2, (26+/-1)%, P=0.005) and the ophthalmic artery (mean reduction in Group 1, (24+/-2)%, P=0.005; mean reduction in Group 2, (16+/-1)%, P=0.005). The percentage reduction in vascular impedance did not differ significantly between the two groups. Our results show that transdermal HRT is effective in reducing plasma viscosity, TXB2 levels and vascular impedance in the peripheral and central vessels both in normotensive and hypertensive postmenopausal patients.


Zoobooth: A portable, open-source and affordable approach for repeated size measurements of live individual zooplankton.
Repeated size measurements of individual animals are valuable data for many research questions, but it is often hard to obtain without causing stress or damage to the animal. We developed a video-based approach called Zoobooth to size individual zooplankton, which involves a low risk of handling accidents and stress. Here we describe the process of assembling the instrument we used to acquire video recordings of single zooplankton and the procedure to obtain size estimates from the recorded videos. Our setup produces accurate size estimates for Daphnia magna (correlation to manual measurements = 0.97), and was also tested with other zooplankton species. Zoobooth is especially advantageous when one needs size measurements of live, individual mesozooplankton. The device is small, portable, and comprised of very affordable and readily available components. It can easily be modified for other purposes, such as studies of coloration or behavior of micro-and macro-plankton. We share all the files to build and use Zoobooth.


Achievement of specified lipid and high-sensitivity C-reactive protein levels with two statins in Chinese patients with hypercholesterolaemia.
Statins reduce plasma low-density lipoprotein cholesterol (LDL-C) and high-sensitivity C-reactive protein (hsCRP) levels. Rosuvastatin 10 mg daily appears to be more potent in reducing LDL-C than simvastatin 40 mg, but the relative effect of these two statin doses on hsCRP is unknown. Chinese hyperlipidaemic patients with high cardiovascular risk or familial hypercholesterolaemia (FH) were treated with rosuvastatin 10 mg and simvastatin 40 mg daily in an open-label crossover study. Lipid profiles were measured off treatment and after at least 4 weeks treatment with each of the two statins and hsCRP levels were measured on treatment with both statins. Both treatments were well tolerated in 247 patients (age 55.7 ± 11.1 years; 100 male; 140 with FH) with good treatment compliance. There were statistically significant differences (P < 0.001) for rosuvastatin versus simvastatin for LDL-C reduction (-52.4 ± 11.9 % vs. -47.7 ± 10.8 %) and on-treatment LDL-C (2.62 ± 0.99 mmol/L vs. 2.86 ± 0.97 mmol/L), respectively, but the on-treatment hsCRP levels (1.33 ± 1.37 mg/L vs. 1.41 ± 1.57 mg/L, P > 0.05) were not significantly different. The lipid target (LDL-C <2.6 mmol/L) was achieved by 52.9 % with rosuvastatin compared with 42.6 % with simvastatin (P < 0.05). The proportions of patients attaining hsCRP targets of < 2 and < 1 mg/L were similar with the two statins (57.1 % and 74.6 % for rosuvastatin vs. 57.1 % and 80.1 % for simvastatin, P > 0.05). A significantly greater proportion of patients achieved LDL-C targets with rosuvastatin 10 mg compared to simvastatin 40 mg in Chinese patients with hypercholesterolaemia, but there was no significant difference in achieving hsCRP target levels with the two statins.


Studies on the isolation, growth and maintenance of Malassezia pachydermatis.
Results related to the isolation, cultivation, culture and maintenance of the opportunistic pathogen Malassezia pachydermatis are reported. A dextrose nutrient medium with 1.5% yeast extract turned out to be the most favourable medium for its development. It permitted identification in 24 hours and maintenance of isolates for three months without subculturing. Addition of Tween 80 (1%) significantly enhanced the isolation of this yeast from clinical materials.


Characterization of the deoxyribonucleic acid polymerase associated with Kilham rat virus.
Purified preparations of the parvovirus, Kilham rat virus, have associated with them a protein with DNA polymerase activity. The enzyme has been separated from the other two or three viral proteins and purified 63-fold. The viral associated enzyme was found in a single peak of DNA polymerase activity after chromatography on DEAE-cellulose, DNA-cellulose, and phosphocellulose columns. It shares some properties in common with the host cellular DNA polymerases, described in the preceding paper (Salzman, L.A., and McKerlie, L. (1975) J. Biol. Chem. 250, 5589-5595), but also has some important distinguishing characteristics. The Kilham rat virus-associated DNA polymerase has increased enzyme activity in the presence of 0.02 M KCl and has a strong preference for a synthetic DNA polymer containing deoxyadenylate and deoxythymidylate. The enzyme has a molecular weight of approximately 75,000 plus or minus 3,000 and appears to contain endonuclease activity.


KLF17 promotes human naive pluripotency through repressing MAPK3 and ZIC2.
The pluripotent state of embryonic stem cells (ESCs) is regulated by a sophisticated network of transcription factors. High expression of KLF17 has recently been identified as a hallmark of naive state of human ESCs (hESCs). However, the functional role of KLF17 in naive state is not clear. Here, by employing various gain and loss-of-function approaches, we demonstrate that KLF17 is essential for the maintenance of naive state and promotes the primed to naive state transition in hESCs. Mechanistically, we identify MAPK3 and ZIC2 as two direct targets repressed by KLF17. Overexpression of MAPK3 or ZIC2 partially blocks KLF17 from promoting the naive pluripotency. Furthermore, we find that human and mouse homologs of KLF17 retain conserved functions in promoting naive pluripotency of both species. Finally, we show that Klf17 may be essential for early embryo development in mouse. These findings demonstrate the important and conserved function of KLF17 in promoting naive pluripotency and reveal two essential transcriptional targets of KLF17 that underlie its function.


N-3, a novel synthetic derivative of bifendate, inhibits metastasis of triple-negative breast cancer via decreasing p38-regulated FOXC1 protein stability.
Triple-negative breast cancer (TNBC) is an aggressive breast cancer subtype with high invasiveness, metastatic potential, and poor prognosis. Epithelial-mesenchymal transition (EMT) is pivotal in TNBC progression, becoming a promising target for TNBC treatment. Our study evaluated N-3, a novel synthetic bifendate derivative, which inhibited the EMT-associated migration and invasion of MDA-MB-231 and 4T1 TNBC cells. The results were consistent with the suppression of FOXC1 expression and transcriptional activity. Additional studies indicated that N-3 reduced the protein stability of FOXC1 by enhancing ubiquitination and degradation. Moreover, N-3 downregulated p-p38 expression and FOXC1 interaction, decreasing the stability of p38-regulated FOXC1. Further, N-3 blocked TNBC metastasis with an artificial lung metastasis model in vivo, related to FOXC1 suppression and EMT. These results highlight the potential of N-3 as a TNBC metastasis treatment. Therefore, FOXC1 regulation could be a novel targeted therapeutic strategy for TNBC metastasis.


Differential regulation of the two xGATA-1 genes during Xenopus development.
Xenopus laevis provides an established developmental system to study the regulation of cell lineage establishment and the generation of tissue-specific patterns of gene expression. We have isolated from Xenopus erythroid cell RNA two distinct cDNA clones encoding the xGATA-1 transcription factor. We have identified, in erythroid nuclear extracts, the sequence-specific DNA-binding protein they encode. By characterizing the expression patterns for RNA derived from two distinct homologues, we find that the two xGATA-1 genes are differentially regulated. The xGATA-1a mRNA predominates in embryos prior to terminal differentiation of erythroid cells, while the differentiated cells contain RNA derived predominantly from the xGATA-1b gene. Both proteins activate a target globin promoter in transient transfection assays. During early development, GATA-1 transcripts are localized to ventral regions of the embryo. GATA-1 should therefore provide a useful early marker for studying signalling pathways which result in the generation of ventral mesoderm. The differentially regulated genes may be distinct markers for targets of ventral mesoderm induction.


Managing Complexity: The Experience of an Italian Internal Medicine Unit During the SARS-CoV-2 Pandemic, Looking to the Future.
The impact of COVID-19 pandemic put the Italian health system into a test. In the period between September 2020 and June 2021, a weekly average of 114 SARS-CoV-2 infections were recorded in Forlì-Cesena province (north of Italy), with a maximum of 330 cases per week in March 2021; in the same months, the Internal Medicine Unit of Cesena M. Bufalini Hospital managed 954 COVID-19 patients. To allow the management of these patients, the ward was divided into areas at different intensity of care, with a maximum of 39-47 beds and 19-24 in ordinary and sub-intensive area, respectively. Patients had an average age of 66 years, and 62% of the total was female; prevalent comorbidities were arterial hypertension (53%), smoking habit (28.7%), obesity (27.9%), uncomplicated (10%), and complicated diabetes (9%). On the total, 339 patients were hospitalized in sub-intensive area, subjected to non-invasive ventilatory support. Hospitalization lasted about 7 days in the ordinary ward and 13 days in the sub-intensive area. One hundred six patients died. In the considered period, the mean percentage of deaths compared to hospitalizations in Italy was equal to 22.21%; in our experience, the overall mortality rate was 11%. Our organizational model, which included different intensity areas in the same ward and various specialist skills, as the ability to manage non-invasive ventilation and bedside ultrasound, allowed flexible management of the "complex" COVID patient. Even the mortality rate may be the result of this model. These features mark what modern Internal Medicine should be like.


Comparison of the outcomes of anterolateral thigh flap and peroneal flap in total laryngectomy reconstruction.
We usually use an ALT or peroneal flap in total laryngectomy reconstruction, depending on the surgeons' preference. No direct comparison of the outcomes of the ALT flap and peroneal flap exists. From 2014 to 2022, we reviewed patients who had total laryngectomy and were reconstructed with an ALT flap and peroneal flap. Patient characteristics and surgical outcomes were collected and compared. The peroneal group had a significantly higher risk of neopharynx leakage (40% vs. 13.2%, p = .020) and late pharyngocutaneous fistula formation (30% vs. 5.3%, p = .009) than the ALT group. Peroneal flap was found to be the only independent risk factor for neopharynx leakage (p = .025, odds ratio [OR] = 5.5) and late pharyngocutaneous fistula formation (p = .02, OR = 7.7) in multivariate logistic regression. In the reconstruction of total laryngectomy, the ALT flap is preferable over the peroneal flap.


Stabilizing the Transcription Factors by E3 Ligase COP1.
Photomorphogenesis is oppositely regulated by two groups of transcription factors. CONSTITUTIVE PHOTOMORPHOGENIC 1 (COP1) degrades the positive factors but stabilizes the negative ones to predominantly repress photomorphogenesis. It is known that COP1 degrades substrates as an E3 ligase. Two recent studies unraveled the long-sought mechanisms of how COP1 stabilizes the negative transcription factors.


Inferior vena cava resection for a leiomyosarcoma: A case report.
Leiomyosarcomas of the inferior vena cava are scarce. Surgery is the only potential curative treatment. Such a surgery is complex and technically demanding, requiring a huge experience in both visceral and vascular surgery. Indeed, enlarged visceral resection may be needed to achieve a free tumor margins as the tumor may invade the duodenum, the head of the pancreas and the liver. Moreover, vascular reconstruction is mandatory which might be complex as both venal reins are usually invaded by the tumor like in our case below. A 53-year-old woman presented with abdominal vague pain. Imaging consisting in abdominal CT-scan with contrast and magnetic resonance angiography were suggestive of the diagnosis of leiomyosarcoma of the inferior vena cava. A surgical complete resection with free margins was conducted. A primary repair of the inferior vena cava and the right renal vein walls. End-to-side anastomosis with a polytetrafluorethylene (PTFE) prosthesis was mandatory to reconstruct the renal left vein to the inferior vena cava. Postoperative outcomes were uneventful. Pathology examination confirmed the diagnosis. The optimal management of leiomyosarcoma of inferior vena cava is controversial. Surgery remains the cornerstone of treatment. However, not all patients are qualified for surgical resection. The prognosis depends on the early diagnosis and histology type. The management of leiomyosarcomas of inferior vena cava requires a multidisciplinary consensus involving experienced surgery and oncology teams.


Modulation of kappa-mediated antitussive activity in rats by a delta-agonist.
When co-administered intracisternally, the selective delta-opioid agonist [D-Pen2,5]enkephalin (DPDPE), which had no significant effect on the cough reflex, consistently and significantly decreased the antitussive potencies of kappa-receptor agonists, U-50,488H and U-62,066E. The decrease in the antitussive effects of these kappa-receptor agonists caused by DPDPE were prevented by selective delta receptor antagonist, naltrindole. These results suggest that delta receptors may play an inhibitory role in antitussive processes that are mediated by the kappa-receptors.


Deletion of Nrf2 impairs functional recovery, reduces clearance of myelin debris and decreases axonal remyelination after peripheral nerve injury.
Oxidative stress is generated in several peripheral nerve injury models. In response to oxidative stress, the transcription factor Nrf2 is activated to induce expression of antioxidant responsive element (ARE) genes. The role of Nrf2 in peripheral nerve injury has not been studied to date. In this study, we used a sciatic nerve crush model to examine how deletion of Nrf2 affects peripheral nerve degeneration and regeneration. Our study demonstrated that functional recovery in the Nrf2(-/-) mice were impaired compared to the wild type mice after sciatic nerve crush. Larger myelin debris were present in the distal nerve stump of the Nrf2(-/-) mice than in the wild type mice. The presence of larger myelin debris in the Nrf2(-/-) mice coincides with less macrophages accumulation in the distal nerve stump. Less accumulation of macrophages may have contributed to slower clearance of myelin and thus resulted in the presence of larger myelin debris. Meanwhile, axonal regeneration is comparatively lower in the Nrf2(-/-) mice than in the wild type mice. Even after 3months post the injury, more thinly myelinated axon fibers were present in the Nrf2(-/-) mice than in the wild type mice. Taken collectively, these data support the concept of therapeutic intervention with Nrf2 activators following nerve injury.


Safety and immunogenicity of co-administered hookworm vaccine candidates Na-GST-1 and Na-APR-1 in Gabonese adults: a randomised, controlled, double-blind, phase 1 dose-escalation trial.
Hookworms cause substantial morbidity in children and women of reproductive age. The control strategy of mass drug administration is suboptimal, hence the need for a vaccine. Necator americanus aspartic protease-1 (Na-APR-1) and N americanus glutathione S-transferase-1 (Na-GST-1) are involved in the digestion and detoxification of haemoglobin in the hookworm digestive tract. In animal models, vaccination against these antigens resulted in protection from challenge infection. Both vaccine candidates were shown to be safe and well tolerated when administered separately to healthy adults. We assessed the safety and immunogenicity of co-administered Na-GST-1 and Na-APR-1 (M74) vaccines in healthy Gabonese adults. This randomised, controlled, double-blind, phase 1, dose-escalation trial was done at the Centre de Recherches Médicales de Lambaréné, in a region of Gabon where N americanus and other helminths are prevalent. Healthy adults aged 18-50 years and living in Lambaréné or the surrounding areas were recruited to the study. Participants were enrolled consecutively into two dose cohorts (30 μg or 100 μg of the experimental vaccines) and randomly assigned in blocks (block size four) to receive three doses of either co-administered Na-GST-1 plus Na-APR-1 (M74; 30 μg or 100 μg of each), adjuvanted with Alhydrogel (aluminium hydroxide gel suspension) together with an aqueous formulation of glucopyranosyl lipid A, or hepatitis B vaccine plus saline (control group). Vaccines were administered intramuscularly on days 0, 28, and 180. The primary endpoint was safety, with immunogenicity a secondary endpoint. The intention-to-treat population was used for safety analyses, whereas for immunogenicity analyses, the per-protocol population was used (participants who received all scheduled vaccinations). Control vaccine recipients for both dose cohorts were combined for the analyses. The trial is registered with ClinicalTrials.gov, NCT02126462. Between Oct 27, 2014, and Jan 31, 2015, 56 individuals were screened for eligibility, of whom 32 were enrolled and randomly assigned to one of the three study groups (12 each in the 30 μg and 100 μg experimental vaccine groups and eight in the control group). Both study vaccines were well tolerated in both dose groups. The most common adverse events were mild-to-moderate injection-site pain, headache, myalgia, and nausea. No severe or serious adverse events related to the vaccines were recorded. 52 unsolicited vaccine-related adverse events occurred during the study, but there was no difference in frequency between vaccine groups. IgG antibodies were induced to each of the vaccine antigens, with mean IgG levels increasing after each vaccination. Vaccination with 100 μg of each vaccine antigen consistently induced IgG seroconversion (IgG levels above the reactivity threshold). Peak IgG responses were observed 2 weeks after the third vaccine dose for both antigens, with all participants who received the 100 μg doses seroconverting at that timepoint. IgG levels steadily declined until the final study visit 6 months after the third vaccination, although they remained significantly higher than baseline in the 100 μg dose group. Vaccination with recombinant Na-GST-1 and Na-APR-1 (M74) in healthy adults living in N americanus-endemic areas of Gabon was safe and induced IgG to each antigen. To our knowledge, this study is the first to report results of Na-APR-1 (M74) co-administered with Alhydrogel in participants from an N americanus-endemic area. Further clinical development of these vaccines should involve efficacy studies. European Union Seventh Framework Programme.


In vitro studies of the carotid rete mirabile of Artiodactyla.
The carotid rete of Artiodactyla, an intracranial arterial plexus which supplies blood to the brain, has intrigued investigators for a long time. This study was designed to examine the responsiveness of isolated retial arteries (250-700 microns in external diameter) of goat, pig, and cattle. The findings in these arteries were compared to those observed in cerebral arteries (250-650 microns in external diameter) of the same animal species. The magnitude of the arterial responses to potassium chloride varied with the resting tension applied to the tissue. The two types of vessels exhibited similar resting tension values (0.3 g) for maximal tension development in response to potassium chloride; however, the ability of retial vessels to contract in the presence of potassium chloride was consistently smaller than that of cerebral arteries. The contractile response of retial arteries to norepinephrine (10(-9) to 10(-4) M), tyramine (10(-8) to 10(-3) M), and field electrical stimulation (2-16 Hz) was negligible. The same retial arteries exhibited dose-dependent contractions in response to 5-hydroxytryptamine (10(-9) to 10(-5) M) and histamine (10(-9) to 3 X 10(-4) M). Cerebral arteries exhibited larger responses to the vasoactive agents than retial arteries. Our findings indicate that retial arteries have a small vasomotor activity in response to adrenergic stimulation or to vasoactive agents. Therefore, the carotid rete of Artiodactyla may have a low ability to change resistance to blood flow under neural or hormonal influences.


(Dis)embodying gender and sexuality in testicular cancer.
Testicular cancer is the most common cancer in men aged 15-34. Although post-treatment prognosis is generally very good, the impact on sexuality, gender identity and fertility is amplified in this age group. A Canadian study of men with testicular cancer explores how men (re)consider questions of sexuality and gender post diagnosis and treatment. Semi-structured interviews with 40 men were analyzed using thematic decomposition, an analytic technique that combines discursive approaches with thematic analysis. The theoretical framework that guides this work relies on material discursive approaches. From an analytic stance, this perspective is concerned with a focus on the ways in which both subjectivity and the body are experienced and constituted in language. In particular, we are concerned with how these men interpret the (altered) male body as a locus of gender signification and gender disruption. Men in this study construct testicular cancer as alternately inhibiting and enhancing masculinity and sexuality. Disruption interpolates with potentiality. A discourse of precarious masculinity predominates these accounts, wherein the link between anatomy and masculinity is simultaneously asserted and disavowed. Constructions of anatomical essentialism (i.e., testicular integrity is equated with masculinity) are juxtaposed against construals of anatomical superfluousness (i.e., other sites of sexuality and male identity are emphasized as being more central).


Fabrication of cellulose acetate/polybenzoxazine cross-linked electrospun nanofibrous membrane for water treatment.
Herein, polybenzoxazine based cross-linked cellulose acetate nanofibrous membrane exhibiting enhanced thermal/mechanical properties and improved adsorption efficiency was successfully produced via electrospinning and thermal curing. Initially, suitable solution composition was determined by varying the amount of the benzoxazine (BA-a) resin, cellulose acetate (CA) and citric acid (CTR) to obtain uniform nanofibrous membrane via electrospinning. Subsequently, thermal curing was performed by step-wise at 150, 175, 200 and 225°C to obtain cross-linked composite nanofibrous membranes. SEM images and solubility experiments demonstrated that most favorable result was obtained from the 10% (w/v) CA, 5% (w/v) BA-a and 1% (w/v) CTR composition and cross-linked nanofibrous membrane (CA10/PolyBA-a5/CTR1) was obtained after the thermal curing. Chemical structural changes (ring opening) occurred by thermal curing revealed successful cross-linking of BA-a in the composite nanofibrous membrane. Thermal, mechanical and adsorption performance of pristine CA and CA10/PolyBA-a5/CTR1 nanofibrous membranes were studied. Char yield of the pristine CA nanofibrous membrane has increased notably from 12 to 24.7% for composite CA10/PolyBA-a5/CTR1 membrane. When compared to pristine CA membrane, CA10/PolyBA-a5/CTR1 nanofibrous membrane has shown superior mechanical properties having tensile strength and Young's modulus of 8.64±0.63MPa and 213.87±30.79MPa, respectively. Finally, adsorption performance of pristine CA and CA10/PolyBA-a5/CTR1 nanofibrous membranes was examined by a model polycyclic aromatic hydrocarbon (PAH) compound (i.e. phenanthrene) in aqueous solution, in which CA10/PolyBA-a5/CTR1 nanofibrous membrane has shown better removal efficiency (98.5%) and adsorption capacity (592μg/g).


It's more than just Tenet. Analysis shows not-for-profit hospitals, including a cluster in New Jersey, also heavily rely on outlier payments.
After using a loophole in the Medicare outlier payment program to fuel growth, Tenet Healthcare Corp. has found itself operating under a financial cloud. But the spotlight hasn't focused on many other hospitals and systems that have relied heavily on outlier payments to boost their bottom lines. At left, Saint Barnabas Medical Center in Livingston, N.J., ranks among hospitals with a high reliance on outlier payments.


GABA and GABA receptors in the central nervous system and other organs.
Gamma-aminobutyrate (GABA) is a major inhibitory neurotransmitter in the adult mammalian brain. GABA is also considered to be a multifunctional molecule that has different situational functions in the central nervous system, the peripheral nervous system, and in some nonneuronal tissues. GABA is synthesized primarily from glutamate by glutamate decarboxylase (GAD), but alternative pathways may be important under certain situations. Two types of GAD appear to have significant physiological roles. GABA functions appear to be triggered by binding of GABA to its ionotropic receptors, GABA(A) and GABA(C), which are ligand-gated chloride channels, and its metabotropic receptor, GABA(B). The physiological, pharmacological, and molecular characteristics of GABA(A) receptors are well documented, and diversity in the pharmacologic properties of the receptor subtypes is important clinically. In addition to its role in neural development, GABA appears to be involved in a wide variety of physiological functions in tissues and organs outside the brain.


Lipoprotein lipase transporter GPIHBP1 and triglyceride-rich lipoprotein metabolism.
Increased plasma triglyceride serves as an independent risk factor for cardiovascular disease (CVD). Lipoprotein lipase (LPL), which hydrolyzes circulating triglyceride, plays a crucial role in normal lipid metabolism and energy balance. Hypertriglyceridemia is possibly caused by gene mutations resulting in LPL dysfunction. There are many factors that both positively and negatively interact with LPL thereby impacting TG lipolysis. Glycosylphosphatidylinositol-anchored high-density lipoprotein-binding protein 1 (GPIHBP1), a newly identified factor, appears essential for transporting LPL to the luminal side of the blood vessel and offering a platform for TG hydrolysis. Numerous lines of evidence indicate that GPIHBP1 exerts distinct functions and plays diverse roles in human triglyceride-rich lipoprotein (TRL) metabolism. In this review, we discuss the GPIHBP1 gene, protein, its expression and function and subsequently focus on its regulation and provide critical evidence supporting its role in TRL metabolism. Underlying mechanisms of action are highlighted, additional studies discussed and potential therapeutic targets reviewed.


Embryonic platelet-activating factor: temporal expression of the ligand and receptor.
Preimplantation embryos synthesize platelet-activating factor (PAF) and this embryo-derived PAF is required for development. PAF's signal transduction is receptor-mediated and PAF-receptor mRNA is present in early embryos. The study objective was to determine the relationship between PAF production and PAF-receptor mRNA expression levels in mouse preimplantation embryos. Embryo-derived PAF levels were determined by radioimmunoassay. Embryonic PAF-receptor mRNA levels were determined by semiquantitated reverse transcription-polymerase chain reaction. Embryonic-PAF increased as time progressed at a relatively constant rate (1.4-1.9 x) except between the eight- and morula-cell stages where levels increased sevenfold. Embryonic PAF-receptor expression was highest at the two-cell stage and decreased steadily until the morula stage before increasing again. Regression analysis of embryo-derived PAF on PAF-receptor expression does not demonstrate a significant relationship. PAF-receptor expression (mRNA) levels decrease, while embryo-derived PAF levels decrease, as the preimplantation embryo develops. Embryonic-PAF and PAF-receptor mRNA expression do not appear related. Therefore, embryonic-PAF does not appear to regulate expression of its own receptor in vitro. The data provide a clue to the complicated cell signaling system involving PAF production and receptor expression that may help our understanding of developmental events.


Transscleral iridotomy using a neodymium: YAG laser operated both with standard equipment and an optical fiber system--a preliminary report: Part I--Optical system and biomicroscopic results.
The effects upon the iris of radiation emitted by a Nd:YAG laser working in its thermal (free-running) mode and transmitted through the sclera were studied. The advantages and disadvantages of a fiber optic system, as compared to a conventional optical system, were studied using eight autopsy eyes. It was shown that the conventional optical system as well as a fiber optic system are both able to perforate the iris completely and to blast away or destroy the pigment epithelium. One merit of the fiber optic system as compared to the conventional system is that it requires considerably less energy for the same effect. This is partially because the sclera, when compressed, becomes more transparent and thus transmits a greater amount of laser energy.


Strategies to assess systemic exposure of chemicals in subchronic/chronic diet and drinking water studies.
Strategies were developed for the estimation of systemically available daily doses of chemicals, diurnal variations in blood levels, and rough elimination rates in subchronic feeding/drinking water studies, utilizing a minimal number of blood samples. Systemic bioavailability of chemicals was determined by calculating area under the plasma concentration curve over 24 h (AUC-24 h) using complete sets of data (> or =5 data points) and also three, two, and one selected time points. The best predictions of AUC-24 h were made when three time points were used, corresponding to Cmax, a mid-morning sample, and C(min). These values were found to be 103 +/- 10% of the original AUC-24 h, with 13 out of 17 values ranging between 96 and 105% of the original. Calculation of AUC-24 h from two samples (Cmax and Cmin) or one mid-morning sample afforded slightly larger variations in the calculated AUC-24 h (69-136% of the actual). Following drinking water exposure, prediction of AUC-24 h using 3 time points (Cmax, mid-morning, and Cmin) was very close to actual values (80-100%) among mice, while values for rats were only 63% of the original due to less frequent drinking behavior of rats during the light cycle. Collection and analysis of 1-3 blood samples per dose may provide insight into dose-proportional or non-dose-proportional differences in systemic bioavailability, pointing towards saturation of absorption or elimination or some other phenomenon warranting further investigation. In addition, collection of the terminal blood samples from rats, which is usually conducted after 18 h of fasting, will be helpful in rough estimation of blood/plasma half-life of the compound. The amount of chemical(s) and/or metabolite(s) in excreta and their possible use as biomarkers in predicting the daily systemic exposure levels are also discussed. Determining these parameters in the early stages of testing will provide critical information to improve the appropriate design of other longer-term toxicity studies.


The WUSCHEL Related Homeobox Protein WOX7 Regulates the Sugar Response of Lateral Root Development in Arabidopsis thaliana.
Sugars promote lateral root formation at low levels but become inhibitory at high C/N or C/P ratios. How sugars suppress lateral root formation is unclear, however. Here we report that WOX7, a member of the WUSCHEL related homeobox (WOX) family transcription factors, inhibits lateral root development in a sugar-dependent manner. The number of lateral root primordia increased in wox7 mutants but decreased in plants over-expressing WOX7. Plants expressing the WOX7-VP16 fusion protein produced even more lateral roots than wox7, suggesting that WOX7 acts as a transcriptional repressor in lateral root development. WOX7 is expressed at all stages of lateral root development, but it is primarily involved in lateral root initiation. Consistent with this, the wox7 mutant had a higher mitotic activity only at early stages of lateral root development. Further studies suggest that WOX7 regulates lateral root development through direct repression of cell cycle genes, particularly CYCD6;1. WOX7 expression was enhanced by sugar, reduced by auxin, but did not respond to salt and mannitol. In the wox7 mutant, the effect of sugar on lateral root formation was mitigated. These results together suggest that WOX7 plays an important role in coupling the lateral root development program and sugar status in plants.


The basis for half-site specificity explored through a non-cognate steroid receptor-DNA complex.
Steroid receptors recognize bipartite targets composed of six base-pair half-sites. There are two canonical types of half-site which differ only in their central two base pairs. The crystal structure of an estrogen receptor-like DNA-binding domain bound to the wrong type of half-site (a glucocorticoid response element) reveals an interface that resembles the specific interfaces of the glucocorticoid receptor or estrogen receptor bound to their correct response elements. The underlying stereochemical defect that weakens the non-cognate interface is a difference in the helical geometry of the incorrect DNA half-site which prevents a side-chain contact and results in a gap which is filled by at least five additional fixed water sites, imposing a potential entropic burden on the stability of the interface.


Rare Case of Persistently Depressed T Lymphocyte Subsets After SARS-CoV-2 Infection.
BACKGROUND On rare occasions, viral infections are known to also depress immune cell lines, further worsening clinical outcomes. We describe a patient who presented 3 weeks after recovery from mild COVID-19 disease with clinical features of an atypical pneumonia and was found to have a low CD4+ T-cell count. CASE REPORT An 82-year-old man with a past medical history of coronary artery disease, rheumatoid arthritis, gout, hypertension, and atrial fibrillation presented with a 1-week history of progressively worsening shortness of breath and cough. He was noted to have recovered from mild SARS-CoV-2 infection 3 weeks prior to his current presentation and had been at his baseline level of health following infection. A T cell subset panel was obtained, which revealed an absolute CD3 count of 92 (reference range 840-3060), absolute CD4 count of 52 (reference range 500-1400), absolute CD8 count of 37 (reference range 180-1170), and a normal CD4: CD8 ratio. He was subsequently started on atovaquone for pneumocystis jiroveci pneumonia prophylaxis. CONCLUSIONS This case highlights the need for a high index of suspicion for lymphocyte depletion in older patients with multiple comorbidities who present during or after SARS-CoV-2 infection with atypical symptoms that are suggestive of immunosuppression. In such instances, there should be a low threshold to start prophylactic therapy for possible opportunistic infections.


Association between Baseline Cortisol Serum Concentrations and the Effect of Prophylactic Hydrocortisone in Extremely Preterm Infants.
To define nomograms of serum cortisol values before 24 hours of postnatal life for extremely preterm infants and determine whether baseline cortisol values affect the benefit/risk ratio of prophylactic hydrocortisone to improve survival without bronchopulmonary dysplasia (BPD). We performed a predefined secondary analysis of the multicenter randomized controlled PREMILOC trial that included inborn infants delivered before 28 weeks of gestation. Nomograms of baseline serum cortisol values measured in 325 enrolled patients were determined for male and female neonates and correlated to perinatal events. BPD-free survival and severe adverse events were analyzed in placebo and hydrocortisone groups according to the cortisol z score in multivariate logistic regression models. Increased cortisol levels measured before 24 hours following birth were associated with a significantly higher chance of BPD-free survival only in placebo-treated infants (aOR [95% CI] 1.57 [1.08-2.27], P = .02) based on sex-specific nomograms for baseline cortisol levels. The cortisol z score for infants treated with prophylactic hydrocortisone predicted a risk of high-grade intraventricular hemorrhage (aOR [95% CI] 1.82 [1.06-3.15], P = .03) and spontaneous intestinal perforation (aOR [95% CI] 4.81 [1.34-17.22], P = .02). We found no predictive value of baseline cortisol levels for BPD-free survival in infants born extremely preterm treated with hydrocortisone. However, high cortisol levels early after birth were associated with a greater risk of severe intraventricular hemorrhage and spontaneous intestinal perforation in infants treated with hydrocortisone and, therefore, a lower benefit/risk ratio for the treatment. EudraCT 2007-002041-20, ClinicalTrial.gov: NCT00623740.


Loss of dipeptidyl peptidase IV immunostaining discriminates malignant melanomas from deep penetrating nevi.
The deep penetrating nevus is a rare variant of benign melanocytic nevus with histologic features mimicking vertical growth phase, nodular malignant melanoma. In this study, we expand on the search for new complementary discriminating markers by analyzing a selection of both cell cycle-related factors, such as retinoblastoma protein and phospho-retinoblastoma protein Ser795 as indicators for retinoblastoma protein activation/inactivation status, and invasion-related factors, such as matrix metalloproteinase-1, matrix metalloproteinase-2, membrane-type matrix metalloproteinase-1 and integrin beta3. MIB-1/Ki-67 was analyzed as an example for a common proliferation marker. Dipeptidyl peptidase IV/CD26 was analyzed as a marker affecting both proliferation and invasion of malignant melanocytic tumors. Semiquantitative assessment of both immunolocalization and immunoreactivity of retinoblastoma protein and phospho-retinoblastoma protein Ser795, MIB-1/Ki-67, matrix metalloproteinase-1, matrix metalloproteinase-2, membrane-type matrix metalloproteinase-1 and integrin beta3 revealed no consistent differences between deep penetrating nevi (n=14) and matched cases of nodular malignant melanomas (n=10). Matrix metalloproteinase-1 and matrix metalloproteinase-2 immunostaining of some deep penetrating nevi even exceeded that of nodular malignant melanomas. Membrane-type matrix metalloproteinase-1 expression scores of nodular malignant melanomas were higher than those of deep penetrating nevi, which was, however, not significantly discriminative. In contrast, immunostaining of dipeptidyl peptidase IV was significantly discriminative due to a consistent lack of dipeptidyl peptidase IV-expression in nodular malignant melanomas. These results add evidence that among the selected markers supposed to be relevant for melanoma progression the presence of dipeptidyl peptidase IV can be used to support diagnosis of deep penetrating nevi in doubtful cases. As loss of dipeptidyl peptidase IV may also be causally linked to the transition of invasive to metastatic phenotypes, the molecular mechanisms downstream of dipeptidyl peptidase IV deserve to be studied in more detail in future investigations.


A sensitive predictor for potential GPI lipid modification sites in fungal protein sequences and its application to genome-wide studies for Aspergillus nidulans, Candida albicans, Neurospora crassa, Saccharomyces cerevisiae and Schizosaccharomyces pombe.
The fungal transamidase complex that executes glycosylphosphatidylinositol (GPI) lipid anchoring of precursor proteins has overlapping but distinct sequence specificity compared with the animal system. Therefore, a taxon-specific prediction tool for the recognition of the C-terminal signal in fungal sequences is necessary. We have collected a learning set of fungal precursor protein sequences from the literature and fungal proteomes. Although the general four segment scheme of the recognition signal is maintained also in fungal precursors, there are taxon specificities in details. A fungal big-Pi predictor has been developed for the assessment of query sequence concordance with fungi-specific recognition signal requirements. The sensitivity of this predictor is close to 90%. The rate of false positive prediction is in the range of 0.1%. The fungal big-Pi tool successfully predicts the Gas1 mutation series described by C. Nuoffer and co-workers, and recognizes that the human PLAP C terminus is not a target for the fungal transamidase complex. Lists of potentially GPI lipid anchored proteins for five fungal proteomes have been generated and the hits have been functionally classified. The fungal big-Pi prediction WWW server as well as precursor lists are available at


African elephants interpret a trunk gesture as a clue to direction of interest.
Orienting to gaze-direction is widespread among animal species, but evidence for spontaneous use of gesture for direction is limited [1]. Remarkably, African elephants (Loxodonta africana) have been found able to follow human pointing, including subtle actions in which the contralateral hand is used, and in which the body silhouette is not broken [2,3]. The natural origin of this ability is puzzling, as the species is not reported to use trunk- or limb-gesture for showing directions [4]. One natural gesture, the 'periscope-sniff' presumed to be used to enhance olfactory sampling by an elephant in circumstances of alarm or curiosity [5], might also betray the elephant's direction of focal attention. Here we investigate what information elephants gain from seeing periscope-sniff. When one elephant in a group gave a periscope-sniff, we recorded the location and orientation of the next periscope-sniff given. Elephants that could not see the first gesturer only gestured themselves if immediately adjacent to the first or closer to the presumed stimulus of interest. In contrast, elephants able to see the first signaller's periscope-sniff were often a considerable distance behind it, further from the stimulus. Focusing on these cases, where making the periscope-sniff was apparently caused by seeing the first gesture, we found its orientation significantly matched the first, suggesting that direction information was gained from seeing the periscope-sniff. Elephants' ability to use a conspecific's periscope-sniff as if it were an ostensive pointing gesture enables them to react to the presence and location of potential dangers.


Green and black tea extracts inhibit HMG-CoA reductase and activate AMP kinase to decrease cholesterol synthesis in hepatoma cells.
Recent studies have demonstrated that green and black tea consumption can lower serum cholesterol in animals and in man, and suppression of hepatic cholesterol synthesis is suggested to contribute to this effect. To evaluate this hypothesis, we measured cholesterol synthesis in cultured rat hepatoma cells in the presence of green and black tea extracts and selected components. Green and black tea decreased cholesterol synthesis by up to 55% and 78%, respectively, as measured by a 3-h incorporation of radiolabeled acetate. Inhibition was much less evident when radiolabeled mevalonate was used, suggesting that the inhibition was mediated largely at or above the level of HMG-CoA reductase. Both extracts directly inhibited HMG-CoA reductase when added to microsomal preparations, although the extent of inhibition was considerably less than the decrease in cholesterol synthesis observed in whole cells. As HMG-CoA reductase activity also can be decreased by enzyme phosphorylation by AMP kinase, the phosphorylation state of HMG-CoA reductase and AMP kinase, which is activated by phosphorylation, was determined in lysates from cells treated with tea extracts. Both extracts increased AMP-kinase phosphorylation and HMG-CoA reductase phosphorylation by 2.5- to 4-fold, but with different time courses: maximal phosphorylation with green tea was evident within 30 min of treatment, whereas with black tea phosphorylation was slower to develop, with maximal phosphorylation occurring > or =3 hours after treatment. These results suggest that both green and black tea decrease cholesterol synthesis in whole cells by directly inhibiting HMG-CoA reductase and by promoting its inactivation by AMP kinase.


Empowering general practitioners in nutrition communication: individual-based nutrition communication strategies in Croatia.
There are a number of factors influencing individual food habits and the prerequisite for effective national strategy development is reorganization of factors aggravating or facilitating this dynamic process. The aims of this paper are to present a Croatian contextual framework influencing individual-based nutrition communication strategies and to identify important factors by general practitioners (GPs) and population. Two Croatian surveys are presented. The first one was carried out among GPs (random 425 GPs, all with office on lease) and the second on an adult population in Croatia (random 9060 respondents) answering the question: 'In the past year, has anyone advised you to change your behaviour?' The Croatian case studies showed that GPs considered smoking and alcohol as more important public health issues than unbalanced nutrition and physical activity. GPs also recognized their role in individual work with patients in secondary and tertiary prevention, less in primary prevention or work with groups and the community. Among the obstacles in individual-based communication in daily work, they reported lack of time, lack of incentives, lack of knowledge and lack of family approach in nutrition consultation. They were more likely to advise the elderly, those with lower education, unemployed, overweight, those who lived alone and those who visited GP's office regularly. Differences in respondents' answers are determined by education and workplace.


Validation of a prediction model for post-discharge nausea and vomiting after general anaesthesia in a cohort of Swedish ambulatory surgery patients.
In ambulatory surgery, post-discharge nausea and vomiting (PDNV) has been identified as a significant problem occurring in more than one-third of patients. To validate a simplified PDNV score in a Swedish population. Prospective observational study. Two county hospitals in Sweden: Sundsvall from June 2012 to May 2013 and Sunderbyn from January to October 2014. Adult patients undergoing ambulatory surgery under general anaesthesia. Postoperative outcomes with a focus on nausea and vomiting were collected at 2, 4, and 6 h after surgery and on the first three postoperative days. The simplified PDNV score, calculated before discharge, included the factors: female sex, age less than 50 years, history of postoperative nausea and vomiting, postoperative nausea and opioids given postoperatively. The prediction performance of the simplified PDNV score was evaluated in terms of discrimination (area under receiver-operating characteristics curve) and calibration plots and was compared with that of the original development study. A total of 559 patients were asked to participate, of which 431 were included in the final study cohort. The overall risk of postoperative nausea and vomiting and PDNV were 18.8 [95% confidence interval (CI), 15.4-22.8]% and 28.1 (95% CI, 24.0-32.5)%, respectively. The discrimination capacity of the simplified PDNV score in our study was similar to that of the original dataset [area under the curve 0.693 (95% CI, 0.638-0.748) vs. 0.706 (0.681-0.731), absolute difference 0.013]. The slope of the calibration curve was 0.893, with a constant of 0.021 (R-square 0.884). In a Swedish cohort of patients, the simplified PDNV score performs well in discriminating between patients who will experience post-discharge nausea and/or vomiting after ambulatory surgery. Our results indicate that the simplified PDNV score is as valid in other cohorts as it was in the original development cohort.


Necessity of D2 lymph node dissection in older patients ≥80years with gastric cancer.
This study analyzed the effect of D2 lymph node (LN) dissection on complications and survival in older patients with gastric cancer. A total of 103 octogenarian patients who underwent curative gastrectomy for gastric cancer were divided into two groups (D2 and D1) according to the extent of LN dissection and analyzed retrospectively for complications and survival. No differences were observed in short-term postoperative outcomes, including complication rates, between the two groups. In a survival analysis, D2 LN dissection did not improve overall survival (OS) in any patient, including advanced cases. A Cox regression analysis revealed that the independent risk factors for OS were history of coronary artery disease (hazard ratio [HR], 11.095), postoperative short-term complications (HR, 9.939), and TNM stage (HR, 6.299). The extent of LN dissection was not an independent risk factor for OS, and D2 or more LN dissection (odds ratio, 10.89) increased the risk independently. D2 or more LN dissection did not improve survival, but rather increased the risk of complications. Thus, LN dissection should be performed sparingly in octogenarian patients with gastric cancer.


Production of Bioactive Human PAX4 Protein from E. coli.
Paired box 4 (PAX4) is a pivotal transcription factor involved in pancreatogenesis during embryogenesis, and in adults, it is key for β-cell proliferation and survival. Additionally, PAX4 also functions as a tumor suppressor protein in human melanomas. The present study demonstrates the production of bioactive recombinant human PAX4 transcription factor. At first, the inserts (PAX4 protein-coding sequence having tags at either ends) were cloned in an expression vector to give rise to pET28a(+)-HTN-PAX4 and pET28a(+)-PAX4-NTH genetic constructs, and these were then transformed into Escherichia coli (E. coli) for their expression. The HTN-PAX4 and PAX4-NTH fusion proteins produced were purified with a yield of ~ 3.15 mg and ~ 0.83 mg, respectively, from 1.2 L E. coli culture. Further, the secondary structure retention of the PAX4 fusion proteins and their potential to internalize the mammalian cell and its nucleus was demonstrated. The bioactivity of these fusion proteins was investigated using various assays (cell migration, cell proliferation and cell cycle assays), demonstrating it to function as a tumor suppressor protein. Thus, this macromolecule can prospectively help understand the function of human PAX4 in cellular processes, disease-specific investigations and direct cellular reprogramming.


Cardiovascular and renal effects of novel nonpeptide nociceptin opioid peptide receptor agonists.
Partial agonists of the nociceptin opioid peptide (NOP) receptor have potential therapeutic use as antihypertensive and water diuretics (aquaretics). To date, peptide NOP receptor ligands have failed to progress in clinical trials due to poor pharmacokinetics and adverse effects. Nonpeptide, small-molecule NOP receptor ligands may be more suitable as therapeutic agents. This study investigated the cardiovascular and renal responses produced by the novel nonpeptide NOP agonists AT-403, AT-090, AT-127, and AT-039. Changes in mean arterial pressure (MAP), heart rate (HR), renal excretory function and occurrence of sedation and hyperphagia were determined before and after i.v. bolus injection or infusion of the NOP agonists in conscious Sprague-Dawley rats. Additional studies involving (i) measurement of renal sympathetic nerve activity (RSNA) and (ii) renal denervation were conducted to investigate the role of the renal nerves in the cardiorenal responses to AT-039. Bolus i.v. injection of AT-403, AT-090, AT-127 and AT-039 produced significant decreases in MAP and HR and a sodium-sparing diuresis. AT-403, AT-090, and AT-127, but not AT-039, induced sedation and hyperphagia at all doses tested. Infusion i.v. of AT-039 produced hypotension and aquaresis without adverse central nervous system effects or change in HR, responses that were also observed in renal denervated rats. Nonpeptide NOP agonists decrease blood pressure and produce aquaresis in conscious rodents. Due to lack of sedation and hyperphagia, AT-039 represents a novel NOP agonist that may be useful for treatment of hypertension and/or volume overload/hyponatraemic states.


Confirmation among college women: the eating disorders not otherwise specified diagnostic profile.
Previously, the researchers proposed and tested a diagnostic framework for women with eating-related concerns who seek college health and mental health treatment. The framework emphasized moderate problems characterized by frequent binging, occasional purging, and frequent exercise; rumination; body image and self-esteem concerns; ambivalence about help seeking; and developmental themes. Several questions remained about the framework's generalizability beyond women in treatment. In the current study, the authors provide new support for the framework's generalizability to college women with clinically significant eating concerns who do not seek treatment. Female students (N = 112) completed a screening instrument providing descriptive and comparative data about primary symptoms, associated features, and help seeking. Results suggested that the framework is specific to women with eating problems and does not suffer the pitfalls of applying over-generally to all college women or all female patients and clients. Implications for practice, research limitations, and remaining questions are discussed.


Elucidation of mode of retroviral-inhibitory effects of imexon through use of immune competent and severe combined immune deficiency (SCID) mice.
Mice infected with various tumor retroviruses have been used as models for evaluating therapeutic substances for the treatment of some cancers, and more recently, for human immunodeficiency virus (HIV) infection, the causative agent of acquired immune deficiency syndrome (AIDS). Consequently, there is a need to determine the ability of biological response modifiers (BRMs) to specifically reduce virus-infected cells, as compared to their non-specific anti-proliferative effects. To address this need, a BRM, imexon, was evaluated in this study using three strains of mice having different Friend virus (FV)-specific immunological capabilities. The first strain, (B10.A x A/WySn)F1, was genetically capable of producing FV-specific neutralization and cytotoxic antibodies, the second, Balb/c, was not, and the third, SCID mice, lacked functional T and B cell immunity. Imexon treatment reduced virally-induced splenomegaly in all 3 strains; however, the concentration of splenic viral infectious centers (IC) were not affected. Since imexon was efficacious in reducing splenomegaly in SCID mice, the mode of action was concluded to not require functional T or B cell immunity. The observation that imexon did not affect splenic IC titers also suggested that imexon did not specifically eliminate virally infected cells, but may have functioned by other mechanisms. This study also demonstrated the use of various mouse strains as a strategy for delineating the modes of action of BRMs against murine retroviral infections.


If a well-stabilized epileptic patient has a subtherapeutic antiepileptic drug level, should the dose be increased? A randomized prospective study.
In an attempt to determine whether the dose of an antiepileptic drug should be increased in epileptic patients who were seizure-free and had subtherapeutic serum levels, 79 patients with idiopathic generalized tonic-clonic seizures treated with monotherapy [phenytoin (PHT) or phenobarbital (PB)] and with a subtherapeutic serum level were prospectively studied. Their last seizure was at least 3 months prior to entry, and no patient had any clinical evidence of toxicity. They were randomized to study arm A (keeping the level in the subtherapeutic range) or study arm B (increasing the dose until the level reached and stayed at the therapeutic range). Over a mean follow-up period of 24 months, there was no significant difference between the two study arms in the occurrence of seizures, but arm B patients had an increased incidence of neurotoxic side effects from the dose increment. These results confirm the clinical impression that it is unnecessary to increase the dose of the antiepileptic drug despite a subtherapeutic serum concentration in a relatively well-stabilized patient, thus minimizing the frequency of dose adjustment and the need for expensive therapeutic drug monitoring.


Anticipatory postural adjustments in conditions of simulated reduced gravity.
The study investigates the role of decreased gravity on anticipatory postural adjustments (APAs). Subjects performed fast bilateral arm-raising movements and load releases while in conditions of normal and reduced gravity. Reduced gravity conditions were simulated by changing the ratio between the body weight and mass. Electromyographic (EMG) activity of dorsal and ventral trunk and leg muscles, as well as ground reaction forces, were recorded and quantified within the time intervals typical of APAs. Anticipatory postural adjustments were seen in normal gravity conditions as well as in simulated reduced gravity conditions. However, in decreased gravity conditions, the magnitudes of the anticipatory integrals of electromyography muscle activity (EMG) were smaller compared to normal gravity. Moreover, there was a linear relation between EMG and simulated decreased gravity and between the displacement of the center of pressure (COP) and simulated gravity. The study provides new data on the effect of gravity in feed-forward postural control and stresses the importance of taking into consideration its role in the control of upright posture.


Pyloric diaphragm, antral web, congenital antral membrane--a surgery rarity?
Three cases of pyloric diaphragm in elderly patients are reported. The finding of these patients within a 2-year period (1976-8) suggests that the condition is not as rare as previously thought. Diagnosis was achieved endoscopically and the condition managed surgically. While there has been controversy in the past over whether the disorder is acquired or congenital, the histological evidence of the cases presented here tends to suggest that it is a late presentation of a congenital abnormality.


Modulation by sodium ascorbate of the effect of chloroquine on low density lipoprotein retention and degradation in cultured human skin fibroblasts.
Human skin fibroblasts in culture were incubated for 48 h with 125I-labelled low density lipoprotein and chloroquine in the presence and absence of sodium ascorbate. Pretreatment of the cells for 3 days with sodium ascorbate and addition of the vitamin during incubation resulted in a decrease in cellular retention and an increase in degradation of the labelled low density lipoprotein. Similar results were obtained when the cells were pretreated for 3 days but the vitamin was not added during the final 48 h of incubation. Pretreatment of the cells with dithiothreitol, butylated hydroxy-toluene, beta-mercaptoethanol and D-alpha-tocopherol had a similar effect to that of ascorbate, i.e. reduction in low density lipoprotein retention and increase in degradation. Neither ascorbate nor the other reducing agents affected low density lipoprotein catabolism in control cells not treated with chloroquine. Sodium ascorbate pretreatment resulted also in a slight but significant alleviation of the chloroquine-induced inhibition of hydrolysis of cholesterol linoleate. It is proposed that sodium ascorbate by virtue of its reducing properties provides some protection to the intralysosomal hydrolases against the inhibitory action of chloroquine. If cholesterol accumulation in human and experimental atheroma is caused by partial inhibition of lysosomal enzymes, sodium ascorbate could play a role in the alleviation of such an inhibition.


Design, synthesis, and evaluation of potential inhibitors of nitric oxide synthase.
Selective inhibitors of neuronal nitric oxide synthase (nNOS) were shown to protect brain and may be useful in the treatment of neurodegenerative diseases. In this context, our purpose has been to design and synthesize a new family of derivatives of thiadiazoles as possible inhibitors of nNOS. To achieve it a supervised artificial neural network model has been developed for the prediction of inhibition of Nitric Oxide Synthase using a dataset of 119 nNOS inhibitors. The definition of the molecules was achieved from a not-supervised neural network using a home made program named CODES. Also, thiadiazole-based heterocycles, previously predicted, were prepared as conformationally restricted analogues of a selective nNOS inhibitor, S-ethyl N-phenylisothiourea.


Gender differences in the epidemiology of Rheumatic Fever/Rheumatic heart disease (RF/RHD) patient population of hill state of northern India; 9 years prospective hospital based, HP-RHD registry.
We report the gender-based differences in the prevalence, severity, pattern of valvular involvement, and complications in patients with Rheumatic Fever/Rheumatic heart disease (RF/RHD). The 2475 consecutive patients with RF/RHD diagnosed using clinical and echocardiographic criteria were registered prospectively from January 2011 till December 2019. The association of gender with the pattern of valvular involvement, nature, and severity of valvular dysfunction and cardiovascular complications was analyzed using a logistic regression model, and odds ratios with 95% CI were estimated. The mitral and tricuspid valve involvement was significantly lower in the male gender, odds ratio with 95% CI of 0.55 (0.44-0.61), and 0.69 (0.58-0.83) respectively, while the aortic valve was affected more frequently than females, odds ratio 1.36 (1.14-1.62). The severity of valvular disease had no significant association with gender, 0.99 (0.82-1.20). The association between gender and cardiovascular complications, heart failure, stroke, and atrial fibrillations were not statistically significant. The prevalence of RF/RHD was more than two-fold higher in female gender than male (71.4% vs. 29.6%, p < 0.0001). RF/RHD is more prevalent in females. Gender has a significant association with the pattern of valvular involvement. The severity of valvular dysfunction and cardiovascular complications had no significant association with gender.


PEG-induced molecular crowding leads to a relaxed conformation, higher thermal stability and lower catalytic efficiency of Escherichia coli β-galactosidase.
Enzymatic activities were historically assayed in dilute solutions where molecular crowding, molecular confinement and their consequences were not taken into account. Here we report how macromolecular crowding tunes catalytic parameters for the tetrameric β-Galactosidase from Escherichia coli, β-Gal. We detected increases in KM (weaker substrate binding) and a nonlinear variation in Vmax, with a minimum at 25% W/P of the crowding agent (polyethyleneglycol molecular mass 6000, PEG(6000)) resulting in a linear decrease in the catalytic efficiency (kcat/KM) within the whole [PEG(6000)] range tested). Presence of crowding agent affected β-Gal structural content and increased its thermal resistance. Steady state fluorescence and Fourier transformed infrared spectroscopic observations are compatible with crowding-induced disordering and restricted internal dynamics as a result of excluded volume and solvent structuring effects. This leads to a non-optimal substrate-binding site and a less conformationally strained protein.


Molecular Advances on Cannabinoid and Endocannabinoid Research.
Since ancient times, cannabis has been used for recreational and medical purposes [...].


Mutation of wrb, a Component of the Guided Entry of Tail-Anchored Protein Pathway, Disrupts Photoreceptor Synapse Structure and Function.
Tail-anchored (TA) proteins contain a single hydrophobic domain at the C-terminus and are posttranslationally inserted into the ER membrane via the GET (guided entry of tail-anchored proteins) pathway. The role of the GET pathway in photoreceptors is unexplored. The goal of this study was to characterize the zebrafish pinball wizard mutant, which disrupts Wrb, a core component of the GET pathway. Electroretinography, optokinetic response measurements (OKR), immunohistochemistry, and electron microscopy analyses were employed to assess ribbon synapse function, protein expression, and ultrastructure in 5-day-old zebrafish larvae. Expression of wrb was investigated with real-time qRT-PCR and in situ hybridization. Mutation of wrb abolished the OKR and greatly diminished the ERG b-wave, but not the a-wave. Ribeye and SV2 were partially mislocalized in both photoreceptors and hair cells of wrb mutants. Fewer contacts were seen between photoreceptors and bipolar cells in wrb-/- mutants. Expression of wrb was observed throughout the nervous system and Wrb localized to the ER and synaptic region of photoreceptors. Morpholino knockdown of the cytosolic ATPase trc40, which targets TA proteins to the ER, also diminished the OKR. Overexpression of wrb fully restored contrast sensitivity in mutants, while overexpression of mutant wrbR73A, which cannot bind Trc40, did not. Proteins Wrb and Trc40 are required for synaptic transmission between photoreceptors and bipolar cells, indicating that TA protein insertion by the TRC pathway is a critical step in ribbon synapse assembly and function.


Determining the impact of alternative splicing events on transcriptome dynamics.
The complete sequencing of the human genome and its subsequent analysis revealed a predominant role for alternative splicing in the generation of proteome diversity. Splice switching oligonucleotides (SSOs) are a powerful and specific tool to experimentally control alternative splicing of endogenous messenger RNAs in living cells. SSOs also have therapeutic potential to treat diseases that are caused by aberrant splicing. The assignment of biological roles to alternative splicing events of currently unknown function promises to provide a largely untapped source of potential new therapeutic targets. Here we have developed a protocol that combines high sensitivity microarrays with the transfection of SSOs to monitor global changes in gene expression downstream of alternate, endogenous splice events. When applied to a well-characterized splicing event in the Bcl-x gene, the application of high sensitivity microarrays revealed a link between the induction of the Bcl-xS isoform and the repression of genes involved in protein synthesis. The strategy introduced herein provides a useful approach to define the biological impact of any given alternative splicing event on global gene expression patterns. Furthermore, our data provide the first link between Bcl-xS expression and the repression of ribosomal protein gene expression.


The effect of manganese and ferric ions on the in vitro formation of dihydrodihydroxy metabolites of benzo(a)pyrene.
The effect of ferric and manganese ions on the in vitro metabolism of benzo(a)pyrene (BP) to dihydrodihydroxy (diol) metabolites by rat liver microsomal preparations was studied. Of the 3 diols separated by high-pressure liquid chromatography (HPLC) and called diols 1, 2 and 3 in order of elution, diol 1 was identified by its U.V. spectrum as the 9,10-diol; diols 2 and 3 have not yet been identified positively but are probably the 4,5- and 7,8-diols respectively. Higher concentrations of both metals altered the diol profile; 10 and 50 mumol Fe3+ per incubation caused the disappearance of diols 1 and 2 and an increase in diol 3; 10 mumol Mn2+ caused a significant decrease in diol 2 while 50 mumol reduced diol 2 to a negligible amount and inhibited the formation of diol 1; both concentrations caused a relative increase in diol 3. If the tentative identification of diol 3 as the 7,8-diol is correct, manganese and ferric ions could be significant in the metabolism of BP to the active metabolite, the 7,8-diol-9,10-epoxide.


Role of monovalent cations in fluid secretion from the exocrine rabbit pancreas.
The role of Na+ in fluid secretion by the isolated rabbit pancreas was investigated. The fluid secretion rate is reduced upon replacement of Na+ in the bathing medium by Li+, K+ or choline. The inhibition depends on the nature of the substituting cation, and is largest with choline. Upon replacement, the substituent cation appears in the secreted fluid, and the Na+ concentration in the secreted fluid is decreased in a mirror-like fashion. When Na+ is replaced by Li+ or choline, the secretory Na+ concentration is decreased, although less than in the bathing medium, and the K+ concentration is increased. When Na+ is replaced by K+, the Na+ and the K+ concentration in the secreted fluid are approximately equal to their bathing medium concentrations. In the Li+ and choline medium, stimulation of the pancreas by carbachol or CCK-8 increases the fluid secretion rate. In addition, it increases the Li+ or choline concentration, and decreases the Na+ and K+ concentrations in the secreted fluid. In normal and K+ medium, stimulation causes only a slight increase in fluid secretion rate, with no change in the secretory Na+ concentration. In normal medium, stimulation leads to a decrease in the secretory K+ concentration. The effects of replacing Na+ appear to be the result of a direct inhibition of the active HCO3- transport underlying secretion, and an indirect inhibition related to the permeability of the pancreas for the various cations. The stimulants are likely to act by increasing the permeability of the tight junctions.


Perception and Challenges of Time Management for Caregivers of People with Heart Failure: A Qualitative Study.
Informal caregivers contribute substantially to the self-care of people with heart failure (HF) by helping with concrete and interpersonal tasks. Time perception and management are essential issues among caregivers. However, investigators have not explored this topic in caregivers of people with HF. The aim of this study was to describe the perceptions and challenges of the time management experience among caregivers who support the self-care efforts of their relatives with HF. Adult informal caregivers of patients with HF, taking care of the patient for at least 3 months and without cognitive limitations, were recruited from Spain, Italy, and the Netherlands. Data were collected using semistructured interviews. Maryring's qualitative content analysis strategy with both a deductive and an inductive approach was used for analysis. We enrolled 50 participants (20 Italians, 19 Spanish, and 11 Dutch). Caregivers had a mean (SD) age of 62.8 (12.8) years and were mostly female (84%). They dedicated 31.2 (SD, 21.7) hours per week to providing caring activities for their patients. After extracting 33 codes from their qualitative interview data, we summarized them into 8 categories and identified 4 main themes: (1) time for yourself, (2) house management, (3) time for the patient (dedicated to directing care), and (4) time for own socialization. Caregivers navigate the complexity of time management by balancing dedicated time for supporting patients with HF and their own personal time.


Engineering of multifunctional gels integrating highly efficient growth factor delivery with endothelial cell transplantation.
Transplantation of Bcl-2-transduced human umbilical vein endothelial cells (ECs) in protein gels into the gastrocnemius muscle improves local reperfusion in immunodeficient mouse hosts with induced hind limb ischemia. We tested the hypothesis that incorporation of local, sustained growth factor delivery could enhance and accelerate this effect. Tissue engineering scaffolds often use synthetic polymers to enable controlled release of proteins, but most synthetic delivery systems have major limitations, most notably hydrophobicity and inefficient protein loading. Here, we report the development of a novel alginate-based delivery system for vascular endothelial growth factor-A(165) (VEGF) that exhibits superior loading efficiency and physical properties to previous systems in vitro. In vivo, VEGF released from alginate microparticles within protein gels was biologically active and, when combined with EC transplantation, led to increased survival of transplanted cells at 28 days. The composite graft described also improved early (14 days) tissue perfusion and late (28 days) muscle myoglobin expression, a sign of recovery from ischemia, compared with EC transplantation and VEGF delivery separately. We conclude that our improved approach to sustained VEGF delivery in tissue engineering is useful in vivo and that the integration of high efficiency protein delivery enhances the therapeutic effect of protein gel-based EC transplantation.


The effect of nerve section on the incidence and distribution of gap junctions in the odontoblast layer of the cat.
Gap junctions are numerous in the odontoblast layer of the dental pulp and may link sensory axons to odontoblasts. If these junctions do link axons and odontoblasts, they, together with the axons, should disappear after cutting the pulpal nerves centrally. Under general anesthesia the inferior alveolar nerve on one side of two young adult cats was sectioned. Under general anesthesia the animals were perfused with fixative 56 hours later and the coronal dental pulp prepared for electron microscopy. Ultrathin sections were examined from the level of the pulpal cornu and levels approximately one, two, and three mm below this. The incidence of cell processes and gap junctions was measured at different distances from the pulp predentin junction, and operated and control sides compared. The odontoblast layer at the level of the cornu differed from elsewhere in having, on the control side, a greater density of cell processes and gap junctions and in having clearly recognizable axons approaching to within 5 to 10 micron of the predentin. The only statistically significant changes after nerve section occurred in this layer and consisted of a decline in the incidence of cell processes and of gap junctions that link one cell process to another. There was no significant difference between the operated and control sides in the number of gap junctions linking cell processes to recognizable cell bodies. The odontoblast layer in the pulpal cornu contained substantial numbers of unsheathed axons, many presumably en route to the dentin. These axons may participate in gap junctions that link them to other cell processes, possibly even other axons.(ABSTRACT TRUNCATED AT 250 WORDS)


[Transient dilatation of the intra- and extra-hepatic bile ducts. A new aspect of bile duct involvement in scarlet fever].
An extra- and intra-hepatic bile duct dilatation has been observed in a child in the course of scarlet fever. Manifestations of cholestasis, cytolysis and inflammation were present. Pruritus disappeared within 2 months, biological abnormalities within 3 months and ultrasonic bile duct abnormalities between 3 and 6 months, with a follow-up of 9 months. This case report suggests a relationship between a transitory obstruction, possibly toxic in origin, of the bile ducts and the scarlet fever.


Successful treatment of methimazole-induced severe aplastic anemia by granulocyte colony-stimulating factor, methylprednisolone, and cyclosporin.
A 52-year-old Japanese woman was examined because of general malaise, weight loss and a lump in her left breast. She was diagnosed with cancer of the left breast and Graves' disease, and was administered methimazole (MMI). A left mastectomy was performed for the breast cancer. She presented with a high fever and peripheral blood examination revealed a severe pancytopenia. She was diagnosed with severe aplastic anemia, and administered G-CSF, however, the treatment was unsuccessful. Thus, oral methyprednisolone and cyclosporin were added. There was a remarkable improvement in the peripheral blood count.


Neoplastic invasion of laryngeal cartilage: reassessment of criteria for diagnosis at MR imaging.
To evaluate whether proposed diagnostic criteria applied to magnetic resonance (MR) images of patients with laryngeal and hypopharyngeal carcinoma may be used to distinguish neoplastic from inflammatory involvement of the laryngeal cartilages. The radiologic and histopathologic data in 121 consecutive patients with primary squamous cell carcinoma of the larynx (n = 63) or hypopharynx (n = 58) who underwent MR imaging before laryngectomy formed the basis of this retrospective study. Patient consent for retrospective chart review was waived by the institutional review board. All laryngectomy specimens were processed with a dedicated histopathologic whole-organ slice technique. MR images were evaluated by two readers according to established ("old") and proposed ("new") diagnostic criteria on the basis of the signal intensity behavior of cartilage on T2-weighted images and contrast material-enhanced T1-weighted images compared with that of the adjacent tumor. Specifically, with the new criteria, T2-weighted or postcontrast T1-weighted cartilage signal intensity greater than that of the adjacent tumor was considered to indicate inflammation, and signal intensity similar to that of the adjacent tumor was considered to indicate neoplastic invasion. The results of the MR image interpretation were compared with the histologic reference standard. The area under the receiver operating characteristic curve for the new criteria (0.94) was nominally but significantly larger than that for the old criteria (0.92) (P = .01). Overall specificity was significantly improved (82% for new vs 74% for old criteria, P < .001) and was greatest for the thyroid cartilage (75% for new vs 54% for old criteria, P < .001) with the new criteria. The sensitivities of the established and the proposed criteria were identical. The proposed MR imaging criteria enable improved differentiation of neoplastic cartilage invasion from peritumoral inflammation.


The ultrastructure and mechanics of elastic ligaments.
The major central part of elastic ligaments is constituted by longitudinally orientated elastic fibres, each surrounded by a sheath of spirally orientated collagen fibrils with an intervening thin layer of microfibrils. In the terminal region of such ligaments the orientation of the collagen fibrils is longitudinal and the elastic fibres end within the ligament without any attachment to bone. The ligament response to a first, unphysiologically high load produces a continuous stress-strain curve. On the other hand the curve becomes biphasic after repeated application of such loads. It is suggested that three factors are involved in this mechanical response, namely, frictional forces between elastic fibres and their sheaths, compressive forces exerted upon elastic fibres by their sheaths, and intre-sheath friction due to rearrangement of the constituent collagen fibrils.


The effect of vitamin A on renal damage following acute pyelonephritis in children.
Animal studies suggest that administration of vitamin A to rats with experimental urinary tract infection decreases the frequency of renal scars (Kavukçu et al., BJU Int 83(9):1055-1059, 1999). The aim of this study was to determine the effect of vitamin A on the rate of permanent renal damage in children with acute pyelonephritis. Fifty children, median age of 24 months (range 2-144), with first-time pyelonephritis verified by an uptake defect on acute dimercaptosuccinic acid (DMSA) scan were included in the study and randomly allocated to the case or control groups. All were given intravenous ceftriaxone for 10 days followed by oral cephalexin for 3 months. Cases in addition were given a single intramuscular dose of vitamin A, 25,000 U for infants below 1 year of age and 50,000 U for older children. At the repeat DMSA scan after 3 months, five of 25 cases (20%) and 17 of 25 controls (68%) had abnormal findings (p = 0.001). In conclusion, administration of vitamin A was associated with a significantly lower rate of permanent renal damage.


Manganese-enhanced MRI reveals multiple cellular and vascular layers in normal and degenerated retinas.
To use manganese-enhanced magnetic resonance imaging (MEMRI) at 25 × 25 × 800 μm(3) to image different retinal and vascular layers in the rat retinas. Manganese-chloride was injected intraocularly in normal (n = 5) and Royal College of Surgeons (RCS, an model of photoreceptor degeneration) (n = 5) rats at postnatal day 90. MEMRI at 4.7 T was performed 24 hours later. MRI was repeated following intravenous Gd-DTPA in the same animals to highlight the vasculatures. Layer assignment and thickness were compared to histology. MEMRI 24 hours after intravitreal manganese-chloride injection revealed seven bands of alternating hyper- and hypointensities, corresponding histologically to the ganglion cell layer, inner plexiform layer, inner nuclear layer, outer plexiform layer, outer nuclear layer, photoreceptor-segment layer, and choroidal vascular layer. Intravenous Gd-DTPA-which does not cross the blood-retinal barrier and the retinal pigment epithelium-further enhanced the two layers bounding the retina, corresponding to the retinal and choroidal vascular layers, but not the avascular outer nuclear layer and the photoreceptor-segment layer. MEMRI of the RCS retinas revealed the loss of the outer plexiform layer, outer nuclear layer, and photoreceptor-segment layer. Histological analysis corroborated the MRI laminar assignments and thicknesses. Lamina-specific retinal structures neurodegenerative changes to structure in retinal diseases can be detected using MEMRI.


Multiscale Biofabrication: Integrating Additive Manufacturing with DNA-Programmable Self-Assembly.
Structure and hierarchical organization are crucial elements of biological systems and are likely required when engineering synthetic biomaterials with life-like behavior. In this context, additive manufacturing techniques like bioprinting have become increasingly popular. However, 3D bioprinting, as well as other additive manufacturing techniques, show limited resolution, making it difficult to yield structures on the sub-cellular level. To be able to form macroscopic synthetic biological objects with structuring on this level, manufacturing techniques have to be used in conjunction with biomolecular nanotechnology. Here, a short overview of both topics and a survey of recent advances to combine additive manufacturing with microfabrication techniques and bottom-up self-assembly involving DNA, are given.


Multiple NADPH-cytochrome P450 reductases from Trypanosoma cruzi suggested role on drug resistance.
Cytochrome P450 hemoproteins (CYPs) are involved in the synthesis of endogenous compounds such as steroids, fatty acids and prostaglandins as well as in the activation and detoxification of foreign compounds including therapeutic drugs. Cytochrome P450 reductase (CPR, E.C.22.214.171.124) transfers electrons from NADPH to a number of hemoproteins such as CYPs, cytochrome c, cytochrome b5, and heme oxygenase. This work presents the complete sequences of three non-allelic CPR genes from Trypanosoma cruzi. The encoded proteins named TcCPR-A, TcCPR-B and TcCPR-C have calculated molecular masses of 68.6kDa, 78.4kDa and 71.3kDa, respectively. Deduced amino acid sequences share 11% amino acid identity, possess the conserved binding domains for FMN, FAD and NADPH and differ in the hydrophobic 27-amino acid residues of the N-terminal extension, which is absent in TcCPR-A. Every T. cruzi CPRs, TcCPR-A, TcCPR-B and TcCPR-C, were cloned and expressed in Escherichia coli. All of the recombinant enzymes reduced cytochrome c in a NADPH absolutely dependent manner with low K(m) values for this cofactor. They all were also strongly inhibited by diphenyleneiodonium, a classical flavoenzyme inhibitor. In addition, TcCPRs could support CYP activities when assayed in reconstituted systems containing rat liver microsomes. Polyclonal antiserum rose against the recombinant enzymes TcCPR-A and TcCPR-B demonstrated its presence in every T. cruzi developmental stages, with a remarkable expression of TcCPR-A in cell-cultured trypomastigotes. Overexpression of TcCPR-B in T. cruzi epimastigotes increased its resistance to the typical chemotherapeutic agents Nifurtimox and Benznidazole. We suggest a participation of TcCPR-B in the detoxification metabolism of the parasite.


Experimental methodologies and evaluations of computer-aided drug design methodologies applied to a series of 2-aminothiophene derivatives with antifungal activities.
Fifty 2-[(arylidene)amino]-4,5-cycloalkyl[b]thiophene-3-carbonitrile derivatives were screened for their in vitro antifungal activities against Candida krusei and Cryptococcus neoformans. Based on experimentally determined minimum inhibitory concentration (MIC) values, we conducted computer-aided drug design studies [molecular modelling, chemometric tools (CPCA, PCA, PLS) and QSAR-3D] that enable the prediction of three-dimensional structural characteristics that influence the antifungal activities of these derivatives. These predictions provide direction with regard to the syntheses of new derivatives with improved biological activities, which can be used as therapeutic alternatives for the treatment of fungal infections.


Monitoring long-term chemical exposome by characterizing the hair metabolome using a high-resolution mass spectrometry-based suspect screening approach.
Hair has recently emerged as a biospecimen for characterizing the long-term chemical exposome in biomonitoring investigations spanning several months, as chemical compounds circulating in the bloodstream accumulate in hair. Although there has been interest in using human hair as a biospecimen for exposome studies, it has yet to be widely adopted compared to blood and urine. Here, we applied a high-resolution mass spectrometry (HRMS)-based suspect screening strategy to characterize the long-term chemical exposome in human hair. Hair samples were collected from 70 subjects and cut into 3 cm segments, which were then mixed to prepare pooled samples. The pooled hair samples underwent a sample preparation procedure, and the hair extracts were further analyzed using an HRMS-based suspect screening approach. An in-house chemical suspect list containing 1227 chemical entries from National Report on Human Exposure to Environmental Chemicals (Report) published by the U.S. CDC and the Exposome-Explorer 3.0 database developed by the WHO was subsequently used to screen and filter the suspect features against the HRMS dataset. Overall, we matched 587 suspect features in the HRMS dataset to 246 unique chemical formulas in the suspect list, and the structures of 167 chemicals were further identified through a fragmentation analysis. Among these, chemicals such as mono-2-ethylhexyl phthalate, methyl paraben, and 1-naphthol, which have been detected in the urine or blood for exposure assessment, were also identified in human hair. This suggests that hair reflects the accumulation of environmental compounds to which an individual is exposed. Exposure to exogenous chemicals may exert adverse effects on cognitive function, and we discovered 15 chemicals in human hair that may contribute to the pathogenesis of Alzheimer's disease. This finding suggests that human hair may be a promising biospecimen for monitoring long-term exposure to multiple environmental chemicals and perturbations in endogenous chemicals in biomonitoring investigations.


Pharmacokinetics of pefloxacin in renal insufficiency.
The kinetics of pefloxacin has been studied after a single intravenous infusion of 8 mg X kg-1 in 15 male patients with various degrees of renal failure. No difference in distribution or elimination of the drug was observed between patients with mild or severe renal impairment. The mean volume of distribution (Vd area) and the mean plasma clearance were 2.03 l X kg-1 and 121.3 ml X min-1, respectively. The mean apparent elimination half-life was 13.5 h. These values are close to those observed in healthy subjects. No accumulation of the active N-desmethylmetabolite was observed in cases of severe failure as compared to mild impairment; its apparent elimination half-life was about twice that of the parent drug. The efficacy of a 4 haemodialysis in 6 additional anuric subjects done to remove pefloxacin from the body was poor.


Pediatric and adolescent gynecology for the primary care pediatrician.
This review represents the author's personal style of practice in a new and evolving field, where the standard academic framework is still in its infancy. When there is no prior established practice to follow, good clinical care needs to come from listening very carefully to patients and empirically judging appropriate treatment by actual outcomes. The information provided here that is not cited represents 14 years of careful clinical listening and observation, as well as public health training in maternal and child care and epidemiology. Health care is more effective, thereby improving its quality, if the following principles are applied consistently to the pediatric or adolescent gynecologic patient. 1. The developmental stage of the patient dictates clinical practice. 2. Confidentiality is paramount in caring for adolescents and may necessitate referral. 3. Autonomy in medical choices leads to responsible self-care.


Clavatol and patulin formation as the antagonistic principle of Aspergillus clavatonanicus, an endophytic fungus of Taxus mairei.
Many endophytic fungi are known to protect plants from plant pathogens, but the antagonistic mechanism has rarely been revealed. In this study, we wished to learn whether an endophytic Aspergillus sp., isolated from Taxus mairei, would indeed produce bioactive components, and if so whether (a) they would antagonize plant pathogenic fungi; and (b) whether this Aspergillus sp. would produce the compound also under conditions of confrontation with these fungi. The endophytic fungal strain from T. mairei was identified as Aspergillus clavatonanicus by analysis of morphological characteristics and the sequence of the internal transcribed spacers (ITS rDNA) of rDNA. When grown in surface culture, the fungus produced clavatol (2',4'-dihydroxy-3',5'-dimethylacetophenone) and patulin (2-hydroxy-3,7-dioxabicyclo [4.3.0]nona-5,9-dien-8-one), as shown by shown by NMR, MS, X-ray, and EI-MS analysis. Both exhibited inhibitory activity in vitro against several plant pathogenic fungi, i.e., Botrytis cinerea, Didymella bryoniae, Fusarium oxysporum f. sp. cucumerinum, Rhizoctonia solani, and Pythium ultimum. During confrontation with P. ultimum, A. clavatonanicus antagonized its growth of P. ultimum, and both clavatol as well as patulin were formed as the only bioactive components, albeit with different kinetics. We conclude that A. clavatonanicus produces clavatol and patulin, and that these two polyketides may be involved in the protection of T. mairei against attack by plant pathogens by this Aspergillus sp.


Mathematical models in quantitative assessment of carcinogenic risk.
Regulatory guidelines have specified default assumptions for the calculation of upper bounds on potential carcinogenic risk. These default "science policy" assumptions are used in the absence of more appropriate biological information, but can, in theory, be overridden by compound-specific data. As a screening tool, the standard LMS procedure provides a useful yardstick for regulators to compare hazard potential across compounds and across exposure scenarios. This standard methodology does not estimate actual risk, however. When specific compounds are being considered for detailed evaluation, methodology for more accurately estimating the true risk is often available. Pharmacokinetic models can be used to more accurately define the high-dose to low-dose extrapolation as well as to provide insight into across route and across species extrapolations. "More central" estimates of the upper bound on risk can be derived, or decision analysis techniques can be used to define the probability distribution of risk estimates, as the assumptions are changed. These data give the risk manager more information as to the relevance of the upper bound, and should be presented in any risk estimate to provide a richer demonstration as to the true risk.