Delusional infestation may be caused by recreational drug usage in some patients, but they may not disclose their habit.
Patients with delusional infestation (DI) believe that their skin is infested with organisms or inanimate particles, despite objective evidence to the contrary. Previous studies indicate a higher rate of recreational drug use among patients with DI. It is known that symptoms of infestation such as formication can be secondary to use of recreational drugs and alcohol. We carried out a pilot study observing the prevalence of recreational drug use in patients with DI. To observe the prevalence of recreational drug use in patients with DI. A retrospective review of 86 consecutive patient records was carried out over a 3-year period (1 March 2012 to 1 March 2015) to determine current or previous recreational drug use. Additionally, 24 consecutive patients with DI attending clinic between 1 December 2014 and 1 March 2015 were prospectively offered a urine test for recreational drugs. Of the 86 patients, 19 had recreational drug use documented in their medical notes (22.1%). Of the 24 patients offered a urine test, 18 accepted (75%); the remaining 6 patients agreed to provide a urine sample immediately after consultation, but did not do so. Of the 18 samples collected, 6 were positive (33.3% of completed tests) for recreational drugs. Cannabis was the most commonly used drug. This study indicates that the prevalence of recreational drug use in patients with DI is high, and that patients do not always disclose their habit. It is hoped that this pilot study will lead to further research into this finding.


Linguistic Stereotyping in Older Adults' Perceptions of Health Care Aides.
The cultural and linguistic diversity of the U.S. health care provider workforce is expanding. Diversity among health care personnel such as paraprofessional health care assistants (HCAs)-many of whom are immigrants-means that intimate, high-stakes cross-cultural and cross-linguistic contact characterizes many health interactions. In particular, nonmainstream HCAs may face negative patient expectations because of patients' language stereotypes. In other contexts, reverse linguistic stereotyping has been shown to result in negative speaker evaluations and even reduced listening comprehension quite independently of the actual language performance of the speaker. The present study extends the language and attitude paradigm to older adults' perceptions of HCAs. Listeners heard the identical speaker of Standard American English as they watched interactions between an HCA and an older patient. Ethnolinguistic identities-either an Anglo native speaker of English or a Mexican nonnative speaker-were ascribed to HCAs by means of fabricated personnel files. Dependent variables included measures of perceived HCA language proficiency, personal characteristics, and professional competence, as well as listeners' comprehension of a health message delivered by the putative HCA. For most of these outcomes, moderate effect sizes were found such that the HCA with an ascribed Anglo identity-relative to the Mexican guise-was judged more proficient in English, socially superior, interpersonally more attractive, more dynamic, and a more satisfactory home health aide. No difference in listening comprehension emerged, but the Anglo guise tended to engender a more compliant listening mind set. Results of this study can inform both provider-directed and patient-directed efforts to improve health care services for members of all linguistic and cultural groups.


Antibacterial activities of lactic acid bacteria isolated from cow faeces against potential enteric pathogens.
The addition of sub therapeutic doses of antibiotics to cattle feed for growth promotion is a contributory factor to antibiotic resistance, thus an alternative to antibiotics is needed in animal feed additives. To determine the antimicrobial activity of cow's intestinal Lactic acid bacteria (LAB) against enteric commensals. Escherichia coli, Klebsiella species (spp) and LAB were isolated from thirty different cow faecal samples and the LAB identified by partial sequencing of 16S rRNA. The antimicrobial activity of the LAB was determined against the test Escherichia coli and Klebsiella spp. Five species of LAB were isolated from thirty cow faecal samples and identified as Enterococcus hirae (8), Enterococcus durans (6), Enterococcus faecium (1), Enterococcus faecalis (1) and Weissella confusa (1). Viable cells and cell free supernatant (CFS) of the LAB were able to inhibit the growth of the test organisms with the largest zone of inhibition by the viable cells being 26mm against Escherichia coli CB6 produced by Enterococcus hirae CO6A while Weissella confusa CO29M and Enterococcus hirae CO2A produced the largest zones of inhibition (26mm) against Klebsiella CB2. This study shows that LAB from cow faeces possess considerable antimicrobial activity against resistant Escherichia coli from the same environment.


Cloning and mapping of the genetic determinants for microcin C51 production and immunity.
Microcin C51 is a small peptide antibiotic produced by Escherichia coli cells harbouring the 38 kb low copy number plasmid pC51, which codes for microcin production and immunity. The genetic determinants for microcin synthesis and immunity were cloned into the vectors pBR325, pUC19 and pACYC184. Physical and phenotypic analysis of deletion derivatives and mutant plasmids bearing insertions of transposon Tn5 showed that a DNA fragment of about 5 kb is required for microcin C51 synthesis and expression of complete immunity to microcin. Partial immunity can be provided by a 2 kb DNA fragment. Mutant plasmids were tested for their ability to complement Mic- mutations. Results of these experiments indicate that at least three plasmid genes are required for microcin production. The host OmpR function is also necessary for microcin C51 synthesis.


The impact of the COVID-19 pandemic and earthquake on mental health of persons with severe mental illness: A survey study among people receiving community mental health care versus treatment as usual in Croatia.
The COVID-19 pandemic led to the disruption of mental health services in most countries. Croatia has been developing and strengthening its mental health system, including the introduction of community mental health teams (CMHT) for persons with severe mental illness (SMI), whose implementation was ongoing during the pandemic through the RECOVER-E project. The aim of this study was to assess the differences in mental health outcomes, perceived social support and healthcare utilization in the group of participants receiving treatment as usual (TAU group) compared to the group receiving TAU and additional care by the CMHT (CMHT group) during the COVID-19 pandemic and two earthquakes. This is a cross-sectional survey administered among 90 participants with SMI at two time points: in May/June 2020 (first COVID-19 wave, earthquake) and in December 2020/January 2021 (second COVID-19 wave, earthquake). A significantly larger proportion of participants from the CMHT group visited the general practitioners in both waves of COVID-19 (first wave: CMHT 72.1%, TAU 44.2%, p = .009; second wave: CMHT 91.1%, TAU 64.1%, p = .003), as well as psychiatric services in the second wave (CMHT 95.3%, TAU 79.5%, p = .028). The use of long-acting injectables was also more frequent in the CMHT group (p = .039). Furthermore, analysis of the first wave showed higher perceived support of significant others (p = .004) in the CMHT group. We did not identify any differences in mental health outcomes between groups in either wave. While mental health outcomes did not differ between TAU and CMHT group, people in CMHT used services and treatments more frequently than those in TAU during the pandemic, which may indicate that CMHT services enable the continuity and accessibility of care for people with SMI under the circumstances where standard care is interruped (for example pandemic, disaster conditions).


Traumatic Dislodgement of Tibial Polyethylene Insert after a High-Flex Posterior-Stabilized Total Knee Replacement.
Many pathologic entities can produce a painful total knee replacement (TKR) that may lead to potential prosthetic failure. Polyethylene insert dissociation from the tibial baseplate has been described most frequently after mobile-bearing and cruciate-retaining TKRs. However, only 3 tibial insert dislocations in primary fixed-bearing High-Flex posterior-stabilized TKRs have been reported. We present a new case of tibial insert dislocation in a High-Flex model that shares similarities and differences with the cases reported, facilitating the analysis of the potential causes, which still remain undefined.


Blind detection of vascular sources and territories using random vessel encoded arterial spin labeling.
The goal of this work is to use vessel encoded arterial spin labeling (VEASL) methods to detect feeding arteries without prior knowledge of their positions, and map the vascular territory of each. Five healthy subjects were scanned, each with four different tagging planes. The VEASL tagging method was modified to use 60 different pairs of encoding steps with random orientation and spacing. A signal model was developed to calculate the theoretical ASL signal resulting from a vessel in any position in the tagging plane. For each voxel, the location of the feeding vessel was estimated by finding the theoretical signal that correlates most closely with the data. The main intracranial arteries, including carotid, vertebral, basilar, and cerebral arteries above the Circle of Willis were identified and localized from the ASL data in all subjects. In addition, external carotid branches were detected in all subjects. Randomly encoded VEASL provides data that allows for blind detection of source vessels. This method simplifies the VEASL prescription process and allows for efficient detection of atypical or collateral circulation.


Enzyme-linked immunosorbent assay for human skin collagenase.
A specific and sensitive indirect inhibition enzyme-linked immunosorbent assay (ELISA) was develop for human skin collagenase. Using an antiserum dilution of 1: 8000, the ELISA could detect 0.2 ng of enzyme, which was approximately 10 times more sensitive than the previously described radioimmunoassay for human skin collagenase. The assay was also highly reproducible. In comparative studies, bacterial, tadpole and crab collagenases did not react in the ELISA. Rat uterine collagenase and collagenases from bovine gingival explants and fibroblasts displayed approximately 0.1% of the reactivity found with human skin collagenase. Human synovial and gingival collagenases and collagenase from skin fibroblasts from patients with recessive dystrophic epidermolysis bullosa showed almost complete identity with the normal human skin fibroblast enzyme.


NIH BRAIN Circuits Programs: An Experiment in Supporting Team Neuroscience.
The NIH BRAIN Initiative is aimed at revolutionizing our understanding of the human brain. Here, we present a discussion of support for team research in investigative neuroscience at different stages and on various scales.


In vitro uptake and therapeutic application of liposome-encapsulated methotrexate in mouse hepatoma 129.
The biological activities of liposome-encapsulated and non-encapsulated methotrexate (MTX) were compared in vitro and in vivo using mouse Hepatoma 129 ascites tumor cells. Under in vitro conditions, cells accumulated up to 29% of [3H]-MTX when the drug was incorporated in the lipid bilayers of positively charged, unilamellar liposomes. There was no significant uptake of non-encapsulated MTX under the same conditions. A single intraperitoneal injection of liposome-encapsulated MTX (3 mg MTX/kg) increased the mean survival time of tumor-bearing mice to 42.5 +/- 11.2 days, compared to 23.5 +/- 2.2 days for untreated controls. Non-encapsulated MTX had no significant effect on survival time. Thus the in vivo treatment studies appear to agree with the in vitro uptake measurements. Addition of galactolipids to the lipid bilayers of liposomes did not increase in vitro uptake of encapsulated MTX and gave no additional improvement in therapeutic effectiveness. Encapsulation of MTX in liposomes might thus be used to increase uptake of the drug in cells which may be deficient in MTX transport.


Long-term blood glucose monitoring with implanted telemetry device in conscious and stress-free cynomolgus monkeys.
Continuous blood glucose monitoring, especially long-term and remote, in diabetic patients or research is very challenging. Nonhuman primate (NHP) is an excellent model for metabolic research, because NHPs can naturally develop Type 2 diabetes mellitus (T2DM) similarly to humans. This study was to investigate blood glucose changes in conscious, moving-free cynomolgus monkeys (Macaca fascicularis) during circadian, meal, stress and drug exposure. Blood glucose, body temperature and physical activities were continuously and simultaneously recorded by implanted HD-XG telemetry device for up to 10 weeks. Blood glucose circadian changes in normoglycemic monkeys significantly differed from that in diabetic animals. Postprandial glucose increase was more obvious after afternoon feeding. Moving a monkey from its housing cage to monkey chair increased blood glucose by 30% in both normoglycemic and diabetic monkeys. Such increase in blood glucose declined to the pre-procedure level in 30 min in normoglycemic animals and >2 h in diabetic monkeys. Oral gavage procedure alone caused hyperglycemia in both normoglycemic and diabetic monkeys. Intravenous injection with the stress hormones, angiotensin II (2 μg/kg) or norepinephrine (0.4 μg/kg), also increased blood glucose level by 30%. The glucose levels measured by the telemetry system correlated significantly well with glucometer readings during glucose tolerance tests (ivGTT or oGTT), insulin tolerance test (ITT), graded glucose infusion (GGI) and clamp. Our data demonstrate that the real-time telemetry method is reliable for monitoring blood glucose remotely and continuously in conscious, stress-free, and moving-free NHPs with the advantages highly valuable to diabetes research and drug discovery.


X-ray sensitivity and single-strand DNA break repair in mutagen-sensitive mutants of Drosophila melanogaster.
Mutants of Drosophila melanogaster that are sensitive to chemical mutagens were analyzed for sensitivity to X-rays and for the capacity to repair single-strand DNA breaks induced by X-rays. Analysis of X-ray sensitivity demonstrated that 74% of the mutants assayed display some X-ray sensitivity, with 75% of the sensitive lines being extremely sensitive. Repair of single-strand breaks was assayed after both high and low doses of irradiation in order to permit detection of repair over a wide range of damage. The results of this investigation fail to show a correlation between X-ray sensitivity and this particular repair process. Repair of single-strand breaks is therefore mediated by repair processes unrelated to those that are disrupted in the current mutant collection.


Crisis and Emergency Risk Messaging in Mass Media News Stories: Is the Public Getting the Information They Need to Protect Their Health?
The mass media provide an important channel for delivering crisis and emergency risk information to the public. We conducted a content analysis of 369 newspaper and television broadcast stories covering natural disaster and foodborne outbreak events and coded for seven best practices in crisis and emergency risk messaging. On average, slightly less than two (1.86) of the seven best practices were included in each story. The proportion of stories including individual best practices ranged from 4.6% for "expressing empathy" to 83.7% for "explaining what is known" about the event's impact to human health. Each of the other five best practices appeared in less than 25% of stories. These results suggest much of the risk messaging the public receives via mass media does not follow best practices for effective crisis and emergency communication, potentially compromising public understanding and actions in response to events.


Membrane depolarization facilitates sperm entry, large fertilization cone formation, and prolonged current responses in sea urchin oocytes.
Depolarization of the sea urchin egg's membrane is required for two processes during fertilization: the entry of the fertilizing sperm and the block to polyspermy which prevents the entry of supernumerary sperm. In an immature sea urchin oocyte, the depolarization is very small in response to the attachment of a sperm. The purpose of this study was to determine whether the depolarization evoked by sperm attaching to an oocyte can facilitate sperm entry or induce the block to polyspermy. Individual oocytes of the sea urchin with diameters which ranged from 86 to 102% that of the average diameter for mature eggs from the same female were examined. The oocytes have a membrane potential of -73 +/- 6 mV (SD, n = 80) and a very low input resistance compared to that of mature eggs. Single sperm, following attachment to an oocyte, elicit a brief, small depolarization with a maximum amplitude of 8 +/- 1.4 mV (SE, n = 15), frequently followed by the formation of tiny filament-like fertilization cones, but the sperm fail to enter. If oocytes are voltage-clamped at membrane potentials more negative than -20 mV, following attachment of the sperm small transient inward currents occur, similar filament-like cones form, and the sperm do not enter. When many sperm attach to an oocyte which is not voltage clamped, the depolarizations sum to create a large depolarization with an amplitude of 60 to 80 mV, which shifts the oocyte's membrane potential to a value between -10 and +5 mV; more positive values are not attained. At such membrane potentials, whether the potential is maintained by the summed depolarizations of many attached sperm or by voltage clamp, large fertilization cones form, the sperm enter, and the oocytes can become highly polyspermic. In oocytes voltage clamped at +20 mV, however, both sperm entry and fertilization cone formation are suppressed. Therefore, both types of voltage-dependence for sperm entry are present in oocytes, although the depolarization caused by a single sperm is not large enough to permit its entry, nor is the depolarization caused by many sperm sufficient to prevent the entry of supernumerary sperm.


Bile acids in tissues: binding of lithocholic acid to protein.
Human liver contains two forms of lithocholic acid. One form is readily extractable by 95% ethanol/0.1% ammonia (soluble lithocholate, SL), while the other remains firmly bound to the residue (tissue-bound lithocholate, TBL). TBL could be hydrolytically released using clostridial cholanoylamino acid hydrolase, suggesting a peptide link between lithocholate and protein. With bovine serum albumin (BSA), lithocholic acid showed spontaneous amino group-modifying activity. When small molecular weight lysine (alpha-t-BOC-1-lysyl-beta-naphthylamide) and arginine peptides (alpha-CBZ-di-arginyl-beta-naphthylamide) were used in place of BSA, lithocholate bound specifically to the lysine peptide. The unusual affinity for lysine suggested that this amino acid might be involved as a residue in TBL. Synthesis of lithocholyl lysines and comparison with products of acid hydrolysis of TBL established epsilon-lithocholyl lysine as the predominant form in which lithocholic acid is found in tissue bound form.


Effects of urea and trimethylamine-N-oxide on enzyme activity and stability.
The interactions of urea, trimethylamine-N-oxide (TMAO), and related solutes on a number of enzymes were examined. Urea inhibited enzymatic activity and accelerated the thermal inactivation of catalase, whereas TMAO activated some enzymes but inhibited others. The effects of urea and of TMAO, whether parallel or in opposition, were exerted independently. Thus, in those cases where TMAO increases enzymatic activity, it did so to the same relative degree, whether or not urea was present. TMAO markedly decreased the rate of thermal inactivation of catalase, indicating that it does favor compact protein structures. The assumption that TMAO factors compaction of protein structure, whereas urea has the contrary effect, does not lead to the expectation that TMAO must always oppose the effect of urea on enzymatic activity, since the most compact form of an enzyme may not always be the most active form.


Oral Hodgkin lymphoma: a wolf in wolf's clothing.
Hodgkin lymphoma typically presents as a nodal lesion and infrequently involves extranodal sites. The English language literature contains only 7 reports of primary Hodgkin lymphoma arising in the oral mucosa in the absence of nodal disease. We report a case of primary, extranodal Hodgkin lymphoma in the palatal mucosa of a 79-year-old white female. An incisional biopsy revealed a diffuse, mixed cellular infiltrate, consisting of benign lymphocytes, plasma cells, histiocytes, and foci rich in eosinophils. Within this background was a scattering of large, atypical cells, including Reed-Sternberg forms that exhibited immunoreactivity for CD30 and CD20 and nonreactivity for CD15 and CD45RO, supporting a diagnosis of classical Hodgkin lymphoma. Positron emission tomography exhibited a single focal area of abnormal hypermetabolic activity involving the left palate area, without involvement of any other site. The clinical stage was Ann Arbor I-A. The primary tumor and submandibular and upper neck lymph nodes were treated with a 6-MV photon beam to a total dose of 4000 cGy. There was no evidence of disease at 15-month follow-up.


Mediating effects of self-directed learning on the relationship between critical thinking and problem-solving in student nurses attending online classes: A cross-sectional descriptive study.
With the increased prevalence of online education due to the coronavirus 2019 pandemic and advancements in information technology, essential competencies, such as critical thinking, self-directed learning, and problem-solving ability, should be examined among student nurses taking online classes. Based on the findings of several studies suggesting that critical thinking does not affect problem-solving, this study aimed to examine the relationship among critical thinking, self-directed learning, and problem-solving in student nurses attending online classes, and to determine whether self-directed learning could mediate the relationship between critical thinking and problem-solving. Cross-sectional, descriptive design. K University in South Korea. In total, 138 junior and senior nursing students were enrolled. The critical thinking disposition scale for nursing students, self-directed learning scale, and problem-solving scale for college students were used to quantify participants' responses. Our results revealed significant positive correlations among critical thinking, self-directed learning, and problem-solving. Furthermore, self-directed learning (β = 0.78, p < 0.001) had a significant mediating effect on the relationship between critical thinking and problem-solving ability (Z = 5.10, p < 0.001). Developing and implementing appropriate self-directed learning programs are critical for improving problem-solving ability affected by critical thinking among student nurses engaged in online education.


Comparison of Dural Venous Sinus Volumes Before and After Flight in Astronauts With and Without Spaceflight-Associated Neuro-Ocular Syndrome.
Spaceflight-associated neuro-ocular syndrome (SANS) occurs in 40% to 60% of National Aeronautics and Space Administration (NASA) International Space Station (ISS) astronauts who present postflight with ophthalmological findings and elevated intracranial pressure. The etiology of SANS is unknown; it is hypothesized that venous outflow congestion from the head and neck occurs because of microgravity, which is supported by the finding of internal jugular vein stagnant flow and thrombosis in some astronauts, but the impact on intracranial dural venous sinus structures remains unknown. To clarify the potential risk of retrograde extension of clot intracranially among astronauts with internal jugular venous thrombosis by evaluating intracranial venous structures following spaceflight and to assess for any association between intracranial venous congestion and SANS. This retrospective cohort study of all NASA astronauts who had undergone magnetic resonance (MR) venography at the time of the study included quantitative and qualitative assessments of the intracranial venous system on preflight and postflight MR venograms. Data were collected a mean (SD) of 525.8 (187.5) days before spaceflight and 2.0 (1.5) days after return to Earth. A semiautomated segmentation of the venogram images was used, which was then compared with a neuroradiologist's assessment. A mean (SD) 184.3 (66.0) days of ISS spaceflight missions. Dural venous sinus volumes before and after spaceflight. A total of 12 astronauts (2 [16.67%] women; 10 [83.33%] men), with a mean (SD) age of 47.8 (5.8) years, were included. Overall, 4 astronauts (33.33%) met the diagnostic criteria for SANS. No dural venous sinus thrombosis was detected for any astronaut. Astronauts with SANS had significantly greater median (range) preflight to postflight increases in volume vs astronauts without SANS for all 3 venous sinus structures: superior sagittal sinus (13.40% [8.70% to 17.47%] vs -2.66% [-15.84% to 5.31%,]; P = .004), right transverse/sigmoid sinus (17.15% [7.63% to 30.08%] vs 0.77% [-14.98% to 15.12%]; P = .02), and left transverse/sigmoid sinus (9.40% [5.20% to 15.50%] vs -1.40% [-14.20% to 12.50%]; P = .03). There was a positive correlation between the neuroradiologist's evaluation and the semiautomated method for the superior sagittal sinus (rpb = 0.64; P = .02) and the right transverse/sigmoid sinus (rpb = 0.58; P = .050). These findings, in conjunction with the growing body of evidence of abnormal blood flow dynamics during spaceflight, suggest an association between intracranial venous congestion and SANS. Thus, there is an implication that individuals with increased venous sinus compliance may be at increased risk of developing SANS. These findings should be confirmed in a larger astronaut population and may contribute to understanding disorders of intracranial venous outflow on Earth.


A clinical evaluation of dry socket in a Nigerian teaching hospital.
We sought to determine the incidence of dry socket in a Nigerian teaching hospital and to evaluate the patients' demographic pattern, predisposing factors, the treatment given, and treatment outcome. A retrospective review of records of dental extractions complicated by dry socket in Obafemi Awolowo University Teaching Hospital, Ile-Ife, between January 1996 and December 2000 was undertaken. Information retrieved included patient sociodemographic data, indications for extraction, tooth extracted, status of attending surgeon, onset of symptoms, relevant findings of the examining clinician, interval before presentation, treatment given, and its outcome. Of the 3,319 dental extractions performed in 3,008 patients, 136 (4.1%) were complicated by dry socket. The patients' mean age was 33.4 (15.4) years and a peak age incidence of 21 to 30 years was found. A slight female preponderance (1.4:1) was observed. The majority of patients were in the low-income group, and presentation in the hospital was prompt in the high-income group. Mandibular teeth were affected 3 times more than maxillary teeth (P =.00080). Most cases of dry socket resulted from extractions performed by undergraduates and house officers. Various underlying systemic conditions were found in 11.0% of cases, none of which included use of oral contraceptives. Treatment was usually the use of zinc oxide eugenol dressing in an irrigated socket, combined with antibiotic therapy in 45.3% of cases. No adverse reaction to zinc oxide eugenol was observed. The incidence of dry socket in our hospital is 4.1%. The mandible was involved 3 times more than the maxilla. With the use of zinc oxide eugenol dressing, 70.6% of patients completed treatment satisfactorily and 29.2% were lost to follow-up.


Distribution and characterization of neuropeptide FF-like immunoreactivity in the rat nervous system with a monoclonal antibody.
Monoclonal antibodies against neuropeptide FF were produced and characterized. The antibodies are directed and highly specific to neuropeptide FF, and reactivity requires the C-terminal dipeptide of neuropeptide FF (Arg-Phe-NH2). Tissue extracts from bovine spinal cord, rat spinal cord and hypothalamus were analysed by high-pressure liquid chromatography coupled with radioimmunoassay using the characterized monoclonal antibody. Only one immunoreactive peptide was detected and it coeluted with authentic neuropeptide FF. Using this highly specific monoclonal antibody, the distribution of neuropeptide FF-like immunoreactivity was further studied by indirect immunohistochemistry. Immunoreactivity was seen in two major cell groups in the rat brain. The largest cell group was located in the medial hypothalamus between the dorsomedial and ventromedial nuclei. The other one was found in the nucleus of the solitary tract. Fibres immunoreactive for neuropeptide FF were located in the lateral septal nucleus, amygdala, different hypothalamic areas, nucleus of the solitary tract, ventral medulla, trigeminal complex and the dorsal horn of the spinal cord. Spinal and sympathetic ganglia were non-reactive. No neuropeptide FF immunoreactivity was seen in the gut autonomic nervous system or endocrine cells. The results show that neuropeptide FF-like immunoreactivity has a clearly more limited distribution in the nervous system than typical brain-gut peptides.


Human-induced pluripotent stem cells as models for rare cardiovascular diseases: from evidence-based medicine to precision medicine.
Rare cardiovascular diseases (RCDs) refer to those cardiovascular diseases that display a low prevalence as well as morbidity. Due to the vast variety of underlying genetic mutations and the relatively low patient population, RCDs present additional challenges for diagnosis. Precision medicine may offer opportunities for designing patient-specific therapies in particular for carriers of variants with undetermined significance. Moreover, precision medicine strategies provide benefit to patients with "common" symptoms but carry in rare genetic variants. Induced pluripotent stem cells (iPSCs) present a state-of-the-art precision medicine approach which recently made contributions to the study of RCDs via patient-specific iPSC-derived cardiomyocytes (iPSC-CMs). Human iPSC-CMs are derived from a patient's somatic cells and thus recapitulate a personalized genomics background, serving as patient-specific disease models. In light of these advantages, iPSC-CMs evolved as an effective tool for modeling cardiac disease phenotypes and accurately evaluating the toxicity of potential therapeutic compounds. This review covers approaches for studying RCDs and iPSC-CM models generated so far for different RCDs, such as long QT syndrome (LQT), short QT syndrome (SQT), Brugada syndrome (BrS), arrhythmogenic right ventricular cardiomyopathy (ARVC), and other rare diseases accomplished by cardiac-related syndromes like Fabry disease (FD) and Marfan syndrome (MFS). This overview aims to aid better understanding of the utility of iPSC-CM models, their various features, and future prospects.


Regulating the Spin State of FeIII by Atomically Anchoring on Ultrathin Titanium Dioxide for Efficient Oxygen Evolution Electrocatalysis.
Ferric oxides and (oxy)hydroxides, although plentiful and low-cost, are rarely considered for oxygen evolution reaction (OER) owing to the too high spin state (eg filling ca. 2.0) suppressing the bonding strength with reaction intermediates. Now, a facile adsorption-oxidation strategy is used to anchor FeIII atomically on an ultrathin TiO2 nanobelt to synergistically lower the spin state (eg filling ca. 1.08) to enhance the adsorption with oxygen-containing intermediates and improve the electro-conductibility for lower ohmic loss. The electronic structure of the catalyst is predicted by DFT calculation and perfectly confirmed by experimental results. The catalyst exhibits superior performance for OER with overpotential 270 mV @10 mA cm-2 and 376 mV @100 mA cm-2 in alkaline solution, which is much better than IrO2 /C and RuO2 /C and is the best iron-based OER catalyst free of active metals such as Ni, Co, or precious metals.


Buffer-dependent regulation of aquaporin-1 expression and function in human peritoneal mesothelial cells.
Biocompatible peritoneal dialysis fluids (PDF) are buffered with lactate and/or bicarbonate. We hypothesized that the reduced toxicity of the biocompatible solutions might unmask specific effects of the buffer type on mesothelial cell functions. Human peritoneal mesothelial cells (HPMC) were incubated with bicarbonate (B-)PDF or lactate-buffered (L-)PDF followed by messenger RNA (mRNA) and protein analysis. Gene silencing was achieved using small interfering RNA (siRNA), functional studies using Transwell culture systems, and monolayer wound-healing assays. Incubation with B-PDF increased HPMC migration in the Transwell and monolayer wound-healing assay to 245 ± 99 and 137 ± 11% compared with L-PDF. Gene silencing showed this effect to be entirely dependent on the expression of aquaporin-1 (AQP-1) and independent of AQP-3. Exposure of HPMC to B-PDF increased AQP-1 mRNA and protein abundance to 209 ± 80 and 197 ± 60% of medium control; the effect was pH dependent. L-PDF reduced AQP-1 mRNA. Addition of bicarbonate to L-PDF increased AQP-1 abundance by threefold; mRNA half-life remained unchanged. Immunocytochemistry confirmed opposite changes of AQP-1 cell-membrane abundance with B-PDF and L-PDF. Peritoneal mesothelial AQP-1 abundance and migration capacity is regulated by pH and buffer agents used in PD solutions. In vivo studies are required to delineate the impact with respect to long-term peritoneal membrane integrity and function.


Non-destructive analysis of pigments and other organic compounds in lichens using Fourier-transform Raman spectroscopy: a study of Antarctic epilithic lichens.
Lichens in Antarctic habitats are subjected to environmental extremes, including UVB radiation, desiccation and low temperatures, as well as to rapid fluctuations in these. Lichens synthesise a variety of chemical compounds in response to their environmental conditions which contribute towards their colour, and which act as protectants against physiological stresses. The fluorescence generated by the lichens at 532 nm can be used in epifluorescence microscopy to identify their presence on substrata but this can severely affect the Raman spectra using visible excitation. The advantage of the near infrared excitation used in FT-Raman spectroscopy in minimising fluorescence emission facilitates the molecular characterisation of lichen encrustations without having to remove the thallus from its substrate or remove or otherwise damage any part of the thallus. Spectroscopic biomarkers are proposed which allow the lichens to be characterised by the identification of characteristic lichen substances; the use of these biomarkers for the preliminary taxonomic identification of Antarctic lichens is examined and some potential pitfalls are described.


A Scoping Review on the Concept of Physician Caring.
Physicians' interest in the health and well-being of their patients is a tenet of medical practice. Physicians' ability to act upon this interest by caring for and about their patients is central to high-quality clinical medicine and may affect burnout. To date, a strong theoretical and empirical understanding of physician caring does not exist. To establish a practical, evidence-based approach to improve health care delivery and potentially address physician burnout, we sought to identify and synthesize existing conceptual models, frameworks, and definitions of physician caring. We performed a scoping review on physician caring. In November 2019 and September 2020, we searched PubMed MEDLINE, Embase, PsycINFO, CINAHL, and CENTRAL Register of Controlled Trials to identify conceptual models, frameworks, and definitions of physician caring. Eligible articles involved discussion or study of care or caring among medical practitioners. We created a content summary and performed thematic analysis of extracted data. Of 11,776 articles, we reviewed the full text of 297 articles; 61 articles met inclusion criteria. Commonly identified concepts referenced Peabody's "secret of care" and the ethics of care. In bioethics, caring is described as a virtue. Contradictions exist among concepts of caring, such as whether caring is an attitude, emotion, or behavior, and the role of relationship development. Thematic analysis of all concepts and definitions identified six aspects of physician caring: (1) relational aspects, (2) technical aspects, (3) physician attitudes and characteristics, (4) agency, (5) reciprocity, and (6) physician self-care. Caring is instrumental to clinical medicine. However, scientific understanding of what constitutes caring from physicians is limited by contradictions across concepts. A unifying concept of physician caring does not yet exist. This review proposes six aspects of physician caring which can be used to develop evidence-based approaches to improve health care delivery and potentially mitigate physician burnout.


Chronic reduction of renal mass: glomerular morphometry by electron microscopy.
Changes of glomerular volume in rats were measured up to 21 weeks following subtotal nephrectomy, using morphometric methods. A linear increase of glomerular volume was observed between 2 and 21 weeks after subtotal nephrectomy. This progressive increase in glomerular volume may reflect compensatory hemodynamic changes leading to an increased single nephron glomerular filtration rate.


Improved Insulin Secretion Response and Beta-cell Function Correlated with Increased Prolactin Levels After Laparoscopic Sleeve Gastrectomy in Morbidly Obese Patients with Acanthosis Nigricans.
To explore insulin secretion patterns, β-cell function, and serum prolactin (PRL) concentrations in Chinese morbidly obese patients with Acanthosis nigricans (AN) and their alterations after laparoscopic sleeve gastrectomy (LSG). A total of 138 morbidly obese subjects undergoing LSG were categorized as simple obesity without AN (OB group, n = 55) and obesity with AN (AN group, n = 83). Oral glucose tolerance test (OGTT), PRL, and related metabolic indices were performed pre- and 12 months post-LSG. Insulin secretion patterns were derived from insulin secretion peak time during OGTT: type I (peak at 30 or 60 min) and type II (peak at 120 or 180 min). Preoperatively, AN group showed significantly higher proportions of type II insulin secretion pattern, fasting insulin (FINS), and homeostatic model assessment of insulin resistance (HOMA-IR) whereas lower oral glucose insulin sensitivity (OGIS), insulinogenic index (IGI), and disposition index (DI) than OB group, which were improved significantly at 12 months postoperatively in both groups, more pronounced in AN group. Intriguingly, serum PRL declined substantially in AN group than OB group at baseline whereas elevated only in the AN group post-LSG. After adjusting for confounding factors, elevated PRL correlated significantly with increased IGI and DI, and decreased HOMA-IR in both genders, as well as increased OGIS in females, which was detected only in the AN group CONCLUSION: Morbidly obese patients with AN presented delayed insulin secretion response, impaired insulin secretion, and β-cell dysfunction, which were significantly improved by LSG and might benefit from elevated PRL.


Prediction and treatment of bleeding in patients under anticoagulant treatment.
Major bleeding is a common complication of anticoagulant treatment. Risk assessment tools are relevant in the management of patients with atrial fibrillation and venous thromboembolism. The combination of clinical, biological and genetic markers is incorporated to build predictive scores to help in the decision process about intensity and duration of treatment. The optimal management of bleeding involves the application of predictive scores in combination with anticoagulant reversal strategies.


Lacrimal androgen-binding proteins protect against Aspergillus fumigatus keratitis in mice.
To clarify the regulatory mechanisms of lacrimal androgen-binding proteins (ABPs) in mice with keratitis caused by Aspergillus fumigatus (A. fumigatus). Mouse models of A. fumigatus keratitis were established. Lacrimal glands were removed after 24 h for general and histological comparison. Lacrimal ABPs were detected by qRT-PCR and quantitative proteomic analysis, or were detected by qRT-PCR after subconjunctival or lacrimal gland injection with dexamethasone. Unique inflammatory factors were detected by qRT-PCR, Western blot and/or immunofluorescence. Interleukin-1β (IL-1β) was injected into the lacrimal gland to explore the relationship between IL-1β and lacrimal ABPs. The lacrimal glands of mice with fungal keratitis were larger than normal mice and these structures became disorganized. Moreover, the expression of ABP ε and ABP δ were increased. Subconjunctival injection with dexamethasone could reduce the size of the lacrimal gland and increase the expression of ABP ε and ABP δ, while lacrimal gland injection with dexamethasone had no obvious effects. The expression of IL-1β in the lacrimal gland of mice with A. fumigatus keratitis was increased. When IL-1β was injected into the lacrimal gland, the lacrimal gland enlarged and the expression of ABP ε and ABP δ decreased. Lacrimal glands contributed to protection in fungal keratitis, which was not due to the involvement of inflammatory cells in mice. ABP δ and ABP ε of mice were involved in reducing the severity of corneal damage in mice with A. fumigatus keratitis. Moreover, the expression of IL-1β and ABP δ and ABP ε were intrinsically linked.


Seven new 3,4-dihydro-furanocoumarin derivatives from Angelica dahurica.
To study the chemical constituents of the roots of Angelica dahurica, a well-known Chinese herbal medicine named Baizhi in Chinese. Compounds were separated by various chromatographies, and the structures of new compounds were elucidated based on the analysis of their spectroscopic and spectrometric data (1D, 2D NMR, HRESI MS, IR, and UV). The absolute configurations of new compounds were determined by the calculated electronic circular dichroism and chemical derivatization. The inhibitory activities of all isolates against nitric oxide (NO) production were evaluated using lipopolysaccharide-activated RAW 264.7 macrophage cells. Seven new 3,4-dihydro-furanocoumarin derivatives (1a/1b, 2a/2b, 3a/3b, 4) together with a known furanocoumarin (5) were isolated from the roots of A. dahurica. The new compounds included three pairs of enantiomers, (4S, 2''R)-angelicadin A (1a)/(4R, 2''S)-angelicadin A (1b), (4S, 2''S)-angelicadin A (2a)/(4R, 2''R)-angelicadin A (2b), and (4S, 2''S)-secoangelicadin A (3a)/(4R, 2''R)-secoangelicadin A (3b), together with (4R, 2''R)-secoangelicadin A methyl ester (4). The known xanthotoxol (5) inhibited the NO production with the half-maximal inhibitory concentration (IC50) value of (32.8 ± 0.8) µmol/L, but all the new compounds showed no inhibitory activities at the concentration of 100 µmol/L. This is the first report of the discovery of 3,4-dihydro-furanocoumarins from A. dahurica. The results are not only meaningful for the understanding of the chemical constituents of A. dahurica, but also enrich the reservoir of natural products.


Inhibition of hepatitis C virus replication by antimonial compounds.
Chronic hepatitis C virus (HCV) infection is a worldwide health problem causing serious complications, such as liver cirrhosis and hepatoma. Alpha interferon (IFN-alpha) or its polyethylene glycol-modified form combined with ribavirin is the only recommended therapy. However, an alternative therapy is needed due to the unsatisfactory cure rate of the IFN-based therapy. Using a modified reporter assay based on the HCV subgenomic-replicon system, we found that sodium stibogluconate (SSG), a compound used for leishmania treatment, suppressed HCV replication. We have previously reported that SSG is effective at inhibiting HCV replication in a cell line permissive for HCV infection/replication and in an ex vivo assay using fresh human liver slices obtained from patients infected with HCV (26). In this study, we show that the SSG 50% inhibitory dose for HCV replication is 0.2 to 0.3 mg/ml (equivalent to 345 to 517 microM of Sb) in the HCV subgenomic-replicon system. We also found that SSG and IFN-alpha exert a strong synergistic anti-HCV effect in both the traditional isobologram analysis and the median effect principle (CalcuSyn analysis). The combination of SSG and IFN-alpha could sustain the antiviral response better than SSG or IFN-alpha alone. The results suggest that SSG may be a good drug candidate for use in combination with other therapeutics, such as IFN-alpha and ribavirin, to treat HCV infection.


Fluorinated triphenylamine silicon phthalocyanine nanoparticles with two-color imaging guided in vitro photodynamic therapy through lysosomal dysfunction.
Lysosome-targeting therapy has emerged as a promising strategy for combating drug-resistant tumors. However, the synthesis of nanodrugs to achieve efficient lysosome targeting remains a challenging task. In this study, a nanoparticle DSPE@TPA-FBPA-SiPc was developed for lysosome targeting therapy. The nanoparticle was prepared by loading 2-[4-(diphenylamino)-1-diphenicacid-1-carbobenzoxy-4-(1,1,1,3,3,3-hexafluoropropane-4-phenoxy) silicon phthalocyanine (TPA-FBPA-SiPc) into 1,2-distearoyl-sn‑glycero-3-phosphoethanolamine-N-[succinyl(polyethyleneglycol)-2000] (DSPE). DSPE@TPA-FBPA-SiPc demonstrated remarkable capabilities such as two-color imaging, lysosome targeting and in vitro photodynamic therapy functions. The results revealed that DSPE@TPA-FBPA-SiPc efficiently accumulated in lysosomes, leading to generation of a high amount of reactive oxygen species upon irradiation. This induced apoptosis in MCF-7 cells by disrupting lysosomal function. Consequently, DSPE@TPA-FBPA-SiPc holds great potential as a photosensitizer for photodynamic therapy, utilizing the lysosomal-mediated cell death pathway.


Establishment of molecular diagnostics targeting the 23S ribosomal RNA gene for the detection of Mycoplasma suis infection in Thai domestic pigs.
Mycoplasma (M.) suis is a pathogenic hemotropic Mycoplasma sp. that causes acute hemolytic anemia or chronic infection in pigs. M. suis infection can be diagnosed using several methods, including molecular diagnosis such as conventional PCR (cPCR) and quantitative PCR (qPCR). In these cases, the common target is the 16S rRNA gene; however, this genetic marker cannot distinguish hemoplasma at the species level owing to high sequence identity. Therefore, the 23S rRNA gene has emerged as another target gene. Other than PCR, the loop-mediated isothermal amplification (LAMP) method can be applied for M. suis. The objective of the present study was to establish cPCR, TaqMan qPCR, and LAMP assays in which the 23S rRNA gene is used to detect M. suis infection in Thai domestic pigs. The analytical sensitivity of cPCR was determined as 7.46 × 104 copies/μl of plasmid DNA, whereas those of qPCR and LAMP were 7.46 × 102 copies/μl. There was no cross reaction with other pathogens in any of the assays. To evaluate the diagnostic performance of the assays, they were tested using 173 samples of genomic DNA. The detection percentage of M. suis infection was 24.86% (43/173; 95% CI: 18.61%-31.89%), 28.32% (49/173; 95% CI: 21.75%-35.66%), and 29.48% (51/173; 95% CI: 22.80%-36.88%) using cPCR, qPCR, and LAMP, respectively. Using qPCR as a reference assay, cPCR showed 81.63% sensitivity, 97.58% specificity, and an almost perfect level of agreement (kappa = 0.823). In comparison, LAMP showed 77.55% sensitivity, 89.52% specificity, and a substantial level of agreement (kappa = 0.662). All assays tested here could be applied in veterinary diagnostic laboratories for monitoring porcine health in the herds. Furthermore, the LAMP assay could be used as a screening test in farm practice without the need for any special equipment.


Synaptosomal susceptibility on global ischaemia caused by cardiac arrest correlated with early and late times after recirculation in rats.
The aim of the study was to assess the sensitivity of brain synaptosomes and their mitochondria to the effects of global cerebral ischaemia caused by temporary cardiac arrest and the early and late consequences. The effects of 10 min of global ischaemia were measured immediately and after 1 h, 24 h and 7 days post-resuscitation. Ischaemia caused a reduction in oxygen consumption by synaptosomes of about 20%, a drop in ATP/ADP ratio of about 40%, a decrease in CrP/Cr ratio at about 45% and a reduction of synaptic vesicles and disturbances in the mitochondrial structure in isolated synaptosomes and in nerve endings of brain specimens. Morphometric analysis showed that ischaemic conditions caused a decrease in synaptic vesicles by about 61% and an increase of mitochondrial damage to 58 and 50% after 1 and 24 h postreperfusion time, respectively. Seven days postresuscitation, all the observed changes returned to normal but small numbers (about 2%) of neurones which were destroyed neurons appeared at that time. It is concluded that global ischaemia with early resuscitation after cardiac arrest may lead to damage of synaptosomes and synaptic mitochondria. This, in turn, modifies substrate oxidation, synthesis of energy variables and affects neurotransmitter function. The observed disturbances return to normal later after resuscitation but the ischaemic events and reoxygenation caused selective morphological injury of certain neurones and this may form the basis for irreversible brain damage.


Determination of the nucleotide sequence for the glutamate synthase structural genes of Escherichia coli K-12.
We have determined the complete nucleotide sequence of a 6.3-kb chromosomal HpaI-EcoRI fragment, that contains the structural genes for both the large and small subunits of the Escherichia coli K-12 glutamate synthase (GOGAT) enzyme, as well as the 5'- and 3'-flanking and intercistronic DNA regions. The Mrs of the two subunits, as deduced from the nucleotide (nt) sequence, were estimated as 166,208 and 52,246. Partial amino acid sequence of the GOGAT enzyme revealed that the large subunit starts with a cysteine residue that is probably generated by a proteolytic cleavage. Northern blotting experiments revealed a transcript of approximately 7300 nt, that at least contains the cistrons for both subunits. A transcriptional start point and a functional promoter were identified in the 5' DNA flanking region of the large subunit gene. The messenger RNA nontranslated leader region has 120 nt and shares identity with the leader regions of E. coli ribosomal operons, in particular around the so-called boxA sequence implicated in antitermination. Other possible regulatory sequences are described.


A hybrid neural network algorithm for on-line state inference that accounts for differences in inoculum of Cephalosporium acremonium in fed-batch fermentors.
One serious difficulty in modeling a fermentative process is the forecasting of the duration of the lag phase. The usual approach to model biochemical reactors relies on first-principles, unstructured mathematical models. These models are not able to take into account changes in the process response caused by different incubation times or by repeated fedbatches. To overcome this problem, we have proposed a hybrid neural network algorithm. Feedforward neural networks were used to estimate rates of cell growth, substrate consumption, and product formation from on-line measurements during cephalosporin C production. These rates were included in the mass balance equations to estimate key process variables: concentrations of cells, substrate, and product. Data from fed-batch fermentation runs in a stirred aerated bioreactor employing the microorganism Cephalosporium acremonium ATCC 48272 were used. On-line measurements strongly related to the mass and activity of the cells used. They include carbon dioxide and oxygen concentrations in the exhausted gas. Good results were obtained using this approach.


Functional MRI of the brain: localisation of eloquent cortex in focal brain lesion therapy.
The aim of this study was to assess the feasibility of functional MRI (fMRI) in a clinical environment on a large patient group, and to evaluate the pretherapeutic value of localisation of eloquent cortex. Forty patients with focal brain lesions of different origin were studied using fMRI. Functional information was obtained using motor, somatosensory, auditory and phonological stimuli depending on the localisation of the lesions. To obtain information about the spatial accuracy of fMRI, the results were compared with postoperative electrocortical stimulation. Two patients with secondary trigeminal neuralgia were scanned using a motor protocol and were implanted with an extradural plate electrode. Imaging was successful in 40 of 42 patients (including the 2 with trigeminal neuralgia). These patients were analysed for strength of activation, the relation of the lesion to activation sites and the presence of mass effect. The correlation between these data and surgical findings provided significant additional clinical information. Functional MRI can be accurately performed in patients with focal brain lesions using a dedicated approach. Functional MRI offers important clinical information as a contribution to a decrease in posttherapeutic morbidity. The accuracy of the technique can be confirmed by other modalities, including invasive cortical electrostimulation.


Metal accumulation in dragonfly nymphs and crayfish as indicators of constructed wetland effectiveness.
Constructed wetland effectiveness is often assessed by measuring reductions of contaminant concentrations in influent versus departing effluent, but this can be complicated by fluctuations in contaminant content/chemistry and hydrology. We assessed effectiveness of a constructed wetland at protecting downstream biota from accumulating elevated metal concentrations-particularly copper and zinc in effluents from a nuclear materials processing facility. Contaminants distributed throughout a constructed wetland system and two reference wetlands were assessed using six dragonfly nymph genera (Anax, Erythemis, Libellula, Pachydiplax, Tramea, and Plathemis) as biomonitors. Additionally, the crayfish, Cambarus latimanus, were analyzed from the receiving and two reference streams. Concentrations of Cu, Zn, Pb, Mn, Cr, Cd, and Al were evaluated in 597 dragonfly nymph and 149 crayfish whole-body composite samples. Dragonfly genera varied substantially in metal accumulation and the ability to identify elevated metal levels throughout components of the constructed wetland. Genera more closely associated with bottom sediments tended to accumulate higher levels of metals with Libellula, Pachydiplax, and Erythemis often accumulating highest concentrations and differing most among sites. This, combined with their abundance and broad distributions make the latter two species suitable candidates as biomonitors for constructed wetlands. As expected, dragonfly nymphs accumulated higher metal concentrations in the constructed wetland than reference sites. However, dragonfly nymphs often accumulated as high of metal concentrations downstream as upstream of the water treatment cells. Moreover, crayfish from the receiving stream near the constructed wetland accumulated substantially higher Cu concentrations than from downstream locations or reference streams. Despite reducing metal concentrations at base flow and maintaining regulatory compliance, metal fluxes from the wetland were sufficient to increase accumulation in downstream biota. Future work should evaluate the causes of downstream accumulation as the next step necessary to develop plans to improve the metal sequestering efficiency of the wetland under variable flow regimes.


Synthesis and anti-HIV evaluation of novel 1,3-disubstituted thieno[3,2-c][1,2,6]thiadiazin-4(3H)-one 2,2-dioxides(TTDDs).
A series of novel 1,3-disubstituted thieno[3,2-c] [1,2,6]thiadiazin-4(3H)-one 2,2-dioxides (TTDDs), designed as non-nucleoside reverse transcriptase inhibitors (NNRTIs), was synthesized, structurally confirmed by spectral analysis and evaluated for their anti-HIV-1 activities by inhibition of HIV-1(IIIB)-induced cytopathogenicity in MT-4 cell culture. The results showed that TTDD analogues exhibited marked potency as anti-HIV-1 agents. The most active and selective compound was 1-(3-cyano)benzyl-3-benzyl-thieno[3,2-c][1,2,6]thiadiazin-4(3H)-one 2,2-dioxide (5f) with a 50% effective concentration (EC(50)) of 4.0 microM and a selectivity index (SI) of >76. The structure-activity relationship (SAR) is discussed.


Crystal structure of MbnF: an NADPH-dependent flavin monooxygenase from Methylocystis strain SB2.
Methanobactins (MBs) are ribosomally produced and post-translationally modified peptides (RiPPs) that are used by methanotrophs for copper acquisition. The signature post-translational modification of MBs is the formation of two heterocyclic groups, either an oxazolone, pyrazinedione or imidazolone group, with an associated thioamide from an X-Cys dipeptide. The precursor peptide (MbnA) for MB formation is found in a gene cluster of MB-associated genes. The exact biosynthetic pathway of MB formation is not yet fully understood, and there are still uncharacterized proteins in some MB gene clusters, particularly those that produce pyrazinedione or imidazolone rings. One such protein is MbnF, which is proposed to be a flavin monooxygenase (FMO) based on homology. To help to elucidate its possible function, MbnF from Methylocystis sp. strain SB2 was recombinantly produced in Escherichia coli and its X-ray crystal structure was resolved to 2.6 Å resolution. Based on its structural features, MbnF appears to be a type A FMO, most of which catalyze hydroxylation reactions. Preliminary functional characterization shows that MbnF preferentially oxidizes NADPH over NADH, supporting NAD(P)H-mediated flavin reduction, which is the initial step in the reaction cycle of several type A FMO enzymes. It is also shown that MbnF binds the precursor peptide for MB, with subsequent loss of the leader peptide sequence as well as the last three C-terminal amino acids, suggesting that MbnF might be needed for this process to occur. Finally, molecular-dynamics simulations revealed a channel in MbnF that is capable of accommodating the core MbnA fragment minus the three C-terminal amino acids.


SEM-EDX study of prepared human dentine surfaces exposed to gingival retraction fluids.
To evaluate the effects of gingival retraction fluids (GRF) on prepared dentine surfaces, and to test the null-hypothesis that prior exposure of dentine surfaces for prolonged period to any of the fluids evaluated does not influence acid-etching of the exposed surfaces. The investigation was carried out using SEM and energy-dispersive X-ray analysis (EDX). The GRF studied were Hemodent (HMDT), Astringedent (AST) and Ultradent buffered 25% Aluminium Chloride (ULTB). Longitudinal sections of 220-grit ground dentine surfaces were exposed to GRF for 0.5-, 1-, 2- and 5-min (n=4 each). Another group of samples was produced by 20s application of 35% phosphoric acid (PA) on GRF-pretreated dentine. Control samples were not exposed to GRF. Differences in etching effect-a function of the Ca-contents detected by EDX, were analysed using Friedman's and Wilcoxon's rank test (P<0.05). The SEM demonstrated the presence of a relatively non-porous amorphous dentine matrix, but many of the dentine tubule orifices remained occluded. Granular precipitates, which remained even after acid-etching, were noted on surfaces exposed to Hemodent for 5 min. Characteristic crystal growth was observed on surfaces exposed to Astringedent for 1- or 2-min prior to acid-etching. The EDX data demonstrated that there were differences in resulting Ca-content; ULTB >AST>HMDT>ULTB+PA>HMDT+PA>AST+PA>PA, but AST and HMDT were not significantly different. The exposure of dentine prepared surfaces to these three GRF altered its morphology and reduced the dentine's susceptibility to acid-etching, thus the null-hypothesis is rejected.


Next generation sequencing of RB1gene for the molecular diagnosis of ethnic minority with retinoblastoma in Yunnan.
Retinoblastoma is a rare intraocular malignancy and typically initiated by inactivating biallelic mutations of RB1 gene. Each year, ~ 8000 children worldwide are diagnosed for retinoblastoma. In high-income countries, patient survival is over 95% while low-income countries is ~ 30%.If disease is diagnosed early and treated in centers specializing in retinoblastoma, the survival might exceed 95% and many eyes could be safely treated and support a lifetime of good vision. In China, approximate 1100 newly diagnosed cases are expected annually and 28 hospitals covering 25 provinces established centers classified by expertise and resources for better treatment options and follow-up. Comparing with other province of eastern China, Yunnan province is remote geographically. This might result that healthcare staff have low awareness of the role of genetic testing in management and screening in families. The patients with retinoblastoma were selected in Yunnan. DNA from blood was used for targeted gene sequencing. Then, an in-house bioinformatics pipeline was done to detect both single nucleotide variants and small insertions/deletions. The pathogenic mutations were identified and further confirmed by conventional methods and cosegregation in families. Using our approach, targeted next generation sequencing was used to detect the mutation of these 12 probands. Bioinformatic predictions showed that nine mutations were found in our study and four were novel pathogenic variants in these nine mutations. It's the first report to describe RB1 mutations in Yunnan children with retinoblastoma. This study would improve role of genetic testing for management and family screening.


[Ultrastructural changes in the neurons and interneuronal synapses of the hypothalamus in the progeny of morphine-dependent rats].
The electron microscopic method has revealed early maturation of the protein-synthesizing apparatus of some cells, on the one hand, and underdevelopment of neuroblast volumes, appearance of artificial contacts, axonal degeneration and destruction in large trunks, on the other hand. Compensatory-restorative processes in the cell on the 14th and 21st days of the postnatal development result in a complete restoration of morphological organization of neurons and interneuronal connections in the hypothalamus of offspring of the morphine-dependent rats.


Geriatric Nephrology.
Older adults receiving critical care have a high risk for acute kidney injury (AKI) for many reasons. It is important that critical care nurses know and have the skills to address the risk factors, conduct a comprehensive geriatric assessment, and implement evidence-based interventions. This article provides a review of this information.


Influence of image analysis strategy, cooling rate, and sample volume on apparent protein cloud-point temperature determination.
The protein cloud-point temperature (TCloud) is a known representative of protein-protein interaction strength and provides valuable information during the development and characterization of protein-based products, such as biopharmaceutics. A high-throughput low volume TCloud detection method was introduced in preceding work, where it was concluded that the extracted value is an apparent TCloud (TCloud,app). As an understanding of the apparent nature is imperative to facilitate inter-study data comparability, the current work was performed to systematically evaluate the influence of 3 image analysis strategies and 2 experimental parameters (sample volume and cooling rate) on TCloud,app detection of lysozyme. Different image analysis strategies showed that TCloud,app is detectable by means of total pixel intensity difference and the total number of white pixels, but the latter is also able to extract the ice nucleation temperature. Experimental parameter variation showed a TCloud,app depression for increasing cooling rates (0.1-0.5 °C/min), and larger sample volumes (5-24 μL). Exploratory thermographic data indicated this resulted from a temperature discrepancy between the measured temperature by the cryogenic device and the actual sample temperature. Literature validation confirmed that the discrepancy does not affect the relative inter-study comparability of the samples, regardless of the image analysis strategy or experimental parameters. Additionally, high measurement precision was demonstrated, as TCloud,app changes were detectable down to a sample volume of only 5 μL and for 0.1 °C/min cooling rate increments. This work explains the apparent nature of the TCloud detection method, showcases its detection precision, and broadens the applicability of the experimental setup.


Usefulness of one-point plasma SN-38G/SN-38 concentration ratios as a substitute for UGT1A1 genetic information after irinotecan administration.
It was recently reported that genetic polymorphisms of UDP glucuronyltransferase-1 polypeptide A1 (UGT1A1), a glucuronidation enzyme, were associated with irinotecan (CPT-11) metabolism. The active metabolite of CPT-11, 7-ethyl-10-hydroxycamptothecin (SN-38) was glucuronidated (SN-38G) by UGT1A1. Genetic polymorphisms of UGT1A1 were associated with potentially serious adverse events, including neutropenia. Several studies have suggested that the dose of CPT-11 should be decreased in patients homozygous for UGT1A1*6 or UGT1A1*28, or double heterozygotes (*6/*28). However, the reference dose for patients with these genetic polymorphisms is unclear. We investigated the relationship between the SN-38G/SN-38 concentration ratio and the dose of CPT-11 in 70 patients with colorectal cancer who received FOLFIRI-based regimens, by measuring the plasma concentrations of CPT-11, SN-38, and SN-38G. The SN-38G/SN-38 concentration ratio was lower in patients who were homozygous for UGT1A1*6, heterozygous for UGT1A1*6 or UGT1A1*28, or were double heterozygotes compared with patients with wild-type genes. The relative decreases in the SN-38G/SN-38 concentration ratio in patients homozygous for UGT1A1*6 and in double heterozygotes were greater than in patients heterozygous for UGT1A1*6 or UGT1A1*28. Interestingly, decreases in the SN-38G/SN-38 concentration ratio were associated with decreases in the neutrophil count and the final infusion dose of CPT-11. Our results suggest that the SN-38G/SN-38 concentration ratio is an important factor for guiding dose adjustments, even in patients with wild-type genes. Therefore, the SN-38G/SN-38 concentration ratio, as an index of the patient's metabolic capacity, is useful for assessing dose adjustments of CPT-11.


Influence of citalopram and chlorimipramine on uptake of 5-HT by isolated perfused guinea pig lungs.
A method is described for the preparation and perfusion in vitro of isolated guinea pig lungs, 5-HT is rapidly taken up from a perfusing solution by the organ (about 70% uptake). In the concentration range 10(-7) to 5 X 10(-6) M citalopram concentration dependently inhibited the uptake of 5-HT by the lung preparation. Chlorimipramine inhibited the uptake of 5-HT only by 40--50%, and the inhibition was not concentration dependent. The method used seems to be a reliable model for studying the uptake of 5-HT.


Partial replacement of red and processed meat with legumes: a modelling study of the impact on nutrient intakes and nutrient adequacy on the population level.
The shift towards plant-based diets with less meat and more legumes is a global target and requires an understanding of the consequences of dietary adequacy on the population level. Our aim was to model the impact of partial replacement of red and processed meat with legumes on nutrient intakes and population shares below dietary reference intakes. Modelling study with three scenarios anchored in meat cut-offs: ≤ 70 g/d (Finnish dietary guideline); ≤ 50 g/d (Danish dietary guideline); ≤ 30 g/d (EAT-Lancet recommendation). In all subjects, the amount of meat in grams over the cut-off was replaced with the same amount of legumes. The SPADE method was used to model usual intake distributions. Meaningful differences in average intakes and in population shares below dietary reference intakes compared to the reference (FinDiet) were evaluated based on non-overlapping 95 % CI. Finnish national food consumption survey (FinDiet 2017). Finnish adults (n 1655) aged18-74 years (47 % men). The scenarios introduced increases in the average intakes of fibre, folate, K, Mg, Cu and Fe, and decreases in intakes of saturated fat, niacin, vitamin B12, Se and Zn. Meaningful shifts of the usual intake distributions of fibre and folate towards improvement in intakes emerged already in 'scenario 70 g'. Overall, distribution shifts towards a higher probability of inadequate intakes of the studied nutrients were not observed. These results support the public health message to partly replace meat with legumes and may benefit nutrition policy actions towards sustainable diets in the Nordic countries and beyond.


Interpenetrated nano-capsule networks based on the alkali metal assisted assembly of p-carboxylatocalix[4]arene-O-methyl ether.
Reaction of p-carboxylatocalix[4]arene-O-methyl ether with either rubidium or caesium hydroxide results in the formation of interpenetrated nano-capsule networks with the calixarene in the 1,3-alternate conformation.


Spectrum of ocular disease in children aged between 0 and 3 years at an Australian paediatric tertiary hospital.
Childhood ocular disease can be a significant health burden to the child, family and society. Previous studies have examined the spectrum of paediatric ocular disease presenting to tertiary hospitals; however, these studies have broader age ranges, smaller sample sizes, and are largely based in developing countries. This study aims to assess the spectrum of ocular disease in the first 3 years of life presenting to the eye department of an Australian tertiary paediatric hospital. The records of 3337 children who had their initial presentation at the eye clinic between the age of 0 and 36 months were reviewed, spanning 6.5 years from 1st July 2012 to 31st December 2018. The most common primary diagnoses overall were strabismic amblyopia (6.0%), retinopathy of prematurity (5.0%) and nasolacrimal duct obstruction (4.5%). Bilateral visual impairment was more common in younger children, while unilateral visual impairment was more common in older children. The proportion of all children presenting with visual impairment was 10.3%, with 5.7% of all children presenting with bilateral visual impairment and 4.6% presenting with unilateral visual impairment. In children with visual impairment, the most common sites of primary abnormality were lens (21.4%), retina (17.3%), and cerebral and visual pathways (12.1%). The most common primary diagnoses in children with visual impairment were cataract (21.4%), strabismic amblyopia (9.3%) and retinoblastoma (6.5%). The spectrum of eye disease and vision impairment presenting in the first 3 years of life facilitates health care planning, greater community education about vision impairment and importance of early intervention, and guidance for appropriate resource allocation. Health systems can apply these findings to aid in early identification and intervention to reduce preventable blindness and institute appropriate rehabilitation services.


Safety of conscious sedation in electroanatomical mapping procedures and cryoballoon pulmonary vein isolation.
Immobilization of patients during electrophysiological procedures, to avoid complications by patients' unexpected bodily motion, is achieved by moderate to deep conscious sedation using benzodiazepines and propofol for sedation and opioids for analgesia. Our aim was to compare respiratory and hemodynamic safety endpoints of cryoballoon pulmonary vein isolation (PVI) and electroanatomical mapping (EAM) procedures. Included patients underwent either cryoballoon PVI or EAM procedures. Sedation monitoring included non-invasive blood pressure measurements, transcutaneous oxygen saturation (tSpO2) and transcutaneous carbon-dioxide (tpCO2) measurements. We enrolled 125 consecutive patients, 67 patients underwent cryoballoon atrial fibrillation ablation and 58 patients had an EAM and radiofrequency ablation procedure. Mean procedure duration of EAM procedures was significantly longer (p < 0.001) and propofol doses as well as morphine equivalent doses of administered opioids were significantly higher in EAM patients compared to cryoballoon patients (p < 0.001). Cryoballoon patients display higher tpCO2 levels compared to EAM patients at 30 min (cryoballoon: 51.1 ± 7.0 mmHg vs. EAM: 48.6 ± 6.2 mmHg, p = 0.009) and at 60 min (cryoballoon: 51.4 ± 7.3 mmHg vs. EAM: 48.9 ± 6.6 mmHg, p = 0.07) procedure duration. Mean arterial pressure was significantly higher after 60 min (cryoballoon: 84.7 ± 16.7 mmHg vs. EAM: 76.7 ± 13.3 mmHg, p = 0.017) in cryoballoon PVI compared to EAM procedures. Regarding respiratory and hemodynamic safety endpoints, no significant difference was detected regarding hypercapnia, hypoxia and episodes of hypotension. Despite longer procedure duration and deeper sedation requirement, conscious sedation in EAM procedures appears to be as safe as conscious sedation in cryoballoon ablation procedures regarding hemodynamic and respiratory safety endpoints.


A meta-analysis: comparison of esomeprazole and other proton pump inhibitors in eradicating Helicobacter pylori.
To assess effects of esomeprazole-based therapies and others (PPIs) on Helicobacter pylori eradication when co-administered with antibiotics. Using the terms 'H. pylori eradication' and 'esomeprazole', we searched Medline, Embase, and the Cochrane Library (2000-2005) and conducted a manual search of related Chinese journals to identify randomized controlled trials (RCTs) that compared the treatment effects of esomeprazole and other PPI therapies in patients with H. pylori infection. Of the 75 articles screened, 11 RCTs including 2,159 subjects were included in the meta-analysis. The mean H. pylori eradication rates (intention-to-treat, ITT) with esomeprazole + antibiotics was 86%, a rate comparable with other PPI therapies, 81% (ITT); the odds ratio (OR) was 1.38 (95% CI = 1.09-1.75). A focused meta-analysis of 6 selected high-quality studies was produced an OR of this comparison that was closer to one (1.17, 95% CI = 0.89-1.54), with the results being statistically homogeneous. Sub-analysis that included only studies comparing different does of esomeprazole with omeprazole or pantoprazole did not reveal significant differences. No additional serious adverse events were reported. Esomeprazole-based triple therapy may effectively eradicate H. pylori infection and promote favorable outcome with good tolerance and offers comparable efficacy to omeprazole-based therapy.


Stem Cell Therapy for Hepatocellular Carcinoma: Future Perspectives.
Hepatocellular carcinoma (HCC) is one of the most common types of cancer and results in a high mortality rate worldwide. Unfortunately, most cases of HCC are diagnosed in an advanced stage, resulting in a poor prognosis and ineffective treatment. HCC is often resistant to both radiotherapy and chemotherapy, resulting in a high recurrence rate. Although the use of stem cells is evolving into a potentially effective approach for the treatment of cancer, few studies on stem cell therapy in HCC have been published. The administration of stem cells from bone marrow, adipose tissue, the amnion, and the umbilical cord to experimental animal models of HCC has not yielded consistent responses. However, it is possible to induce the apoptosis of cancer cells, repress angiogenesis, and cause tumor regression by administration of genetically modified stem cells. New alternative approaches to cancer therapy, such as the use of stem cell derivatives, exosomes or stem cell extracts, have been proposed. In this review, we highlight these experimental approaches for the use of stem cells as a vehicle for local drug delivery.


Intermediates in the barnol biosynthesis in Penicillium baarnense.
By using appropriate 14C-labelled phenolic substances as precursors in feeding experiments the following steps in barnol biosynthesis have been established: acetate + malonate leads to 2,4-dihydroxy-6-ethyl-5-methylbenzaldehyde leads to 1,3-dihydroxy-4,6-dimethyl-2-ethylbenzene leads to barnol. P. baarnense can also reduce orcylaldehyde and 2,4-dihydroxy-5,6-dimethylbenzaldehyde to dimethylresorcinol and trimethylresorcinol, respecitively.


Modification of TPN-dependent isocitrate dehydrogenase by the 2', 3'-dialdehyde derivatives of TPNH and TPN.
Catalytic reaction of the 2', 3'-dialdehyde analog of TPN (oTPN) with pig heart TPN-dependent isocitrate dehydrogenase in the presence of the substrate manganous isocitrate results in the formation of the dialdehyde derivative of TPNH (oTPNH). In the absence of the substrate, modification by oTPN leads to a progressive inactivation of the enzyme. The dependence of the pseudo-first order rate constants on the reagent concentration indicates the formation of a reversible complex with the enzyme prior to covalent modification (kmax = 5.5 X 10(-2) min-1; K1 = 290 microM). Reaction of [14C]oTPN with the enzyme results in the incorporation of 2 mol of oTPN/mol of peptide chain. No appreciable protection against either inactivation or incorporation by the natural ligands TPN and TPNH was obtained, suggesting different modes of binding of the analog in the presence and absence of the substrate isocitrate. Enzymatically synthesized oTPNH has been isolated and demonstrated to act as an affinity label for a TPNH-binding site of isocitrate dehydrogenase. The inactivation process exhibits saturation kinetics (kmax = 2.67 X 10(-3) min-1; K1 = 33 microM). Protection against activity loss, as well as a decrease in incorporation from 2 to 1 eq of [14C]oTPNH bound/peptide chain was observed in the presence of 1 mM TPNH. From the TPNH concentration dependence of the inactivation rate by oTPNH, a dissociation constant of 3.4 microM is calculated for TPNH, indicating binding of the analog to a specific TPNH-binding site on the enzyme. Although dialdehyde derivatives are frequently assumed to form Schiff bases with proteins, the evidence presented suggests the formation of morpholino derivatives as the products of the covalent reaction of isocitrate dehydrogenase with the dialdehyde derivatives of TPN and TPNH. The new reagent, oTPNH, may serve as an affinity label for other dehydrogenases.


Brain processing of rectal sensation in adolescents with functional defecation disorders and healthy controls.
Decreased sensation of urge to defecate is often reported by children with functional constipation (FC) and functional nonretentive fecal incontinence (FNRFI). The aim of this cross-sectional study was to evaluate cerebral activity in response to rectal distension in adolescents with FC and FNRFI compared with healthy controls (HCs). We included 15 adolescents with FC, 10 adolescents with FNRFI, and 15 young adult HCs. Rectal barostat was performed prior to functional magnetic resonance imaging (fMRI) to determine individual pressure thresholds for urge sensation. Subjects received 2 sessions of 5 × 30 seconds of barostat stimulation during the acquisition of blood oxygenation level-dependent fMRI. Functional magnetic resonance imaging signal differences were analyzed using SPM8 in Matlab. Functional constipation and FNRFI patients had higher thresholds for urgency than HCs (P < .001). During rectal distension, FC patients showed activation in the anterior cingulate cortex, dorsolateral prefrontal cortex, inferior parietal lobule, and putamen. No activations were observed in controls and FNRFI patients. Functional nonretentive fecal incontinence patients showed deactivation in the hippocampus, parahippocampal gyrus, fusiform gyrus (FFG), lingual gyrus, posterior parietal cortex, and precentral gyrus. In HCs, deactivated areas were detected in the hippocampus, amygdala, FFG, insula, thalamus, precuneus, and primary somatosensory cortex. In contrast, no regions with significant deactivation were detected in FC patients. Children with FC differ from children with FNRFI and HCs with respect to patterns of cerebral activation and deactivation during rectal distension. Functional nonretentive fecal incontinence patients seem to resemble HCs when it comes to brain processing of rectal distension.


Defining Clinical Meaningfulness in Huntington's Disease.
Minimal clinically important difference (MCID) represents the smallest within-person change on an outcome measure considered meaningful to the patient. Anchor-based MCID methods evaluate the relationship between changes in an outcome measure and the patient-reported clinical importance of that change. This study aims to estimate longitudinal MCID for clinically relevant outcome measures for individuals who have Stages 2 or 3 disease as measured by the Huntington's Disease Integrated Staging System (HD-ISS). Data were drawn from Enroll-HD, a large global longitudinal, observational study and clinical research platform for HD family members. We analyzed HD participants (N = 11,070) by staging group using time frames ranging from 12 to 36 months. The anchor was the physical component summary score of the 12-item short-form health survey. HD-relevant motor, cognitive, and functional outcome measures were independent, external criterion outcomes. Complex analysis was conducted using multiple, independent, linear mixed effect regression models with decomposition to calculate MCID for each external criterion by group. MCID estimates varied by progression stage. MCID estimates increased as stage progression increased and as the time frame increased. MCID values for key HD measures are provided. For example, starting in HD-ISS stage 2, meaningful group change over 24 months equals an average increase of 3.6 or more points on the Unified Huntington's Disease Rating Scale Total Motor Score. This is the first study to examine MCID estimation thresholds for HD. The results can be used to improve clinical interpretation of study outcomes and enable treatment recommendations to support clinical decision-making and clinical trial methodology. © 2023 International Parkinson and Movement Disorder Society.


Correction: Hao et al. Processing Aspectual Agreement in a Language with Limited Morphological Inflection by Second Language Learners: An ERP Study of Mandarin Chinese. Brain Sci. 2022, 12, 524.
In the published publication [...].


Variability of serum concentrations of cystatin C and urinary retinol-binding protein, neutrophil gelatinase-associated lipocalin, immunoglobulin G, and C-reactive protein in dogs.
Markers of kidney dysfunction and damage have potential to detect chronic kidney disease (CKD) in early stages. However, data on long-term variation of these markers in healthy dogs is lacking and is crucial for the interpretation of results. To determine temporal variations of serum cystatin C (sCysC) and urinary retinol-binding protein (uRBP), neutrophil gelatinase-associated lipocalin (uNGAL), immunoglobulin G (uIgG), and C-reactive protein (uCRP) in healthy dogs. Eight clinically healthy adult Beagles were evaluated. Longitudinal observational study. Serum cystatin C was determined by particle-enhanced nephelometric immunoassay. Urinary retinol-binding protein, uNGAL, uIgG and uCRP were determined by ELISA and concentrations were indexed to urinary creatinine. Within- and between-dog variance components (VC) and within-dog coefficients of variation (CV) were determined from blood and urine collected at eight time points over 1.5 years. Urinary C-reactive protein (uCRP) concentrations were consistently below the detection limit (5.28 ng/mL). Mean ± within-dog standard deviation for sCysC, uRBP/c, uNGAL/c and uIgG/c was 0.15 ± 0.01 mg/L, 0.09 ± 0.03 mg/g, 2.32 ± 2.03 μg/g and 12.47 ± 10.98 mg/g, respectively. Within-dog CV for sCysC, uRBP/c, uNGAL/c and uIgG/c was 8.1%, 33.7%, 87.2% and 88.1%, respectively. Serum cystatin C, uRBP/c, uNGAL/c and uIgG/c exhibit a wide range of long-term within-dog variability. Researchers and veterinarians might need to take this into account when interpreting their results. To assess their diagnostic and predictive ability, future studies need to establish reference ranges for healthy dogs and dogs with CKD.


Effects of nano-chitosan coatings incorporating with free/nano-encapsulated essential oil of Golpar (Heracleum persicum L.) on quality characteristics and safety of rainbow trout (Oncorhynchus mykiss).
This study investigated the effect of nano-chitosan coating containing free/nano forms of Heracleum persicum L. essential oil (HPEO) on microbial, chemical and sensory properties of rainbow trout (Oncorhynchus mykiss) at 4 °C during storage. In this study, nanoliposomes were prepared using different soy lecithin/cholesterol ratios (30:30, 40:20, 50:10 and 60:0) and sonication-hydration techniques with sizes of 78.70-123 nm. Liposome with a ratio of 50:10 had the greatest EE%. The zeta potantial was between -5.62 ± 0.28 to -3.71 ± 0.31 mV. The index of polydispersity and encapsulation efficiency (EE) were 0.452-0.481 and 51.93-68.13 %, respectively. Based on the results of microbiological and chemical analyses, samples treated with with nanochitosan/nano essential oil (NCS-NEO) had a better effect at the end of the study than other samples (NCS-EO, NCS and control). For NCS-NEO treatment at the end of storage time, analysis the value of total volatile basic nitrogen (TVB-N), pH, thiobarbituric acid reactive substances (TBARs) and peroxide value (PV) were, 27.95 (mg/100 g), 6.9, 0.37 (mEq/kg), 5.13 (mg MDA/kg), respectively, and the total pseudomonads count (TPC), total viable counts (TVC) and lactic acid bacteria (LAB) were, 8, 6.8, and 6.5 CFU/g, respectively. Further, the NCS-NEO treatment (at the end of study) achieved the highest sensory characteristics. HPEO encapsulation decreases diffusion rates, so improving activity of antioxidant and antimicrobial, and characteristics of sensory. This study suggested that coating nano forms of HPEO with NCS could be used to enhance shelf life of rainbow trout fish.


Accuracy and digital screw path design of TiRobot-assisted pedicle screw placement for lumbar spondylolisthesis.
To investigate lumbar spondylolisthesis screw placement assisted by TiRobot in terms of digital screw path design, accurate implementation, and accuracy evaluation method. In this study, we enrolled 40 patients with lumbar spondylolisthesis between December 2020 and August 2021 who underwent spine surgery at the Affiliated Hospital of PuTian University. Pre-operative computed tomography position and screw path designation, intra-operative pedicle screw placement according to pre-operative planning, and post-operative evaluation of the accuracy of screw placement were performed. 3D coordinates of the entry and exit points before and after the operation were collected. The qualified points at different levels of accuracy were counted. The screw placement accuracy was based on the absolute difference using the Chi-squared test. In total, 194 screws were successfully implanted with no screws penetrating the cortex. The absolute difference of entry points X, Y, and Z coordinates before and after the operation was 0.425 ± 0.294 mm, 0.417 ± 0.310 mm, and 0.466 ± 0.327 mm, respectively. The corresponding values in terms of exit points were 0.702 ± 0.470 mm, 0.963 ± 0.595mm, and 0.983 ± 0.566 mm, respectively. No obvious differences in coordinates before and after the operation were observed with an entry point degree of accuracy of ≥ 1.2 mm and exit point degree of accuracy of ≥ 2.1 mm. Therefore, the real surgery was consistent with the design. TiRobot-assisted lumbar spondylolisthesis surgery achieved optimal path designation and precise surgery.


Engineering Saccharomyces cerevisiae to produce odd chain-length fatty alcohols.
Fatty aldehydes and alcohols are valuable precursors used in the industrial manufacturing of a myriad of specialty products. Herein, we demonstrate the de novo production of odd chain-length fatty aldehydes and fatty alcohols in Saccharomyces cerevisiae by expressing a novel biosynthetic pathway involving cytosolic thioesterase, rice α-dioxygenase and endogenous aldehyde reductases. We attained production titers of ∼20 mg/l fatty aldehydes and ∼20 mg/l fatty alcohols in shake flask cultures after 48 and 60 h respectively without extensive fine-tuning of metabolic fluxes. In contrast to prior studies which relied on bi-functional fatty acyl-CoA reductase to produce even chain-length fatty alcohols, our biosynthetic route exploits α-oxidation reaction to produce odd chain-length fatty aldehyde intermediates without using NAD(P)H cofactor, thereby conserving cellular resource during the overall synthesis of odd chain-length fatty alcohols. The biosynthetic pathway presented in this study has the potential to enable sustainable and efficient synthesis of fatty acid-derived chemicals from processed biomass.


Scalene muscle activity during progressive inspiratory loading under pressure support ventilation in normal humans.
We hypothesized that (1) in healthy humans subjected to intermittent positive pressure non-invasive ventilation, changes in the ventilator trigger sensitivity would be associated with increased scalene activity, (2) if properly processed - through inspiratory phase-locked averaging - surface electromyograms (EMG) of the scalenes would reliably detect and quantify this, (3) there would be a correlation between dyspnea and scalene EMG. Surface and intramuscular EMG activity of scalene muscles were measured in 10 subjects. They breathed quietly through a face mask for 10min and then were connected to a mechanical ventilator. Recordings were performed during three 15-min epochs where the subjects breathed against an increasingly negative pressure trigger (-5%, -10% and -15% of maximal inspiratory pressure). With increasing values of the inspiratory trigger, inspiratory efforts, dyspnea and the scalene activity increased significantly. The scalene EMG activity level was correlated with the esophageal pressure time product and with dyspnea intensity. Inspiration-adjusted surface EMG averaging could be useful to detect small increases of the scalene muscles activity during mechanical ventilation.


Cloning and restriction endonuclease mapping of the genome of an equine herpesvirus 4 (equine rhinopneumonitis virus), strain 405/76.
Purified virion DNA of an Australian isolate of equine herpesvirus 4(EHV 4.405/76) was digested with restriction enzymes and the DNA fragments were cloned into pUC19. The resulting recombinant plasmid library, representing 92% of the virus genome, was used in hybridization analyses to construct restriction maps for BamHI, EcoRI, and SalI for the EHV4 genome. The results show that the genome of EHV 4.405/76 was approximately 145 kb and comprised a unique long (UL) region of 112 kb and a unique short (US) region of 12.4 kb. US is flanked by an internal and terminal repetitive sequence (IRS and TRS) of about 10.3 kb. The BamHI and EcoRI restriction maps are similar to those previously published for an English isolate EHV 4.1942 strain although some differences such as location of an additional fragment and changes in positions of two other small fragments were found.


Drugs activating hypoxia-inducible factors correct erythropoiesis and hepcidin levels via renal EPO induction in mice.
The erythroid growth factor erythropoietin (EPO) is mainly produced by the kidneys in adult mammals and induces expansion of erythroid cells and iron use for hemoglobin synthesis. The liver also produces EPO at a lower level than the kidneys. Renal and hepatic EPO production is fundamentally regulated by hypoxia-inducible transcription factors (HIFs) in a hypoxia/anemia-inducible manner. Recently, small compounds that activate HIFs and EPO production in the kidneys by inhibiting HIF-prolyl hydroxylases (HIF-PHIs) have been launched to treat EPO-deficiency anemia in patients with kidney disease. However, the roles of the liver in the HIF-PHI-mediated induction of erythropoiesis and iron mobilization remain controversial. Here, to elucidate the liver contributions to the therapeutic effects of HIF-PHIs, genetically modified mouse lines lacking renal EPO-production ability were analyzed. In the mutant mice, HIF-PHI administration marginally increased plasma EPO concentrations and peripheral erythrocytes by inducing hepatic EPO production. The effects of HIF-PHIs on the mobilization of stored iron and on the suppression of hepatic hepcidin, an inhibitory molecule for iron release from iron-storage cells, were not observed in the mutant mice. These findings demonstrate that adequate induction of EPO mainly in the kidney is essential for achieving the full therapeutic effects of HIF-PHIs, which include hepcidin suppression. The data also show that HIF-PHIs directly induce the expression of duodenal genes related to dietary iron intake. Furthermore, hepatic EPO induction is considered to partially contribute to the erythropoietic effects of HIF-PHIs but to be insufficient to compensate for the abundant EPO induction by the kidneys.


Alemtuzumab-induced lung injury in multiple sclerosis: Learning from adversity in three patients.
Respiratory alemtuzumab-related adverse events are clinically heterogeneous and include respiratory infections, infusion-related dyspnea, hypoxia and secondary autoimmune disorders. Here we report three cases of drug-induced lung disease following treatment with alemtuzumab in multiple sclerosis patients. First case was diagnosed as a non-specified intestitial pneumonitis associated with organizing pneumonia with subacute onset, second case was an acute respiratory distress syndrome with onset during second cycle, third case was a diffuse acute alveolar hemorrhage during first cycle infusion. All patients developed acute respiratory failure, reversible after steroid therapy. Clinicians should be aware to early recognize acute and subacute respiratory adverse events for a promptly management. In these patients re-treatment is challenging.


The combination of a left aortic arch, a right-sided descending thoracic aorta with a left-sided arterial duct.
A nine-year-old male child presented with a history of recurrent chest infections and breathlessness. On investigation, he was found to have a left aortic arch with a right-sided descending thoracic aorta along with a left-sided arterial duct. He also had mild aortic stenosis with regurgitation. The duct was successfully ligated under controlled hypotensive anesthesia through a left posterolateral thoracotomy.


Radionuclide scintigraphy in the evaluation of ventricular septal defect complicating acute myocardial infarction.
A 67-year-old man with an autopsy-proven ventricular septal defect (VSD) following an acute inferoposterior myocardial infarction underwent bedside evaluation by radionuclide imaging techniques. The presence of the VSD was confirmed, and the size of the shunt determined by qualitative and quantitative radionuclide angiocardiography. In addition, radionuclide ventriculography permitted assessment of global as well as regional left ventricular function.


Dynein's Life in the Slow Lane.
Dynein synthesized in neuronal cell bodies is conveyed into the axon by slow transport, but underlying mechanisms are unclear. In this issue, Twelvetrees et al. (2016) propose a model where dynein is transported by direct-but transient-interactions with kinesin.


Development of mice with brain-specific deletion of floxed glud1 (glutamate dehydrogenase 1) using cre recombinase driven by the nestin promoter.
In the brain, Glud1-encoded glutamate dehydrogenase plays a major role in the recycling of the neurotransmitter glutamate. We recently reported a new model of brain-specific Glud1 null mice (Cns-Glud1 (-/-)) lacking glutamate dehydrogenase in the central nervous system. Cns-Glud1 (-/-) mice exhibit reduced astrocytic glutamate breakdown and redirection of glutamate pathways without altering synaptic transmission. Cns-Glud1 (-/-) mice were generated using LoxP and Nestin-Cre technology. Nestin-Cre mice are widely used to investigate gene deletion in the central nervous system. However, the Nes-Cre transgene itself was reported to induce a phenotype related to body weight gain. Here, we review the potential side-effects of Nes-Cre and analysed Cns-Glud1 (-/-) body weight gain. Overall, Nestin-Cre mice may exhibit transient and moderate growth retardation during the few weeks immediately following weaning. Pending appropriate controls and homogenization of the genetic background, Nestin-Cre technology is a valuable tool enabling disruption of genes of interest in the central nervous system.