Isolation of plant nuclei.
The isolation of plant nuclei is a useful first step in many experiments concerned with the mechanism and control of gene expression in plants. For example, isolated nuclei can be used for the isolation of nuclear components such as chromosomal proteins (1), for the study of the processing of primary transcripts (2,3), for the assay and characterization of RNA polymerase activities (4-7), or for the measurement of transcription rates of specific genes (8) (see Chapter 37 ). A method is described here for the isolation of a crude preparation of intact plant nuclei, with an additional protocol for nuclei purification on a discontinuous gradient of Percoll (9) (see Note 1 in section 4). Centrifugation of crude nuclei preparations through Percoll gradients removes much of the contaminating cytoplasmic material such as starch grains (10), and the Percoll step appears to reduce the ribonuclease activity associated with nuclei (10). It is therefore recommended for transcription experiments. The crude nuclei preparation alone may be adequate for some work, for example, when attempting RNA polymerase assays for the first time, when very small amounts of tissue are involved, for preliminary experiments on the characterization of enzyme activities, or for the isolation of nuclear components that may be lost during purification.


Heritability of nociception. III. Genetic relationships among commonly used assays of nociception and hypersensitivity.
We and others have previously demonstrated that nociception in the mouse is heritable. A genetic correlation analysis of 12 common measures of nociception among a common set of inbred strains revealed three major clusters (or 'types') of nociception in this species. In the present study, we re-evaluated the major types of nociception and their interrelatedness using ten additional assays of nociception and hypersensitivity, including: three thermal assays (tail withdrawal from 47.5 degrees C water or -15 degrees C ethanol; tail flick from radiant heat), two chemical assays of spontaneous nociception (bee venom test; capsaicin test) and their subsequent thermal hypersensitivity states (including contralateral hypersensitivity in the bee venom test), a mechanical nociceptive assay (tail-clip test), and a mechanical hypersensitivity assay (intrathecal dynorphin). Confirming our earlier findings, the results demonstrate distinct thermal and chemical nociceptive types. It is now clear that mechanical hypersensitivity and thermal hypersensitivity are genetically dissociable phenomena. Furthermore, we now see at least two distinct types of thermal hypersensitivity: afferent-dependent, featuring a preceding significant period of spontaneous nociceptive behavior associated with afferent neural activity, and non-afferent-dependent. In conclusion, our latest analysis suggests that there are at least five fundamental types of nociception and hypersensitivity: (1) baseline thermal nociception; (2) spontaneous responses to noxious chemical stimuli; (3) thermal hypersensitivity; (4) mechanical hypersensitivity; and (5) afferent input-dependent hypersensitivity.


Degenerative changes in the A375 melanoma line induced by transforming growth factor beta 1.
A375 human melanoma cell cultures grown in the presence of TGF beta contained greatly reduced cell numbers and exhibited drastic alterations in cell morphology compared to the control cultures. Preincubation of the cells with the cytokine for only 18 h was sufficient to induce these changes irreversibly. Examination of TGF beta-treated cells in the electron microscope revealed large numbers of lipid-filled vacuoles in the cytoplasm, greatly contracted nuclei and some loss of the otherwise abundant microvilli. Thus TGF beta may have a direct toxic effect on the A375 melanoma cells.


STAT3 serine phosphorylation is required for TLR4 metabolic reprogramming and IL-1β expression.
Detection of microbial components such as lipopolysaccharide (LPS) by Toll-like receptor 4 (TLR4) on macrophages induces a robust pro-inflammatory response that is dependent on metabolic reprogramming. These innate metabolic changes have been compared to aerobic glycolysis in tumour cells. However, the mechanisms by which TLR4 activation leads to mitochondrial and glycolytic reprogramming are unknown. Here we show that TLR4 activation induces a signalling cascade recruiting TRAF6 and TBK-1, while TBK-1 phosphorylates STAT3 on S727. Using a genetically engineered mouse model incapable of undergoing STAT3 Ser727 phosphorylation, we show ex vivo and in vivo that STAT3 Ser727 phosphorylation is critical for LPS-induced glycolytic reprogramming, production of the central immune response metabolite succinate and inflammatory cytokine production in a model of LPS-induced inflammation. Our study identifies non-canonical STAT3 activation as the crucial signalling intermediary for TLR4-induced glycolysis, macrophage metabolic reprogramming and inflammation.


Validation of a laboratory-developed test of human sperm capacitation.
Sperm must undergo capacitation to become fertilization competent. Here we validated that monosialotetrahexosylganglioside (GM1 ) localization patterns, which were assessed in the Cap-Score™ Sperm Function Test, reflect a capacitated state in human sperm. First, we defined patterns representing sperm that do or do not respond to stimuli for capacitation. Sperm with "capacitated" patterns had exposed acrosomal carbohydrates and underwent acrosome exocytosis in response to calcium ionophore (A23187). Precision was evaluated by percent change of the Cap-Score measured for 50, 100, 150, and 200 sperm. Changes of 11%, 6%, and 5% were observed (n ≥ 23); therefore, we counted ≥150 sperm per condition. Variance within and between readers was evaluated using 20 stitched image files generated from unique ejaculates. Two trained readers randomly resampled each image 20 times, reporting an average standard deviation of 3 Cap-Score units and coefficient of variation of 13% when rescoring samples, with no difference between readers. Semen liquefaction times ≤2 hr and mechanical liquefaction with Pasteur or wide-orifice transfer pipettes did not alter Cap-Score values. However, liquefaction with chymotrypsin (p = 0.002) and bromelain (p = 0.049) reduced response to capacitating stimuli and induced membrane damage, while counterintuitively improving sperm motility. Together, these data validate the Cap-Score assay for the intended purpose of providing information on sperm capacitation and male fertility. In addition to its clinical utility as a diagnostic tool, this test of sperm function can reveal the impact of common practices of semen handling on the ability of sperm to respond to capacitation stimuli.


Functional analysis and expression patterns of members of the FLOWERING LOCUS T (FT) gene family in Lilium.
Lilium is an important commercial flowering species, and there are many varieties and more than 100 species of wild Lilium. Lilium × formolongi is usually propagated from seedlings, and the flowering of these plants is driven mainly by the photoperiodic pathway. Most of the other lily plants are propagated via bulblets and need to be vernalized; these plants can be simply divided into pretransplantation types and posttransplantation types according to the time at which the floral transition occurs. We identified three Lilium FLOWERING LOCUS T (LFT) family members in 7 Lilium varieties, and for each gene, the coding sequence of the different varieties was identical. Among these genes, the LFT1 gene of Lilium was most homologous to the AtFT gene, which promotes flowering in Arabidopsis. We analyzed the expression patterns of LFT genes in Lilium × formolongi seedlings and in different Lilium varieties, and the results showed that LFT1 and LFT3 may promote floral induction. Compared with LFT3, LFT1 may have a greater effect on floral induction in Lilium, which is photoperiod sensitive, while LFT3 may play a more important role in the floral transition of lily plants, which have a high requirement for vernalization. LFT2 may be involved in the differentiation of bulblets, which was verified by tissue culture experiments, and LFT1 may have other functions involved in promoting bulblet growth. The functions of LFT genes were verified by the use of transgenic Arabidopsis thaliana plants, which showed that both the LFT1 and LFT3 genes can promote early flowering in Arabidopsis. Compared with LFT3, LFT1 promoted flowering more obviously, and thus, this gene could be an important promoter of floral induction in Lilium.


Early clinical experience with anidulafungin at a large tertiary care medical center.
To evaluate early clinical experience with anidulafungin. Retrospective cohort study. Large, university-affiliated, tertiary care medical center. All patients receiving anidulafungin between July 15, 2006, and January 15, 2007. Thirty-five patients received at least one dose of anidulafungin. Safety and tolerability were evaluated in all patients; efficacy outcomes were assessed in 13 patients who had a documented fungal infection and received anidulafungin for a minimum of 5 days. Common conditions at baseline were hepatic dysfunction (25 patients [71%]), severe sepsis (17 patients [49%]), and solid organ or hematopoietic stem cell transplantation (10 patients [29%]). Eight patients (23%) were receiving drugs with the potential to interact with echinocandins other than anidulafungin. Seventeen (49%) of the 35 patients received anidulafungin as empiric antifungal therapy. Anidulafungin was used to treat invasive candidiasis in seven patients (20%) and candidemia in 10 patients (29%); Candida albicans or Candida glabrata was isolated most frequently in these two infections combined (7 isolates each [41%]/17 infections). A favorable efficacy outcome was noted in 10 (77%) of 13 evaluable patients. One patient developed breakthrough Candida parapsilosis fungemia while receiving anidulafungin. Overall, anidulafungin was well tolerated, with only one patient having an infusion- related reaction. Anidulafungin was also well tolerated among patients receiving concomitant metronidazole. Anidulafungin was well tolerated and produced favorable outcomes in the majority of the patients evaluated. The availability of anidulafungin makes it a feasible option for antifungal therapy, particularly in patients who have hepatic dysfunction and in those who are receiving drugs than can interact with other echinocandins.


Cholesterol-dependent transcriptome remodeling reveals new insight into the contribution of cholesterol to Mycobacterium tuberculosis pathogenesis.
Mycobacterium tuberculosis (Mtb) is an obligate human pathogen that can adapt to the various nutrients available during its life cycle. However, in the nutritionally stringent environment of the macrophage phagolysosome, Mtb relies mainly on cholesterol. In previous studies, we demonstrated that Mtb can accumulate and utilize cholesterol as the sole carbon source. However, a growing body of evidence suggests that a lipid-rich environment may have a much broader impact on the pathogenesis of Mtb infection than previously thought. Therefore, we applied high-resolution transcriptome profiling and the construction of various mutants to explore in detail the global effect of cholesterol on the tubercle bacillus metabolism. The results allow re-establishing the complete list of genes potentially involved in cholesterol breakdown. Moreover, we identified the modulatory effect of vitamin B12 on Mtb transcriptome and the novel function of cobalamin in cholesterol metabolite dissipation which explains the probable role of B12 in Mtb virulence. Finally, we demonstrate that a key role of cholesterol in mycobacterial metabolism is not only providing carbon and energy but involves also a transcriptome remodeling program that helps in developing tolerance to the unfavorable host cell environment far before specific stress-inducing phagosomal signals occur.


Impact of pregnancy and lactation on GABA(A) receptor and central-type and peripheral-type benzodiazepine receptors.
The effect of pregnancy and lactation on GABA(A) receptor and central- and peripheral-type benzodiazepine receptors (CBR and PBR, respectively) was studied in female Sprague-Dawley rats. Pregnancy was associated with increased CBR density (on day 19) in the hippocampus and with decreased [3H]Ro 15-1788-specific binding in the hypothalamus during pregnancy and lactation. A similar decrease in [3H]PK 11195-specific binding was observed in the hypothalamus and pituitary. An increase in PBR density in the ovary and uterus was observed during pregnancy, while adrenal PBR density was down-regulated during pregnancy and lactation. It seems that the hormonal changes occurring during pregnancy and lactation play a role in the regulation of CBR and PBR in discrete tissues.


Binding of polylysine and ethidium bromide to nucleosomal DNA: comparison of biochemical and electron microscopical results.
Ethidium bromide and polylysine interact with nucleosomal DNA and lead to change of biochemical properties and to morphological changes as to the distance between the two core particles of a nucleosome dimer. With increasing polylysine concentration, the buoyant density of nucleosomes decrease and the accessibility of the nucleosomal DNA to micrococcal nuclease is lowered. Electron microscopy of polylysine treated nucleosome dimers reveals a shortening of the internucleosomal distance as compared with controls. Treatment of nucleosomes with ethidium bromide leads to an enhanced accessibility of the nucleosomal DNA to micrococcal nuclease. Electron microscopy reveals an increase in length of the DNA connecting the two nucleosome cores in the presence of the dye. Both the binding of polylysine and the treatment with ethidium bromide apparently do not affect the histone arrangement within the nucleosome core as suggested by chemical cross-linking of histones and DNA with formaldehyde, and no obvious morphological change of the nucleosome cores can be observed.


Effects of a disrupted blood-brain barrier on cholesterol homeostasis in the brain.
The presence of the blood-brain barrier (BBB) is critical for cholesterol metabolism in the brain, preventing uptake of lipoprotein-bound cholesterol from the circulation. The metabolic consequences of a leaking BBB for cholesterol metabolism have not been studied previously. Here we used a pericyte-deficient mouse model, Pdgfb(ret/ret), shown to have increased permeability of the BBB to a range of low-molecular mass and high-molecular mass tracers. There was a significant accumulation of plant sterols in the brains of the Pdgfb(ret/ret) mice. By dietary treatment with 0.3% deuterium-labeled cholesterol, we could demonstrate a significant flux of cholesterol from the circulation into the brains of the mutant mice roughly corresponding to about half of the measured turnover of cholesterol in the brain. We expected the cholesterol flux into the brain to cause a down-regulation of cholesterol synthesis. Instead, cholesterol synthesis was increased by about 60%. The levels of 24(S)-hydroxycholesterol (24S-OHC) were significantly reduced in the brains of the pericyte-deficient mice but increased in the circulation. After treatment with 1% cholesterol in diet, the difference in cholesterol synthesis between mutants and controls disappeared. The findings are consistent with increased leakage of 24S-OHC from the brain into the circulation in the pericyte-deficient mice. This oxysterol is an efficient suppressor of cholesterol synthesis, and the results are consistent with a regulatory role of 24S-OHC in the brain. To our knowledge, this is the first demonstration that a defective BBB may lead to increased flux of a lipophilic compound out from the brain. The relevance of the findings for the human situation is discussed.


The impact of strict COVID-19 lockdown in Spain on glycemic profiles in patients with type 1 Diabetes prone to hypoglycemia using standalone continuous glucose monitoring.
Spain has been one of the worst affected countries by the COVID-19 pandemic. A very strict lockdown at home was imposed with a tough restriction of mobility. We aimed to evaluate the impact of this exceptional scenario on glucose profile of patients with T1D prone to hypoglycemia using standalone continuous glucose monitoring. Patients with T1D prone to hypoglycemia using multiple daily injections and either a Dexcom G5® or a Free Style Libre® CGM systems for at least 6 months under the funding of National Health Service were included in an observational, retrospective study. Data were collected in two periods: pre-lockdown (PL), February 23rd-March 7th and within lockdown (WL), April 1st-14th 2020. The primary outcome was the difference in the proportion of time in target glucose range of 70-180 mg/dL (TIR). Additional glucometric data were also analysed. 92 patients were included: 40 women, age 42.8 ± 3.9 years, disease duration of 23.1 ± 12.6 years. Seventeen patients used Dexcom G5® and 75 Free Style Libre®. TIR 70-180 mg/dL (59.3 ± 16.2 vs 62.6 ± 15.2%), time > 180 (34.4 ± 18.0 vs 30.7 ± 16.9%), >250 (11.1 ± 10.6 vs 9.2 ± 9.7%) and Glucose Management Indicator (7.2 ± 0.8 vs 7.0 ± 0.8%) significantly improved (PL vs WL, respectively, p < 0.05). Time in hypoglycemia remained unchanged. Lockdown conditions imposed by the COVID-19 pandemic may be managed successfully in terms of glycemic control by population with T1D prone to hypoglycemia using CGM. The strict daily routine at home could probably explain the improvement in the time in glycemic target without increasing the time in hypoglycemia.


Gating and conduction properties of a sodium-activated cation channel from lobster olfactory receptor neurons.
The gating and conduction properties of a channel activated by intracellular Na+ were studied by recording unitary currents in inside-out patches excised from lobster olfactory receptor neurons. Channel openings to a single conductance level of 104 pS occurred in bursts. The open probability of the channel increased with increasing concentrations of Na+. At 210 mm Na+, membrane depolarization increased the open probability e-fold per 36.6 mV. The distribution of channel open times could be fit by a single exponential with a time constant of 4.09 msec at -60 mV and 90 mm Na+. The open time constant was not affected by the concentration of Na+, but was increased by membrane depolarization. At 180 mm Na+ and -60 mV, the distribution of channel closed times could be fit by the sum of four exponentials with time constants of 0.20, 1.46, 8.92 and 69.9 msec, respectively. The three longer time constants decreased, while the shortest time constant did not vary with the concentration of Na+. Membrane depolarization decreased all four closed time constants. Burst duration was unaffected by the concentration of Na+, but was increased by membrane depolarization. Permeability for monovalent cations relative to that of Na+ (PX/PNa), calculated from the reversal potential, was: Li+ (1.11) > Na+ (1.0) > K+ (0.54) > Rb+ (0.36) > Cs+ (0.20). Extracellular divalent cations (10 mm) blocked the inward Na+ current at -60 mV according to the following sequence: Mn2+ > Ca2+ > Sr2+ > Mg2+ > Ba2+. Relative permeabilities for divalent cations (PY/PNa) were Ca2+ (39.0) > Mg2+ (34.1) > Mn2+ (15.5) > Ba2+ (13.8) > Na+ (1.0). Both the reversal potential and the conductance determined in divalent cation-free mixtures of Na+ and Cs+ or Li+ were monotonic functions of the mole fraction, suggesting that the channel is a single-ion pore that behaves as a multi-ion pore when the current is carried exclusively by divalent cations. The properties of the channel are consistent with the channel playing a role in odor activation of these primary receptor neurons.


Progressive multifocal leukoencephalopathy in an elderly immunocompetent-appearing patient: Relevance with L-selectin (CD62L) expression and immunosenescence.
Progressive multifocal leukoencephalopathy (PML) is attributed to reactivation of the John Cunningham virus (JCV), in the central nervous system as a result of immunosuppression. Low L-selectin (CD62L) expression on cryopreserved T-cells has been advocated as a biomarker for natalizumab related PML in patients with Relapsing-Remitting Multiple Sclerosis. A rare case of PML in an elderly patient without known factors of immunosuppression or immunomodulation is hereby presented. T-cell L-selectin expression levels and serum anti-JCV antibody index were evaluated in order to explore mechanistic insight to the pathways that presumably contribute towards PML development in this rare clinical setting.


Intermittent social stress produces different short- and long-term effects on effort-based reward-seeking behavior.
Previous studies show that intermittent social defeat (ISD) stress increases self-administration of psychostimulants, which suggests that ISD promotes reward-seeking behavior and, ultimately, increases vulnerability to develop drug abuse. The present study investigates whether ISD alters cost/benefit evaluations to promote reward-seeking behavior and whether these alterations are time-dependent. Male rats performed two different tasks that assessed their motivation to seek and consume food rewards. An effort-discounting task in which rats chose between less and more effortful options (i.e., 1 lever-press versus 2, 5, 10 or 20 lever-presses) associated with low- and high-reward (i.e., 1 sugar pellet versus 3 sugar pellets), respectively; and a progressive ratio task in which rats had to increase their effort (more lever presses) to obtain a sugar pellet. ISD consisted of exposing animals to social defeat once every three days for ten days (4 stress episodes). Rats were tested 24-48 h after stress episodes, and 1 week and 6 weeks after the last stress episode. In the effort-discounting task, stressed animals showed a decrease in their preference for high rewards associated with more effort (i.e., 10 and 20 lever-presses). These effects were transient and not maintained one week after stress. In the progressive ratio task, stressed animals showed an increase in the number of lever presses to obtain rewards that emerged six weeks after the last stress episode. These results suggest different short- and long-term effects on the motivation for rewards after ISD and indicate temporal dynamic adaptations in the function of the brain reward system.


Interfering with Wnt signalling alters the periodicity of the segmentation clock.
Somites are embryonic precursors of the ribs, vertebrae and certain dermis tissue. Somite formation is a periodic process regulated by a molecular clock which drives cyclic expression of a number of clock genes in the presomitic mesoderm. To date the mechanism regulating the period of clock gene oscillations is unknown. Here we show that chick homologues of the Wnt pathway genes that oscillate in mouse do not cycle across the chick presomitic mesoderm. Strikingly we find that modifying Wnt signalling changes the period of Notch driven oscillations in both mouse and chick but these oscillations continue. We propose that the Wnt pathway is a conserved mechanism that is involved in regulating the period of cyclic gene oscillations in the presomitic mesoderm.


Validation of an educational game to promote cardiovascular health in children.
The aim of this study was to validate the content and appearance of an educational game for children aged 7 to 10 years, focusing mainly on cardiovascular health promotion. The study used methodological research, with a quantitative approach. The validation process included the participation of 17 specialists in children and/or cardiovascular health. A concordance index of at least 0.80 was considered for content validation and 0.75 for the appearance of the proposed educational material. The proposal of the educational game was considered valid, through some suggestions, in view of the purpose of sensitizing children in relation to the promotion of cardiovascular health while still in childhood. As a relevant factor in the use of games as strategies to improve health education, we emphasize that the participant is the active agent and the protagonist of the health-disease process.


Evaluating and improving residential treatment during group leisure situations: a program replication and refinement.
The authors attempted to replicate and refine a program for evaluating and improving residential treatment services during group leisure periods in living units. In Study 1, an active treatment program was implemented using a consultant model in two living units serving persons with severe handicaps. Similar to previous research, the program was accompanied by decreases in nonadaptive client behavior in both units during leisure periods. In addition, a time-efficient monitoring system was developed which suggested that two staff-related variables, provision of leisure materials, and interactions with clients were related to the program's success. Study 2 evaluated more thoroughly the relationship of these variables to client nonadaptive behavior in 20 residential units in four states. Significant predictive relationships again were found between nonadaptive behavior and material availability, and between nonadaptive behavior and staff interactions with clients. Results of Study 2 also indicated that three living unit supervisors readily learned to use the evaluation system. Results are discussed regarding the importance of simple and efficient monitoring systems for routinely evaluating and improving residential services.


Simple models for the effect of aliphatic alcohol additives on the retention in reversed-phase liquid chromatography.
Four retention models for the effect of aliphatic alcohol additives on the retention of analytes in reversed-phase liquid chromatography have been developed following either a semi-thermodynamic treatment or an empirical approach. Their performance was tested using the experimental retention times of six non-polar analytes (alkylbenzenes) and ten o-phthalaldehyde derivatives of amino acids under different isocratic chromatographic runs when a small amount of ethanol, 1-propanol, 1-butanol, 1-pentanol, 1-hexanol or 1-heptanol was added to methanol/water mixtures containing a constant amount of methanol. It was shown that for the structurally simple alkylbenzenes all the models can be adopted for retention prediction with good results. In contrast, just one out of four models, that with the fewest approximations, predicts satisfactorily the retention properties of amino acids derivatives. However, the most interesting feature is that this model can predict the effect of an alcohol-additive on the retention properties of solutes, even if this additive was not used in chromatographic runs done for the fitting procedure, provided that it belongs to the same homologous series of alkanols. This feature is also observed in all models described the retention of alkylbenzenes.


Imaging neuroinflammation in Alzheimer's disease and other dementias: Recent advances and future directions.
Alzheimer's disease (AD), dementia with Lewy bodies, frontotemporal dementia (FTD), and Huntington's disease (HD) are the main neurodegenerative causes of dementia. Causes and mechanisms of these diseases remain elusive. Neuroinflammation is increasingly emerging as an important pathological factor in their development. Positron emission tomography (PET) using [11C]PK11195 represents a method of visualizing the microglial component of neuroinflammation via the translocator protein (TSPO) and we discuss the valuable insights this has yielded in neurodegenerative diseases. We discuss the limitations of this method and the development of second generation TSPO PET ligands which hope to overcome these limitations. We also discuss other methods of visualizing neuroinflammation and review the state of current dementia treatments targeted at neuroinflammation. It is our view that a multimodal investigation into neuroinflammation in AD, Parkinson's disease dementia, FTD and HD will yield valuable pathological insights which will usefully inform development of therapeutic targets and biomarkers.


Comparison of the effectiveness of slow-release propranolol (Inderal LA) 80, 160 and 320 mg orally daily in the treatment of angina pectoris.
Doses of 80, 160 and 320 mg of Inderal LA (slow-release propranolol) were compared in 12 patients with stable angina pectoris. There was an unpredictable and variable individual response to treatment and hence no differences were detected between the effects of the three doses on mean treadmill exercise duration, time to angina, frequency of angina or glyceryl/trinitrate consumption. There was a marked deterioration in two patients whilst on 320 mg Inderal LA. In these patients, the time to angina fell by 2 minutes 50 seconds and 4 minutes 20 seconds on 320 mg daily and 160 mg daily respectively. Given the inter-patient variability in response to Inderal LA, the dose for each patient should be individually titrated according to response. Two patients reported here suggest that some subjects may benefit from lower doses, although further work is required to clarify this point. Possible explanations for this finding are discussed.


Malignant transformation of a mature cystic ovarian teratoma into thyroid carcinoma, mucinous adenocarcinoma, and strumal carcinoid: a case report and literature review.
Malignant transformation of a mature cystic teratoma (MCT) is an infrequent, often asymptomatic event. We report the first example of a struma ovarii with a focus of follicular variant of papillary thyroid carcinoma (a), mucinous adenocarcinoma (b), and strumal carcinoid tumor (c)-all three arising in one mature cystic teratoma of the ovary. From our reviews, we found limited data to guide management when these malignant foci occur within an MCT. Consideration should be given to thyroidectomy followed by total-body scanning and serum studies for foci of thyroid carcinoma and adjuvant therapy with thyroidectomy and radioablation if residual disease is identified (a). Additionally, extrapolating from data for mucinous adenocarcinomas, consideration could be given to adjuvant chemotherapy after appropriate staging (b). Strumal carcinoid tumors should be treated as tumors of low malignant potential. Observation is appropriate if after complete staging, no invasive implants are noted (c).


Lignin and lignans in plant defence: insight from expression profiling of cinnamyl alcohol dehydrogenase genes during development and following fungal infection in Populus.
Cinnamyl alcohol dehydrogenase (CAD) catalyses the final step in the biosynthesis of monolignol, the main component of lignin. Lignins, deposited in the secondary cell wall, play a role in plant defence against pathogens. We re-analysed the phylogeny of CAD/CAD-like genes using sequences from recently sequenced genomes, and analysed the temporal and spatial expression profiles of CAD/CAD-like genes in Populus trichocarpa healthy and infected plants. Three fungal pathogens (Rhizoctonia solani, Fusarium oxysporum, and Cytospora sp.), varying in lifestyle and pathogenicity, were used for plant infection. Phylogenetic analyses showed that CAD/CAD-like genes were distributed in classes represented by all members from angiosperm lineages including basal angiosperms and Selaginella. The analysed genes showed different expression profiles during development and demonstrated that three genes were involved in primary xylem maturation while five may function in secondary xylem formation. Expression analysis following inoculation with fungal pathogens, showed that five genes were induced in either stem or leaves. These results add further evidence that CAD/CAD-like genes have evolved specialised functions in plant development and defence against various pest and pathogens. Two genes (PoptrCAD11 and PoptrCAD15), which were induced under various stresses, could be treated as universal markers of plant defence using lignification or lignan biosynthesis.


The impact of regular diving on the condition of the middle ear.
It is generally held that exposure to both high-pressure and long-lasting contact with water makes diving a potentially hazardous sports activity as far as the ears are concerned. There is a number of research investigating the condition of the middle ear in a short period following diving; however, the knowledge regarding the long-term effects of regularly repeated diving remains limited. The aim of this study is to evaluate the function of the middle ear after a diving season in a group of 31 adults diving regularly (1-17 years) by means of the following methods: 1) interview, 2) otoscopy, 3) pure tone audiometry, 4) classic tympanometry, and 5) wideband tympanometry. Periodic problems with pressure equalization in the middle ear were observed in 12 individuals (38.7%). In all the analyzed cases, the authors found a normal condition of the external auditory canal and the tympanic membrane in otoscopy, normal hearing in pure tone audiometry, curve type A, and normal gradient in both classic and wideband tympanometry. Safe diving (according to safety precautions) does not have any long-term negative effects on the condition of the middle ear. However, these observations should be verified in a larger group of divers. Int J Occup Med Environ Health. 2021;34(6):779-88.


Physicochemical characterization of the reassembled dimer of an integral membrane protein OmpF porin.
The in vitro reassembled species of OmpF porin, which was renatured from its denatured monomer using n-octyl-beta-D-glucopyranoside, was characterized by low-angle laser light scattering photometry, circular dichroism spectroscopy and synchrotron radiation small-angle X-ray scattering measurements. The light scattering measurement reconfirmed that the reassembled species was the dimer of the protein. Circular dichroism spectra of the reassembled dimer showed a native-like beta-structure. A small-angle X-ray scattering measurement indicated that the size of the reassembled dimer was nearly equal to that of the native trimer under the present experimental conditions. In a thermal denaturation experiment followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the reassembled dimer was less stable than the native trimer.


Protocol for Site-Specific Photo-Crosslinking Proteomics to Identify Protein-Protein Interactions in Mammalian Cells.
Protein-protein interactions (PPIs) play essential roles in almost all aspects of cellular processes. However, PPIs remain challenging to study due to their substoichiometry, low affinity, dynamic nature, and context dependence. Here, we present a protocol for the capture and identification of PPIs in live mammalian cells, which relies on site-specific photo-crosslinking in live cells, affinity purification, and quantitative proteomics. The protocol facilitates efficient and reliable identification of the interacting proteins of a given protein of interest in live cells. For complete details on the use and execution of this protocol, please refer to Wu et al. (2020).


The Histone Variant MacroH2A1 Is a BRCA1 Ubiquitin Ligase Substrate.
The breast- and ovarian-cancer-specific tumor suppressor BRCA1 and its heterodimeric partner BARD1 contain RING domains that implicate them as E3 ubiquitin ligases. Despite extensive efforts, the bona fide substrates of BRCA1/BARD1 remain elusive. Here, we used recombinant GST fused to four UBA domains to enrich ubiquitinated proteins followed by a Lys-ε-Gly-Gly (diGly) antibody to enrich ubiquitinated tryptic peptides. This tandem affinity purification method coupled with mass spectrometry identified 101 putative BRCA1/BARD1 E3 substrates. We identified the histone variant macroH2A1 from the screen and showed that BRCA1/BARD1 ubiquitinates macroH2A1 at lysine 123 in vitro and in vivo. Primary human fibroblasts stably expressing a ubiquitination-deficient macroH2A1 mutant were defective in cellular senescence compared to their wild-type counterpart. Our study demonstrates that BRCA1/BARD1 is a macroH2A1 E3 ligase and implicates a role for macroH2A1 K123 ubiquitination in cellular senescence.


Estimated Costs and Outcomes Associated With Use and Nonuse of Medications for Opioid Use Disorder During Incarceration and at Release in Massachusetts.
Most prisons and jails in the US discontinue medications for opioid use disorder (MOUD) upon incarceration and do not initiate MOUD prior to release. To model the association of MOUD access during incarceration and at release with population-level overdose mortality and OUD-related treatment costs in Massachusetts. This economic evaluation used simulation modeling and cost-effectiveness with costs and quality-adjusted life-years (QALYs) discounted at 3% to compare MOUD treatment strategies in a corrections cohort and an open cohort representing individuals with OUD in Massachusetts. Data were analyzed between July 1, 2021, and September 30, 2022. Three strategies were compared: (1) no MOUD provided during incarceration or at release, (2) extended-release (XR) naltrexone offered only at release from incarceration, and (3) all 3 MOUDs (naltrexone, buprenorphine, and methadone) offered at intake. Treatment starts and retention, fatal overdoses, life-years and QALYs, costs, and incremental cost-effectiveness ratios (ICERs). Among 30 000 simulated incarcerated individuals with OUD, offering no MOUD was associated with 40 927 (95% uncertainty interval [UI], 39 001-42 082) MOUD treatment starts over a 5-year period and 1259 (95% UI, 1130-1323) overdose deaths after 5 years. Over 5 years, offering XR-naltrexone at release led to 10 466 (95% UI, 8515-12 201) additional treatment starts, 40 (95% UI, 16-50) fewer overdose deaths, and 0.08 (95% UI, 0.05-0.11) QALYs gained per person, at an incremental cost of $2723 (95% UI, $141-$5244) per person. In comparison, offering all 3 MOUDs at intake led to 11 923 (95% UI, 10 861-12 911) additional treatment starts, compared with offering no MOUD, 83 (95% UI, 72-91) fewer overdose deaths, and 0.12 (95% UI, 0.10-0.17) QALYs per person gained, at an incremental cost of $852 (95% UI, $14-$1703) per person. Thus, XR-naltrexone only was a dominated strategy (both less effective and more costly) and the ICER of all 3 MOUDs compared with no MOUD was $7252 (95% UI, $140-$10 018) per QALY. Among everyone with OUD in Massachusetts, XR-naltrexone only averted 95 overdose deaths over 5 years (95% UI, 85-169)-a 0.9% decrease in state-level overdose mortality-while the all-MOUD strategy averted 192 overdose deaths (95% UI, 156-200)-a 1.8% decrease. The findings of this simulation-modeling economic study suggest that offering any MOUD to incarcerated individuals with OUD would prevent overdose deaths and that offering all 3 MOUDs would prevent more deaths and save money compared with an XR-naltrexone-only strategy.


Supervised machine learning enables non-invasive lesion characterization in primary prostate cancer with [68Ga]Ga-PSMA-11 PET/MRI.
Risk classification of primary prostate cancer in clinical routine is mainly based on prostate-specific antigen (PSA) levels, Gleason scores from biopsy samples, and tumor-nodes-metastasis (TNM) staging. This study aimed to investigate the diagnostic performance of positron emission tomography/magnetic resonance imaging (PET/MRI) in vivo models for predicting low-vs-high lesion risk (LH) as well as biochemical recurrence (BCR) and overall patient risk (OPR) with machine learning. Fifty-two patients who underwent multi-parametric dual-tracer [18F]FMC and [68Ga]Ga-PSMA-11 PET/MRI as well as radical prostatectomy between 2014 and 2015 were included as part of a single-center pilot to a randomized prospective trial (NCT02659527). Radiomics in combination with ensemble machine learning was applied including the [68Ga]Ga-PSMA-11 PET, the apparent diffusion coefficient, and the transverse relaxation time-weighted MRI scans of each patient to establish a low-vs-high risk lesion prediction model (MLH). Furthermore, MBCR and MOPR predictive model schemes were built by combining MLH, PSA, and clinical stage values of patients. Performance evaluation of the established models was performed with 1000-fold Monte Carlo (MC) cross-validation. Results were additionally compared to conventional [68Ga]Ga-PSMA-11 standardized uptake value (SUV) analyses. The area under the receiver operator characteristic curve (AUC) of the MLH model (0.86) was higher than the AUC of the [68Ga]Ga-PSMA-11 SUVmax analysis (0.80). MC cross-validation revealed 89% and 91% accuracies with 0.90 and 0.94 AUCs for the MBCR and MOPR models respectively, while standard routine analysis based on PSA, biopsy Gleason score, and TNM staging resulted in 69% and 70% accuracies to predict BCR and OPR respectively. Our results demonstrate the potential to enhance risk classification in primary prostate cancer patients built on PET/MRI radiomics and machine learning without biopsy sampling.


Characteristics and driving conditions of flash drought in different grassland ecosystems.
At present, flash droughts are poorly understood. Mature prevention and control measures are not available. This study aims to thoroughly explore the evolution characteristics of flash droughts in grassland ecosystems and to determine the meteorological driving conditions for inducing and relieving flash droughts. We propose the concept of the strong evapotranspiration flash drought (SEFD), a new type of flash drought in mid-temperate grasslands. The frequency of SEFDs is lower than that of heat wave flash droughts (HWFDs), but the intensity and impact of SEFDs are greater than those of HWFDs. Flash droughts in grasslands exhibit a high outbreak period from May to August, with the intensity basically above that of moderate drought. HWFDs occur most frequently in June and SEFDs in May. Meadow grasslands are the type of grassland with the highest risk of flash drought. Typical grasslands are more prone to HWFDs, while desert grasslands are more prone to SEFD outbreaks. In this study, a multifactor method was established to quantitatively evaluate the key influencing factors inducing flash droughts of different intensities in different time periods and to quantitatively predict the evolution of flash droughts into seasonal droughts. The temperature, water vapor pressure, precipitation, and wind speed were the key influencing factors of flash droughts and that the most important period in terms of inducing flash droughts is one pentad before the onset of flash drought. The outbreak rate and threshold of key influencing factors are the driving conditions for inducing flash droughts, and the recovery rate can be used as the basis for predicting whether an flash drought will evolve into a seasonal drought. The results showed that the characteristics and driving conditions of flash droughts are different in different types of grasslands and different time periods and are different for different types of flash droughts. In the future, the impact of flash droughts will become more severe.


Burkitt-like lymphoma in a pediatric patient with familial adenomatous polyposis.
Familial adenomatous polyposis (FAP) is an autosomal dominant condition that predisposes to multiple malignancies, most commonly colorectal carcinoma, but has rarely been associated with lymphoma. We discuss one patient found to have Burkitt-like Lymphoma (BLL) with 11q aberration in the setting of previously undiagnosed FAP. We review the literature of FAP and associated malignancies and the provisional WHO classification of Burkitt-like lymphoma with 11q aberration. Both FAP and Burkitt-like lymphoma with 11q aberration involve perturbation of the MYC network and this may provide insight into a connection between these two diagnoses. However, further study is needed to elucidate if there is an increased risk of BLL and other subtypes of lymphoma among patients with FAP in order to provide optimal counseling and surveillance for patients with FAP.


Urethral pressure profile during the spinal shock stage in man: a preliminary report.
The functional state of the proximal urethra in the spinal shock stage in man is not fully understood. We studied patients with spinal cord injuries during spinal shock and found that the urethral pressure profile had a normal configuration, the peak profile pressure increased with bladder filling and phentolamine (10 mg. intravenously) reduced the peak pressure, with empty and full bladders.


Paeonol Suppresses Vasculogenesis Through Regulating Vascular Smooth Muscle Phenotypic Switching.
Smooth muscle cell (SMC) phenotypic switching is associated with development of a variety of occlusive vascular diseases. Paeonol has been reported to be involved in suppressing SMC proliferation. However, it is still unknown whether paeonol can regulate SMC phenotypic switching, and which eventually result in suppressing vasculogenesis. Murine left common carotid artery was injured by completely ligation, and paeonol was administrated by intraperitoneal injection. Hematoxylin and eosin (H&E) staining was performed to visualize vascular neointima formation. Rat aortic SMCs were used to determine whether paeonol suppresses cell proliferation and migration. And murine hind limb ischemia model was performed to confirm the function role of paeonol in suppressing vasculogenesis. Complete ligation of murine common carotid artery successfully induced neointima formation. Paeonol treatment dramatically reduced the size of injury-induced neointima. Using rat aortic primary SMC, we identified that paeonol strongly suppressed cell proliferation, migration, and decreased extracellular matrix deposition. And paeonol treatment dramatically suppressed vasculogenesis after hind limb ischemia injury. Paeonol could regulate SMC phenotypic switching through inhibiting proliferation and migration of SMC, which results in inhibiting ischemia-induced vasculogenesis.


Prediction of fetal RR intervals from maternal factors using machine learning models.
Previous literature has highlighted the importance of maternal behavior during the prenatal period for the upbringing of healthy adults. During pregnancy, fetal health assessments are mainly carried out non-invasively by monitoring fetal growth and heart rate (HR) or RR interval (RRI). Despite this, research entailing prediction of fHRs from mHRs is scarce mainly due to the difficulty in non-invasive measurements of fetal electrocardiogram (fECG). Also, so far, it is unknown how mHRs are associated with fHR over the short term. In this study, we used two machine learning models, support vector regression (SVR) and random forest (RF), for predicting average fetal RRI (fRRI). The predicted fRRI values were compared with actual fRRI values calculated from non-invasive fECG. fRRI was predicted from 13 maternal features that consisted of age, weight, and non-invasive ECG-derived parameters that included HR variability (HRV) and R wave amplitude variability. 156 records were used for training the models and the results showed that the SVR model outperformed the RF model with a root mean square error (RMSE) of 29 ms and an average error percentage (< 5%). Correlation analysis between predicted and actual fRRI values showed that the Spearman coefficient for the SVR and RF models were 0.31 (P < 0.001) and 0.19 (P < 0.05), respectively. The SVR model was further used to predict fRRI of 14 subjects who were not included in the training. The latter prediction results showed that individual error percentages were (≤ 5%) except in 3 subjects. The results of this study show that maternal factors can be potentially used for the assessment of fetal well-being based on fetal HR or RRI.


Cervical pathology with intrauterine contraceptive devices--a cyto-colpo-pathological study.
One-hundred Lippes loop and 100 Cu T-200 intrauterine contraceptive device (IUCD) users, for more than one year, and 200 control non-users were examined clinically, cytologically, colposcopically and histopathologically for associated cervical pathological lesions. Nonspecific infection increased significantly in both Lippes loop and Cu T-200 IUCD users, compared to non-users (P less than 0.05). On the other hand, there was no statistically significant difference in the incidence of total, or individual, specific infections, dyskaryosis or CIN between both groups (P greater than 0.05). Colposcopically, there was a significant increase in inflammatory changes with both Lippes loop and Cu T-200 IUCD, compared to controls (P less than 0.05), but there was no difference in other colposcopic findings (P greater than 0.05).


Evolution of energetic metabolism: the respiration-early hypothesis.
The main energy-transducing metabolic systems originated and diversified very early in the evolution of life. This makes it difficult to unravel the precise steps in the evolution of the proteins involved in these processes. Recent molecular data suggest that homologous proteins of aerobic respiratory chains can be found in Bacteria and Archaea, which points to a common ancestor that possessed these proteins. Other molecular data predict that this ancestor was unlikely to perform oxygenic photosynthesis. This evidence, that aerobic respiration has a single origin and may have evolved before oxygen was released to the atmosphere by photosynthetic organisms, is contrary to the textbook viewpoint.


The influence of body core temperature and peripheral temperatures on oxygen consumption in the pig.
1. The rate of oxygen consumption was measured in young pigs exposed to different ambient temperatures and the effect on metabolic rate of changing the temperature of thermodes implanted in the hypothalamus and over the spinal cord was determined. In some experiments the temperature of the skin over the trunk was changed by means of a water-perfused coat.2. Cooling the hypothalamus or the spinal cord in a warm ambient did not change the rate of oxygen consumption. At a thermoneutral ambient temperature, cooling either thermode increased oxygen consumption. In a cold environment, cooling either thermode increased the rate of oxygen consumption more than at a thermoneutral temperature. The increase in the rate of oxygen consumption was greatest during cooling of the spinal cord and it appeared that the pigs also shivered more violently. Heating either thermode tended to decrease oxygen consumption in a cold environment.3. In pigs with thermodes both in the hypothalamus and over the spinal cord, cooling both thermodes was accompanied by a greater increase in oxygen consumption than cooling either thermode alone. The increase in oxygen consumption on cooling one thermode could be reduced by heating the other.4. The skin temperature (fixed by the water perfused coat, or the ambient temperature) at which the rate of oxygen consumption increased, was lowered during heating of the thermodes, but the rate of increase in oxygen consumption appeared not to change as a function of falling skin or ambient temperature.


NIRF Nanoprobes for Cancer Molecular Imaging: Approaching Clinic.
Near-IR fluorescence imaging (NIRFI) is a highly promising technique for improving cancer theranostics in the era of precision medicine. Through the combination with cutting-edge bionanotechnologies, the potential of NIRFI can be greatly broadened. A variety of novel NIRF nanoprobes has been developed with ultimate goals of addressing unmet medical needs. Here, we present recent breakthroughs on the fundamental aspects of NIRFI, such as imaging at long wavelengths (1000-1700 nm), and the use of new approaches (X-rays, chemiluminescence, radioluminescence, etc.) for the excitation of novel nanoprobes. Within two decades, research on NIRF nanoprobes has translated to clinical trials and it will further translate to cancer management.


Olive leaf extract supplementation improves the vascular and metabolic alterations associated with aging in Wistar rats.
Olive leaves are rich in bioactive substances which exert anti-inflammatory, antioxidant, insulin-sensitizing and antihypertensive effects. The aim of this study was to analyze the possible beneficial effects of an olive leaf extract (OLE) rich in secoiridoids and phenolic compounds on the aging-induced metabolic and vascular alterations. Three experimental groups of rats were used: 3-month-old rats, 24-month-old rats and 24-month-old rats supplemented 21 days with OLE (100 mg/kg). Administration of OLE to aged rats decreased the weight of adrenal glands and prevented the aging-induced loss of body weight and muscle mass. In the serum, OLE reduced the circulating levels of LDL-cholesterol and IL-6 and increased the concentrations of leptin and adiponectin. In the liver OLE attenuated the decreased gene expression of SOD-1, GSR, GCK and GSK-3β and reduced the aging-induced overexpression of NOX-4, Alox-5, iNOS and TNF-α. In aorta segments, OLE prevented endothelial dysfunction and vascular insulin resistance and improved vasoconstriction in response to KCl and NA. Improvement in vascular function was associated with the attenuation of the alterations in the gene expression of COX-2, IL-6, GPx, NOX-1 and IL-10. In conclusion, OLE exerts anti-inflammatory and antioxidant effects in aged rats and attenuates the alterations in vascular function associated with aging.


HER2 Overexpression as a Poor Prognostic Determinant in Resected Biliary Tract Cancer.
HER2 overexpression has been investigated as a potential biomarker and therapeutic target in biliary tract cancer (BTC), but a prognostic role of such alteration has not been demonstrated yet. We retrospectively evaluated HER2 protein expression by immunohistochemistry (IHC) in 100 patients with radically resected BTC. HER2 gene amplification was assessed by fluorescence in situ hybridization (FISH) in 2+ and 3+ cases at IHC. High HER2 protein expression was defined as either IHC 3+ or 2+ associated with FISH positivity. The primary objective of the study was to evaluate the prognostic role of HER2 overexpression in terms of disease-free survival (DFS) and overall survival (OS). Secondary endpoints were the prevalence of HER2 overexpression and the possible correlation with other clinicopathological features. HER2 overexpression was identified in 11 patients and was not related to other clinicopathological factors. DFS was significantly shorter in HER2-positive compared with HER2-negative patients (10.6 vs. 20.9 months, log-rank p = .017). HER2 confirmed its prognostic value for DFS at multivariate analysis (hazard ratio 2.512; 95% confidence interval, 1.232-5.125; p = .011) together with nodal stage (p < .001), resection margin (p = .027), and tumor site (p = .030). There was no difference in OS between HER2-positive and -negative patients (p = .068). HER2 overexpression represents an independent prognostic factor for disease recurrence in patients with BTC treated with potentially curative surgery. HER2 overexpression may play an independent role in promoting an aggressive behavior in resectable biliary tract cancer. This evidence could be helpful in improving prognostic stratification after resection and, primarily, should endorse the rationale to investigate HER2 as a therapeutic target in biliary tract cancer.


[Prognostic value of histological rare breast cancer (author's transl)].
Among 835 cases of female breast cancer in the years 1951-1971 we found 44 histological rare cancer (comedo-carcinoma 21, carcinoma cribosum 1, gelatinous carcinoma 8, squamous cell carcinoma 3, mixed carcinoma 11). In agreement with the literature we are able to underline the favourable prognosis of the slow growing forms of the neoplasms like gelatinous carcinoma, comedo-carcinoma and carcinoma cribrosum opposite to squamous cell carcinoma and mixed carcinoma.


Helicobacter pylori eradication in a clinical setting: success rates and the effect on the quality of life in peptic ulcer.
Helicobacter pylori eradication for peptic ulcer has been widely taken up. Evidence for the efficacy of different regimens is often derived from small series in clinical trials but there is little reporting of everyday practice with unselected patients. Freedom from ulcer relapse has been demonstrated, but not whether this equates with clinical success. We report on a series of 706 patients with H. pylori infection who, between January 1991 and April 1995, received eradication therapy followed by assessment of H. pylori status. Two-hundred and seven of these patients were followed-up by postal questionnaire, validated by parallel questionnaires to their general practitioners, covering clinical outcome measures. The overall eradication rate was 81.7%, and a 1-week course of omeprazole plus two antibiotics was significantly better than a 2-week course of standard triple therapy (85.0% vs. 78.0%, P < 0.05). Amongst 21 first-time failures, a 7-day course of a clarithromycin-containing triple therapy succeeded in 18. The questionnaire replies indicate that, following successful H. pylori eradication, ulcer patients are less likely to consult with ulcer symptoms (P < 0.0005), take medication (P < 0.0005), require further prescription (P < 0.0005), or lose work-time because of their ulcer (P < 0.005). They are more likely to have a subjective sense of ulcer cure (P < 0.0005). In addition to clear cost savings, social benefits are now demonstrated when H. pylori is eradicated. A well-tolerated 1 week regimen is genuinely effective in everyday practice.


Weight Loss After Stroke Through an Intensive Lifestyle Intervention (Group Lifestyle Balance-Cerebrovascular Accident): Protocol for a Randomized Controlled Trial.
Weight gain can be a consequence of stroke, or cerebrovascular accident (CVA), because of impaired mobility, behavioral and emotional disorders, and sensory losses. Weight gain increases the patient's risk of recurrent stroke and chronic diseases, such as diabetes, metabolic syndrome, and pulmonary and heart disease. Approaches to weight loss in this population are lacking, although necessary because of the unique physiological and cognitive needs of persons after a stroke. Evidence shows that intensive behavioral therapy interventions that address both physical activity and diet offer the greatest potential for weight loss. The Group Lifestyle Balance (GLB) intervention is a 12-month, evidence-based weight loss program that has been used extensively with the general population; this program was modified to meet the needs of people who have had a stroke (GLB-CVA). This randomized controlled trial (RCT) aims to examine the efficacy of the GLB-CVA on weight and secondary outcomes, compared with that of a waitlist control group. This RCT will enroll and randomize 64 patients over an 18-month period. Currently, 51 people are waitlisted, with 23 out of 51 screened and 16 out of 23 eligible. It is anticipated that the findings from this RCT will contribute to the evidence base regarding weight loss strategies for people living with stroke. ClinicalTrials.gov NCT03873467; https://clinicaltrials.gov/ct2/show/NCT03873467.


Albuminuria Is Affected by Urinary Tract Infection: A Comparison between Biochemical Quantitative Method and Automatic Urine Chemistry Analyzer UC-3500.
The automated urine reagent strip test is a cost-effective tool for detecting albuminuria in patients. However, prior research has not investigated how urinary tract infections (UTIs) affect the test's accuracy. Therefore, this study aims to assess the impact of UTIs on albuminuria diagnosis using both the biochemical quantitative method and the test strip method of the Fully Automatic Urine Chemistry Analyzer, UC-3500 (Sysmex, Kobe, Japan). From March to December 2019, we prospectively collected midstream urine from adult female UTI patients before and after one week of cephalexin treatment. The urine samples were subjected to culture, routine urinalysis, and albuminuria diagnosis using the biochemical quantitative method and UC-3500. Albuminuria was defined as a urine albumin to creatinine ratio (UACR) ≥ 30 mg/g in the biochemical quantitative method. The results were compared between the two methods. Among fifty-four female patients (average age: 50.5 ± 4.4 years) with UTIs, 24 (44.44%) had transient albuminuria. The quantitative UACR significantly decreased after one week of antibiotic treatment (median: 53 mg/g to 9 mg/g; median difference: -0.54, p < 0.0001). UC-3500 exhibited a higher false positive rate for diagnosing albuminuria during UTIs (42%) compared to after treatment (19%). Its agreement with the biochemical quantitative method was moderate during UTI (κ = 0.49, 95% confidence interval [CI]: 0.24-0.73) and good after treatment (κ = 0.65, 95% CI: 0.45-0.86). UC-3500's accuracy in diagnosing albuminuria is influenced by UTIs, leading to either transient albuminuria or a false positive reaction of the test strip. UTI should be excluded or treated before its application in albuminuria screening.


Characterization and identification of three novel aldo-keto reductases from Lodderomyces elongisporus for reducing ethyl 4-chloroacetoacetate.
Lodderomyces elongisporus LH703 isolated from soil samples contained three novel aldo-keto reductases (AKRs) (LEAKR 48, LEAKR 49, and LEAKR 50). The three enzymes were cloned, expressed, and purified to homogeneity for characterization. These three AKRs shared <40% amino acid identity with each other. LEAKR 50 was identified as a member of AKR3 family, whereas the other two LEAKRs were identified as members of two novel AKR families, respectively. All the three AKRs required nicotinamide adenine dinucleotide phosphate as a cofactor. However, they showed diverse characteristics, including optimum catalyzing conditions, resistance to adverse reaction conditions, and substrate specificity. LEAKR 50 was estimated to be a promising biocatalyst that could reduce ethyl 4-chloroacetoacetate with high enantiomeric excess (98% e. e.) and high activity residue under adverse conditions.


Fast and Broadband Photoresponse of a Few-Layer GeSe Field-Effect Transistor with Direct Band Gaps.
Few-to-monolayer germanium selenide, a new IV-VI group layered material recently fabricated by mechanical exfoliation and subsequent laser thinning, is promising in very fast and broadband optoelectronic applications for its excellent stability, complicated band structures, inert surface properties, and being a natural p-type semiconductor. However, large-scale production of such few-layer GeSe devices with superior performance is still in early stages. In this study, field-effect transistors made of few-layer GeSe with direct band gaps are fabricated. Transistor performance with Schottky contact characteristics is measured at room temperature. A field-effect mobility of 4 cm2/(V s) and drain currents modulated both by holes and electrons are measured. Photoresponses as a function of illumination wavelength, power, and frequency are characterized. The few-layer GeSe transistor shows photoresponse to the illumination wavelengths from visible up to 1400 nm and a photoresponse rise (fall) time of 13 μs (19 μs), demonstrating very broadband and fast detection. The ambipolar behavior and the photoresponse characteristics demonstrate great potential of few-layer GeSe for applications in highly stable, very fast, and very broadband optoelectronic devices.


Sacroiliac Joint Anatomy.
The sacroiliac joint can be a source of low back pain. This review article summarizes current anatomic evidence of the innervation of the intraarticular and extraarticular parts of the sacroiliac joint relative to bony landmarks identifiable with fluoroscopy and ultrasound. This article aims to provide clinicians with an anatomic basis for clinical application to diagnostic blocks and radiofrequency ablation for sacroiliac pain to optimize clinical outcomes.


Tunable 7-12 μm picosecond optical parametric amplifier based on a LiInSe2 mid-infrared crystal.
Mid-infrared (MIR) nonlinear optical crystals of LiInSe2 (LISe) were grown by a modified Bridgman technique on a (001)-seed. A 7-12 μm widely tunable picosecond (ps) MIR optical parametric amplifier (OPA) based on a LISe crystal was demonstrated for the first time, to the best of our knowledge. The MIR OPA was pumped by a 30 ps 1064 nm Nd:YAG laser and injected by a KTiOPO4 (KTP)-based widely tunable near-infrared seed. The idler operating at 7.5 μm with the highest pulse energy of 170 μJ was obtained under a pump energy of 14 mJ. The corresponding energy conversion efficiency is ∼1.21%, and the photon conversion efficiency is 8.6%. The output energies were measured to be ∼121 μJ at 7 μm and ∼21 μJ at 12 μm.


Infantile achalasia. Brief clinical report.
Achalasia of the esophagus is an unusual lesion in childhood and is quite rare under the age of one year. The case of a 3-month-old child with esophageal achalasia treated with a Heller esophagomyotomy is illustrated. Seven previously documented cases of achalasia in children under the age of one year are reviewed and the difficulties of diagnosing this lesion in childhood are discussed.


Virtual reality goes to war: a brief review of the future of military behavioral healthcare.
Numerous reports indicate that the incidence of posttraumatic stress disorder (PTSD) in returning OEF/OIF military personnel is creating a significant healthcare challenge. These findings have served to motivate research on how to better develop and disseminate evidence-based treatments for PTSD. Virtual Reality delivered exposure therapy for PTSD has been previously used with reports of positive outcomes. This article details how virtual reality applications are being designed and implemented across various points in the military deployment cycle to prevent, identify and treat combat-related PTSD in OIF/OEF Service Members and Veterans. The summarized projects in these areas have been developed at the University of Southern California Institute for Creative Technologies, a U.S. Army University Affiliated Research Center, and this paper will detail efforts to use virtual reality to deliver exposure therapy, assess PTSD and cognitive function and provide stress resilience training prior to deployment.


A new approach to patients with suspected anaerobic postpartum pelvic infections. Transabdominal uterine aspiration for culture and metronidazole for treatment.
Oral metronidazole was given as therapy in 25 women with the clinical diagnosis of a postpartum anaerobic soft tissue pelvic infection following a vaginal delivery. There was a good clinical response in 80 per cent of these women. No anaerobic organisms resistant to metronidazole were recovered in the treatment failures. Different specimen collection techniques were evaluated, with anaerobes recovered in 4 per cent of the blood cultures, 26.3 per cent of the transabdominal endometrial aspirations, and 88 per cent of the transcervical endometrial samples. The aerobic recovery of organisms was 4 per cent from blood cultures, 15.8 per cent from transabdominal endometrial aspirations, and 96 per cent from the transcervical approach. The significance of these findings is discussed.


Therapy-Induced Tumor Cell Senescence: Mechanisms and Circumvention.
Plasticity of tumor cells (multitude of molecular regulation pathways) allows them to evade cytocidal effects of chemo- and/or radiation therapy. Metabolic adaptation of the surviving cells is based on transcriptional reprogramming. Similarly to the process of natural cell aging, specific features of the survived tumor cells comprise the therapy-induced senescence phenotype. Tumor cells with this phenotype differ from the parental cells since they become less responsive to drugs and form aggressive progeny. Importance of the problem is explained by the general biological significance of transcriptional reprogramming as a mechanism of adaptation to stress, and by the emerging potential of its pharmacological targeting. In this review we analyze the mechanisms of regulation of the therapy-induced tumor cell senescence, as well as new drug combinations aimed to prevent this clinically unfavorable phenomenon.


Effects of low and high resistance training intensities on clinical outcomes in patients with COPD - a randomized trial.
To compare the effects of two different intensities of combined resistance and aerobic training on physical-functional aspects and perceptual aspects of health status in patients with Chronic Obstructive Pulmonary Disease (COPD). This was a randomized parallel group trial. Thirty-one patients were assessed regarding: symptoms (COPD Assessment Test - CAT), quality of life (Saint George's Respiratory Questionnaire - SGRQ); exercise capacity (six-minute walk test and constant-workload resistance test - CWRT); and muscle strength (one-repetition maximum test and isometric muscle strength for knee extensors - Kext and elbow flexors - EFlex). Patients were randomized in two groups to perform a combined training: 1) LL/HR = Low-load/high-repetition resistance training (n = 16; 68 ± 9.3 years; FEV1 = 50.0 ± 15.7%pred); and 2) HL/LR = High-load/low-repetition resistance training (n = 15; 70 ± 6.5 years; FEV1 = 46.8 ± 14.5%pred), and underwent 36 sessions. Symptoms, quality of life and exercise capacity presented significant improvement (p < .05) in both groups. Both intensities of resistance training improved symptoms (∆CAT: LL/HR = -3.9; HL/LR = -2.8) and exercise capacity (∆ tolerance time in CWRT: LL/HR = +450.9s; HL/LR = +583.4s) above minimal clinically important difference value. Nevertheless, improvement in quality of life (∆SGRQ: LL/HR = -3.8; HL/LR = -10) and in isometric muscle strength (∆KExt: LL/HR = +15.8 N; HL/LR = +37.7 N and (∆EFlex: LL/HR = +9.5 N; HL/LR = +29.9 N) was observed only in the HL/LR group, which also presented a larger number of responders considering isometric muscle strength. High-intensity resistance training in a combined training was more effective in ameliorating several aspects of patients with COPD, responding more and better to physical-functional aspects and perceptual aspects of health status.


Circulating prolactin level in systemic lupus erythematosus and its correlation with disease activity: a meta-analysis.
Objective This study aimed to evaluate the relationship between circulating prolactin level and systemic lupus erythematosus (SLE), and to establish a correlation between plasma/serum prolactin levels and SLE activity. Methods We performed a meta-analysis comparing the plasma/serum prolactin levels in patients with SLE to controls, and examined correlation coefficients between circulating prolactin level and SLE disease activity. Results Twenty-five studies with a total of 1056 SLE patients and 426 controls were included. Prolactin levels were significantly higher overall in the SLE group than in the control group (standardized mean difference (SMD) = 0.987, 95% CI = 0.512-1.463, p = 4.7 × 10-5). Stratification by ethnicity showed significantly elevated prolactin levels in the SLE group in Asian, Latin American, and mixed populations (SMD = 0.813, 95% CI = 0.137-1.490, p = 0.018; SMD = 0.981, 95% CI = 0.307-1.655, p = 0.004; SMD = 1.469, 95% CI = 0.443-2.495, p = 0.005, respectively), but not in the European population. Subgroup analysis by sample size showed significantly higher prolactin levels in the SLE group by small ( n < 30) and large sample numbers ( n > 30). Meta-analysis of correlation coefficients showed a significantly positive correlation between circulating prolactin level and SLE activity (correlation coefficient = 0.379, 95% CI = 0.026-0.487, p = 4.0 × 10-9). Circulating prolactin levels were positively associated with SLE activity in European, Asian, and mixed populations (SMD = 0.532, 95% CI = 0.443-0.609 p < 1.0 × 10-8; SMD = 0.427, 95% CI = 0.240-0.583, p = 2.4 × 10-5; SMD = 0.433, 95% CI = 0.212-0.591, p = 2.7 × 10-5, respectively). Conclusions Our meta-analysis demonstrated that circulating prolactin levels are higher in patients with SLE, and that a significantly positive correlation exists between prolactin levels and SLE activity.


[The use of botulinum toxin A in the treatment of focal dystonias].
22 patients with focal dystonias were treated with local injections of botulinum toxin A (Botox) into muscles with involuntary, sustained movements. The method is highly effective, practically without serious side effects and now is considered the treatment of choice in this group of patients what has been also confirmed in our own material.


Catalytic wet peroxide oxidation of phenol by pillared clays containing Al-Ce-Fe.
A Colombian bentonite was successfully pillared with mixed polyhydroxocationic solutions of Al-Fe or Al-Ce-Fe, with the iron in small quantities, to generate a modified Fenton process on the surface of the pillared clays with a possible activation of the hydrogen peroxide and the organic substances on the iron sites of the solids in order to obtain active and stable catalysts for the wet peroxide oxidation of organic pollutants from industrial wastewaters. The catalysts were very efficient in the reaction of phenol oxidation in diluted aqueous media under mild experimental conditions (298 K and atmospheric pressure) and in the elimination of the different intermediary compounds of the reaction reaching high-mineralization levels. The solids showed high stability in the reaction media due to the strong interaction between the iron species and the catalyst support. The incorporation of Ce in the solids showed a favorable effect in the pillaring of the materials allowing the increase of the basal spacing and enhancing the catalytic activity of the solids.


Laparoscopic Roux-en-Y gastric bypass for the treatment of type II diabetes mellitus in Chinese patients with body mass index of 25-35.
Laparoscopic Roux-en-Y gastric bypass (LRYGB) can dramatically ameliorate type 2 diabetes mellitus (T2DM) in morbidly obese patients. However, there is little evidence supporting the effectiveness of LRYGB in low body mass index (BMI) patients. The study was designed to evaluate the safety and results of LRYGB for achieving T2DM remission in patients with BMI in the range of 25-35 kg/m(2). Twenty-two patients (two men and 20 women) with T2DM underwent LRYGB. Data on patient demographics, BMI, co-morbidities, and details of diabetes mellitus, including disease duration, family history, medication use, and remission, were prospectively collected and analyzed. The mean age was 47 years (range, 28-63 years), mean BMI was 30.81 (range, 25.00-34.80 kg/m(2)), and mean duration of T2DM onset was 6.57 years (range, 1-20 years). Sixteen (72.27%) patients had a family history of T2DM. There was no mortality, but two (9%) patients experienced complications: an early gastrojejunostomy hemorrhage and frequent loose stools that required revision surgery. At 12 months, 14 (63.6%) patients showed T2DM remission, six (27.3%) showed glycemic control, and two (9.1%) showed improvement. The group achieving remission had a higher BMI (p = 0.001), younger age (p = 0.002), and shorter duration of diabetes (p = 0.001). These three factors may be predictors of diabetes resolution at 12 months. Early intervention in low-BMI patients yields better remission rates because age, BMI, and duration of T2DM predict glycemic outcomes.


Evaluating motor performance with the Bruininks-Oseretsky Test of motor proficiency in impoverished Pakistani children.
To evaluate the sensitivity of the modified Brief Form of Bruininks Oseretsky Test in identifying motor differences secondary to malnutrition and poverty. This longitudinal cohort study was conducted at Nowshero Feroze, Sindh from 2013 to 2014 and comprised data drawn from children who participated in a randomised controlled trial, that assessed responsive stimulation and nutrition interventions in the first two years of life. Outcome measures included motor development assessed using Brief Form of Bruininks Oseretsky Test, child anthropometry and household economic and demographic information. Data was analysed using SPSS 15 and STATA 12. Of the 1058 children, 570(53%) were boys. Moderate-severe stunting was reported in 171(16.12%) subjects, while moderate-severe underweight was reported in 117(11.1%). Also, 591(56%) subjects belonged to poor families, 343(32%) had illiterate mothers, and 392(37%) were food-insecure. Malnutrition, socio-economic status and maternal literacy were significantly associated with a 6-item motor composite of the Brief Form of Bruininks Oseretsky Test (p<0.05). The 6-item motor composite of the Brief Form of Bruininks Oseretsky Test was found to be a reliable tool to measure motor performance in Pakistani pre-school children.


A comparison of the effects of symmetry and magnetoanisotropy on paramagnetic relaxation in related dysprosium single ion magnets.
Dysprosium complexes of the tmtaa(2-) ligand were synthesized and characterized by X-band EPR and magnetism studies. Both complexes demonstrate magnetoanisotropy and slow paramagnetic relaxation. Comparison of these compounds with the seminal phthalocyanine complex [Dy(Pc)(2)](-) shows the azaannulide complexes are more susceptible to relaxation through non-thermal pathways.


'Womb for work' experiences of Thai women and gestational surrogacy practice in Thailand.
Risks to gestational surrogates became a concern for public health. In commercial gestational surrogacy arrangements, gestational surrogates are commonly recruited from low- to middle-income countries. Thailand is well known as a surrogacy hub in this regard. However, little is known concerning Thai surrogacy practice and the risks that Thai gestational surrogates experiences. The semi-structured interviews with fifteen Thai women who had been gestational surrogates were conducted over the telephone in Thai between March and May 2020 and lasted approximately 30 minutes. Thematic analysis was applied to analyse the translated interviews. The findings indicated that 'womb for work' was perceived as a surrogacy career among Thai women. 'Womb for work' was defined as a superordinate theme that consisted of three subthemes: (i) gestational surrogacy arrangements in Thailand; (ii) the business model of gestational surrogacy arrangements in Thailand; and (iii) risk experiences of gestational surrogates. Clear deficiencies in surrogacy practice and regulations were identified, which put gestational surrogates at risk, including those associated with embryo transfer, transnational gestational surrogacy, and unsupported pregnancies. This study shows the urgent need to introduce regulations to protect women's health transnationally in this domain more effectively.


Patients' reports of the factors influencing medication adherence in bipolar disorder - an integrative review of the literature.
As with other long-term conditions patients with bipolar disorder are rarely totally adherent or non-adherent. Rates of non-adherence have not changed since the first introduction of psychotropic medications in the 1950s despite vast numbers of new compounds being marketed. Non-adherence with medication in bipolar disorder is associated with affective relapse and consequently poor quality of life. The reasons that patients are non-adherent with medication are not well understood by clinicians who often assume it is related to the illness itself. To identify patients' perceptions of medication adherence in bipolar disorder. An integrated review of the literature published between 1999 and 2010. Ovid (Medline, CINAHL, Embase, PsycINFO) and manual searching. An integrative review of the literature was conducted which included: (a) problem formation, (b) literature search and initial screening, (c) gathering data from studies, (d) evaluating study quality, (e) data analysis and integration, (f) data interpretation, and (g) presentation of the findings. Thirteen articles met criteria for inclusion in the review. These articles identified how patients reported their perceptions on medication and were integrated into four categories: illness factors, personal attitudes and beliefs, medication factors and environmental factors. These findings suggest a need to address adherence from the full range of influencing factors (patient, illness, medication and environmental). Clinicians need to utilise a collaborative approach to working together with patients in order to identify the meaning that patients attribute to the symptoms, diagnosis, prognosis and medication. Understanding patients' perceptions and accepting these may facilitate greater medication adherence and the consequent improved clinical outcomes for patients with bipolar disorder.


Evolution of hominoids and the search for a genetic basis for creating humanness.
The phylogenetic relationship of human and apes are reviewed. The history of molecular phylogenetic studies in this field is then discussed, as is the role of natural selection at the molecular level. It is argued that approximately 10,000 genetic changes are responsible for creating human specific phenotypes. A genome-wide comparison is necessary to decipher those changes.


Maintaining Access to Orthopaedic Surgery During Periods of Operating Room Resource Constraint: Expanded Use of Wide-Awake Surgery During the COVID-19 Pandemic.
Wide-awake local anesthesia no tourniquet (WALANT) presents a nonstandard anesthetic approach initially described for use in hand surgery that has gained interest and utilization across a variety of orthopaedic procedures. In response to operating room resource constraints imposed by the COVID-19 pandemic, our orthopaedic service rapidly adopted and expanded its use of WALANT. A retrospective review of 16 consecutive cases performed by 7 surgeons was conducted. Patient demographics, surgical details, and perioperative outcomes were assessed. The primary end point was WALANT failure, defined as intraoperative conversion to general anesthesia. No instances of WALANT failure requiring conversion to general anesthesia occurred. In recovery, one patient (6%) required narcotics for pain control, and the average postoperative pain numeric rating scale was 0.6. The maximum pain score experienced was 4 in the patient requiring postoperative narcotics. The average time in recovery was 42 minutes and ranged from 8 to 118 minutes. The WALANT technique was safely and effectively used in 16 cases across multiple orthopaedic subspecialties, including three procedures not previously described in the literature. WALANT techniques hold promise for use in future disaster scenarios and should be evaluated for potential incorporation into routine orthopaedic surgical care.


Preventive Health Care for Working Dogs.
The goal of preventive care is to maintain and optimize health by averting preventable problems. Effective preventive care programs for working dogs must incorporate standard procedures applicable to dogs in general with additional elements pertinent to the more specific characteristics of breed, geographic location, living and working conditions, and physical and mental tasks required of the working dog. This article covers the basic essential preventive health guidelines for all working dogs as well as the specific breed, occupational, and regional considerations to be taken into account.


The role of negativity bias in political judgment: a cultural neuroscience perspective.
Hibbing et al. provide a comprehensive overview of how being susceptible to heightened sensitivity to threat may lead to conservative ideologies. Yet, an emerging literature in social and cultural neuroscience shows the importance of genetic and cultural factors on negativity biases. Promising avenues for future investigation may include examining the bidirectional relationship of conservatism across multiple levels of analysis.


Blow-up and control of marginally separated boundary layers.
Interactive solutions for steady two-dimensional laminar marginally separated boundary layers are known to exist up to a critical value Gamma(c) of the controlling parameter (e.g. the angle of attack of a slender airfoil) Gamma only. Here, we investigate three-dimensional unsteady perturbations of such boundary layers, assuming that the basic flow is almost critical, i.e. in the limit Gamma(c)-Gamma-->0. It is then shown that the interactive equations governing such perturbations simplify significantly, allowing, among others, a systematic study of the blow-up phenomenon observed in earlier investigations and the optimization of devices used in boundary-layer control.


Cepharanthine hydrochloride inhibits the Wnt/β‑catenin/Hedgehog signaling axis in liver cancer.
Cepharanthine, a biscoclaurine alkaloid isolated from the roots of Stephania cephalantha Hayata, has been reported to demonstrate antitumor activity across multiple cancer types; however, the mechanisms are still under investigation. High transcriptional responses by both the Hedgehog and Wnt pathways are frequently associated with specific human cancers, including liver cancer. To investigate whether these signaling pathways are involved in the pharmaceutical action of cepharanthine, we investigated Hedgehog and Wnt signaling in models of liver cancer treated with a semi‑synthetic cepharanthine derivative, cepharanthine hydrochloride (CH), in vitro and in vivo. By using MTT cytotoxic, scratch, Transwell, colony formation and flow cytometry assays, the pharmaceutical effect of CH was assessed. The compound was found to inhibit cellular proliferation and invasion, and promote apoptosis. Subsequent mechanistic investigations revealed that CH suppressed the Hedgehog/Gli1 signaling pathway by inhibiting Gli1 transcription and its transcriptional activity. CH also inhibited Wnt/β‑catenin signaling, and the pathway was found to be an upstream regulator of Hedgehog signaling in CH‑treated liver cancer cells. Finally, the antitumor effects of CH were demonstrated in an in vivo xenograft tumor model. Immunohistochemical analysis indicated that Gli1 protein levels were diminished in CH‑treated xenografts, compared with that noted in the controls. In summary, our results highlight a novel pharmaceutical antitumor mechanism of cepharanthine and provide support for CH as a clinical therapy for refractory liver cancer and other Wnt/Hedgehog‑driven cancers.


A Risk-Based Approach to Evaluate and Control Elemental Impurities in Therapeutic Proteins.
Control of elemental impurities in the drug products evolved from the generic visual testing of heavy metals as their sulfides to specific elements of toxicological concern in the final drug products by instrumental analysis. The International Council for Harmonisation (ICH) Q3D (R1) guideline for elemental impurities describes a risk-based approach to identify, assess, and control the potential elemental impurities in drug products within the established permitted daily exposures (PDE). Challenges to this approach include how to assess the risks associated with contributing sources such as utilities, manufacturing equipment, container-closure systems, and excipients. Defining at what stage of development that such assessment should be performed to identify the risk levels can be equally challenging. In this article, we report an approach to control elemental impurities of toxicological concern, compliant to the Q3D (R1) guideline, and a summary of results obtained on multiple protein therapeutic products. This approach follows the elements of Process Validation, i.e., Design, Qualification, and Continuous Verification. The design includes the selection of excipients and their suppliers that meet the Option 1 requirement of Q3D (R1). It also comprises the selection of manufacturing equipment, container-closure systems, and utilities. The qualification includes the testing of the potential sources of elemental impurities, i.e., excipients, utilities, and leachables/extractables from the manufacturing equipment and container-closure systems. The Continuous Verification comes from the testing of representative batches at the initiation of stability studies of clinical or commercial drug product batches and at the end of shelf-life expiry of the drug product, and when changes are made to the manufacturing equipment, sources of excipients and container closure systems, and any formulation changes. Our experience shows that the risk associated with the impurity levels of the ten elements of toxicological concern in the therapeutic protein drug products, parenterally administered, is well below the control threshold (30% PDE) in the drug product recommended by the ICH Guideline. Although our focus is on the injectable therapeutic proteins, this approach can be applied to the products administered via other routes as well.


Venous reflux in the great saphenous vein is driven by a suction force provided by the calf muscle pump in the compression-decompression maneuver.
The gravitational pressure gradient is considered the driving force of venous reflux. The results from our previous study demonstrated that gravitational force is not a necessary condition for the occurrence of venous reflux. We hypothesized that a force exists in addition to gravity that drives venous reflux. The present study was designed to test this hypothesis by measuring the acceleration of blood flow during venous reflux in a clinical study and by simulating reflux ex vivo in physical models. A total of 80 lower extremities of 80 patients with primary incompetence of the great saphenous vein were included in the present study. The cross-sectional area of the great saphenous vein, peak velocity of venous reflux (PV), and time required to achieve the PV (Δt, seconds) were measured on duplex ultrasound scans taken with the patient in the standing rest position. Noncycling operator-dependent distal cuff inflation-deflation was used as the reflux provoking maneuver. The acceleration of venous reflux (areflux) was calculated as areflux = PV/Δt in m/s2. Physical models were used to demonstrate the difference in acceleration between the free-fall stream and the flow forced by suction. The magnitude of areflux was greater than gravity in 24 of 80 extremities (30%), with a range of 9.83 to 24.13 m/s2. The maximum observed value of areflux was approximately 2.5g (24.13 m/s2). The areflux weakly, but statistically significant inversely, correlated with the subject height (r = -0.26; P = .001). The difference in water flow acceleration was 2.5 times between the free-fall model and suction model (9.07 ± 0.2 m/s2 vs 23.32 ± 2.6 m/s2, respectively). The acceleration of blood flow during reflux exceeded the value of gravitational acceleration, suggesting the action of an additional nongravitational force. The calf muscle pump might create such force by negative pressure during muscle diastole.


Kinetics of electron transfer reactions by humic substances: Implications for their biogeochemical roles and determination of their electron donating capacity.
Humic substances (HS) possess redox active groups covering a wide range of potentials and are used by facultative anaerobic microorganisms as electron acceptors. To serve as suitable electron shuttles for anaerobic respiration, HS should be able to re-oxidize relatively quickly to prevent polarization of the surrounding medium. Mediated electrochemical oxidation and decolorization assays, based on the reduction of the radical ion of 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate) (ABTS•-) allow to determine the electron donating capacity (EDC) of HS, but uncertainties remain about the reaction time that should be allowed to obtain environmentally meaningful EDC values. In this work, we performed a kinetic analysis of the time trend of the reduction of ABTS•- by HS by Vis and Electron Paramagnetic Resonance (EPR) spectroscopies and by cyclic voltammetry. We found evidences of two concomitant separate mechanisms of electron exchange: a fast and a slow transfer processes which may have different environmental roles. These results can set a base to identify the appropriate conditions for the spectrophotometric determination of the fast and slow components of the EDC of HS.


A literature review and case reports series on the use of phosphodiesterase inhibitors in the treatment of female sexual dysfunction.
The term 'female sexual dysfunction' (FSD) encompasses a number of different disorders, and while their aetiologies are not fully understood, the sub-classifications of this broad umbrella term are increasingly becoming more established and accepted. However, there is less consensus regarding the optimal treatment of these conditions. While it is known that phosphodiesterase (PDE5) is involved in the female sexual response, the clinical and research evidence supporting the unlicensed use of PDE5 inhibitors (PDE5i) in women is inconclusive and at times contradictory. In this article we explore this further by means of a comprehensive literature review on the use of PDE5i in the treatment of FSD and we also present our clinical experience of using these drugs in this context.


Cardiopulmonary bypass in the presence of uncorrected coarctation.
We performed correction of intracardiac lesions in 10 patients with coarctation. All cardiovascular lesions were corrected except coarctation. Six patients had a large left-to-right shunt though a ventricular septal defect (VSD), two patients had left ventricular outflow obstruction, two patients had mitral insufficiency, and one patient had mitral stenosis. Arterial perfusion was through the ascending aorta. Urine output was 6.0 +/- 1.5 cc/kg/hr during cardiopulmonary bypass. There was no morbidity related to cardiopulmonary bypass or inadequate perfusion of the lower body, and left ventricular failure did not occur. One patient with Taussig-Bing abnormality and pulmonary vascular disease died postoperatively of right ventricular failure. Intracardiac repair with aortic perfusion in the presence of coarctation is a safe alternative to coarctation repair or combined procedures.


Examining Specificity of Neural Correlates of Childhood Psychotic-like Experiences During an Emotional n-Back Task.
Psychotic-like experiences (PLEs) during childhood are associated with greater risk of developing a psychotic disorder in adulthood, highlighting the importance of identifying neural correlates of childhood PLEs. Furthermore, impairment of cognitive functions, such as working memory and emotion regulation, has also been linked to psychosis risk as well as to disruptions in several brain regions. However, impairments in these domains have also been linked to other disorders, including depression. Therefore, the aim of the current study was to examine whether neural impairments in regions associated with working memory and implicit emotion regulation impairments are specific to PLEs versus depression. The current study used an emotional n-back task to examine the relationship between childhood PLEs and neural activation of regions involved in both working memory and implicit emotion regulation using data from 8805 9- to 11-year-olds in the Adolescent Brain Cognitive Development (ABCD) Study 2.0 release. To examine specificity, we also analyzed associations with depressive symptoms. Our results indicated that increased PLEs during middle childhood were associated with decreased activation of the dorsolateral prefrontal cortex, striatum, and pallidum during trials requiring working memory. In contrast, increased activation of the parahippocampus, caudate, nucleus accumbens, and rostral anterior cingulate during face-viewing trials was associated with increased depressive symptoms. These results support the dimensional view of psychosis across the lifespan, providing evidence that neural correlates of PLEs, such as decreased activation during working memory, are present during middle childhood. Furthermore, these correlates are specific to psychotic-like symptoms as compared with depressive symptoms.


Cell-type-specific expression of STAT transcription factors in tissue samples from patients with lymphocytic thyroiditis.
Expression of cytokine-regulated signal transducer and activator of transcription (STAT) proteins was histochemically assessed in patients diagnosed as having Hashimoto's disease or focal lymphocytic thyroiditis (n = 10). All surgical specimens showed histological features of lymphocytic thyroiditis, including a diffuse infiltration with mononuclear cells and an incomplete loss of thyroid follicles, resulting in the destruction of glandular tissue architecture. Immunohistochemical analysis demonstrated differential expression patterns of the various members of the STAT transcription factors examined, indicating that each member of this conserved protein family has its distinct functions in the development of the disease. Using an antibody that specifically recognized the phosphorylated tyrosine residue in position 701, we detected activated STAT1 dimers in numerous germinal macrophages and infiltrating lymphocytes as well as in oncocytes. In contrast, STAT3 expression was restricted to epithelial cells and showed a clear colocalization with the antiapoptotic protein Bcl-2. Moreover, expression of phospho-STAT3 was associated with low levels of stromal fibrosis, suggesting that STAT3 serves as a protective factor in the remodeling of the inflamed thyroid gland. Phospho-STAT5 immunoreactivity was detected in numerous infiltrating cells of hematopoietic origin and, additionally, in hyperplastic follicular epithelia. This tissue distribution demonstrated that activated STAT5 molecules participate in both lymphocytopoiesis and possibly also in the buildup of regenerating thyroid follicles. Taken together, the cell-type-specific expression patterns of STAT proteins in human lymphocytic thyroiditis reflect their distinct and partially antagonistic roles in orchestrating the balance between degenerating and regenerating processes within a changing cytokine environment.


Satisfaction, stress and burnout of nurse managers and care nurses in Primary Health Care.
To correlate the indicators of job satisfaction, factors of occupational stress and professional exhaustion among nurse managers and care nurses in Primary Health Care. Descriptive, exploratory field study conducted in 45 units of Primary Health Care in Brazil. The following instruments were used for data collection: Sociodemographic Questionnaire, Work Stress Scale, Burnout Characterization Scale and the Job Satisfaction Questionnaire - S20/23. Participation of 122 nurses (47.5% managers; 62.5% care), 32% had considerable stress levels, indicators of emotional exhaustion, dehumanization and disappointment at work in moderate to high levels. The indicator of satisfaction with the physical work environment did not show significance with variables of professional exhaustion, while satisfaction with hierarchical relations and intrinsic job satisfaction are strongly correlated. There is an association between organizational problems and working conditions that hinder nurses' work. Regardless of the position, job satisfaction is inversely proportional to burnout.