Fat cycling in the mosquitofish (Gambusia affinis): fat storage as a reproductive adaptation.
We argue, based on reviewed literature covering reptiles, amphibians, birds, and fish, that fat storage may represent a life history adaptation because it enables an organism to shift in time when resources are allocated to reproduction. We applied these arguments to fat and population cycles in three populations of the mosquito fish, Gambusia affinis. For males, there appeared to be a constant size at maturation during the reproductive season. Mature males became scarce late in the summer. At the same time, immature males delayed maturity and attained much larger sizes; they matured in large numbers in the fall. The amount of stored fat tended to be equal for immature and mature males at all times except in the late summer. In the August samples, when mature males were relatively rare, they also had the lowest level of fat reserves. It appears that the older generation of mature males did not store fat and did not overwinter. At the same time, immature males registered a two to three fold increase in fat reserves. These differences in fat content between mature and immature males disappeared by September, probably because of the recruitment of a new generation of mature males. The reserves were gradually utilized during the winter. Females reproduced from the late spring through mid- to late-summer. They stopped reproducing in the late summer, when there was ample time to produce an additional litter of young. There was an inverse relationship between resources devoted to reproduction and fat reserves. As reproductive allotment decreased in the late summer, fat reserves increased. The magnitude of the change in fat reserves was similar to that displayed by males. The reserves were depleted over the winter. Significant reserves remained at the beginning of the reproductive season the following spring. Reproducing females utilized the remaining reserves significantly more rapidly than non-reproducing females. An analysis of resource availability revealed an overall decrease in food availability in the late summer, coincident with the increase in fat reserves. These cycles are therefore not attributable to changes in resource availability. They instead indicate a change in how resources are allocated by the fish. The trends in the data indicate that fat reserves are used to shift investment in reproduction from the late summer to the following spring. In males, deferring maturity, rather than maturing in August, allows them to store the necessary reserves to survive the winter so that they can mate the following spring. In females, a subset of the fat reserves is intended for producing the first clutch of eggs the following spring. The female pattern corresponds to those reported for a diversity of organisms. The possible advantages of shifting reproductive effort from the fall to the following spring include higher fecundity and higher offspring fitness. The limitations of the methodology and potential directions for future research are discussed.


Impact of different post-remission strategies on quality of life in patients with acute myeloid leukemia.
The impact on quality of life of allogeneic stem cell transplantation or conventional chemotherapy in patients with acute myeloid leukemia remains unclear, mainly because of a lack of studies with long-term follow-up. The German AML-Intergroup, therefore, initiated a survey on quality of life of patients with a relapse-free survival of at least 5 years after first-line treatment. The EORTC Quality of Life Core Questionnaire (QLQ-C30), supplemented by information on self-assessed concomitant diseases, late treatment effects, and demographics was used. The questionnaire was returned by 419 of 818 patients (51.2%) identified by six study groups. The patients' median age at diagnosis was 42 years, and the median follow-up period was 8 years. One hundred and seventy patients were treated with stem cell transplantation (121 allogenic, 49 autologous) in first complete remission; the other 249 patients were treated with conventional chemotherapy. The ECOG activity index revealed normal activity in 45% vs. 60% of the patients in the allogeneic stem cell transplantation vs. conventional chemotherapy groups, respectively and disabled person status in 60% vs. 35%. All QLQ-C30 functions, except physical functioning and pain, were poorer in allogeneic stem cell transplantation patients. Problems in leisure-time activities, social life, and financial management, sexual limitations and adverse effects were significantly more frequent in patients after allogeneic stem cell transplantation than after conventional chemotherapy. Multivariate logistic regression models on global health status revealed concomitant disease, age > 45 years, and allogeneic stem cell transplantation as significant risk factors. These results indicate that, compared to conventional chemotherapy, allogeneic stem cell transplantation has a significantly worse long-term impact on quality of life. This needs to be considered when treatment options are discussed.


Transcription factor substitution during the evolution of fungal ribosome regulation.
Coordinated ribosomal protein (RP) gene expression is crucial for cellular viability, but the transcriptional network controlling this regulon has only been well characterized in the yeast Saccharomyces cerevisiae. We have used whole-genome transcriptional and location profiling to establish that, in Candida albicans, the RP regulon is controlled by the Myb domain protein Tbf1 working in conjunction with Cbf1. These two factors bind both the promoters of RP genes and the rDNA locus; Tbf1 activates transcription at these loci and is essential. Orthologs of Tbf1 bind TTAGGG telomeric repeats in most eukaryotes, and TTAGGG cis-elements are present upstream of RP genes in plants and fungi, suggesting that Tbf1 was involved in both functions in ancestral eukaryotes. In all Hemiascomycetes, Rap1 substituted Tbf1 at telomeres and, in the S. cerevisiae lineage, this substitution also occurred independently at RP genes, illustrating the extreme adaptability and flexibility of transcriptional regulatory networks.


Disappearance of calcium-induced phase separation in phosphatidylserine-phosphatidylcholine membranes caused by protonation and by electric current.
Disappearance of Ca2+-induced phase separation in phosphatidylserine-phosphatidylcholine membrane has been studied under several conditions by monitoring electron spin resonance spectrum of spin-labeled phosphatidylcholine. The membranes were prepared in Millipore filters. Electron micrographs of the pre parations showed formation of multilayered structures lined on the pore surface. The phase separation was disappeared when the membrane was soaked in non-buffered salt solution (100 ml KCl, pH 5.5). It was markedly contrasting that when the bathing salt solution was buffered no disappearance was observed. Disappearance of the phase separation was also observed when the Ca2+-treated membrane was transferred to acidic salt solutions (less than or equal to pH 2.5) or to low ionic strength media (less than or equal to mM) buffered at pH 5.5, and then to the buffered salt solution (100 mM KCl, pH 5.5). These are due to replacement of Ca2+ by proton, proton-induced separation, followed by disappearance of the phase separation in the buffered salt solution. Biological significance of the competition between Ca2+ and proton for the phase separation or domain formation in the membranes was emphasized.


Crystallization and preliminary crystallographic studies of AAL-2, a novel lectin from Agrocybe aegerita that binds nonreducing terminal N-acetylglucosamine.
AAL-2 is a recently discovered lectin from the mushroom Agrocybe aegerita that specifically recognizes nonreducing terminal acetylglucosamine (GlcNAc) and that could be used as a probe in studies of protein O-linked β-N-acetylglucosamination (O-GlyNAcylation). In order to illustrate the mechanism of how this protein specifically recognizes nonreducing terminal GlcNAc and to evaluate the efficacy of AAL-2 as a macromolecular probe in O-GlyNAcylation studies, expression and crystallization studies of AAL-2 were performed and a diffraction data set was collected to 2.0 Å resolution. Preliminary crystallographic studies revealed that the AAL-2 crystals belonged to space group P2(1)2(1)2(1), with unit-cell parameters a = 52.60, b = 111.70, c = 135.97 Å.


Imaging retinal pigment epithelial proliferation secondary to PASCAL photocoagulation in vivo by polarization-sensitive optical coherence tomography.
To image the retinal pigment epithelium (RPE) after macular laser and to monitor healing responses over time in vivo in patients with diabetic maculopathy using polarization-sensitive optical coherence tomography (OCT). Prospective, nonrandomized clinical trial. In this single-center trial (Department of Ophthalmology and Optometry, Medical University of Vienna, Vienna, Austria), 13 patients (13 eyes) underwent grid photocoagulation for diabetic maculopathy. Retinal healing processes were continuously followed over the course of 3 months. A polarization-sensitive OCT prototype was used, allowing detection and measurement of the RPE changes based on their specific polarization-scrambling qualities. After 1 day, the intraretinal photocoagulation lesions were sharply demarcated, whereas RPE changes were rather subtle. At 1 week, all lesions exhibited traction of the inner retinal layers toward the RPE and loss of photoreceptor cells. In tissue-sensitive polarization-sensitive OCT imaging, polarization-scrambling columns were found at the level of the RPE. During follow-up, different healing responses were seen in the polarization-scrambling RPE layer, ranging from hyperproliferation to focal atrophy. Because of the properties of the polarization state of backscattered light, polarization-sensitive OCT revealed specific morphologic changes in the RPE and outer retinal layers secondary to retinal laser treatment undetectable with intensity-based spectral-domain OCT. The increase in polarization-scrambling tissue over the course of 3 months indicates a more intense healing reaction and proliferation of RPE cells than previously characterized in rodent studies.


Conformation affinity of cross-linked polyacrylic gels towards nucleobases, nucleosides and nucleotides.
By radical copolymerisation of monofunctional acrylic derivatives with 1,4-tetramethylene dimethacrylate, cross-linked polyacrylic gels which show a high affinity towards nucleic acid residues have been synthesised. Using these polyacrlic gels, mixtures of nucleobases, nucleosides and nucleotides can be chromatographically separated to differing extents, in some cases quantitatively. The elution of nucleobases and nucleosides from the gel in the order Cyt, Gua, Thy, Ade (or dC, dG, dT, dA and C, G, U, A, respectively) shows that Ade (dA, A) is retarded to the highest degree from the gel matrix, Cyt (dC, C) to the lowest degree. Further, the results of the separations prove that the affinity of the polyacrylic gels is much stronger towards deoxyribonucleosides than towards ribonucleosides and nucleobases. The affinity of the polyacrylic gels towards nucleobases and nucleosides depends mainly upon their tertiary structure whereas the functional side groups of the polyacrylate matrix do not contribute significantly to the interaction of the gels.


Antibacterial cotton fabric with enhanced durability prepared using silver nanoparticles and carboxymethyl chitosan.
Carboxymethyl chitosan (CMCTS) and silver nanoparticles (Ag NPs) were successfully linked onto a cotton fabric surface through a simple mist modification process. The CMCTS binder was covalently linked to the cotton fabric via esterification and the Ag NPs were tightly adhered to the fiber surface by coordination bonds with the amine groups of CMCTS. As a result, the coating of Ag NPs on the cotton fabric showed excellent antibacterial properties and laundering durability. After 50 consecutive laundering cycles, the bacterial reduction rates (BR) against both S. aureus and E. coli remained over 95%. It has potential applications in a wide variety of fields such as sportswear, socks, and medical textile.


Anticoagulation in hospitalized patients with COVID-19.
Coronavirus disease-19 (COVID-19) includes a thromboinflammatory syndrome that may manifest with microvascular and macrovascular thrombosis. Patients with COVID-19 have a higher incidence of venous thromboembolism than other hospitalized patients. Three randomized control trials suggesting benefit of therapeutic heparin in hospitalized noncritically ill patients with COVID-19 have led to conditional guideline recommendations for this treatment. By contrast, prophylactic-dose heparin is recommended for critically ill patients. Unprecedented collaboration and rapidly funded research have improved care of hospitalized patients with COVID-19.


Nd:YAG laser in treatment of rectal cancer. Are there features predicting a curative result?
In the present study we report the results of Nd:YAG laser treatment in 36 patients with rectal carcinoma in whom negative biopsies were obtained at the end of the treatment. The laser (100-W maximal power output) was applied through a flexible endoscope during 20- to 40-min sessions repeated every three days until the lesion was destroyed completely. Follow-up examinations, including endoscopy with biopsies, liver and endorectal ultrasonography and chest x-ray were performed every three months during the first year and thereafter once a year. Between 1980 and 1991, 272 patients were treated. All were unfit for surgery because of metastasis (78), recurrence after an other procedure (54), associated conditions, or old age (140). No circumferential tumors of any size were obliterated, but among the 139 patients presenting with a noncircumferential lesion less than 7 cm in diameter, negative biopsies were obtained after laser treatment in 36 patients (26%). Of these 36 patients, eight had been treated previously by surgery (5) or radiotherapy (3). Mean follow-up is 37 months (range 12-71). Recurrences were observed in four cases. Seven patients died during the study but only one death was related to the cancer (pelvic extension 19 months after treatment). Endorectal ultrasonography was performed prior to treatment in 15 patients and showed no invasion of the rectal wall deeper than the submucosa. After treatment, endorectal ultrasonography in 22 patients showed significant changes corresponding to cicatricial pattern in 60% of controlled patients.(ABSTRACT TRUNCATED AT 250 WORDS)


Studies in heterochromatin DNA: characterization of transcripts synthesized in situ from C-banded preparations.
The transcript of in situ RNA synthesis using Peromyscus eremicus cells, which had been treated with HCl followed by NaOH, proved to be rich in heterochromatin specifying sequences. Such transcript, referred to as heterochromatin RNA, can be characterized biochemically to probe the organization of heterochromatin DNA. It was found that the heterochromatin RNA of P. eremicus contained non-repetitive sequences covalently linked to repetitive sequences.


Predictive value of the second-trimester fibronectin concentration for severe preeclampsia: A prospective nested case-control study in China.
To evaluate the value of the second-trimester fibronectin concentration, alone and in combination with other markers (e.g., mean arterial pressure, inhibin A), in the identification of women who subsequently develop severe preeclampsia. For this prospective nested case-control study, serum from pregnant women (gestational age 15-22 weeks) who underwent routine Down syndrome screening was analyzed. The women were tracked to delivery and assigned to the severe preeclampsia or control group, according to whether they developed severe preeclampsia. Each woman who later developed severe preeclampsia was paired with five healthy women with pregnancies of similar gestational age (± 1 week). Fibronectin, inhibin A, human chorionic gonadotropin, placental growth factor, cysteine, and homocysteine concentrations were measured in 44 cases in the severe preeclampsia group and 220 cases in the control group. The body mass index and mean arterial pressure were calculated. All results were compared between the two groups. Logistic regression analysis and receiver operating characteristic curve construction were conducted for markers differing significantly between two groups. The second-trimester fibronectin value was positively correlated with severe preeclampsia and predicted 67.7% of severe preeclampsia cases. The combination of fibronectin, inhibin A, and mean arterial pressure predicted 76.7% of severe preeclampsia cases; predictive values for combinations of fibronectin with mean arterial pressure or inhibin A were 75.4% and 74.6%, respectively. Combination with these other markers increased the predictive value of fibronectin. In addition, fibronectin was more powerful for the late severe preeclampsia and severe preeclampsia without fetal growth restriction subgroups. The second-trimester fibronectin concentration can be used to predict severe preeclampsia.


Decreased phagocytosis of apoptotic cells in diseased SLE mice.
Antibodies against nucleosomes are a serological hallmark of systemic lupus erythematosus (SLE). Apoptotic cells are the unique source of nucleosomes, which are formed through cleavage of chromatin by nucleases. These nucleosomes and other autoantigens targeted in SLE are expressed in apoptotic blebs or at the surface of apoptotic cells. Therefore, it is conceivable that circulating antibodies can influence apoptotic cell clearance. Using an in vitro phagocytosis assay, we analysed the phagocytic efficacy for apoptotic cells of resident peritoneal macrophages from pre-morbid and diseased lupus mice. The assay was carried out in the presence of autologous serum, using autologous apoptotic thymocytes as targets. Under these conditions macrophages from diseased MRL/lpr and NZBxNZW(F1) lupus mice, and from age-matched NZB mice showed a decreased phagocytic efficacy (decrease 47%, 48% and 37%, respectively compared to measurements in pre-morbid mice). The cause of this decrease resides in the serum, and is not due to an acquired defect of macrophages. In conclusion, during disease progression in murine SLE, apoptotic cell clearance becomes impaired, which might amplify further disease progression.


Parietal area 5 neuronal activity encodes movement kinematics, not movement dynamics.
A previous study reported that proximal-arm related area 5 neurons showed continuously-graded changes in activity during unloaded arm movements in different directions (Kalaska et al. 1983), which resembled the responses of primary motor cortex cells in several respects (Georgopoulos et al. 1982). We report here that loading the arm reveals an important difference between cell activity in the two areas. Loads were continuously applied to the arm in different directions. The loads produced large continuously-graded changes in muscle activity but did not alter the handpath or joint angle changes of the arm during the movements. The activity of most area 5 cells was only weakly affected by the loads, and the overall pattern of population activity was virtually unaltered under all load conditions. This indicates that area 5 activity encodes the invariant spatial parameters (kinematics) of the movements. In contrast, many motor cortex cells showed large changes in activity during loading, and so signal the changing forces, torques or muscle activity (movement dynamics; Kalaska et al. 1989).


Genome Sequences of the Novel Porcine Parvovirus 3, Identified in Guangxi Province, China.
Porcine parvovirus 3 is a novel parvovirus that infects pigs. Here, we report two genome sequences of porcine parvovirus 3 strains GX1 and GX2, which are highly prevalent in Guangxi province. It will help in understanding the epidemiology and molecular characteristics of the porcine parvovirus 3.


A Patient-Centred Approach to Early Pregnancy Loss: The First 18 Months of a Canadian Outpatient Program for Early Pregnancy Loss (OPEL).
Manual vacuum aspiration is a safe surgical option for the management of early pregnancy loss. To provide rapid, patient-centred access to MVA, an Outpatient Program for Early pregnancy Loss ("OPEL") was established at our institution. Objectives were to (1) assess complete uterine evacuation rates; (2) assess complication rates, and (3) assess patient satisfaction with the program. With REB approval, a retrospective study was performed. Patient records from the first 18 months of OPEL (November 2015 to April 2017) were reviewed. Anonymous patient satisfaction questionnaires were completed immediately post-procedure. A total of 162 patients received treatment. Missed abortions accounted for 94 cases (58%). Median delay from referral to clinic appointment was 4.0 (interquartile range [IQR] 2.0-6.0) days. Average length of stay was 3.0 (IQR 2.5-3.0) hours. Efficacy of the procedure was 95.1%. Complication rate (immediate and delayed) was 14.2% and included intraoperative hemorrhage (3.1%; 5/162), Asherman's syndrome (1.9%; 3/162), retained products of conception requiring further treatment (4.9%; 8/162), and postoperative infection requiring antibiotic therapy (1.9%; 3/162). A total of 151 post-procedure satisfaction surveys were completed (93%); 100% agreed/strongly agreed that the nursing staff and physicians provided professional and compassionate care; 99.3% were satisfied with their care overall. Qualitative feedback was positive. Pregnant patients experiencing early pregnancy loss benefit from safe, timely, accessible, patient-centred care in the outpatient OPEL program. Similar models should be adopted nationally to ensure women experiencing this common pregnancy complication receive safe and compassionate care.


Raman microprobe spectrometer installed in a super-conducting magnet.
A Raman microprobe spectrometer that could be installed in the bore of a cryogen free super-conducting magnet (10 T) was designed and constructed for the investigation of the external magnetic field effect on the Raman spectra of molecular aggregates in solutions and at interfaces. The performance of the present instrument was demonstrated by measuring the magnetic field effect (0 - 10 T) on the resonance Raman spectra of diprotonated meso-tetra-(sulfonatophenyl)porphine aggregates in an acidic solution. The Raman shifts of the aggregates were not significantly influenced even in 10 T. However, the relative intensity of 1123 cm(-1) peak (nu(C(a)-N)) was interestingly enhanced about 20% under the magnetic fields higher than 2.5 T.


[Endangered domestic animal breeds in German-speaking countries. III. Sheep].
Sheep are mainly kept extensively. They therefore have to tolerate the local climatic conditions. By these conditions many traditional breeds still exist, those are hardy and modest enough to find an ecological niche. Because of the local occurrence the number of individuals of many breeds is rather small, so that these breeds are endangered. The hope lies on hobby-breeders, who do not exclusively follow economical reasons and present breeding aims. In the care of the countryside and landscape and in using natural grassland the importance of sheep is growing.


Osteoarthrosis following insufficiency of the cruciate ligaments in man. A clinical study.
Based on a series of 48 patients with old untreated ruptures of one or both cruciate ligaments, the development of osteoarthrotic changes was demonstrated by radiography and at operation. The changes were similar to those seen in animal experiments and developed in the same sequence. Osteophytes occurred in knees with anterior cruciate insufficiency after about 2 years; significant osteoarthrotic changes occurred after longer periods, and particularly in knees with damage to both cruciate ligaments.


Establishment of a nude mouse transplantable model of a human malignant fibrous histiocytoma of the mandible with high metastatic potential to the lung.
Malignant fibrous histiocytoma (MFH) is one of the highest-grade sarcomas arising in bone and soft tissue. Its prognosis is poor because of chemoresistance and high metastatic potential to various organs. Few cases arising of MFH of the mandible or oral cavity have been documented. We established a tumor line in nude mice (MFH-N), which was derived from human MFH of the mandible and examined the characteristics of this tumor line. Histologically, MFH-N was identical to the original tumor and showed a storiform-pleomorphic pattern, but had low metastatic potential. Immunohistochemically, both the original and xenografted tumors expressed vimentin, S-100, alpha-SMA, and histiocytic marker CD68. Lysozyme was expressed by the original tumor, but only sporadically by the xenografted tumor. RT-PCR analysis demonstrated human beta-actin in this tumor line, indicating the human origin. In a parallel experiment, we established a new MFH cell line (MFH-NC) from MFH-N. Tumor cells inoculated into the flanks and submandibular region of nude mice developed into tumors histologically similar to MFH-N and the original tumor; multiple lung metastases were detected approximately 5 months after inoculation. The expression levels of various metastasis-related molecules differed between MFH-N and MFH-NC on Western blotting. In MFH-NC, the expressions of MMP7, MMP9, MT1-MMP, CXCR4, COX-2 and integrin alpha4 were up-regulated, while those of MMP2 and TIMP1 were down-regulated. Expression of TIMP2, integrinalphaL and sialyl lewis X were not detected in either line. Our findings suggest that the MFH-N tumor line transplantable in nude mice is a useful model for studying the biological behavior of MFH.


Effect of transamniotic administration of epidermal growth factor on fetal rabbit small intestinal nutrient transport and disaccharidase development.
As fetal swallowing is documented in utero, supplementation of the ingested amniotic fluid with nutrients or hormones has been postulated as a potential prenatal treatment for intrauterine growth retardation (IUGR). To study the effect of epidermal growth factor (EGF) on the developing fetal small intestine, 12 pregnant rabbits underwent operation on day 24 of a normal 31-day gestation. Bilateral ovarian end fetuses underwent catheterization of their respective amniotic cavities with attachment to a miniosmotic pump. Study fetuses received recombinant human EGF at approximately 300 micrograms/kg/d for 1 week; controls received carrier solution only at an equivalent rate. On gestational day 31, fetuses were delivered by cesarean section and somatic measurements were recorded. The small intestine was harvested and proximal, middle, and distal regions were analyzed for lactase and maltase enzyme activity. Additionally, the uptake of radiolabeled glucose and proline was measured by a standard everted mucosal sleeve technique for each segment. Results were analyzed by Student's paired t test and reported as mean +/- SEM. Nine fetal pairs survived (75%). Small intestinal (SI) length was increased in EGF fetuses (54.8 +/- 1.9 cm) versus control (50.4 +/- 2.7 cm) (P = .02). Lactase activity, reported as UE/g protein, was significantly increased in the proximal segments in the EGF-infused fetuses; maltase was significantly increased in both the proximal and middle segments (P < .05).(ABSTRACT TRUNCATED AT 250 WORDS)


Central scotoma associated with intraocular silicone oil tamponade develops before oil removal.
Unexplained sudden visual loss after removal of silicone oil from the eye has recently been described. We report the occurrence and features of unexplained central scotoma developing with silicone oil in situ in the vitreous cavity. A retrospective case series of five patients (from two centres) who reported a central scotoma commencing during silicone oil tamponade was studied. All patients had vitrectomy for macula-on retinal detachment, with ultra-purified silicone oil tamponade (four out of five had giant retinal tear). Investigations included visual acuity, intraocular pressure, optical coherence tomography, fluorescein angiography, visual fields and electrophysiology. All patients reported a central scotoma that appeared during oil tamponade. Visual acuity fell by a mean of 0.93 LogMAR units after onset of the scotoma. After cataract extraction and oil removal, vision remained reduced by a mean of 0.8 units. The mean duration of oil in the eye was 2.7 months when the scotoma was noted by the patient. Investigations were performed after removal of oil. Fluorescein angiography (FFA) was performed in two cases and optical coherence tomography (OCT) in five patients. No abnormality was demonstrated. Electrophysiology was performed in five patients with pattern electroretinography suggestive of macular dysfunction in four patients. This is the first case series describing central scotoma associated with silicone oil in situ. Electrophysiology indicated macular dysfunction in most cases. We suggest that early removal of oil in cases with good visual potential should be considered to avoid this sight-threatening complication.


Molecular insights on the conformational dynamics of a P76C mutant of human cytochrome c and the enhancement on its peroxidase activity.
In apoptotic pathway, the interaction of Cytochrome c (Cytc) with cardiolipin in vivo is a key process to induce peroxidase activity of Cytc and trigger the release of Cytc in the inner mitochondria into cytosol. The peroxidase active form of Cytc occurs due to local conformational changes that support the opening of the heme crevice and the loss of an axial ligand between Met80 and heme Fe. Structural adjustments at the Ω-loop segments of Cytc are required for such process. To study the role of the distal Ω-loop segments comprising residues 71-85 in human Cytc (hCytc), we investigated a cysteine mutation at Pro76, one of the highly conserved residues in this loop. The effect of P76C mutant was explored by the combination of experimental characterizations and molecular dynamics (MD) simulations. The peroxidase activity of the P76C mutant was found to be significantly increased by ∼13 folds relative to the wild type. Experimental data on global denaturation, alkaline transition, heme bleaching, and spin-labeling Electron Spin Resonance were in good agreement with the enhancement of peroxidase activity. The MD results of hCytc in the hexacoordinate form suggest the important changes in P76C mutant occurred due to the unfolding at the central Ω-loop (residues 40-57), and the weakening of H-bond between Tyr67 and Met80. Whereas the experimental data implied that the P76C mutant tend to be in equilibrium between the pentacoordinate and hexacoordinate forms, the MD and experimental information are complementary and were used to support the mechanisms of peroxidase active form of hCytc.


[Heterogeneous expression of human CYP and its application in early stage of new drug research and development].
Human cytochrome P450 (CYP) has a pivotal role on metabolism of xenobiotics and endogenous substances in clinical practice. Since the CYP from human tissue is very complex, and the human tissue itself is not easy to obtain, investigators begin to use all kinds of expression system to heterogeneously express the CYP. The single CYP expressed was then used for drug metabolism and drug-drug interaction research, to improve the efficiency of high-throughput drug screening greatly. Besides, since the polymorphism of drug-metabolizing enzymes makes efficacy variance for some drugs in different population, the heterogeneous expression and drug metabolizing research of certain CYP mutants will be helpful to guide the optimization of therapeutic regimen and conduct the personalized medication.


Immediate transthoracic pacing for cardiac asystole in an emergency department setting.
This study was conducted to prospectively evaluate immediate transthoracic pacing in the emergency department for cardiac arrest patients presenting with asystole. All adult patients presenting over an 11-month period to a university teaching hospital with asystole following nontraumatic cardiopulmonary arrest received immediate transthoracic cardiac pacing. In these 48 patients, electrical capture was achieved in 23% and mechanical capture in 17%. With subsequent intraventricular administration of epinephrine and sodium bicarbonate, the percentage of responders increased to 48% and 33%, respectively. This is a statistically significant improvement in both electrical and mechanical capture rates (P less than 0.001) as compared with historical controls in whom transthoracic pacemakers were employed several minutes into the resuscitation. In mechanical responders, blood pressure never exceeded 50 mm Hg and could not be sustained for over 2 minutes. Immediate transthoracic pacing was temporarily effective at restoring myocardial electrical and mechanical activity in a substantial number of asystolic patients. Although there were no survivors, the improved electrical and mechanical capture rates with early use of transthoracic pacing is encouraging. Future studies of transthoracic pacing in the prehospital setting appear warranted.


Oblique wedge osteotomy for correction of asymmetric pectus excavatum in young children.
The procedure modified by Ravitch is usually used to correct pectus excavatum. Young children with asymmetric sternal depression need more modification of the procedure to maintain the sternum in the elevated flat position. In this report we describe a modification of the sternal elevation procedure, including an oblique wedge osteotomy through the anterior table, to correct the asymmetric sternal depression in young children.


Comparing T- and B-cell responses to COVID-19 vaccines across varied immune backgrounds.
The emergence of adapted variants of the SARS-CoV-2 virus has led to a surge in breakthrough infections worldwide. A recent analysis of immune responses in people who received inactivated vaccines has revealed that individuals with no prior infection have limited resistance to Omicron and its sub-lineages, while those with previous infections exhibit a significant amount of neutralizing antibodies and memory B cells. However, specific T-cell responses remain largely unaffected by the mutations, indicating that T-cell-mediated cellular immunity can still provide protection. Moreover, the administration of a third dose of vaccine has resulted in a marked increase in the spectrum and duration of neutralizing antibodies and memory B cells in vivo, which has enhanced resistance to emerging variants such as BA.2.75 and BA.2.12.1. These results highlight the need to consider booster immunization for previously infected individuals and the development of novel vaccination strategies. The rapid spread of adapted variants of the SARS-CoV-2 virus presents a significant challenge to global health. The findings from this study underscore the importance of tailoring vaccination strategies based on individual immune backgrounds and the potential need for booster shots to combat emerging variants. Continued research and development are crucial to discovering new immunization strategies that will effectively protect public health against the evolving virus.


Mitochondrial complex I subunit deficiency promotes pancreatic α-cell proliferation.
There is strong evidence that mitochondrial DNA mutations and mitochondrial dysfunction play a role in diabetes pathogenesis. The homozygous knock-in mtDNA mutator mouse is a model of premature aging due to the accumulation of mitochondrial DNA mutations. We used this mouse model to investigate the relationship between mitochondrial subunit expression and pancreatic islet cell composition. Quadruple immunofluorescence was used to quantify mitochondrial subunit expression (complex I and IV) and cell composition in pancreatic islets from mitochondrial DNA mutator mice (PolgAmut/mut) and control C57BL/6 mice at 12 and 44 weeks of age. Mitochondrial complex I subunit expression was decreased in islets from 12 week PolgAmut/mut mice. This complex I deficiency persisted with age and was associated with decreased insulin staining intensity at 44 weeks. Complex I deficiency was greater in α-cells compared with β-cells in islets from 44 week PolgAmut/mut mice. Islet cell composition was normal in 12 week PolgAmut/mut mice, but the β: α cell ratio was decreased in islets from 44 week PolgAmut/mut mice. This was due to an increase in α-cell number linked to an increase in α-cell proliferation. Complex I deficiency promotes α-cell proliferation and alters islet cell composition.


The role of fibrinogen plasma levels, the -455G>A fibrinogen and the factor XIII A subunit (FXIII-A) Val34Leu polymorphism in cancer-associated venous thrombosis.
Venous thromboembolism (VTE) is a life-threatening complication in cancer patients. Identification of risk factors has been in focus in the past years. Functional single nucleotide polymorphisms (SNP) of coagulation factors known to influence the concentration or function may be considered to influence the risk of VTE in cancer patients. We evaluated the influence of fibrinogen plasma levels, the -455G>A SNP in the fibrinogen beta gene and the Val34Leu (163G>T) SNP in the factor XIII A-subunit (FXIII-A) gene on the risk of VTE. In 1,079 tumour patients recruited for the prospective Vienna Cancer and Thrombosis Study (CATS) fibrinogen levels were determined by the Clauss method. The FXIII-A Val34Leu and the fibrinogen -455G>A SNPs were tested by allele-specific PCR. The median follow-up time was 604 days, 83 thrombotic events occurred. The median fibrinogen level was 381 mg/dl (25th-75th percentile: 312 to 467). In a multivariable Cox model adjusted to chemotherapy, surgery, radiotherapy, age and sex, neither the fibrinogen concentration (hazard ratio [HR] =1.05, confidence interval [CI] 0.839-1.310 p=0.68), nor the -455G>A SNP (HR=0.77, 95%CI 0.491-1.197; p=0.24), nor the Val34Leu SNP (HR=0.99, 95%CI 0.646-1.542 p=0.99) were associated with occurrence of VTE. The fibrinogen concentration was not significantly different among the fibrinogen -455G or A genotype carriers (p = 0.33). Disseminated intravascular coagulation was observed in only five patients, none of these developed VTE. In conclusion, fibrinogen plasma levels, the fibrinogen -455G>A and the FXIII-A Val34Leu polymorphisms were not associated with VTE in our study.


Regulation and function of CMTR1-dependent mRNA cap methylation.
mRNA is modified co-transcriptionally at the 5' end by the addition of an inverted guanosine cap structure which can be methylated at several positions. The mRNA cap recruits proteins involved in gene expression and identifies the transcript as being cellular or 'self' in the innate immune response. Methylation of the first transcribed nucleotide on the ribose 2'-O position is a prevalent cap modification which has roles in splicing, translation and provides protection against the innate immune response. In this review, we discuss the regulation and function of CMTR1, the first transcribed nucleotide ribose 2'-O methyltransferase, and the molecular interactions which mediate methylated 2'-O ribose function. WIREs RNA 2017, 8:e1450. doi: 10.1002/wrna.1450 For further resources related to this article, please visit the WIREs website.


The Current State of Newborn Screening in the United States.
Newborn screening has evolved since its introduction in 1963. The disorders that are being screened for continue to evolve as new treatments and new technologies advance. In this review, the authors discuss the current state of newborn screening in the United States, including the disorders currently being screened for and how newborn screening is performed. They also discuss the special considerations and limitations of newborn screening in sick and premature infants and as well as some ethical issues related to newborn screening. Finally, new disorders being considered for testing and new technologies that may be used in the future of newborn screening are discussed.


Nurse Education to Reduce Harmful Medication Use in Assisted Living Facilities: Effects of a Randomized Controlled Trial on Falls and Cognition.
Psychotropic and anticholinergic medications may increase the risk of falls and impair cognition. The aim of the study was to investigate whether educating nursing staff in assisted living facilities about harmful medication use has effects on the incidence of falls and cognition. This was a secondary analysis of a cluster randomized controlled trial (N = 227 residents, ≥65 years) in 20 wards in assisted living facilities in Helsinki, Finland. Wards were randomized to those in which staff received two 4-h interactive training sessions to recognize potentially harmful medications (intervention group) and a control group. Cognition (verbal fluency, clock-drawing test) was assessed at baseline and 6 and 12 months. The number of falls per resident over the 12-month follow-up was recorded. The prevalence of harmful medication use declined in the intervention group {-11.7% [95% confidence interval (CI) -20.5 to -2.9]; p = 0.009}, but remained constant in the control group [+3.4% (95% CI -3.7 to 10.6); p = 0.34]. There were 171 falls in the intervention group (2.25 falls/person year, 95% CI 1.93-2.62) and 259 falls in the control group (3.25 falls/person year, 95% CI 2.87-3.67) [incidence rate ratio 0.72 (95% CI 0.59-0.88); p < 0.001]. Residents in the intervention group with a Mini-Mental State Examination (MMSE) score ≥10 had significantly less falls compared with respective residents in the control group (p < 0.001). Changes in verbal fluency or clock drawing test were not significantly different between the groups. Educating nurses using activating learning methods can reduce the prevalence of harmful medications and the incidence of falls among residents in institutional settings.


Physico-chemical characteristics and fibril-forming capacity of carp swim bladder collagens and exploration of their potential bioactive peptides by in silico approaches.
This study explores the sustainable valorization of carp swim bladder by-products. The high-value molecule collagens were successfully extracted from carp swim bladder with a yield of ∼60% (dry weight basis) and characterized by sodium dodecyl sulfate polyacrylamide gel electrophoresis, UV-spectrum and Fourier transform infrared spectroscopy. The extracted collagens showed the fibril-forming ability and high denaturation temperature (38-39°C). Furthermore, the extracted collagens subjected to preparing bioactive collagen hydrolysates with potential antioxidant activities. In vitro and In silico approaches (PepDraw, BIOPEP, PeptideRanker, Pepcalc and ToxinPred) were employed to evaluate the potential of carp collagens as a potential source of bioactive peptides. Furthermore, primary structure, biological potential, physicochemical, sensory and toxicity characteristics of the theoretically release antioxidative collagen peptides were predicted. Overall, the present study highlights the carp collagens hydrolysate could be a promising precursor of bioactive peptides for developing functional food or nutraceutical products.


A new parameter for discriminating malignantly transformed cell lines from nontransformed counterparts: culture in liquid medium containing neutral protease.
Growth properties of malignantly transformed cell lines and their nontransformed counterparts were studied in the presence of a bacterial neutral protease (Dispase I). Among 10 transformed cell lines, 7 lines could proliferate in the presence of the protease in a floating state. Nontransformed cell lines and non-established cell strains failed to grow in this culture condition. The ability to grow in the presence of the protease was well correlated with their ability for anchorage-independent growth and tumorigenicity, and this provides a convenient primary test for evaluating cellular tumorigenicity.


Immunolocalization of farnesoid X receptor (FXR) in mouse tissues using tissue microarray.
Farnesoid X receptor (FXR) is a member of the nuclear receptor family and is known to play important roles in bile acid homeostasis, and lipid and glucose metabolism. In this study, to elucidate the systemic physiological functions of FXR, comprehensive immunohistochemical analysis of cell/subcellular localization of FXR and its heterodimer partner, retinoid X receptor (RXR)-alpha, in adult mice tissues was performed using tissue microarray (TMA)-based immunohistochemistry. FXR immunolabeling was observed in the enterohepatic system--including absorptive epithelium in the intestines, hepatocytes and gall bladder epithelium, several epithelial lineage cells including the basal cells of stratified epithelium in the tongue, esophagus, forestomach--skin, corneal epithelium and ciliary body epithelium in the eye and adrenocortical cells--including glandular cells in the zona reticularis/fasciculata. In these FXP-positive cells, FXR was preferentially localized to the nucleus. RXR-alpha was ubiquitously distributed in the nucleus of most cell types, including FXR-positive cell types in the examined tissues. These data suggest that FXR might have various physiological roles, not only in bile acid homeostasis, and lipid and glucose metabolism, but also in the epithelial cell barrier, visual and urinary function through multiple organ systems.


A U-shaped anterior capsulotomy and modified J-loop posterior chamber intraocular lens with new positioning holes.
The procedure used for an anterior capsulotomy is extremely important for successful in-the-bag intraocular lens insertion. An incision line in the extreme periphery may obscure the margin of the residual anterior capsule and make it difficult to confirm the position of the implanted lens. I describe a U-shaped anterior capsulotomy that facilitates in-the-bag insertion, discuss the importance of the location of the positioning holes for proper rotation, and introduce a modified J-loop posterior lens with the positioning holes in an alternative position.


Regional mobility during the Covid-19 pandemic: Analysis of trends and repercussions using mobile phones data across the EU.
The analysis of mobile phones data at regional level in the EU reveals varying patterns in mobility trends during the Covid-19 pandemic. These depend on the temporal evolution of the pandemic in each EU Member State, the measures taken at local or national level to limit the growth of the pandemic, as well as the level of urbanization and type of economic activity in each region. During the first phase of the pandemic (March- April 2020) the decrease in mobility was in general uniform among regions in the same Member State, especially in Italy, Spain and France, where national level measures were adopted. A relaxation of the measures and a resulting rebound of mobility was evident during the summer period (July- August 2020). At the same time, a shift from urban to rural areas during the summer vacation period is evident, with especially touristic areas increasing the number of movements in the same Member State. The variance in mobility trends during the second wave of the pandemic (October- November 2020) was higher, a result of the predominantly local and regional level measures applied in each Member State. Those insights suggest a certain correlation between the level of mobility and the evolution of the pandemic at regional level. The association with high levels of Covid-19 prevalence is particularly strong in urban regions with high mobility levels.


Visible light-responsive vanadium-based metal-organic framework supported pepsin with high oxidase mimic activity for food spoilage monitoring.
A photo-induced metal-organic framework-enzyme hybrid nanosystem was developed via a controllable physical embedding method that displays dual enzymatic and photo-non-enzymatic strategy which cause high stability and cascade catalytic performance to oxidation of o-phenylenediamine and generate a UV-Vis signal at 450 nm for the tracing and sensitive detection of putrescine (Put). Under optimal conditions, the present bioassay provides a wide detection range from 0.02 to 10 µM and 20-80 µM with a detection limit of 5.5 nM, which is more desirable than numerous previous reports. In addition, the established colorimetric photo-bioassay can selectively and accurately identify Put in the presence of other distributing species. The present work provides an elegant strategy to merged photo-nanozymes' and enzyme capabilities and also broadened the sensing strategies of photo-nanozymes with promising potential in the realm of cancer diagnosis and food quality monitoring as well as its potential in various bioassays and heterogeneous catalysis fields.


Selective regulation of YB-1 mRNA translation by the mTOR signaling pathway is not mediated by 4E-binding protein.
The Y-box binding protein 1 (YB-1) is a key regulator of gene expression at the level of both translation and transcription. The mode of its action on cellular events depends on its subcellular distribution and the amount in the cell. So far, the regulatory mechanisms of YB-1 synthesis have not been adequately studied. Our previous finding was that selective inhibition of YB-1 mRNA translation was caused by suppression of activity of the mTOR signaling pathway. It was suggested that this event may be mediated by phosphorylation of the 4E-binding protein (4E-BP). Here, we report that 4E-BP alone can only slightly inhibit YB-1 synthesis both in the cell and in vitro, although it essentially decreases binding of the 4F-group translation initiation factors to mRNA. With inhibited mTOR kinase, the level of mRNA binding to the eIF4F-group factors was decreased, while that to 4E-BP1 was increased, as was observed for both mTOR kinase-sensitive mRNAs and those showing low sensitivity. This suggests that selective inhibition of translation of YB-1 mRNA, and probably some other mRNAs as well, by mTOR kinase inhibitors is not mediated by the action of the 4E-binding protein upon functions of the 4F-group translation initiation factors.


Rehabilitation and nursing home admission after hospitalization in acute geriatric patients.
This study explored the effect of multimorbidity, cognitive and physical impairment, and sociodemographic factors on the choice of allocation of geriatric patients at hospital discharge. Cross-sectional study Eight hundred thirty patients 65 years or older admitted into an acute geriatric ward in Italy were evaluated (1998-2000). Social characteristics before hospitalization, multimorbidity, physical functioning, and cognitive status were related to allocation of living place after hospitalization (home, rehabilitation unit, or nursing home). Most patients were discharged to their homes (85%); 7% of the younger patients (65-74 years) and 11% of the very old (75+ years) were referred to a rehabilitation unit, while only among the very old 4% were discharged to a nursing home. Worse functional status, longer hospitalization, and being affected by dementia and cerebrovascular diseases (CVD) were associated with the admission to both rehabilitation and nursing home. Cognitive impairment and multimorbidity played a role in discharge destination, but only in functionally impaired patients. Living alone before hospitalization was correlated only with being discharged to a rehabilitation unit. In geriatric patients, both medical and sociodemographic characteristics are key factors for referral to rehabilitation or nursing home at discharge.


Trichoderma agriamazonicum sp. nov. (Hypocreaceae), a new ally in the control of phytopathogens.
The genus Trichoderma comprises more than 500 valid species and is commonly used in agriculture for the control of plant diseases. In the present study, a Trichoderma species isolated from Scleronema micranthum (Malvaceae) has been extensively characterized and the morphological and phylogenetic data support the proposition of a new fungal species herein named Trichoderma agriamazonicum. This species inhibited the mycelial growth of all the nine phytopathogens tested both by mycoparasitism and by the production of VOCs, with a highlight for the inhibition of Corynespora cassiicola and Colletotrichum spp. The VOCs produced by T. agriamazonicum were able to control Capsicum chinense fruit rot caused by Colletotrichum scovillei and no symptoms were observed after seven days of phytopathogen inoculation. GC-MS revealed the production of mainly 6-amyl-α-pyrone, 1-octen-3-ol and 3-octanone during interaction with C. scovillei in C. chinense fruit. The HLPC-MS/MS analysis allowed us to annotate trikoningin KBII, hypocrenone C, 5-hydroxy-de-O-methyllasiodiplodin and unprecedented 7-mer peptaibols and lipopeptaibols. Comparative genomic analysis of five related Trichoderma species reveals a high number of proteins shared only with T. koningiopsis, mainly the enzymes related to oxidative stress. Regarding the CAZyme composition, T. agriamazonicum is most closely related to T. atroviride. A high protein copy number related to lignin and chitin degradation is observed for all Trichoderma spp. analyzed, while the presence of licheninase GH12 was observed only in T. agriamazonicum. Genome mining analysis identified 33 biosynthetic gene clusters (BGCs) of which 27 are new or uncharacterized, and the main BGCs are related to the production of polyketides. These results demonstrate the potential of this newly described species for agriculture and biotechnology.


Defoliation increases nutritional quality and allelochemics of pine seedlings.
The concentration of phenols, procyanidins, and protein in pine foliage was determined at two and eight weeks following mechanical defoliation of 0, 25, 50, and 75%. Changes in both mature and immature ponderosa pine, Pinus ponderosa Dougl. ex Laws, foliage were monitored. The lowest level of defoliation (25%) produced the largest increase in phenols; 75% defoliation caused the largest increase in procyanidins. The number of phenolic compounds in mature foliage doubled following 25% defoliation. Maximum protein production also occurred at moderate defaliation levels. Mechanical defoliation had a significant effect on the production of all compounds tested. The implications of these results for induced resistance research are discussed.


Wilms' tumor: a new cause for urinary extravasation.
We report a case of Wilms' tumor associated with urinary extravasation due to tumor invasion through the renal pelvis and anterior renal capsule. Extravasation of urine exposed to tumor may lead to upstaging of the tumor and the requirement for more intensive therapy.


Reproductive organ transplantation: advances and controversies.
To review the advances and controversies in the field of reproductive organ transplantation. Although many cancer treatments can damage female reproductive organs, leading to temporary or permanent infertility, recent advances in cancer management have translated into improved patient survival, making preservation of reproductive function a more important priority than ever before. Reproductive organ transplantation represents a new potential treatment for patients facing infertility as a result of cancer treatment. Pregnancies have now been reported from several centers after transplanting fresh and frozen ovarian tissue pieces. The longevity of the graft and its potential long-term complications are unknown. However, it seems clear that the ovarian tissue graft has a shortened life span. Allogeneic reproductive organ transplants are under investigation. Patients must take immunosuppressive medications following heterologus transplantation, and these medications can cause adverse side effects. The effects on the fetus are largely unknown. Despite the recent biologic and clinical advances, reproductive organ transplantation remains experimental and controversial.


AuI-Catalyzed Hydroalkynylation of Haloalkynes.
The AuI-catalyzed reaction between terminal alkynes and aromatic haloalkynes proceeds through divergent pathways depending on the nature of the catalyst counteranion. Thus, cationic complexes containing strongly basic NHC ligands and noncoordinating anions such as BArF4 catalyze the cis haloalkynylation of the terminal alkyne, whereas introduction of a weakly basic triflate counteranion results in the stereoselective hydroalkynylation of the haloalkyne, yielding haloenyne products in good yields and complete trans selectivity. Experimental and computational studies suggest that the hydroalkynylation reaction takes place via nucleophilic attack of the terminal alkyne to the C2 carbon of the activated haloalkyne, assisted by a concerted proton abstraction by the triflate, and that the protodeauration is the turnover-limiting step, in agreement with an observed primary kinetic isotope effect.


Identification and transgenic analysis of a murine promoter that targets cholinergic neuron expression.
Choline acetyltransferase (ChAT) is a specific phenotypic marker of cholinergic neurons. Previous reports showed that different upstream regions of the ChAT gene are necessary for cell type-specific expression of reporter genes in cholinergic cell lines. The identity of the mouse ChAT promoter region controlling the establishment, maintenance, and plasticity of the cholinergic phenotype in vivo is not known. We characterized a promoter region of the mouse ChAT gene in transgenic mice, using beta-galactosidase (LacZ) as a reporter gene. A 3,402-bp segment from the 5'-untranslated region of the mouse ChAT gene (from -3,356 to +46, +1 being the translation initiation site) was sufficient to direct the expression of LacZ to selected neurons of the nervous system; however, it did not provide complete cholinergic specificity. A larger fragment (6,417 bp, from -6,371 to +46) of this region contains the requisite regulatory elements that restrict expression of the LacZ reporter gene only in cholinergic neurons of transgenic mice. This 6.4-kb DNA fragment encompasses 633 bp of the 5'-flanking region of the mouse vesicular acetylcholine transporter (VAChT), the entire open reading frame of the VAChT gene, contained within the first intron of the ChAT gene, and sequences upstream of the start coding sequences of the ChAT gene. This promoter will allow targeting of specific gene products to cholinergic neurons to evaluate the mechanisms of diseases characterized by dysfunction of cholinergic neurons and will be valuable in design strategies to correct those disorders.


A novel gene, Translin, encodes a recombination hotspot binding protein associated with chromosomal translocations.
We have identified a novel gene, Translin, encoding a protein which specifically binds to consensus sequences at breakpoint junctions of chromosomal translocations in many cases of lymphoid malignancies. The encoded protein, Translin, is a previously undescribed type with no significant similarity to known proteins. In the native form, Translin polypeptides form a multimeric structure which is responsible for its DNA binding activity. Nuclear localization of Translin is limited to lymphoid cell lines, raising the intriguing possibility that nuclear transport of Translin is regulated in a physiologically significant way such that active nuclear transport is associated with the lymphoid specific process known as Ig/TCR gene rearrangement.


Headache after substance abuse: a diagnostic dilemma.
An 18-year-old man inhaled a substance containing synthetic cannabinoids and 1 hour later developed a severe global headache. Imaging revealed a perimesencephalic subarachnoid haemorrhage. An angiogram suggested that a small superior cerebellar artery aneurysm was the culprit. This report discusses the, as yet undefined, relationship between "herbal highs" and intracranial haemorrhage.


Neurological prognostication after cardiac arrest and targeted temperature management 33°C versus 36°C: Results from a randomised controlled clinical trial.
The reliability of some methods of neurological prognostication after out-of-hospital cardiac arrest has been questioned since the introduction of induced hypothermia. The aim of this study was to determine whether different treatment temperatures after resuscitation affected the prognostic accuracy of clinical neurological findings and somatosensory evoked potentials (SSEP) in comatose patients. We calculated sensitivity and false positive rate for Glasgow Coma Scale motor score (GCS M), pupillary and corneal reflexes and SSEP to predict poor neurological outcome using prospective data from the Target Temperature Management after Out-of-Hospital Cardiac Arrest Trial which randomised 939 comatose survivors to treatment at either 33 °C or 36 °C. Poor outcome was defined as severe disability, vegetative state or death (Cerebral Performance Category scale 3-5) at six months. 313 patients (33%) were prognostically assessed; 168 in the 33 °C, and 145 in the 36 °C group. A GCS M ≤ 2 had a false positive rate of 19.1% to predict poor outcome due to nine false predictions. Bilaterally absent pupillary reflexes had a false positive rate of 2.1% and absent corneal reflexes had a false positive rate of 2.2% due to one false prediction in each group. The false positive rate for bilaterally absent SSEP N20-peaks was 2.6%. Bilaterally absent pupillary and corneal reflexes and absent SSEP N20-peaks were reliable markers of a poor prognosis after resuscitation from out-of-hospital cardiac arrest but low GCS M score was not. The reliability of the tests was not altered by the treatment temperature.


A calcium flux at the termination of replication triggers cell division in Escherichia coli. Hypothesis.
Cell division in Escherichia coli is coupled to chromosome replication. Even in the absence of known inducible division inhibitors, perturbations of chromosome replication affect cell division. Early studies suggested that a signal at the termination of replication might trigger subsequent division. Although later studies have suggested that fork encounter during termination is an active process involving specific termination sites and the tus protein, the coupling mechanism between termination and cell division remains to be elucidated. Recently it has been shown that the chromosome of a bacterium, Pseudomonas tabaci, contains a high proportion of calcium. E. coli maintains an intracellular concentration of free calcium identical to that of higher organisms and in dividing cells of E. coli a twenty-fold increase in the level of total calcium in the cytoplasm, a flux, occurs. In this article I propose that during the replication of the chromosome calcium entry balances calcium binding to DNA. At the termination of replication, there is a brief interval between the end of calcium binding to the chromosome and the end of calcium entry or release into the cytoplasm. During this interval the level of free calcium therefore rises. This rise may result in the observed flux by triggering the entry of calcium directly via voltage-gated calcium channels or indirectly via changes in phospholipid configurations. Mechanisms whereby these changes in calcium levels might be coupled to cell division and to a phospholipid control of the cell cycle are discussed.


Pathophysiology of autonomic dysreflexia: long-term treatment with terazosin in adult and paediatric spinal cord injury patients manifesting recurrent dysreflexic episodes.
Spinal cord injury (SCI) results in disruption of synaptic influences on the sympathetic preganglionic neurones. Remodelling of spinal cord circuits takes place in spinal neurones caudal to cord injury. There is an increased vascular alpha-adrenoceptor responsiveness, and peripheral afferent (bladder) stimulation in SCI subjects induces a marked noradrenaline spillover below the level of spinal lesion. These neurophysiological changes possibly contribute to the development of autonomic dysreflexia, a condition of sympathetic hyper-excitability that develops after cervical, or upper dorsal cord injury with resultant paroxysmal rise in arterial pressure, and provide the scientific basis for the use of terazosin, a once-a-day, selective alpha-one adrenergic blocking drug. The use of terazosin, a long-acting, alpha 1-selective blocking agent was investigated in SCI patients who developed recurrent symptoms of autonomic dysreflexia, eg headache, sweating flushing of the face together with an increase in the arterial pressure. An open, prospective study of the efficacy of terazosin in controlling recurrent autonomic dysreflexia in traumatic tetraplegic/paraplegic patients manifesting clinical features of dysreflexia in the absence of an acute precipitating cause such as a blocked catheter. The initial assessment and treatment were carried out in the Spinal Injuries Centre. Subsequently, the patients were followed-up in the community. They were monitored by telephonic interviews, follow-up visits by the patients to the hospital, and home-visits by the staff of the spinal unit. Eighteen adults with tetraplegia (female: 1; male: 17), three children with ventilator-dependent tetraplegia and three adult male patients with paraplegia who exhibited recurrent features of autonomic dysreflexia in the absence of an acute predisposing factor for dysreflexia eg performance of an invasive procedure such as cystoscopy, digital evacuation of bowels, or acute urinary retention, were enrolled in this study. After discussion with the patients and their carers, terazosin was prescribed with a starting dose of 1 mg in an adult and 0.5 mg in a child administered nocte. The patients were observed for (1) drug-induced hypotension; (2) clinical symptoms due to side effects of terazosin; and (3) continued occurrence of dysreflexic symptoms. Step-wise increments of the dose of terazosin (1 mg in case of adults, and 0.5 mg in a child) was carried out at intervals of 3-4 days, if a patient continued to develop dysreflexia but did not manifest any serious side effect. Complete subsidence of dysreflexic symptoms, or development of an adverse event necessitating termination of the terazosin therapy was the clinical end point. The dysreflexic symptoms subsided completely with the terazosin therapy in all the patients. The twenty-one adult patients required a dose varying from 1-10 mg, whereas the paediatric patients required only 1-2 mg of terazosin. The side effects of postural hypotension and drowsiness were transient, and mild. One tetraplegic patient developed persistent dizziness and therefore, the drug therapy was discontinued. In 21 adult and three paediatric spinal cord injury patients manifesting recurrent episodes of autonomic dysreflexia in the absence of an acute predisposing cause, the use of terazosin, a once-a-day, specific alpha-one blocker resulted in complete subsidence of the dysreflexic symptoms. However, one tetraplegic patient required termination of terazosin therapy because of persistent dizziness.


Infantile and adolescent idiopathic scoliosis in the same individual.
A patient with an infantile, resolving type of idiopathic scoliosis is presented. By the age of 10 years, 4 years after complete spontaneous correction, the girl had developed a structural, progressive idiopathic scoliosis to the opposite side.


Molecular cloning and analysis of the ghrelin/GHSR system in Xenopus tropicalis.
Ghrelin is a gut-derived peptide with several physiological functions, including feeding, gastrointestinal motility, and hormonal secretion. Recently, a host defense peptide, liver-expressed antimicrobial peptide-2 (LEAP2), was reported as an endogenous antagonist of growth hormone secretagogue receptor (GHS-R). The physiological relevance of the molecular LEAP2-GHS-R interaction in mammals has been explored; however, studies on non-mammals are limited. Here, we report the identification and functional characterization of ghrelin and its related molecules in Western clawed frog (Xenopus tropicalis), a known model organism. We first identified cDNA encoding X. tropicalis ghrelin and GHS-R. RT-qPCR revealed that ghrelin mRNA expression was most abundant in the stomach. GHS-R mRNA was widely distributed in the brain and peripheral tissues, and a relatively strong signal was observed in the stomach and intestine. In addition, LEAP2 was mainly expressed in intestinal tissues at higher levels than in the liver. In functional analysis, X. tropicalis ghrelin and human ghrelin induced intracellular Ca2+ mobilization with EC50 values in the low nanomolar range in CHO-K1 cells expressing X. tropicalis GHS-R. Furthermore, ghrelin-induced GHS-R activation was antagonized with IC50 values in the nanomolar range by heterologous human LEAP2. We also validated the expression of ghrelin and feeding-related factors under fasting conditions. After 2 days of fasting, no changes in ghrelin mRNA levels were observed in the stomach, but GHS-R mRNA levels were significantly increased, associated with significant downregulation of nucb2. In addition, LEAP2 upregulation was observed in the duodenum. These results provide the first evidence that LEAP2 functions as an antagonist of GHS-R in the anuran amphibian X. tropicalis. It has also been suggested that the ghrelin/GHS-R/LEAP2 system may be involved in energy homeostasis in X. tropicalis.


Targeting Aryl hydrocarbon receptor for next-generation immunotherapies: Selective modulators (SAhRMs) versus rapidly metabolized ligands (RMAhRLs).
Aryl Hydrocarbon Receptor (AhR) constitutes a major network hub of genomic and non-genomic signaling pathways, connecting host's immune cells to environmental factors. It shapes innate and adaptive immune processes to environmental stimuli with species-, cell- and tissue-type dependent specificity. Although an ever increasing number of studies has thrust AhR into the limelight as attractive target for the development of next-generation immunotherapies, concerns exist on potential safety issues associated with small molecule modulation of the receptor. Selective AhR modulators (SAhRMs) and rapidly metabolized AhR ligands (RMAhRLs) are two classes of receptor agonists that are emerging as interesting lead compounds to bypass AhR-related toxicity in favor of therapeutic effects. In this article, we discuss SAhRMs and RMAhRLs reported in literature, covering concepts underlying their definitions, specific binding modes, structure-activity relationships and AhR-mediated functions.


Comparative pharmacokinetic profiles of picrosides I and II from kutkin, Picrorhiza kurroa extract and its formulation in rats.
Picrosides I and II are the active chemical constituents, present in the roots and rhizomes of Picrorhiza kurroa Royle (family: Scrophulariaceae). The plant is ethnomedically claimed for the treatment of liver and upper respiratory tract infection, fever, dyspepsia and scorpion sting. This study attempts to determine the in vivo pharmacokinetic profile of picrosides I and II in rats after oral administration of three different preparations namely, kutkin (a mixture of picrosides I and II), P. kurroa extract and Picrolax® capsule (marketed formulation). A simple, precise, specific and sensitive method was developed for simultaneous quantification of picrosides I and II in rat plasma and was applied for the pharmacokinetic study. Pharmacokinetic parameters were calculated from the observed plasma concentration of picrosides I and II. The results showed a significant difference (p≤0.05) in oral bioavailability of picrosides I and II from different preparations. Both the compounds were found to be more bioavailable from P. kurroa extract followed by Picrolax® capsule and kutkin. Moreover, we also developed a novel method for isolation of kutkin from roots of P. kurroa with a high yield of 2.4% w/w. The information gained from this study provides a meaningful basis for clinical application and mechanistic study of such phytochemicals.


Platelet-activating factor (Paf) antagonist, WEB 2086, protects against Paf-induced hypotension in Macaca fascicularis.
1. The actions of intravenously administered platelet-activating factor (Paf) (0.1-3.33 nmol kg-1) and the effect of a recently described Paf antagonist, WEB 2086, were investigated in the anaesthetized open-chest monkey, Macaca fascicularis. 2. Paf dose-dependently reduced blood pressure, left ventricular pressure (LVP) and its first differential LV dP/dt. 3. Mean pulmonary artery pressure, recorded in three animals, was essentially unchanged by any dose of Paf. 4. WEB 2086 (0.22 mumol kg-1, i.v.) attenuated the Paf-induced changes in BP, LVP and LV dP/dt. The dose-response curve for fall in BP was shifted to the right by one order of magnitude. 5. Histamine-induced cardiovascular changes (systemic hypotension and tachycardia) were not affected by prior administration of WEB 2086. 6. WEB 2086 should be of value in assessing the role of Paf in pathophysiological conditions.


Evaluation of patients with CNS involvement in SLE.
Patients with systemic lupus erythematosus (SLE) may present with a wide array of neuropsychiatric (NP) clinical features. This may either be a primary manifestation of SLE, the result of a complication of the disease or its therapy, or a concurrent disease process. As there is no single diagnostic gold standard for NP-SLE, the assessment of individual patients is heavily dependent upon clinical evaluation in addition to information from studies of autoantibodies, brain structure and function. Despite their lack of diagnostic sensitivity and specificity, these tests frequently provide information that can be used to support or refute the clinical impression. They may also provide a basis for the prospective evaluation of the efficacy of therapeutic interventions in individual patients with NP-SLE.


Quality over quantity? The role of social contact frequency and closeness in suicidal ideation and attempt.
Social support has been identified as a protective factor against suicidal thoughts and behaviors. Research has not conclusively identified the component of social support most implicated in suicidal thoughts and behaviors: (1) frequency of social contact or (2) closeness of relationships. This study examined the relationships between these facets of social support and suicidal thoughts and behaviors in two nationally representative samples, as well as subsamples with social anxiety disorder (SAD). Study 1 variables for lifetime and past-year suicide ideation and attempt, social contact frequency, and closeness were calculated and examined within the National Comorbidity Survey-Replication (NCS-R). Study 2 examined the independent contributions of social contact frequency and closeness to only lifetime suicide attempt in the National Epidemiologic Survey on Alcohol and Related Conditions-III (NESARC-III). In the NCS-R, lower social closeness but not contact frequency was uniquely associated with suicidal ideation and attempt in the general sample and those with SAD. In the NESARC-III, both components of social support were associated with lifetime suicide attempt in the general sample, while only social closeness was uniquely associated with suicide attempt in the SAD subsample. This study utilized cross-sectional data and was limited in the validity and specificity of the variables assessed. Lower social closeness was more strongly associated with suicidality than social contact frequency and merits attention as a potential target for suicide-related interventions. Social closeness may be especially relevant in populations experiencing high rates of suicidal thoughts and behaviors and decreased social support.


Contribution of the Semiological Approach to Deixis-Anaphora in Sign Language: The Key Role of Eye-Gaze.
We address the issue of deixis-anaphora in sign language (SL) discourse, focusing on the role of eye-gaze. According to the Semiological Approach, SL structuring stems from a maximum exploitation of the visuo-gestural modality, which results in two modes of meaning production, depending on the signer's semiotic intent. Involving both non-manual and manual parameters, the first mode, expressing the intent to say while showing, uses constructions based on structures, the termed "transfer structures." The second one, expressing the intent to say without showing, involves lexical, pointing and fingerspelling units. In order to situate our descriptive concepts with respect to those used by SL linguists who, like us, adopt a cognitive-functionalist perspective, we expose a specific theoretical foundation of our approach, the "enunciation theories." The concept of "enunciation" is decisive for understanding the role of eye-gaze, as being at the foundation of deixis and the key vector of referential creation and tracking in SL discourse. "Enunciation" entails the opposition between "Enunciation" and "Utterance" Domains. The first links, as co-enunciators, the signer/speaker and his/her addressee, establishing them by the very "act of enunciation" as 1st and 2nd person. The second is internal to the discourse produced. Grounding on corpora of narratives in several SLs (some with no historical link), we illustrate this crucial role of eye-gaze and the diversity of functions it fulfills. Our analyses, carried out in this perspective, attest to the multiple structural similarities between SLs, particularly with regard to transfer structures. This result strongly supports the typological hypothesis underlying our approach, namely, that these structures are common to all SLs. We thus show that an enunciative analysis, based on the key role of eye-gaze in these visual languages that are SLs, is able to give the simplest account of their own linguistic economy and, in particular, of deixis-anaphora in these languages.


Investigation into the species-specific deacylation of penta-acetyl-gitoxin.
Penta-acetyl-gitoxin (PAG) shows species-specific deacylation to 16-acetyl-gitoxin (16 AG; I and III) or gitoxin (II and IV) by homogenates of liver and intestinal mucosa of man (I), rabbit (II), guinea-pig (III) and rat (IV), whereas it is degraded into tri- and tetra-acetates by homogenates of guinea-pig myocardium as well as by human blood and serum. The identity of the principal and chloroform-extractable metabolities in human urine after PAG administration with 16-AG has been demonstrated by mass spectrometry.


Rapidly Progressive and Catastrophic Calciphylaxis After Mechanical Valve Replacement: An Anticoagulation Dilemma.
Warfarin is the only approved anticoagulant after mechanical valve replacement, but it is a well described risk factor for calciphylaxis among patients with end-stage kidney disease. Our patient with end-stage kidney disease rapidly developed calciphylaxis after dual mechanical valve replacement in association with warfarin initiation, posing significant challenges in clinical management and a fatal outcome. (Level of Difficulty: Intermediate.).


The Cytosolic NADPH Oxidase Subunit NoxO1 Promotes an Endothelial Stalk Cell Phenotype.
Reactive oxygen species generated by nicotinamide adenine dinucleotide phosphate (NADPH) oxidases contribute to angiogenesis and vascular repair. NADPH oxidase organizer 1 (NoxO1) is a cytosolic protein facilitating assembly of constitutively active NADPH oxidases. We speculate that NoxO1 also contributes to basal reactive oxygen species formation in the vascular system and thus modulates angiogenesis. A NoxO1 knockout mouse was generated, and angiogenesis was studied in cultured cells and in vivo. Angiogenesis of the developing retina and after femoral artery ligation was increased in NoxO1(-/-) when compared with wild-type animals. Spheroid outgrowth assays revealed greater angiogenic capacity of NoxO1(-/-) lung endothelial cells (LECs) and a more tip-cell-like phenotype than wild-type LECs. Usually signaling by the Notch pathway switches endothelial cells from a tip into a stalk cell phenotype. NoxO1(-/-) LECs exhibited attenuated Notch signaling as a consequence of an attenuated release of the Notch intracellular domain on ligand stimulation. This release is mediated by proteolytic cleavage involving the α-secretase ADAM17. For maximal activity, ADAM17 has to be oxidized, and overexpression of NoxO1 promoted this mode of activation. Moreover, the activity of ADAM17 was reduced in NoxO1(-/-) LECs when compared with wild-type LECs. NoxO1 stimulates α-secretase activity probably through reactive oxygen species-mediated oxidation. Deletion of NoxO1 attenuates Notch signaling and thereby promotes a tip-cell phenotype that results in increased angiogenesis.


Dendritic cells from HIV-1 infected individuals are less responsive to toll-like receptor (TLR) ligands.
We compared TLR responsiveness in PBMC from HIV-1-infected and uninfected individuals using the TLR agonists: TLR7 (3M-001), TLR8 (3M-002), and TLR7/8 (3M-011). Activation and maturation of plasmacytoid dendritic cells (pDC) were measured by evaluating CD86, CD40, and CD83 expression and myeloid dendritic cell (mDC) activation was measured by evaluating CD40 expression. All agonists tested induced activation and maturation of pDC in PBMC cultures of cells from HIV+ and HIV- individuals. The TLR7 agonist induced significantly less pDC maturation in cells from HIV+ individuals. Quantitative assessment of secreted IFN-alpha and pro-inflammatory cytokines at the single cell level showed that pDC from HIV+ individuals stimulated with TLR7 and TLR7/8 induced IFN-alpha. TLR8 and TLR7/8 agonists induced IL-12 and COX-2 expression in mDC from HIV+ and HIV- individuals. Understanding pDC and mDC activation and maturation in HIV-1 infection could lead to more rational development of immunotherapeutic strategies to stimulate the adaptive immune response to HIV-1.


Comparing the separation performance of poly(ethyleneimine) embedded butyric and octanoic acid based chromatographic stationary phases.
Poly(ethyleneimine) (PEI) modified silica spheres were used to graft butyric acid and octanoic acid onto their surfaces, forming two stationary phases named Sil-PEI-BAD and Sil-PEI-CAD, respectively. Characterized methods including fourier transform infrared spectroscopy (FT-IR), elemental analysis (EA) and thermogravimetric analysis (TGA) were utilized to determine the successful synthesis of these two stationary phase materials. The chromatographic performance of these two stationary phases was analyzed with hydrophobic and hydrophilic compounds as analytes. Compared with Sil-PEI-CAD column, Sil-PEI-BAD column was more effective in separating hydrophilic compounds including nucleosides, alkaloids and vitamins. Hydrophobic substances including polycyclic aromatic hydrocarbons (PAHs) and alkylbenzenes obtained excellent separation results on Sil-PEI-CAD column than Sil-PEI-BAD column. Additionally, according to the separation of phenols, Sil-PEI-CAD column can be used in HILIC/RPLC mixed-mode. The results showed that the properties and retention mechanisms of the prepared stationary phases depended on the length of the alkyl chains bonded on the silica surface.


The effect of low dietary inflammatory index score formula on inflammatory, metabolic, and clinical outcomes in critically ill traumatic brain injury patients: A single-blind randomized controlled pilot study.
In traumatic brain injury (TBI) patients, a complex cascade of inflammatory responses are frequently observed following trauma. Numerous dietary agents have long been found to have potential in modulating inflammatory responses. This pilot study, designed an enteral formula with low inflammatory properties based on the dietary inflammatory index (DII®) and evaluated its effect on inflammatory and metabolic factors in critically ill TBI patients. This single-blind randomized controlled pilot study was conducted at the Neurosurgical ICU of Shahid Kamyab Hospital (Mashhad, Iran). A total of 20 TBI patients were randomly assigned to receive either low-DII score or standard formula at the intensive care unit (ICU). The primary outcomes of the study included clinical status, inflammatory biomarkers, APACHE II, SAPS II, SOFA, and NUTRIC scores. The trial groups did not differ significantly in baseline values. Following 14 days of intervention, there was a statistically significant decrease in the APACHE II, SAPS II, and NUTRIC scores and a significant increase in the GCS score in the low-DII score formula group compared to the standard formula group. Over 2 weeks, high sensitivity C-reactive protein (hs-CRP) values of -2.73 (95% CI: -3.67, -1.79) mg/dL in the low-DII score formula group versus 0.65 (95% CI: -0.29, 1.58) mg/dL in controls were obtained. Moreover, the length of hospital stay was longer for the standard formula group than for the low-DII score formula group. The low-DII score formula improves inflammatory factors (serum hs-CRP) and metabolic biomarkers (LDL-c and FBS). Furthermore, clinical outcomes, including the length of hospital stay and disease severity, appear to be enhanced.


Disparities in gastric cancer screening among people with disabilities: a national registry-linkage study in South Korea.
Using the national disability registration linked to the cancer screening database in Korea, we examined (1) trends in the gastric cancer screening rate among people with disabilities over time, and (2) whether gastric cancer screening participation and modalities differed according to presence, severity, and type of disability. We examined gastric cancer screening participation rates among individuals with registered disability, from 2006 to 2015. The age- and sex-adjusted rate for gastric cancer screening in people with disabilities increased from 25.9% in 2006 to 51.9% in 2015 (change: + 26.0%). During the same period, screening rates among people without disability improved from 24.7 to 56.5% (change: + 31.8%). Disability was associated with a screening rate [adjusted odds ratio (aOR) 0.89, 95% confidence interval (CI), 0.88-0.89]. Screening rates were markedly lower among people with severe disabilities (aOR 0.58, 95% CI 0.57-0.58) and people with autism (aOR 0.36, 95% CI 0.25-0.52), renal failure (aOR 0.39, 95% CI 0.38-0.39), brain injury (aOR 0.41, 95% CI 0.40-0.41), ostomy problems (aOR 0.53, 95% CI 0.51-0.55), intellectual disabilities (aOR 0.54, 95% CI 0.53-0.54), or mental disorders (aOR 0.55, 95% CI 0.54-0.56). The use of gastroscopy as the initial screening modality in people with disabilities was lower than in people without a disability. In spite of the availability of national gastric cancer screening program, we found significant disparities in gastric cancer screening participation, especially among people with severe disabilities and those with renal failure or brain-related/mental disabilities.


Salt and water homeostasis during oral rehydration therapy.
Changes in sodium balance and urinary and stool output during orally administered rehydration therapy were studied in 22 well-nourished Turkish infants, aged 2 to 13 months, with acute diarrhea mainly of viral origin. The infants randomly received a rehydration solution containing either 90 mmol Na/L (ORS90) or 40 mmol Na/L (ORS40). Slight transient hypernatremia was noted in a few infants receiving ORS90, and slight transient hyponatremia in a few infants receiving ORS40. In both groups, sodium balance increased most rapidly during the first 12 hours of rehydration, and then more slowly because of increased urinary as well as stool sodium output. Sodium balance was always more positive after ORS90 than after ORS40, but the difference did not change much from 12 to 36 hours after therapy was started. Changes in fractional sodium excretion, urinary K/Na quotient, and urinary aldosterone-creatinine quotient were used as indexes of changes in sodium balance. All values were interpreted to indicate that the sodium deficit on admission was corrected within 12 to 18 hours after ORS90 and, in most cases, after 24 to 36 hours after ORS40. Both groups of infants responded well to orally administered rehydration therapy from the clinical viewpoint.


15-deoxy-Δ12,14-prostaglandin J2 relieved acute liver injury by inhibiting macrophage migration inhibitory factor expression via PPARγ in hepatocyte.
15-deoxy-Δ12,14-prostaglandin J2 (15d-PGJ2) exhibited potential to alleviate liver inflammation in chronic injury but was less studied in acute injury. Acute liver injury was associated with elevated macrophage migration inhibitory factor (MIF) levels in damaged hepatocytes. This study aimed to investigate the regulatory mechanism of hepatocyte-derived MIF by 15d-PGJ2 and its subsequent impact on acute liver injury. In vivo, mouse models were established by carbon tetrachloride (CCl4) intraperitoneal injection, with or without 15d-PGJ2 administration. 15d-PGJ2 treatment reduced the necrotic areas induced by CCl4. In the same mouse model constructed using enhanced green fluorescent protein (EGFP)-labeled bone marrow (BM) chimeric mice, 15d-PGJ2 reduced CCl4 induced BM-derived macrophage (BMM, EGFP+F4/80+) infiltration and inflammatory cytokine expression. Additionally, 15d-PGJ2 down-regulated liver and serum MIF levels; liver MIF expression was positively correlated with BMM percentage and inflammatory cytokine expression. In vitro, 15d-PGJ2 inhibited Mif expression in hepatocytes. In primary hepatocytes, reactive oxygen species inhibitor (NAC) showed no effect on MIF inhibition by 15d-PGJ2; PPARγ inhibitor (GW9662) abolished 15d-PGJ2 suppressed MIF expression and antagonists (troglitazone, ciglitazone) mimicked its function. In Pparg silenced AML12 cells, the suppression of MIF by 15d-PGJ2 was weakened; 15d-PGJ2 promoted PPARγ activation in AML 12 cells and primary hepatocytes. Furthermore, the conditioned medium of recombinant MIF- and lipopolysaccharide-treated AML12 respectively promoted BMM migration and inflammatory cytokine expression. Conditioned medium of 15d-PGJ2- or siMif-treated injured AML12 suppressed these effects. Collectively, 15d-PGJ2 activated PPARγ to suppress MIF expression in injured hepatocytes, reducing BMM infiltration and pro-inflammatory activation, ultimately alleviating acute liver injury.


Preclinical toxicology of pirmenol hydrochloride.
Pirmenol hydrochloride, a novel pyridinemethanol derivative, is a long-acting class Ia antiarrhythmic agent. Preclinical toxicology data were obtained in rat, mouse, dog and rabbit. In acute toxicity studies by oral and intravenous routes, no pathologic changes were observed in surviving mice, rats and dogs. In repeated dose toxicity studies, no drug-related pathologic changes were evident; dryness of the oral mucosa in dogs and body weight gain reductions in rodents were the only significant clinical signs. In a chronic (52 week) toxicity study in rats, pirmenol given in the diet was tolerated clinically at doses up to 100 mg/kg/day. No drug-related aberrations in clinical laboratory parameters or ophthalmic or pathologic findings were evident. In a similar study in beagle dogs, pirmenol was tolerated clinically at a dosage up to 30 mg/kg/day. No significant changes in biochemical, hematologic, urinary or bone marrow determinations were found in either species. In reproductive toxicology studies in rats, pirmenol had no significant effect on litter size or embryonic viability. In rabbits pirmenol had no effect on average litter size, embryonic viability or fetal wastage. Given to male rats, pirmenol had no overt effects on fertility. Pirmenol failed to elicit deoxyribonucleic acid damage or induce cytogenetic alterations. Pirmenol appears to be without significant limiting toxicologic properties.


A question of professionalism: leading forward the surgical team.
Surgical practice must be driven by wise professional judgements, involving complex deliberation, in the interests of ensuring the safe care of patients. Large amounts of energy are now being concentrated in training doctors in skills (competencies) in the belief that this will reduce risk to patients. Little thought is being given to developing in the young, wise professional judgement which is at the heart of being a good doctor and a good surgeon.


Chemo-enzymatic functionalized sustainable cellulosic membranes: Impact of regional selectivity on ions capture and antifouling behavior.
Most of the polymeric membranes synthesized for decentralization of polluted water use fossil-based components. Thus, there is an urgent need to create robust and tunable nano/micro materials for confidently designing efficient and selective polymeric water filters with guaranteed sustainability. We have chosen a robust high-grade microfibrillated cellulose (MFC) as the functional material and selectively tuned it via enzymatic catalysis, which led to the attachment of phosphate group at the C6 position, followed by esterification (fatty acid attachment at C2 and C3 carbon), which led to the increase in its antifouling properties. We have demonstrated the robustness of the functionalization by measuring the separation of various metal ions, and the antifouling properties by adding foulants, such as Bovine Serum Albumin (BSA) and cancerous cells to the test solutions. These prototype affinity MFC membranes represent the most promising type of next-generation high-performance filtration devices for a more sustainable society.


Expression and function of the bile acid receptor TGR5 in Kupffer cells.
Kupffer cells are resident macrophages in the liver and play a central role in the hepatic response to injury. Bile acids can impair macrophage function leading to decreased cytokine release. TGR5 is a novel, membrane-bound bile acid receptor, and it has been suggested that the immunosuppressive effect of bile acids can be mediated by TGR5. However, the function of TGR5 in Kupffer cells has not been studied and a direct link between TGR5 and cytokine production in macrophages has not been established. The present study demonstrates that TGR5 is localized in the plasma membrane of isolated Kupffer cells and is responsive to bile acids. Furthermore, bile acids inhibited LPS-induced cytokine expression in Kupffer cells via TGR5-cAMP dependent pathways. TGR5-immunoreactivity in Kupffer cells was increased in rat livers following bile-duct ligation, suggesting that TGR5 may play a protective role in obstructive cholestasis preventing excessive cytokine production thereby reducing liver injury.


Treatment of caffeine toxicity with esmolol.
We present the case of a 58-year-old woman who ingested more than 35 g of caffeine in a suicide attempt. She manifested all the clinical symptoms and signs of caffeine toxicity. Esmolol, an ultrashort-acting beta 1-selective antagonist, controlled her multiple dysrhythmias and symptoms of sympathetic nervous system hyperstimulation. We suggest the use of esmolol for treatment of dysrhythmias secondary to caffeine toxicity; to the best of our knowledge, the use of esmolol has not been reported for this purpose.


Clinical significance of local control of primary tumour in definitive radiotherapy for scalp angiosarcomas.
Scalp angiosarcoma is a rare and aggressive cancer. Definitive radiotherapy is a treatment option for localised scalp angiosarcoma patients. Although definitive surgical resection reportedly prolongs overall survival (OS), whether initial local treatment effect affects OS when definitive radiotherapy is administered is unclear. Therefore, this study analysed whether local recurrence within 6 months of irradiation correlates with OS and cancer-specific survival (CSS). Furthermore, how local control affects patients' quality of life was investigated. Thirty-one localised scalp angiosarcoma patients who had received definitive radiotherapy at our institution between October 2010 and July 2021 were analysed retrospectively. The most commonly used dose fractionation was 70 Gy in 35 fractions (83.9%). Local recurrence within 6 months of radiotherapy and other clinical factors were examined in univariate and subsequent multivariate analyses for correlation with OS and CSS. The median follow-up period was 16 months (range, 6-45 months). Local recurrence was detected in 16 patients (51.6%), 12 of whom had recurrence within 6 months. In multivariate analyses, the presence of local recurrence within 6 months of radiotherapy was significantly associated with OS and CSS (p = 0.003, 0.0001, respectively). Ten of the 16 patients with local recurrence had severe symptoms such as bleeding, pain, difficulty opening the eye and malodour. The initial local treatment effect was significantly associated with OS and CSS after definitive radiotherapy. Furthermore, local recurrence after radiotherapy resulted in a variety of symptoms, including bleeding and pain, which reduced the patient's quality of life.


High aspect ratio diamond nanosecond laser machining.
Laser processing of diamond has become an important technique for fabricating next generation microelectronic and quantum devices. However, the realization of low taper, high aspect ratio structures in diamond remains a challenge. We demonstrate the effects of pulse energy, pulse number and irradiation profile on the achievable aspect ratio with 532 nm nanosecond laser machining. Strong and gentle ablation regimes were observed using percussion hole drilling of type Ib HPHT diamond. Under percussion hole drilling a maximum aspect ratio of 22:1 was achieved with 10,000 pulses. To reach aspect ratios on average 40:1 and up to 66:1, rotary assisted drilling was employed using > 2 M pulse accumulations. We additionally demonstrate methods of obtaining 0.1° taper angles via ramped pulse energy machining in 10:1 aspect ratio tubes. Finally, effects of laser induced damage are studied using confocal Raman spectroscopy with observation of up to 36% increase in tensile strain following strong laser irradiation. However, we report that upon application of 600 °C heat treatment, induced strain is reduced by up to ~ 50% with considerable homogenization of observed strain. The online version contains supplementary material available at 10.1007/s00339-023-06755-2.


Protein insertion into the endoplasmic reticulum of permeabilized cells.
We have established efficient translocation of newly synthesized proteins into the endoplasmic reticulum of permeabilized Mel Juso cells. By site-specific photo-crosslinking we show that translocating polypeptide chains contact the same components of permeabilized cells ER as in dog pancreas rough microsomes. This cellular assay system has the potential to overcome the limitations of isolated microsomes in investigating the molecular environment of a newly synthesized protein after they have left the ER translocation site.