Role of C-terminal regions of the C-terminal fragment of Clostridium perfringens enterotoxin in its interaction with claudin-4.
Claudin family proteins, which contain 4 transmembrane domains, play a pivotal role in the barrier function of tight junctions (TJs) in epithelial sheets. We previously found that a modulator of claudin-4, the C-terminal fragment of Clostridium perfringens enterotoxin (C-CPE), is a potent enhancer of jejunal drug absorption in rats. But the effects of C-CPE on the barrier function of TJs have never been fully understood. In the present study, we investigated the effects of C-CPE on the barrier function of TJs in Caco-2 monolayer and characterized the functional domain of C-CPE that is responsible for interaction with claudin-4. To evaluate the effects of C-CPE on the barrier function of TJs, we measured transepithelial electric resistance (TER) in Caco-2 monolayer cells seeded onto polycarbonate filters. Treatment of Caco-2 cells with C-CPE resulted in a decrease in TER. But, deletion of the 30 C-terminal amino acids of C-CPE, which is the putative binding domain for claudin, attenuated the decrease in TER values. Moreover, ablation of the 16 C-terminal amino acids of C-CPE also resulted in attenuation of the decrease in TER values. The C-terminal-deleted C-CPEs did not interact with claudin-4 or the extracellular domain 2 of claudin-4, which is the C-CPE binding site. These results suggest that the 16 C-terminal amino acids of C-CPE are responsible for the interaction of C-CPE and claudin-4 following the disruption of TJ barrier function.


Phytotoxic, antibacterial, and antioxidant activities of mycotoxins and other metabolites from Trichoderma sp.
A new natural mycotoxin was isolated from the fermentation broth of Trichoderma sp. Jing-8 and the structure was determined as alternariol 1'-hydroxy-9-methyl ether (1), together with twelve known compounds. The structures were elucidated on the basis of their 1D, 2D NMR spectra and mass spectrometric data. Compounds 1, 8 and 9 indicated inhibitions against germination of the seeds of cabbage with MICs < 3 μg/mL. The compound 1 showed the antibacterial activity against Bacillus subtilis and Staphylococcus aureus with MICs at 64 μg/mL. Compound 1 and 3 showed significant DPPH radical-scavenging activities with IC50 at 12 μg/mL, respectively. The OH at C-1' in compound 1 decreased the cytotoxicity of these mycotoxins. A primary structure-activity relationship about the alternariol derivatives was discussed. Compounds 2-7 and 8 were the first time to be isolated from the Trichoderma.


Delocalisation of photoexcited triplet states probed by transient EPR and hyperfine spectroscopy.
Photoexcited triplet states play a crucial role in photochemical mechanisms: long known to be of paramount importance in the study of photosynthetic reaction centres, they have more recently also been shown to play a major role in a number of applications in the field of molecular electronics. Their characterisation is crucial for an improved understanding of these processes with a particular focus on the determination of the spatial distribution of the triplet state wavefunction providing information on charge and energy transfer efficiencies. Currently, active research in this field is mostly focussed on the investigation of materials for organic photovoltaics (OPVs) and organic light emitting diodes (OLEDs). As the properties of triplet states and their spatial extent are known to have a major impact on device performance, a detailed understanding of the factors governing triplet state delocalisation is at the basis of the further development and improvement of these devices. Electron Paramagnetic Resonance (EPR) has proven a valuable tool in the study of triplet state properties and both experimental methods as well as data analysis and interpretation techniques have continuously improved over the last few decades. In this review, we discuss the theoretical and practical aspects of the investigation of triplet states and triplet state delocalisation by transient continuous wave and pulse EPR and highlight the advantages and limitations of the presently available techniques and the current trends in the field. Application of EPR in the study of triplet state delocalisation is illustrated on the example of linear multi-porphyrin chains designed as molecular wires.


Opportunities for Interaction : Natural Observations of Children's Social Behavior in Five Societies.
We examine the opportunities children have for interacting with others and the extent to which they are the focus of others' visual attention in five societies where extended family communities are the norm. We compiled six video-recorded datasets (two from one society) collected by a team of anthropologists and psychologists conducting long-term research in each society. The six datasets include video observations of children among the Yasawas (Fiji), Tanna (Vanuatu), Tsimane (Bolivia), Huatasani (Peru), and Aka (infants and children 4-12 years old; Central African Republic). Each dataset consists of a series of videos of children ranging in age from 2 months to 12 years in their everyday contexts. We coded 998 videos and identified with whom children had opportunities to interact (male and female adults and children) as well as the number of individuals and the proportion of observed time that children spent with these individuals. We also examined the proportion of time children received direct visual gaze (indicating attention to the child). Our results indicate that children less than 5 years old spend the majority of their observed time in the presence of one female adult. This is the case across the five societies. In the three societies from which we have older children (Aka, Yasawa, Peru), we find a clear shift around 5 years of age, with children spending the majority of their time with other children. We also coded the presence or absence of a primary caregiver and found that caregivers remained within 2 ft of target children until 7 years of age. When they were in the company of a primary caregiver, children older than seven spent the majority of their time more than 2 ft from the caregiver. We found a consistent trend across societies with decreasing focal attention on the child with increasing child age. These findings show (1) remarkable consistency across these societies in children's interaction opportunities and (2) that a developmental approach is needed to fully understand human development because the social context is dynamic across the lifespan. These data can serve as a springboard for future research examining social development in everyday contexts.


Hydrogel-mediated delivery of microRNA-92a inhibitor polyplex nanoparticles induces localized angiogenesis.
Localized stimulation of angiogenesis is an attractive strategy to improve the repair of ischemic or injured tissues. Several microRNAs (miRNAs) such as miRNA-92a (miR-92a) have been reported to negatively regulate angiogenesis in ischemic disease. To exploit the clinical potential of miR-92a inhibitors, safe and efficient delivery needs to be established. Here, we used deoxycholic acid-modified polyethylenimine polymeric conjugates (PEI-DA) to deliver a locked nucleic acid (LNA)-based miR-92a inhibitor (LNA-92a) in vitro and in vivo. The positively charged PEI-DA conjugates condense the negatively charged inhibitors into nano-sized polyplexes (135 ± 7.2 nm) with a positive net charge (34.2 ± 10.6 mV). Similar to the 25 kDa-branched PEI (bPEI25) and Lipofectamine RNAiMAX, human umbilical vein endothelial cells (HUVECs) significantly internalized PEI-DA/LNA-92a polyplexes without any obvious cytotoxicity. Down-regulation of miR-92a following the polyplex-mediated delivery of LNA-92a led to a substantial increase in the integrin subunit alpha 5 (ITGA5), the sirtuin-1 (SIRT1) and Krüppel-like factors (KLF) KLF2/4 expression, formation of capillary-like structures by HUVECs, and migration rate of HUVECs in vitro. Furthermore, PEI-DA/LNA-92a resulted in significantly enhanced capillary density in a chicken chorioallantoic membrane (CAM) model. Localized angiogenesis was substantially induced in the subcutaneous tissues of mice by sustained release of PEI-DA/LNA-92a polyplexes from an in situ forming, biodegradable hydrogel based on clickable poly(ethylene glycol) (PEG) macromers. Our results indicate that PEI-DA conjugates efficiently deliver LNA-92a to improve angiogenesis. Localized delivery of RNA interference (RNAi)-based therapeutics via hydrogel-laden PEI-DA polyplex nanoparticles appears to be a safe and effective approach for different therapeutic targets.


Histopathological differences of myotonic dystrophy type 1 (DM1) and PROMM/DM2.
Muscle biopsy findings in DM2 have been reported to be similar to those in DM1. The authors used myosin heavy chain immunohistochemistry and enzyme histochemistry for fiber type differentiation on muscle biopsies. Their results show that DM2 patients display a subpopulation of type 2 nuclear clump and other very small fibers and, hence, preferential type 2 fiber atrophy in contrast to type 1 fiber atrophy in DM1 patients.


Mechanistic implications of alterations in HL-60 cell nascent DNA after exposure to 1-beta-D-arabinofuranosylcytosine.
To improve our understanding of the mechanism of 1-beta-D-arabinofuranosylcytosine (ara-C) incorporation into DNA, we investigated the physical properties (size, position of nucleoside incorporation) of small fragments of nascent DNA (nDNA) obtained by pH-step alkaline elution of intact HL-60 cells following their exposure to ara-C. In the pH-step alkaline elution procedure, the smallest fragments of nDNA elute at pH 11. Anion-exchange high-performance liquid chromatography (HPLC) of nDNA obtained by 1 h elution at pH 11.0 of lysed HL-60 cells revealed a preponderance of nDNA fragments ranging from 0.5 to 40 kb in control ([3H]-dThd-labeled) cells. Exposure of cells to ara-C (0.8-1 microM) resulted in a loss of the preponderance of radiolabel in fragments of 0.5-40 kb along with redistribution of the radiolabel (from [3H]-dThd or [3H]-ara-C) into smaller nDNA fragments (predominantly < 100 bases in length) as determined by HPLC. We used the ability of pH-step alkaline elution to provide these small nDNA fragments produced by ara-C to investigate the paradoxical behavior of ara-C as a chain terminator in cell-free DNA synthetic systems while being incorporated into an internucleotide position in intact cells. Following the digestion of purified nDNA with micrococcal nuclease and spleen phosphodiesterase II, the proportion of radiolabel in 3'-dNMP (indicating an internucleotide position) or free nucleoside (indicating a chain terminus position) was determined by reverse-phase HPLC. In digests of prelabeled genomic DNA, as expected, > 90% of the radiolabel from [14C]-dThd or [3H]-ara-C was found to exist in an internucleotide position (as determined by co-chromatography with authentic 3'-dTMP or 3'-ara-CMP). In contrast, digests of nDNA that eluted at pH 11.0 revealed a significantly higher proportion of radiolabel in the chain terminus position (29%-35%) when the nDNA was obtained from cells exposed to 1 microM [3H]-ara-C as compared with cells exposed to [3H]-dThd or [3H]-dCyd alone (< 10%). These data obtained from pH-step alkaline elution of intact cells suggest that by causing the inhibition of chain elongation while failing to inhibit the formation of new nDNA replication intermediates, ara-C exposure leads to the production of very small nDNA fragments. This relative chain-terminating effect of ara-C is most apparent in the small nDNA replication fragments that elute at pH 11.0.(ABSTRACT TRUNCATED AT 400 WORDS)


Assessment of uterine cavity by hysteroscopy in assisted reproduction programme and its influence on pregnancy outcome.
The aim of the study was to evaluate if the diagnosis and treatment of uterine cavity abnormalities by hysteroscopy in patients undergoing IVF programme is of any value in improving clinical pregnancy outcome. 520 patients participated in this prospective randomized study and were classified into two groups. Group I (n = 265) without office hysteroscopy. Group II (n = 255) had office hysteroscopy and was sub classified into Group II a and Group II b. Group II a (n = 160) had normal hysteroscopic findings whereas Group II b (n = 95) had abnormal office hysteroscopy findings, which were corrected at the same time. There was no difference in the mean number of oocytes retrieved, fertilization rate, and number of embryos transferred among the patients in different groups. Statistically significant difference was observed in terms of clinical pregnancy rates between Group I and Group II a (26.2 and 44.44%, P < 0.05), and Group I and Group II b (26.2 and 39.55%, P < 0.05), respectively. Patients with recurrent IVF embryo transfer failures after normal hysterosalpingography findings should also be reevaluated using hysteroscopy prior to further commencing IVF-embryo transfer cycles in order to enhance the clinical pregnancy rates.


The disclosure of the diagnosis of cancer.
Before significant efforts relative to patient education and subsequent informed decision making can occur, it is essential to assess the patient's perceptions and emotional responses to the diagnosis. The direction and depth of this exploration are likely to be determined by such factors as quality of the previous relationship with the patient, the patient's belief and support system, and the patient's previous experience with trauma. Other determining factors are the sense of urgency with which medical decisions need to be made and the interpersonal skills of the physician. It is important at the time of diagnosis to understand and respond to the patient's cognitive and emotional responses to the diagnosis. The sensitive physician allows time to acknowledge and legitimate those responses. It is often important to allow the ventilation of those reactions before further education and decision making can occur. By accepting and acknowledging the patient's reactions, the physician can reduce the patient's feelings of isolation and helplessness. The quality of such a relationship then becomes the foundation on which the patient and family and physician can begin to attend to the issues before them.


Veratridine delays apoptotic neuronal death induced by NGF deprivation through a Na(+)-dependent mechanism in cultured rat sympathetic neurons.
Superior-cervical ganglion (SCG) cells dissociated from newborn rats depend on nerve growth factor (NGF) for survival. Membrane depolarization with elevated K+ is known to prevent neuronal death following NGF deprivation and/or to promote survival via a Ca(2+)-dependent mechanism. Here we have exploited the possibility of whether or not a Na(+)-dependent pathway for neuronal survival is present in these cells. Veratridine (EC50 = 40 nM), a voltage-dependent Na+ channel activator, significantly delayed the onset of apoptotic cell death in NGF-deprived SCG neurons that had been cultured for 7 days in the presence of NGF. This effect was blocked completely by Na+ channel blockers including tetrodotoxin (TTX, 1 microM), benzamil (25 microM) and flunarizine (1 microM), but was not attenuated by nimodipine (1 microM), an L-type Ca2+ channel blocker. The saving effect of veratridine on cultured neurons was observed even in low Ca2+ media (0-1.0 mM), but was completely abolished in a low Na+ medium (38 mM). Sodium-binding benzofuran is isophthalate was employed as a fluorescent probe for monitoring the level of cytoplasmic free Na+, which revealed a sustained increase in its level (12.9 mM, 307% of that of control) in response to veratridine (0.75 microM). The TTX or flunarizine completely blocked veratridine-induced Na+ influx in these cultured neurons. Moreover, no appreciable increase in intracellular Ca2+ was detected under these conditions. Though Na+ channels were effectual in SCG neurons which were freshly isolated from newborn rats, the Na(2+)-dependent saving effect of veratridine was not observed in these young neurons. These lines of evidence suggest that the death-suppressing effect of veratridine on cultured SCG neurons depends on the Na+ influx via voltage-dependent Na+ channels, and suggest the presence of Na(+)-dependent regulatory mechanism(s) in neuronal survival.


Subglottic stenosis induced by extranodal mucosa-associated lymphoid tissue lymphoma.
Mucosa-associated lymphoid tissue (MALT) lymphoma is usually associated with a chronic inflammatory disease or autoimmune disorders from which lymphoid tissue of MALT type arises as a prerequisite for lymphoma proliferation. Primary hematopoietic neoplasms of the larynx are rare. MALT lymphomas of the larynx are believed to arise from preexisting or acquired lymphoid tissue of the upper airway which is documented in the supraglottic region. Therefore, these are mainly located in the supraglottic and glottic areas, with only a few reported in the subglottic region. We report on a 50-year-old woman with a hoarseness, stridor, and developing exertional dyspnea. On indirect laryngoscope, multiple nodular lesions with smooth surface over the subglottis with subglottic steonsis was found. The biopsy confirmed the diagnosis of a MALT lymphoma. We hope to promote awareness and consideration of MALT lymphoma in the differential diagnosis in subglottic steonsis.


Comparisons between older and usual-aged medical school graduates on the factors influencing their choices of primary care specialties.
To examine whether there are differences between older (30+ years) and usual-aged graduates in the factors that influence their decisions to enter primary care. A national survey of primary care physicians was conducted by mail in early 1993. The survey population comprised physicians who had graduated from U.S. allopathic medical schools in 1983 and 1984. The questionnaire consisted of four parts: practice characteristics, 19 variables influencing the physicians' decisions to enter primary care (rated on a scale from 5 = very strong influence to 1 = no influence), the timing of the decision to enter primary care, and demographic and personal information. Data were analyzed through several statistical methods. In all, 355 (22%) older and 1,241 (78%) usual-aged graduates who were practicing in primary care specialties responded. Compared with the usual-aged graduates, the older graduates were more likely to have grown up in rural or inner-city areas, to have obtained a second academic degree, and to have made the decision to enter primary care earlier. The older graduates' decisions to enter primary care had been more influenced by children and familial responsibilities, whereas the usual-aged graduates had been more influenced by internship and residency experiences and by parents and role models before medical school. This study provides empirical evidence to support the notion that nontraditional students (i.e., older ones) are more likely to commit themselves earlier to a career decision and less likely to be influenced by the socialization process during medical school. In this regard, age-specific factors should receive more attention in the analysis of the physician workforce.


Natural killer cell dysfunction in cancer and new strategies to utilize NK cell potential for cancer immunotherapy.
The continuous, in-depth exploration of the occurrence and development of cancer has shown that immune cell dysfunction is closely associated with tumor progression and poor clinical prognosis. The inhibition of the effector functions of immune cells by numerous immunosuppressive factors in the tumor microenvironment (TME) promotes the progression and metastasis of malignant tumors. Natural killer (NK) cells are the main effector cells in the anti-tumor innate immune system. Dysfunctional NK cells, characterized as weakened proliferation capacity and reduced production of effector cytokines, have limited ability to kill malignant cells and inhibit tumor progression. The reversal of the dysfunctional state of NK cells and enhancement of their effector functions is a promising strategy that could improve the effectiveness of cancer immunotherapy. In order to fully use of the cytotoxic effects of NK cells and revitalize the anti-tumor potential of NK cells in tumor patients, it is necessary to learn more about the characteristics of NK cell dysfunction in TME. This will provide valuable information for the development of personalized strategies to restore anti-tumor immunity. Here, we reviewed the characteristics of dysfunctional NK cells in the TME and latest progress in research, and discussed promising immunotherapy strategies that could utilize NK cell potential for cancer immunotherapy.


"First Come, First Served" and Threshold Effects in a Central-to-Planar-to-Helical Hierarchical Chiral Induction.
Perylene diimide tethered pillar[5]arene derivatives form aggregates in non-polar organic solvents, and the complexation of cationic amino acid ethyl ester (cAA-OEt) with the aggregates induce a central-to-planar-to-helical chirality transfer, leading to intensive circular dichroism (CD) signals having dissymmetric factors (gabs ) of up to 3.67×10-2 . The hierarchical chiral induction exhibited an intriguing threshold dose effect, namely, the chiral induction does not occur in the low concentration range of cAA-OEt but is triggered when cAA-OEt exceeds a threshold concentration. The inhibited interconversion between the Rp and Sp conformers of pillar[5]arene, which is further restricted in the aggregation, plays a crucial role in the threshold effect. When adding enantiopure cAA-OEt first to the threshold concentration and then adding an equal amount of the antipodal cAA-OEt to give cAA-OEt in racemic form, CD spectra having the same sign as the CD induced by first adding pure cAA-OEt were induced, thus showing an unprecedented "first come, first served" effect.


Toxicological evaluation of cyclosporin A.
Toxicological studies in a variety of different animal species have shown that cyclosporin A (CS-A) is a unique immunosuppressant, since it does not cause myelotoxic, teratogenic, mutagenic, or carcinogenic effects. This is undoubtedly due to the fact CS-A produces its immunosuppressive effects by a specific action on T lymphocytes rather than by a cytotoxic mechanism. The incidence of acute inflammatory lesions was also not significantly higher in treated animals than in controls. Clear-cut organ toxicity only occurred in rats and cynomolgus monkeys, where at high doses nephro- and hepatotoxicity were observed.


General and molecular microbiology and microbial genetics in the IM CAS.
This review summarizes the main results obtained in the fields of general and molecular microbiology and microbial genetics at the Institute of Microbiology of the Academy of Sciences of the Czech Republic (AS CR) [formerly Czechoslovak Academy of Sciences (CAS)] over more than 50 years. Contribution of the founder of the Institute, academician Ivan Málek, to the introduction of these topics into the scientific program of the Institute of Microbiology and to further development of these studies is also included.


Epitope mapping of a monoclonal antibody against human thrombin by H/D-exchange mass spectrometry reveals selection of a diverse sequence in a highly conserved protein.
The epitope of a monoclonal antibody raised against human thrombin has been determined by hydrogen/deuterium exchange coupled to MALDI mass spectrometry. The antibody epitope was identified as the surface of thrombin that retained deuterium in the presence of the monoclonal antibody compared to control experiments in its absence. Covalent attachment of the antibody to protein G beads and efficient elution of the antigen after deuterium exchange afforded the analysis of all possible epitopes in a single MALDI mass spectrum. The epitope, which was discontinuous, consisting of two peptides close to anion-binding exosite I, was readily identified. The epitope overlapped with, but was not identical to, the thrombomodulin binding site, consistent with inhibition studies. The antibody bound specifically to human thrombin and not to murine or bovine thrombin, although these proteins share 86% identity with the human protein. Interestingly, the epitope turned out to be the more structured of two surface regions in which higher sequence variation between the three species is seen.


Needle core biopsy characteristics identify patients at risk of compromised margins in breast conservation surgery.
Selection of patients for breast-conserving surgery relies on inexact parameters such as the preoperative estimation of lesion size. This study investigates the value of needle core biopsy findings, in particular, the relative quantity of DCIS, in improving patient selection for breast conservation. Patients undergoing breast-conserving surgery for invasive ductal carcinoma from 1999 to 2004 were identified. Only patients who had a preoperative diagnosis of carcinoma (DCIS and invasive) on core biopsy were included. All core biopsies were reviewed by a breast histopathologist to document the quantity and characteristics of the DCIS component. Of a total of 281 patients, 46% (n=129) had invasive disease on core biopsy (group 1) and 54% (n=152) had either invasive disease with an accompanying DCIS component or DCIS only on core biopsy (group 2). The compromised margin rate for group 1 was 23% compared to 39% for group 2 (P=0.004). The rate of compromised margins increased progressively as the core biopsy DCIS component increased until a rate of 75% (n=18/24) was reached in patients with DCIS only on core biopsy. In patients with a DCIS component on core biopsy, the presence of necrosis (P=0.002), solid type architecture (P=0.008), high grade DCIS (P=0.007), calcification (P=0.003), and the relative proportion of DCIS present (P<0.001) were associated with compromised margins on univariate analysis. On multivariate analysis of this subgroup, the proportion of DCIS in this subgroup (P=0.048) was an independent predictor of compromised margins. The presence and relative proportion of DCIS on core biopsy provides important information as to whether patients are at risk of compromised margins. Documentation of these parameters may assist patient selection for breast-conserving surgery or identify patients who may benefit from wider margins at the time of initial operation.


Development and validation of the Scale for Staff-Family Partnership in Long-term Care (SSFPLC).
A partnership between staff and families is crucial to maintain nursing home residents' health and promote quality of care, and currently, there is a need for a measurement tool to assess the partnership. This study aimed to develop a tool for assessing the partnership between staff and the families of older adult nursing home residents from the perspective of staff members and to verify its reliability and validity. The instrument was developed in the following four stages: (1) generation of an item pool via literature review and focus group interview, (2) experts' content validity analysis of 32 items, (3) a preliminary survey on 30 staff members and (4) validity and reliability tests of the instrument on 346 staff members in nursing homes between July and October 2018. The final instrument consists of 17-items in four categories (family's trust and support, collaborative relationship and communication, encouragement to participate in care and professional care). Each item was rated on a 4-point Likert scale, and the higher scores indicating better partnership between staff and families of nursing home residents. The reliability of the instrument was 0.90, and the test-retest intra-class correlation coefficient was 0.96. The Scale for Staff-Family Partnership in Long-term Care (SSFPLC) showed acceptable reliability and validity as an instrument to assess the partnership between nursing home staff and families. This tool can be used for evaluating staff and family partnerships within nursing homes.


Restorative effect of total parenteral nutrition on natural killer cell activity in malnourished cancer patients.
Decreased natural killer cell activity (NKCA) is associated with malnutrition in both cancer and non-cancer patients. We have studied the effect of total parenteral nutrition (TPN) on NKCA in 9 malnourished cancer patients, candidates for surgery. TPN was administered for a median of 10 days (range 7-11), providing 1.5-fold the estimated resting energy expenditure, with 30% as fat. Calorie:nitrogen ratio was 150:1. Basal human recombinant interferon-alpha 2a (rIFN-alpha 2a) and human recombinant IL-2 rIL-2) activated NKCA were measured, as were the main nutritional parameters, prior to and after TPN. NKCA increased in all patients and reached the normal range in 5, 3 and 4 subjects, respectively, for basal, rIFN-alpha 2a and rIL-2 activated NKCA. As regards nutritional assessment, body weight and IgM levels significantly increased from 47.7 to 50.1 kg and from 174 to 237 mg/dl, respectively. This study demonstrates that a 10-day TPN course increases and sometimes restores normal NKCA. Such effect was constant and preceded nutritional changes.


Effects of gossypol on human and monkey sperm motility in vitro.
The effects of gossypol acetic acid on human and monkey sperm motility in vitro were studied by using multiexposure photography technique. Human and monkey spermatozoa were inhibited by gossypol to different degrees. Monkey sperm were absolutely immotile within 15 min after 50 microM of gossypol was added, but motility of human spermatozoa was not completely suppressed by gossypol even at the highest concentration used and longest duration of exposure.


Effect of the Substrate Structure and Metal Ions on the Hydrolysis of Undamaged RNA by Human AP Endonuclease APE1.
Human apurinic/apyrimidinic (AP) endonuclease APE1 is one of the participants in the DNA base excision repair. The main biological function of APE1 is to hydrolyze the phosphodiester bond on the 5'-side of the AP sites. It has been shown recently that APE1 acts as an endoribonuclease and can cleave mRNA, thereby controlling the level of some transcripts. The sequences of CA, UA, and UG dinucleotides are the cleavage sites in RNA. In the present work, we performed a comparative analysis of the cleavage efficiency of model RNA substrates with short hairpin structures in which the loop size and the location of the pyrimidine-purine dinucleotide sequence were varied. The effect of various divalent metal ions and pH on the efficiency of the endoribonuclease reaction was analyzed. It was shown that site-specific hydrolysis of model RNA substrates depends on the spatial structure of the substrate. In addition, RNA cleavage occured in the absence of divalent metal ions, which proves that hydrolysis of DNA- and RNA substrates occurs via different catalytic mechanisms.


High-grade glioma in a patient with breast cancer.
Breast cancer is one of the most common origins of metastatic lesions in the central nervous system. Many patients with a breast cancer and concurrent brain tumor(s) were diagnosed to have a metastatic lesion or lesions in the brain, based exclusively on their image findings without further pathologic verification, and received radiotherapy alone thereafter. It is, however, possible that a different pathology such as primary brain malignancy, which actually warrants a specific treatment modality, may occur in such patients with an already known malignancy. We, herein, reported a 61-year-old female patient who suffered from an anaplastic oligodendroglioma 1 year after her diagnosis of breast cancer. Demographic data, characteristic imaging findings, treatment, and outcome of the patient were discussed.


Metapath-Based Deep Convolutional Neural Network for Predicting miRNA-Target Association on Heterogeneous Network.
Predicting the interactions between microRNAs (miRNAs) and target genes is of great significance for understanding the regulatory mechanism of miRNA and treating complex diseases. The emergence of large-scale, heterogeneous biological networks has offered unprecedented opportunities for revealing miRNA-associated target genes. However, there are still some limitations about automatically learn the feature information of the network in the existing methods. Since network representation learning can self-adaptively capture structure information of the network, we propose a framework based on heterogeneous network representation, MDCNN (Metapath-Based Deep Convolutional Neural Network), to predict the associations between miRNAs and target genes. MDCNN samples the paths between the node pairs in the form of meta-path based on the heterogeneous information network (HIN) about miRNAs and target genes. Then the node feature and the path feature which is learned by the Deep Convolutional Neural Network (DCNN) are spliced together as the representation of the miRNA-target gene, to predict the miRNA-target gene interactions. The experiment results indicate that the performance of MDCNN outperforms other methods in multiple validation metrics by fivefold cross validation. We set an ablation study to identify the necessity of miRNA similarity and target gene similarity for improving the prediction ability of MDCNN. The case studies on hsa-miR-26b-5p and CDKN1A further demonstrates that MDCNN can successfully predict potential miRNA-target gene interactions.


The impact of increasing workloads and role expansion on female community pharmacists in the United Kingdom.
Evidence from United Kingdom and the United States indicates that community pharmacists' workloads have increased in recent years for 2 reasons. First, because of social and demographic changes there has been a greater demand for pharmaceutical services. Second, the community pharmacists' role has expanded. This article explores the effect of increased workloads on female community pharmacists. Face-to-face interviews (n=30) were conducted with women over the age of 30 years in the North West of England who worked as community pharmacists. The interview schedule was designed to explore factors underpinning female working patterns in community pharmacy. Specifically, interviewees were asked about perceptions of working conditions, positive and negative aspects of community pharmacy working, views of recent changes in pharmacy, and future career plans. Findings suggest that although community pharmacists enjoy aspects of their new roles, their work environment has become increasingly pressurized, resulting in decreased job satisfaction. Additionally, this study found some evidence that increasing workloads resulted in decreased health and well-being. The role of the community pharmacist has significantly altered in recent years, and this has occurred following a decade of increasing workloads. Consideration of the factors shaping community pharmacy points to high-pressure working environments becoming common place. This is likely to have a negative impact on pharmacists and conceivably the services they provide.


Effect of COVID-19 lockdown on regional pollution in Ireland.
This study examines the regional impact of the COVID-19 lockdown restrictions on pollution in Ireland by comparing the 2020 measurements of ozone (O3), nitrogen dioxide (NO2), and particulate matter (PM) from monitoring stations around the country to the previous 3-year average. Results indicate that O3 was 5.6% lower and 13.7% higher than previous years during the lockdown at rural and suburban sites, respectively. NO2 decreased by 50.7% in urban areas, but increased slightly in agricultural regions, consistent with satellite observations. PM concentrations did not change significantly compared to previous years; however, a reduction in the signal variability in the smaller size particle measurements may be the result of different emission sources. The reduction in NO2 likely increased the ratio of volatile organic compounds (VOCs) to NOx (nitrogen oxides), creating a NOx limited environment, which resulted in an initial increase in O3 in suburban areas, and the lower than usual levels observed at rural sites. Meteorology showed higher than average wind speeds prior to lockdown, which likely acted to disperse PM and NO2.


Retinoids, breast cancer and NK cells.
N-(4-hydroxyphenyl) retinamide (4-HPR) is a synthetic retinoid which reduces the incidence of experimental tumours in animals and has been chosen for its weak toxicity to be tested as a chemopreventive agent in humans. The mechanism of antineoplastic action is still unknown but a possible immunoenhancing effect may be postulated. We investigated the NK activity of PBMC from a group of women treated with 4-HPR as a part of a large scale randomised phase III trial on chemoprevention of contralateral disease in mastectomised women. After 180 days of treatment the NK activity was augmented 1.73 times as compared to that of patients given a placebo. The NK activity of PBMC from 4-HPR treated women is maximised, being higher than the basal and even the rIL-2 or alfa-rIFN stimulated activity of controls. For this reason in the majority of cases it cannot be further augmented by incubation with either rIL-2 or alfa-rIFN in vitro. The increased NK activity of 4-HPR treated women is not due to an enhanced production of endogenous IL-2, because PBMC cultures from patients treated with 4-HPR or placebo, incubated in vitro with a panel of different stimulators (recall antigens, PHA, allogeneic and xenogeneic cells) produce similar amounts of IL-2. The functional activity, but not the number of NK cells is increased in 4-HPR treated women. The mechanism by which 4-HPR stimulates NK activity is not a function of direct action on NK cells. Indeed incubation of PBMC from blood donors with 4-HPR or its major metabolite N-(4-methoxyphenyl) retinamide (4-MPR) does not modify their natural cytotoxicity.


Unfavorable Treatment Outcome and Its Predictors Among Patients with Multidrug-Resistance Tuberculosis in Southern Ethiopia in 2014 to 2019: A Multi-Center Retrospective Follow-Up Study.
According to the 2017 global report, Ethiopia is among the top 30 high tuberculosis (TB) and multidrug-resistant tuberculosis (MDR-TB) burden countries. However, studies on MDR-TB treatment outcomes in Southern Ethiopia was very limited. Therefore, the study was aimed at determining the unfavorable treatment outcome and its predictors among patients with multidrug-resistant tuberculosis in Southern Ethiopia MDR-TB treatment centers. A retrospective follow-up study was conducted in Southern Ethiopia MDR-TB treatment initiating centers. Three hundred sixty-three patients were included in the study. Kaplan-Meier failure curve, median time, and Log rank test were used to present the descriptive findings. Then, a Cox regression analysis was used to identify predictors of unfavorable treatment outcome. The strength of the association was reported using an adjusted hazard ratio (AHR) and a 95% confidence interval (CI). Finally, the Cox Snell residual test was used to check the goodness of fit. For the entire cohort, the unfavorable treatment outcome was 23.68% (19.29, 28.09). Hospitalization for care (AHR = 2.07; 95% CI = 1.21, 3.63), male sex (AHR = 1.85; 95% CI = 1.002, 3.42), attending tertiary education (AHR = 0.31; 95% CI = 0.11, 0.91), and those with low hemoglobin (AHR = 2.89; 95% CI = 1.55, 5.38) were predictors for unfavorable treatment outcome. The unfavorable treatment outcome was higher compared with the national goal of END-TB by 2020. Hospitalizations for care, male sex, and low hemoglobin level increased the hazard of the unfavorable treatment outcome. On the other hand, attending territory education decreased the hazard of the unfavorable treatment outcome.


Differential Performance of CoronaCHEK SARS-CoV-2 Lateral Flow Antibody Assay by Geographic Origin of Samples.
We assessed the performance of CoronaCHEK lateral flow assay on samples from Uganda and Baltimore to determine the impact of geographic origin on assay performance. Serum samples from SARS-CoV-2 PCR+ individuals (Uganda: 78 samples from 78 individuals and Baltimore: 266 samples from 38 individuals) and from pre-pandemic individuals (Uganda 1077 and Baltimore 532) were evaluated. Prevalence ratios (PR) were calculated to identify factors associated with a false-positive test. After first positive PCR in Ugandan samples the sensitivity was: 45% (95% CI 24,68) at 0-7 days; 79% (95%CI 64,91) 8-14 days; and 76% (95%CI 50,93) >15 days. In samples from Baltimore, sensitivity was: 39% (95% CI 30, 49) 0-7 days; 86% (95% CI 79,92) 8-14 days; and 100% (95% CI 89,100) 15 days post positive PCR. The specificity of 96.5% (95% CI 97.5,95.2) in Ugandan samples was significantly lower than samples from Baltimore 99.3% (95% CI 98.1,99.8), p<0.01. In Ugandan samples, individuals with a false positive result were more likely to be male (PR 2.04, 95% CI 1.03,3.69) or individuals who had a fever more than a month prior to sample acquisition (PR 2.87, 95% CI 1.12,7.35). Sensitivity of the CoronaCHEK was similar in samples from Uganda and Baltimore. The specificity was significantly lower in Ugandan samples than in Baltimore samples. False positive results in Ugandan samples appear to correlate with a recent history of a febrile illness, potentially indicative of a cross-reactive immune response in individuals from East Africa.


Monitoring offshore oil pollution using multi-class convolutional neural networks.
Oil and gas production operations are a major source of environmental pollution that expose people and habitats in many coastal communities around the world to adverse health effects. Detecting oil spills in a timely and precise manner can help improve the oil spill response process and channel required resources more effectively to affected regions. In this research, convolutional neural networks, a branch of artificial intelligence (AI), are trained on a visual dataset of oil spills containing images from different altitudes and geographical locations. In particular, a VGG16 model is adopted through transfer learning for oil spill classification (i.e., detecting if there is oil spill in an image) with an accuracy of 92%. Next, Mask R-CNN and PSPNet models are used for oil spill segmentation (i.e., pixel-level detection of oil spill boundaries) with a mean intersection over union (IoU) of 49% and 68%, respectively. Lastly, to determine if there is an oil rig or vessel in the vicinity of a detected oil spill and provide a holistic view of the oil spill surroundings, a YOLOv3 model is trained and used, yielding a maximum mean average precision (mAP) of ~71%. Findings of this research can improve the current practices of oil pollution cleanup and predictive maintenance, ultimately leading to more resilient and healthy coastal communities.


High-Resolution Characterization of KIR Genes in a Large North American Cohort Reveals Novel Details of Structural and Sequence Diversity.
The KIR (killer-cell immunoglobulin-like receptor) region is characterized by structural variation and high sequence similarity among genes, imposing technical difficulties for analysis. We undertook the most comprehensive study to date of KIR genetic diversity in a large population sample, applying next-generation sequencing in 2,130 United States European-descendant individuals. Data were analyzed using our custom bioinformatics pipeline specifically designed to address technical obstacles in determining KIR genotypes. Precise gene copy number determination allowed us to identify a set of uncommon gene-content KIR haplotypes accounting for 5.2% of structural variation. In this cohort, KIR2DL4 is the framework gene that most varies in copy number (6.5% of all individuals). We identified phased high-resolution alleles in large multi-locus insertions and also likely founder haplotypes from which they were deleted. Additionally, we observed 250 alleles at 5-digit resolution, of which 90 have frequencies ≥1%. We found sequence patterns that were consistent with the presence of novel alleles in 398 (18.7%) individuals and contextualized multiple orphan dbSNPs within the KIR complex. We also identified a novel KIR2DL1 variant, Pro151Arg, and demonstrated by molecular dynamics that this substitution is predicted to affect interaction with HLA-C. No previous studies have fully explored the full range of structural and sequence variation of KIR as we present here. We demonstrate that pairing high-throughput sequencing with state-of-art computational tools in a large cohort permits exploration of all aspects of KIR variation including determination of population-level haplotype diversity, improving understanding of the KIR system, and providing an important reference for future studies.


Fat storage is partially dependent on vagal activity and insulin secretion of hypothalamic obese rat.
Hypothalamic MSG-obese rats show hyperinsulinemia and tissue insulin resistance, and they display intense parasympathetic activity. Current analysis investigates whether early subdiaphragmatic vagotomy prevents tissue insulin sensitivity impairment in adult obese MSG-rats. Hypothalamic obesity was induced by MSG (4 mg/g BW), daily, from birth up to 5 days. Control animals receiving saline solution. On the 30th day rats underwent bilateral subdiaphragmatic vagotomy or sham surgery. An intravenous glucose tolerance test (i.v.GTT) was performed when rats turned 90 days old. Total white fat tissue (WAT) from rat carcass was extracted and isolated; the interscapular brown fat tissue (IBAT) was weighed. Rather than blocking obesity, vagotomy reduced WAT and IBAT in MSG-obese rats when the latter were compared to sham MSG-rats. High blood fasting insulin and normal glucose levels were also observed in MSG-obese rats. Although glucose intolerance, high insulin secretion, and significant insulin resistance were recorded, vagotomy improved fasting insulinemia, glucose tolerance and insulin tissue sensitivity in MSG-obese rats. Results suggest that increased fat accumulation is caused, at least in part, by high blood insulin concentration, and enhanced parasympathetic activity on MSG-obese rats.


Superoxide dismutase, glutathione peroxidase and thiobarbituric acid-reactive compounds in erythrocytes in Duchenne muscular dystrophy.
The activities of erythrocyte superoxide dismutase and glutathione peroxidase in patients with Duchenne muscular dystrophy were normal compared with healthy male controls, as were the levels of thiobarbituric acid-reactive compounds produced during incubation of erythrocytes in the presence of 10 mmol.1-1 hydrogen peroxide. These results suggest that the chief mechanisms for protecting the erythrocyte membrane from peroxidative damage are unimpaired and do not support the idea that the reported erythrocyte abnormalities in this disease result from increased susceptibility to lipid peroxidation.


Spatiotemporal variation in near-surface CH4 concentrations in China over the last two decades.
Methane is one of the main greenhouse trace gases and seriously affects the radiation balance of Earth systems due to its strong heat absorption capacity and long atmospheric retention time. Based on the methane stratification data simulated by the community atmospheric model with chemistry (CAM-chem), near-surface methane concentrations were estimated by utilizing the Gaussian function, and the spatiotemporal variation in the near-surface methane concentration in China from 2001 to 2019 was discussed in this research. The results show that (1) based on the methane stratification concentration data simulated by the atmospheric chemical model, the near-surface CH4 concentration estimated by Gaussian function model is reliable, which provides a new method to estimate the near-surface CH4 concentration over China; (2) from 2001 to 2019, the near-surface methane concentration in China showed an increasing trend with an annual growth rate of 7.20±0.23 ppb·a-1. The annual maximum near-surface methane concentration was measured in winter, and the minimum was measured in summer; (3) the spatial distribution differences are obvious: the methane concentration in the east was higher than that in the west, and the methane concentration in the north was higher than that in the south. Moreover, the distributions of methane in the east and west are consistent with the division of Hu Huanyong population line.


Porous polymer film formation by water droplet templating using polystyrene.
Studies show that the formation of breath figures over polystyrene is not clearly understood-sometimes the patterns are regular and sometimes they are barely formed. In an attempt to understand this process a little more, breath figures over polystyrene of three molecular weights and on the smooth and grooved DVD surfaces are prepared and studied. The microporous films are prepared by the evaporation of the chloroform solution of the polymers in a humid environment. The thus formed breath figure patterns are studied under a confocal laser scanning microscope and the images are analyzed. Breath figures were formed for (a) three molecular weights of the polymer (b) two casting techniques, and (c) on smooth and grooved surfaces (of a commercial DVD). The wetting of the breath figures formed by water is also reported here. The pore diameters were found to increase with increase in molecular weight and also with concentration of the polymer used. Only drop-casting method yield breath figures. Voronoi entropy, calculated from the images, indicates ordered pores on the grooved surface compared to smooth surfaces. Contact angle studies indicate a hydrophobic nature of the polymer, with the hydrophobicity increasing by the patterning.


Multiple cranial neuropathy as the initial presentation of metastatic prostate adenocarcinoma: case report and review of literature.
The skull base is an atypical metastatic site for prostate carcinoma. It is usually encountered late in the disease process in patients with known advanced disease. However, skull base involvement causing cranial nerve palsies may rarely be the presenting sign of prostate carcinoma. Such patients may present to a number of specialties including neurosurgery and can pose a diagnostic challenge in the absence of lower urinary tract symptoms. Here, we describe an unusual case of prostate adenocarcinoma presenting as a central skull base tumour with multiple cranial neuropathy.


Aggregation induced emission-active two-photon absorption zwitterionic chromophore for bioimaging application.
The fabrication of two-photon absorption material is a versatile approach to achieve high resolution bioimaging with low phototoxicity yet remain sophisticated. Herein, a zwitterionic chromophore, MF, with D-π-A configuration has been rational designed and synthesized. Remarkably, MF exhibited enhanced one- and two-photon fluorescent in the aggregation states. Additionally, the obtained MFNPs encapsulated by Pluronic F-127, could be employed as a two-photon fluorescent probe for bioimaging. The results reveal that MFNPs could target mitochondria by using two-photon confocal microscopy and stimulated emission depletion nanoscopy methods.


Evaluation of sperm damage: beyond the World Health Organization criteria.
Current knowledge of the molecular basis of male infertility that is associated with oxidative stress, apoptosis, and chromatin damage calls for changes in the laboratory assessment of the fertility potential of semen samples.


Systematic identification and replication of factors associated with human papillomavirus vaccine initiation among adolescents in the United States using an environment-wide association study approach.
Human papillomavirus (HPV) vaccination coverage is low among adolescents in the USA. Identification of factors associated with HPV vaccine initiation (receipt of ≥1 dose) is critical for improving uptake. Our objective was to systematically investigate all eligible factors available in a nationally representative sample of adolescents to identify drivers of HPV vaccine initiation using a novel methodological approach. We performed multiple cross-sectional analyses using data from the adolescent component of the National Immunization Surveys (NIS)-Teen between 2014 and 2019. Study participants were parents or caregivers of adolescents aged 13-17 years. Exposure variables measured sociodemographic and geographical characteristics, health conditions and healthcare provision. We tested the association between each factor and HPV vaccine initiation using univariate logistic regression and multivariate logistic regression adjusted for mother's age, mother's education level, mother's marital status, poverty status and adolescent's sex. We validated findings for each type of analysis within surveys, between surveys (across years 2014-2019) and across several subgroups (age, sex, poverty status and race/ethnicity). Six factors were replicated in the multivariate analysis. Most replicated factors characterised the role of healthcare providers and healthcare-seeking behaviours. After adjustment, provider HPV recommendation remained the most strongly associated with HPV vaccine initiation (2019 NIS-Teen: OR 13.4, 95% CI 11.3 to 17.3, p<0.001). The variance explained by a full model including replicated factors was 0.39. This is the first study to explore the association between all available factors in the NIS-Teen and HPV vaccine initiation in a systematic manner. Our study suggests that healthcare-seeking behaviours and interactions with the health system may be drivers of HPV vaccine initiation and warrant further study. Addressing these factors could improve the rate of HPV vaccine initiation among adolescents in the USA.


Evaluating the efficacy of medical treatment: possibilities and limitations.
This review outlines the possible uses of subjective health status measures in the evaluation of medical treatment, and possible limitations and pitfalls that potential users of such measures must be aware. Whilst much has been written about the requirements of measures in terms of reliability, validity and responsiveness, many existing measures which have gained widespread use have serious shortcomings. In this paper, the limitations of some existing questionnaires are outlined. It is argued that whilst outcomes measurement has the promise to positively influence medical care, done without due consideration of its limitations and weaknesses it could lead to inaccurate or inappropriate data. In the current climate, in which health outcomes data is viewed as central to monitoring and evaluation, inappropriate or unconsidered use of measures could have drastic effects.


High central venous oxygen saturation in the latter stages of septic shock is associated with increased mortality.
Current guidelines recommend maintaining central venous oxygen saturation (ScvO2) higher than 70% in patients with severe sepsis and septic shock. As high levels of ScvO2 may reflect an inadequate use of oxygen, our aim was to evaluate the relation between maximal ScvO2 levels (ScvO2max) and survival among intensive care unit (ICU) patients with septic shock. We retrospectively analyzed data from all admissions to our ICU between January 2008 and December 2009. All septic shock patients in whom the ScvO2 was measured were included. The measures of ScvO2max within the first 72 hours after the onset of shock were collected. A total of 1,976 patients were screened and 152 (7.7%) patients met the inclusion criteria. The level of ScvO2max was 85% (78 to 89) in the non-survivors, compared with 79% (72 to 87) in the survivors (P = 0.009). Our findings raise concerns about high levels of ScvO2 in patients with septic shock. This may reflect the severity of the shock with an impaired oxygen use. Future strategies may target an optimization of tissue perfusion in this specific subgroup of patients.


Improving function in people with hip-related pain: a systematic review and meta-analysis of physiotherapist-led interventions for hip-related pain.
To report the effectiveness of physiotherapist-led interventions in improving pain and function in young and middle-aged adults with hip-related pain. Systematic review and meta-analysis. A comprehensive, reproducible search strategy was performed on five databases in May 2019. Reference lists and grey literature were also searched. Population: people aged ≥18 years with hip-related pain (with or without a diagnosis of femoroacetabular impingement syndrome). physiotherapist-led interventions for hip pain. Comparators: sham treatment, no treatment or other treatment (eg, hip arthroscopic surgery). primary outcomes included patient-reported hip pain and function. Secondary outcomes included physical function measures. 1722 papers were identified. After exclusion criteria were applied, 14 studies were included for analysis. They had varied risk of bias. There were no full-scale placebo-controlled randomised controlled trials (RCTs) of physiotherapist-led treatment. Pooled effects ranged from moderate effects (0.67 (95% CI 0.07 to 1.26)) favouring physiotherapist-led intervention over no treatment post-arthroscopy, to weak effects (-0.32 (95% CI 0.57 to 0.07)) favouring hip arthroscopy over physiotherapist-led treatment. Physiotherapist-led interventions might improve pain and function in young and middle-aged adults with hip-related pain, however full-scale high-quality RCT studies are required. CRD42018089088.


A double-blind comparison of single doses of etidocaine 1 % and bupivacaine 0.5 % during continuous lumbar epidural block in obstetrics.
A double-blind investigation of some of the effects of etidocaine 1 % and bupivacaine 0.5 % was undertaken in patients in established labour. The results showed that the duration of bupivacaine was significantly longer than that of etidocaine, while the number of patients with good analgesia was equal in both groups. Motor blockade was less obvious in those patients receiving bupivacaine, but in neither group was this a finding of importance. At 5 minutes the Apgar scores of neonates from both groups were similar and any differences seen at 1 minute may be attributable to factors other than the drugs on trial.


Contemporary kidney transplantation has a limited impact on bone microarchitecture.
Bone microarchitecture is an important component of bone quality and disturbances may reduce bone strength and resistance to trauma. Kidney transplant recipients have an excess risk of fractures, and bone loss affecting both trabecular and cortical bone compartments have been demonstrated after kidney transplantation. The primary aim of this study was to investigate the impact of kidney transplantation on trabecular and cortical bone microarchitecture, assessed by histomorphometry and micro computed tomography (μCT). Iliac crest bone biopsies, analyzed by bone histomorphometry and μCT, were performed at time of kidney transplantation and 12 months post-transplantation in an unselected cohort of 30 patients. Biochemical markers of mineral metabolism and bone turnover were measured at both time-points. At 12 months post-transplantation, bone turnover was low in 5 (17%) and normal in 25 (83%) patients. By histomorphometry, bone remodeling normalized, with decreases in eroded perimeters (4.0 to 2.1%, p = 0.02) and number of patients with marrow fibrosis (41 to 0%, p < 0.001). By μCT, trabecular thickness (134 to 125 μM, p = 0.003) decreased slightly. Other parameters of bone volume and microarchitecture, including cortical thickness (729 to 713 μm, p = 0.73) and porosity (10.2 to 9.5%, p = 0.15), remained stable. We conclude that kidney transplantation with current immunosuppressive protocols has a limited impact on bone microarchitecture.


Phosphorus nitride nano-dots as a versatile and metal-free support for efficient photoelectrochemical water oxidation.
Phosphorus nitride dots (PNDs) are employed as a metal-free and versatile support over a range of metal oxide-based photoanodes for efficient photoelectrochemical (PEC) water oxidation. PNDs have the ability to form various heterojunctions by virtue of their favorable band positions for enhanced charge separation leading to improved photocurrent densities.


Assessing covalent binding of reactive drug metabolites by complete protein digestion and LC-MS analysis.
Covalent binding by reactive drug metabolites represents a poorly understood cause of drug toxicity. Currently, assessing protein covalent binding usually entails the use of radioactive drug and therefore has limited applicability in drug discovery. Several marketed drugs are known to form reactive metabolites and have been shown to covalently bind to proteins. In this article, we describe a new method for the analysis of reactive metabolite-protein binding by MS using a strategy of complete digestion of microsomal proteins into free amino acids. Immobilized pronase was found to be the best method for complete digestion in terms of stability of amino acid modifications as well as minimized spectral background. Modified cysteine residues were identified for four tested drug compounds known to form reactive metabolites following in vitro microsomal incubations and accurate mass measurements by LC-MS analysis.


A comprehensive and systematic review on potential anticancer activities of eugenol: From pre-clinical evidence to molecular mechanisms of action.
Eugenol (1-allyl-4-hydroxy-3-methoxybenzene) is an important simple phenolic compound mainly derived from Syzygium aromaticum and many other plants. It is traditionally used in ayurveda and aromatherapy for the healing of many health problems. It also has significant applications in dentistry, agriculture, and flavour industry. This simple phenol has an eclectic range of pharmacological properties, such as antioxidant, anti-inflammatory, and anticancer activities. It is regarded as safe by the Food and Agricultural Organization of the United Nations due to its non-carcinogenic and non-mutagenic properties. The aim of this comprehensive review is to present a critical and systematic assessment of the antitumor ability of eugenol and its associated molecular targets in various cancers. It was carried out following the preferred reporting items for systematic reviews and meta-analysis guidelines. Risk of bias assessment was performed using the SYstematic review centre for laboratory animal experimentation guidelines. The literature search was performed in standard databases such as Science Direct, PubMed, Google Scholar, Scopus, and Web of Science using the keywords 'eugenol' or 'eugenol essential oil' and 'anti-cancer properties of eugenol'. The scientific information from fifty-three studies was encompassed in the present review work. Eugenol exhibits significant anticancer effects in a variety of biological pathways, namely apoptosis, autophagy, cell cycle progression, inflammation, invasion, and metastasis. Eugenol-induced apoptosis has been noticed in osteosarcoma, skin tumors, melanoma, leukemia, gastric and mast cells. It decreases the expression of cyclin D1, cyclin B, proliferating cell nuclear antigen, nuclear factor-ƙB, inhibitor of nuclear factor ƙB, and B-cell lymphoma-2. Eugenol increases the expression of B-cell lymphoma-2 (BCL-2) associated X, BH3-interacting domain death agonist, BCL-2 associated agonist of cell death, apoptotic protease activating factor 1, cytochrome c, p21, and p53. The anticancer potential exhibited by eugenol is mainly attributed to its anti-metastatic, anti-proliferative, anti-angiogenic, anti-inflammatory, cell cycle arrest, apoptotic, and autophagic effects. Hence, the use of eugenol alone or along with other chemotherapeutic anticancer agents is found to be very effective in cancer therapy.


The effect of radio frequency heating on the inactivation and structure of horseradish peroxidase.
The effects of radio frequency (RF) heating on horseradish peroxidase (HRP) activity and its structure were investigated in this paper. The HRP was heated to 50 °C, 70 °C and 90 °C at different electrode gaps (100, 110 and 120 mm). The relative enzyme activity was 105.33 %-113.73 % at 50 °C, 91.11 %-93.05 % at 70 °C and 47.05 %-68.17 % at 90 °C. Ultraviolet-visible, circular dichroism and fluorescence spectra were used to monitor the variation in secondary and tertiary structure. The results showed that RF heating at the electrode gaps of 120 mm contributed to more severe enzyme inactivation and conformational destruction, which can be explained by the changes in Soret band, secondary structure content and tryptophan fluorescence intensity. This study revealed that enzyme inactivation by RF heating was associated with loss of helical structure, unfolding of enzyme protein and ejection of heme group.


Enhancement of adherence of Helicobacter pylori to host cells by virus: possible mechanism of development of symptoms of gastric disease.
It remains unclear why gastric disease does not develop in all cases of Helicobacter pylori infection. In this study, we analyzed whether simian virus 5 (SV5) enhanced adherence of H. pylori to adenocarcinoma epithelial cells (AGS). H. pylori in AGS (harboring SV5) and SV5-infected Vero cells, and an agglutination of H. pylori mixed with SV5 were observed by light microscopy, scanning and transmission electron microscopies. The adherent rate of H. pylori to SV5-infected Vero cells and treated with an anti-SV5 antibody was determined. H. pylori adhered to the surface of AGS cells near SV5 particles, as shown by scanning and transmission electron microscopies. The adherence of H. pylori to SV5-infected Vero cells was significantly enhanced compared with that to Vero cells. In contrast, the adherence of H. pylori to Vero cells was decreased by treatment with the anti-SV5 antibody. Agglutination of H. pylori mixed with SV5 was observed by scanning and transmission electron microscopies. Agglutination did not occur when SV5 was treated with the anti-SV5 antibody before mixing. These findings demonstrated that SV5 enhanced the adherence of H. pylori to host cells, suggesting that a persistently infected virus may be a factor enhancing the pathogenicity of H. pylori in humans.


Enantioselectivity of indoxacarb enantiomers in Bombyx mori larvae: toxicity, bioaccumulation and biotransformation.
Indoxacarb is a chiral insecticide with excellent insecticidal activity against lepidopterous insects. Because of their enantioselectivity, chiral pesticides' environmental behavior at the enantiomeric level has been highlighted. The chiral stability, enantioselective bioaccumulation, biotransformation behavior of indoxacarb to a non-target insect silkworm are still unclear. A chiral analysis method for indoxacarb and its metabolite DCJW in silkworm was developed using liquid chromatography and high-resolution mass spectrometry (HPLC/HRMS). In silkworms, the recoveries of indoxacarb and DCJW were 86.06%-104.52% with relative standard deviation (RSD) < 9.01%. The 96-h lethal concentration (LC50 ) values of R-indoxacarb, S-indoxacarb, and enriched S-indoxacarb (2.333 S/1R) were 1.08 × 102 , 1.92, and 6.89 mg a.i. L-1 , respectively, according to the acute toxicity test results. When silkworm larvae were exposed to 1/50 of the LC50 concentration, the bioconcentration factor (BCF) of R-indoxacarb was 0.0296-0.318, and the BCF of S-indoxacarb was 0.0125-0.211. In silkworm larvae, the amount of R-DCJW produced by R-indoxacarb was 0.00610 to 2.34 times that of the parent R-indoxacarb, and the amount of S-DCJW produced by S-indoxacarb was 0.125-36.9 times that of the parent S-indoxacarb. There was no chiral transition from S-indoxacarb to R-indoxacarb or a transformation from R-indoxacarb to S-indoxacarb. Indoxacarb was preferentially bioaccumulated in silkworm larva, while S-indoxacarb bioconversion into the metabolite S-DCJW was much greater than R-indoxacarb bioconversion into R-DCJW. This study could improve understanding of the indoxacarb accumulation and transformation process in insects, as well as provide more scientific data for indoxacarb environmental and ecological risk assessment. © 2023 Society of Chemical Industry.


Disruption of perlecan binding and matrix assembly by post-translational or genetic disruption of dystroglycan function.
Dystroglycan is a cell-surface matrix receptor that requires LARGE-dependent glycosylation for laminin binding. Although the interaction of dystroglycan with laminin has been well characterized, less is known about the role of dystroglycan glycosylation in the binding and assembly of perlecan. We report reduced perlecan-binding activity and mislocalization of perlecan in the LARGE-deficient Large(myd) mouse. Cell-surface ligand clustering assays show that laminin polymerization promotes perlecan assembly. Solid-phase binding assays provide evidence for the first time of a trimolecular complex formation of dystroglycan, laminin and perlecan. These data suggest functional disruption of the trimolecular complex in glycosylation-deficient muscular dystrophy.


Mechanism of signal transduction of the LOV2-Jα photosensor from Avena sativa.
Fusion proteins containing blue-light-activable protein domains possess great potential as molecular switches in cell signalling. This has recently been impressively demonstrated by connecting the light oxygen voltage LOV2-Jα-protein domain of A. sativa (AsLOV2-Jα) with the Rac1-GTPase, responsible for regulating the morphology and motility of metazoan cells. However, a target-oriented development of fusion proteins in conjunction with this photosensor is still very challenging, because a detailed understanding of its signal transduction pathway on a molecular level is still lacking. Here, we show through molecular dynamics simulation that, after formation of the cysteinyl-flavin mononucleotide (FMN) adduct, the signalling pathway begins with a rotational reorientation of the residue glutamine 1029 adjacent to the FMN chromophore, transmitting stress through the Iβ strand towards the LOV2-Jα interface. This then results in the breakage of two H-bonds, namely, glutamic acid 1034-Gln995 and aspartic acid (Asp) 1056-Gln1013, at opposite sides of the interface between the Jα helix and the LOV2 domain, ultimately leading to a disruption of Jα helix from the LOV2 core.


MiR-375: A novel multifunctional regulator.
MiR-375, a primitively described beta cell-specific miRNA, is confirmed to function as multi-functional regulator in diverse typical cellular pathways according to the follow-up researches. Based on the existing studies, miR-375 can regulate many functional genes and ectopic expressions of miR-375 are usually associated with pathological changes, and its expression regulation mechanism is mainly related to promoter methylation or circRNA. In this review, the regulatory functions of miR-375 in immunity, such as its relevance with macrophages, T helper cells and autoimmune diseases were briefly discussed. Also, the functions of miR-375 involved in inflammation, development and virus replication were reviewed. Finally, the mechanisms and application prospects of miR-375 in cancers were analyzed. Studies show that the application of miR-375 as therapeutic target and biomarker has a broad developing space in future. We hope this paper can provide reference for its further study.


Primary structure of porcine pepsin. II. Amino acid sequence of two cyanogen bromide fragments, CB3 and CB4.
The amino acid sequences of two cyanogen bromide fragments from porcine pepsin have been determined. Fragment CB3 which represents the NH2-terminal 80 residues of pepsin was assembled from the peptides purified from proteolytic digests of this fragment using alpha-chymotrypsin, thermolysin, and staphylococcal protease. Two chymotryptic peptides were isolated from the NH2-terminal region of this fragment. One of these contains 2 extra residues, Ala-Leu-, at the NH2 terminus. This peptide is apparently derived from a different cleavage site of pepsinogen in its conversion to pepsin. The second cyanogen bromide fragment, CB4, contains 47 residues. The sequence was established from the peptides resulting from proteolytic digests using alpha-chymotrypsin, alpha-lytic protease, and thermolysin. An isoleucyl residue at position 29 of fragment CB4 appears to be absent in some molecules. This represents a structural variant of pepsin.


On the life cycle of Brachymeria podagrica (Fabricius, 1787) (Hymenoptera: Chalcididae) - a parasitoid of sacrophagid, calliphorid, and muscid flies.
The lifecycle of Brachymeria podagrica, a parasitic wasp with a worldwide distribution, was studied under laboratory conditions using the flesh fly, Sarcophaga dux, as a host. Two hundred parasite-free 3rd instars of S. dux were exposed for 24 h to 20 female B. podagrica. In daily intervals, maggots and later pupae were examined for developmental stages of the parasitoid. The whole pre-imaginal development at a temperature of 26 °C lasted 21 to 26 days. Three morphologically different instars, followed by a prepupal and a pupal stage, were described using light and scanning electron microscopy. In a second experiment with 100 3rd stage Sarcophaga larvae and 10 parasitoids, a total of 70 wasps emerged 20 to 25 days after exposure. Two fly larvae did not pupate and dried out, while 28 pupae contained a dry or caseous content, dead wasp imagos, or their larval stages. No fly imagines emerged from exposed groups, while all 100 unexposed larvae pupated and adults eclosed between day 12 and day 14 after the start of the experiment, while the imagoes of the parasitoids appeared 8 to 12 days later.


School Nurses on the Front Lines of Healthcare: Red Flags and Red Herrings: Improving the Recognition of Bruises and Burns Associated With Physical Abuse in School-Age Children.
Through a series of case scenarios, this article describes the assessment and management of suspected physical child abuse presenting as bruises and burns. Although it is not uncommon for school-age children to have accidental injuries, recognizing patterns associated with physical child abuse and understanding red flags for abuse is vital. Failure to recognize injury patterns suspicious for physical child abuse, and thus failure to intervene appropriately, may leave children at risk for more serious injury or death.


Effects of psoralen from Psoralea corylifolia on quinone reductase, ornithine decarboxylase, and JB6 cells transformation promotion.
The cancer chemopreventive effect of psoralen isolated from the seeds of Psoralea corylifolia was investigated in the induction of quinone reductase (QR) activity, intracellular detoxification enzyme, inhibition of 12-O-tetradecanoylphorbol 13-acetate (TPA)-induced ornithine decarboxylase (ODC) activity, a key regulatory enzyme for polyamine metabolism, and tumor promotion in mouse epidermal JB6 cells, sensitive to tumor promoters (clone 415a P+ cells), which are related to suppress multistage carcinogenesis including initiation and promotion. Psoralen was isolated and identified from the ethyl acetate-soluble fraction of the methanolic extract from the seeds. Psoralen was active in induction of QR activity, the concentration of psoralen required to induce 1.5 fold QR activity was 14.8 μg/mL. Also, this pure compound inhibited TPA-induced ODC activity by 50% (designated IC(50)) at the concentration 15.6 μg/mL and exhibited inhibition of TPA-induced tumor promotion in mouse epidermal JB6 cells with an IC(50) value of 17.1 μg/mL. Therefore, it is extrapolated that psoralen has the potential capable of inhibiting the initiation and/or promotion stage of carcinogenesis by induction of QR activity, inhibition of TPA-induced ODC activity and mouse epidermal JB6 cells tumor promotion.


Efficacy and Safety of Intravenous Pamidronate for Parathyroid Hormone-dependent Hypercalcemia in Hospitalized Patients.
Bisphosphonates are effective for hypercalcemia of malignancy (HOM). Efficacy and safety data for bisphosphonates in parathyroid hormone-related hypercalcemia (PTHRH) are rare, including pamidronate (Pam), which is not indicated for this condition. This work aims to evaluate the efficacy and safety of Pam for moderate-to-severe PTHRH. This retrospective case-control study was conducted at a tertiary care medical center. Patients included adults hospitalized with serum calcium levels greater than 12 mg/dL, from October 29, 2013 to December 17, 2019. Etiology was categorized as PTHRH or PTH-independent. Clinical and laboratory data of PTHRH patients treated with Pam (PTHRH-Pam+) were compared to Pam-untreated counterparts (PTHRH-Pam-). Thirty-four patients with 37 hospitalizations for PTHRH (Pam-treated and -untreated) met the inclusion criteria. Pam was given in 24 of 37 cases (64.8%). Admission serum calcium levels for the PTHRH-Pam+ group were higher than for PTHRH-Pam- group (14.4 mg/dL vs 13.0 mg/dL, P = .005). Median total Pam dose was 60 mg (range, 30-180 mg) in the treated group. Serum calcium decreased 3.5 mg/dL for PTHRH-Pam+ vs 1.6 mg/dL for PTHRH-Pam- (P = .003). No PTHRH-Pam+ patients developed hypocalcemia or acute kidney injury. Nadir serum phosphorus levels were lower in the PTHRH-Pam+ vs PTHRH-Pam- group (1.7 mg/dL vs 2.4 mg/dL, respectively, P = .004). Three PTHRH-Pam+ patients developed severe hypophosphatemia; all resolved with intravenous and oral supplementation. Seventeen patients underwent parathyroidectomy, of whom 10 received Pam within 28 days preoperatively. Postoperatively, 4 developed hypocalcemia and 3 hypophosphatemia. This study demonstrates that Pam is effective and safe for treating PTHRH, while ensuring close laboratory monitoring of calcium and phosphorus metabolism. Larger, prospective studies are needed to establish the role of Pam and other potent bisphosphonates in moderate-to-severe PTHRH.


An epidemiologic investigation of amyotrophic lateral sclerosis in Jefferson County, Missouri, 1998-2002.
Amyotrophic lateral sclerosis (ALS) cases diagnosed between 1998 and 2002 were identified to study ALS prevalence and spatial clustering in Jefferson County, Missouri, where an active lead smelter is located. The study used the El Escorial criteria for ALS diagnosis, the capture-recapture analysis for ALS case ascertainment, and the spatial scan statistic for cluster analysis. The estimated crude prevalence of ALS in Jefferson County was 3.9 per 100,000 population (95% CI, 1.7-7.7) at the time point on December 31, 2002. After age-adjustment to the 2002 U.S. population, the prevalence was 4.2 per 100,000 (95% CI, 1.9-6.6). This prevalence estimate was comparable to recent prevalence estimates from Western Europe. A small but significant cluster (p=0.04) was detected around the smelter area. An ALS registry utilizing outpatient, inpatient, and death certificate data is needed to provide comprehensive information for ALS case ascertainment. Etiologic studies are needed to assess whether living in proximity to a lead smelter is associated with the development of ALS.


Phagocytosis of unopsonized zymosan particles by trypsin-sensitive and beta-glucan-inhibitable receptors on bone marrow-derived murine macrophages.
Murine bone marrow cells, plated at 4 X 10(4) cells/well and cultured in 50% fibroblast CM, yielded pure populations of large, individual, adherent cells that were phagocytic and morphologically indistinguishable from macrophages. Adherent macrophages appeared in small numbers with 24 h of culture, increased to maximal cell numbers within 10 days of culture, and remained at these cell densities for at least 11 weeks in culture. The capacities of adherent macrophages to ingest unopsonized zymosan particles and EsIgG, at inputs of 1.25 X 10(7) targets, were expressed by 7 and 40% of the cells derived from 24-hour cultures, respectively, were increased at nearly identical rates to comparable maximal levels within 10-14 days of culture and were exhibited by essentially all adherent cells derived from 2-11-week cultures. The percentage of adherent macrophages from twelve 3-6-week cultures ingesting greater than or equal to 1, greater than or equal to 6 and greater than or equal to 10 zymosan particles was 89 +/- 5, 47 +/- 11 and 14 +/- 9% (mean +/- SD, n = 12), respectively, and the percentage ingesting greater than or equal to 1, greater than or equal 6 and greater than or equal to 10 EsIgG was 86 +/- 5, 49 +/- 10 and 14 +/- 8%, respectively. Incubation of adherent macrophages with mannan-free ss-glucan particles at inputs of 5 X 10(5)-5 X 10(7)/ml initiated a phagocytic response comparable to that obtained with the same doses of zymosan particles which contained mannan and beta-glucan. Preincubation of adherent macrophages with 100 micrograms/ml of a fully soluble beta-glucan, laminarin, and solubilized barley beta-glucan reduced subsequent macrophage phagocytosis of greater than or equal to 6 zymosan particles by 53 and 40%, respectively. In contrast, yeast alpha-mannan was less than 1% as active, and 10 mg/ml reduced the number of adherent macrophages ingesting greater than or equal to 1 zymosan particles by 64%. At concentrations as high as 2 mg/ml, laminarin and barley beta-glucan had no effect on Fc receptor-mediated ingestion of EsIgG, and mannan at 20 mg/ml also failed to inhibit EsIgG ingestion. Pretreatment of adherent macrophages with 20 micrograms/ml of trypsin reduced the number of cells ingesting greater than or equal to 1 zymosan particles from 89 to 10% and those ingesting greater than or equal to 6 zymosan from 43 to 0%, whereas pretreatment with as much as 100 micrograms/ml of trypsin failed to decrease macrophage ingestion of EsIgG.(ABSTRACT TRUNCATED AT 400 WORDS)


An intervention study through education: The change in attitudes and knowledge of educators about the COVID-19 vaccine.
Reliable and culturally informed health communication is vital in developing positive health behaviors. The aim of this study was to investigate the change in knowledge and attitudes about COVID-19 vaccines by providing training to educators in a province in the east of Turkey. A questionnaire developed by the researchers on the basis of the literature was used in this intervention study. It consisted of three parts: socio-demographic information, before education knowledge (BEK) and Attitudes Towards the COVID-19 Vaccine (BEATV), and after education knowledge (AEK) and Attitudes Towards the COVID-19 Vaccine (AEATV). There were 99 teachers, 32 public education teachers, and 144 religious officials who participated in the study. The data were analyzed using SPSS Statistics Pack version 22.0 (IBM, Armonk, NY, USA). Statistical significance value was accepted as p < 0.05. Among the participants, the number of correct answers to all questions increased as a result of the study. There was a positive increase in most of the ATV items after the training. Education on COVID-19 vaccines increased awareness as shown by the total number of correct answers and the improved attitude of educators.


Interinstitutional variation in management decisions for treatment of 4 common types of cancer: A multi-institutional cohort study.
When clinical practice is governed by evidence-based guidelines and there is consensus about their validity, practice variation should be minimal. For areas in which evidence gaps exist, greater variation is expected. To systematically assess interinstitutional variation in management decisions for 4 common types of cancer. Multi-institutional, observational cohort study of patients with cancer diagnosed between July 2006 through May 2011 and observed through 31 December 2011. 18 cancer centers participating in the formulation of treatment guidelines and systematic outcomes assessment through the National Comprehensive Cancer Network. 25 589 patients with incident breast cancer, colorectal cancer, lung cancer, or non-Hodgkin lymphoma. Interinstitutional variation for 171 binary management decisions with varying levels of supporting evidence. For each decision, variation was characterized by the median absolute deviation of the center-specific proportions. Interinstitutional variation was high (median absolute deviation >10%) for 35 of 171 (20%) oncology management decisions, including 9 of 22 (41%) decisions for non-Hodgkin lymphoma, 16 of 76 (21%) for breast cancer, 7 of 47 (15%) for lung cancer, and 3 of 26 (12%) for colorectal cancer. Forty-six percent of high-variance decisions involved imaging or diagnostic procedures and 37% involved choice of chemotherapy regimen. The evidence grade underpinning the 35 high-variance decisions was category 1 for 0%, 2A for 49%, and 2B/other for 51%. Physician identifiers were unavailable, and results may not generalize outside of major cancer centers. The substantial variation in institutional practice manifest among cancer centers reveals a lack of consensus about optimal management for common clinical scenarios. For clinicians, awareness of management decisions with high variation should prompt attention to patient preferences. For health systems, high variation can be used to prioritize comparative effectiveness research, patient-provider education, or pathway development. National Cancer Institute and National Comprehensive Cancer Network.


Network analysis of Contingencies of Self-Worth Scale in 680 university students.
This study investigates the Contingencies of Self-Worth Scale (CSWS) in a sample of 680 university students from a network perspective. We estimated regularized partial correlations among seven CSWS domains: family support, competition, appearance, God's love, academic competence, virtue and other's approval. Competition - academic competence and competition - appearance represent the strongest connections in the network. Mean node predictability (shared variance with surrounding nodes) is 0.25. Appearance and academic competence were the most central (i.e., interconnected) domains in the network. Future studies should explore the network structure of self-worth in other healthy adult samples, and also in people with psychopathology. We provide the anonymized dataset as well as the full code in the supplementary materials to ensure complete reproducibility of the results.


Long term survival and cardiopulmonary outcome in children with Hurler syndrome after haematopoietic stem cell transplantation.
Premature death in untreated children with Hurler syndrome (HS) in the first decade of life is largely due to life-threatening cardiopulmonary complications. We examined the long-term survival and cardiopulmonary outcome in 54 children undergoing haematopoietic stem cell transplantation (HSCT) at the Royal Manchester Children's Hospital from 1985 to 2008. The median age at first HSCT was 15.1 months. Eighteen had graft failure and nine died after first HSCT. Of 18 patients with graft failure, 17 underwent second HSCT and the remaining one was lost to follow-up (LOF). Twelve were alive-and-engrafted after second HSCT. The overall survival at one year and 20-years was the same at 73.7%. Six children were followed up at the referral centers and excluded from cardiopulmonary endpoint review. Of the 33 evaluable children for the cardiopulmonary endpoints, nine (27.3%) had normal cardiac assessment. Of the four children on angiotensin-converting-enzyme inhibitors, two had mild cardiomyopathy and two had aortic valvular replacement. Twenty (60%) had mild/moderate mitral and/or aortic insufficiencies. Two had overnight hypoxia needing nocturnal non-invasive support. Enzyme level and donor chimerism are important predictors of long-term cardiac outcome.


petiteFinder: an automated computer vision tool to compute Petite colony frequencies in baker's yeast.
Mitochondrial respiration is central to cellular and organismal health in eukaryotes. In baker's yeast, however, respiration is dispensable under fermentation conditions. Because yeast are tolerant of this mitochondrial dysfunction, yeast are widely used by biologists as a model organism to ask a variety of questions about the integrity of mitochondrial respiration. Fortunately, baker's yeast also display a visually identifiable Petite colony phenotype that indicates when cells are incapable of respiration. Petite colonies are smaller than their Grande (wild-type) counterparts, and their frequency can be used to infer the integrity of mitochondrial respiration in populations of cells. Unfortunately, the computation of Petite colony frequencies currently relies on laborious manual colony counting methods which limit both experimental throughput and reproducibility. To address these problems, we introduce a deep learning enabled tool, petiteFinder, that increases the throughput of the Petite frequency assay. This automated computer vision tool detects Grande and Petite colonies and computes Petite colony frequencies from scanned images of Petri dishes. It achieves accuracy comparable to human annotation but at up to 100 times the speed and outperforms semi-supervised Grande/Petite colony classification approaches. Combined with the detailed experimental protocols we provide, we believe this study can serve as a foundation to standardize this assay. Finally, we comment on how Petite colony detection as a computer vision problem highlights ongoing difficulties with small object detection in existing object detection architectures. Colony detection with petiteFinder results in high accuracy Petite and Grande detection in images in a completely automated fashion. It addresses issues in scalability and reproducibility of the Petite colony assay which currently relies on manual colony counting. By constructing this tool and providing details of experimental conditions, we hope this study will enable larger-scale experiments that rely on Petite colony frequencies to infer mitochondrial function in yeast.


Linearization of the Disposition Index equation allows evaluation of secretion-sensitivity coupling slopes.
The Disposition Index (DI) is widely used in clinical studies of β-cell function. However, direct physiologic interpretation of the DI value and the inverse exponential slope relating insulin secretion and insulin sensitivity terms is difficult. We evaluated a linearization of the relationship that allows separate evaluation of the DI term and the slope. Insulin secretion and sensitivity indices were derived from standardized oral glucose tolerance testing, including commonly used terms and model-derived terms. The population included participants with normoglycemia, dysglycemia or Type 2 diabetes. Logarithmic transformation of the DI equation to linearize the secretion-sensitivity relationship was performed, and the resulting secretion-sensitivity relationships were evaluated using standard linear regression methods. Simple logarithmic transformation linearized the secretion-sensitivity relationships available from a variety of OGTT-derived metrics. In normoglycemic subjects the slopes approximated -1 in insulin-basedsecretion-sensitivity pairs, and approximated -0.6 in C-peptide based secretion-sensitivity pairs. Group differences in DI terms were observed as expected. These analyses also revealed differing secretion-sensitivity slopes, with IGT and T2D demonstrating progressively impaired coupling. Linearization of the secretion-sensitivity relationship provides simplified interpretation of the DI value and allows simple analysis and meaningful interpretation of the secretion-sensitivity slope. This linear relationship is amenable to standard statistical evaluations for comparisons of insulin secretion responses and of secretion-sensitivity coupling across groups.


Awake prone positioning for patients with COVID-19-induced acute hypoxemic respiratory failure.
Whereas prone positioning of intubated patients suffering from acute respiratory distress syndrome represents the standard of care, proning non-intubated patients, so-called "awake prone positioning (APP)," has only recently gained popularity and undergone scientific evaluation. In this review, we summarize current evidence on physiological and clinical effects of APP on patients' centered outcomes, such as intubation and mortality, the safety of the technique, factors and predictors of success, practical issues for optimal implementation, and future areas of research. Current evidence supports using APP among patients suffering from acute hypoxemic respiratory failure due to COVID-19 and undergoing advanced respiratory support, such as high-flow nasal cannula, in an intensive care unit setting. Healthcare teams should aim to prone patients at least 8 h daily. Future research should focus on optimizing the tolerance of the technique and comprehensively evaluating benefits in other patient populations.


Transcriptomic analysis of the liver of cholesterol-fed rabbits reveals altered hepatic lipid metabolism and inflammatory response.
Rabbits are a suitable animal model for atherosclerosis due to their sensitivity to dietary cholesterol. Moreover, rabbits have lipoprotein profiles that are more similar to humans than those of other laboratory animals. However, little is known about the transcriptomic information related to atherosclerosis in rabbits. We aimed to determine the changes in the livers of rabbits fed a normal chow diet (control) or high cholesterol diet (HCD) by histological examinations and RNA sequencing analysis. Compared with the control group, the lipid levels and small LDL subfractions in plasma were increased, and aortic atherosclerotic plaques were formed in the HCD group. Most importantly, HCD resulted in lipid accumulation and inflammation in the livers. Transcriptomic analysis of the liver showed that HCD induces 1183 differentially expressed genes (DEGs) that mainly participate in the regulation of inflammation and lipid metabolism. Furthermore, the signaling pathways involved in inflammation and lipid metabolism were enriched by KEGG pathway analysis. In addition, hepatic DEGs of the HCD group were further validated by real-time PCR. These results suggest that HCD causes liver lipid accumulation and inflammatory response. Although the relationships between these hepatic changes and atherogenesis need further investigation, these findings provide a fundamental framework for future research on human atherosclerosis using rabbit models.


Feedback on research results to healthcare professionals in the context of HIV.
Reporting the feedback on the results of a multicenter research for healthcare professionals that attend people living with HIV and AIDS in a Specialized Healthcare Center at Belém, state of Pará, Brazil. Case report of the presentation and interpretation of research results, in 2015, totaling 25 participating professionals. The visual resource used was a data projector. Participants reported the importance of the initiative, since, even though it is their right, feedback on research results is uncommon. Among the discussed issues, emerged the need for a culturally appropriate device for adequately strengthening educational practices carried out by healthcare professionals. Feedback on the research results enabled exchange of knowledge about the findings and production of new knowledge. In addition to achieving the social and ethical commitment of research, by promoting dissemination of results, enabling reflections on the reality in which they work.


Role of MRI to confirm healing in complex high cryptoglandular anal fistulas: long-term follow-up of 151 cases.
Complex fistula-in-ano can recur even after complete clinical healing has occurred. 'Radiological healing' of fistula on MRI correlates well with long-term healing rates but no study has yet objectively quantified this. The aim of this study was to assess the accuracy of anal fistula healing as documented on MRI and to correlate it with long-term healing as evidenced on long-term follow-up. Patients with clinically healed anal fistulas who also had radiological healing checked by postoperative MRI were included in the study. Three hundred and twenty-five patients operated for high complex fistula-in-ano were followed up for 14-68 months (median 38 months). Postoperative MRI was done to assess radiological healing of the fistula in 151 patients, and they were included in the study. The mean age was 39.4 ± 10.5 years (116 men). Five patients were lost to follow-up. The fistulas did not heal radiologically (on MRI) in 20 patients and recurred in all these patients. The fistulas healed radiologically (on MRI) in 126 patients. On long-term follow-up, 124/126 patients remained healed while 2/126 had a recurrence. In the first patient, the fistula recurred 40 months after complete radiological healing. In the second patient, the fistula recurred 10 months after complete radiological healing but pus from the fistula tested positive for tuberculosis (by real-time polymerase chain reaction) and he was excluded from the analysis. Thus, there was only one (1/125) recurrence on long-term follow-up. Radiological healing on MRI correlates well with long-term healing in complex fistula-in-ano.


Calcium-dependent slow outward current in visceral primary afferent neurones of the rabbit.
Slow outward currents were recorded from voltage-clamped neurones in nodose ganglia excised from rabbits. In the majority of Type C neurones, a short depolarizing command pulse evoked a slow outward tail current (ISAH) with a decay time constant ranging from 0.5 to 2 s. The ISAH was due to an increase in membrane conductance to K+ because its reversal potential was approximately equal to the Nernst potential for K+. The ISAH was reversibly blocked by removal of external Ca2+ or by Ca2+ antagonists. A Ca2+ ionophore, A23187, produced an outward current which was similar to the ISAH. The ISAH was resistant to tetraethylammonium and depressed by Ba2+, whereas it was not affected by Cs+ and 4-aminopyridine. The ISAH was initially augmented and subsequently depressed by apamin (1-10 nM) and (+)-tubocurarine (100-600 microM). It is concluded that the ISAH in visceral primary neurones may be due to a long-lasting increase in K+ conductance caused by an increase in the concentration of intracellular Ca2+, resulting from Ca2+ entry during the depolarizing command pulse.