Pain persists in mice lacking both Substance P and CGRPα signaling.
The neuropeptides Substance P and CGRPα have long been thought important for pain sensation. Both peptides and their receptors are expressed at high levels in pain-responsive neurons from the periphery to the brain making them attractive therapeutic targets. However, drugs targeting these pathways individually did not relieve pain in clinical trials. Since Substance P and CGRPα are extensively co-expressed we hypothesized that their simultaneous inhibition would be required for effective analgesia. We therefore generated Tac1 and Calca double knockout (DKO) mice and assessed their behavior using a wide range of pain-relevant assays. As expected, Substance P and CGRPα peptides were undetectable throughout the nervous system of DKO mice. To our surprise, these animals displayed largely intact responses to mechanical, thermal, chemical, and visceral pain stimuli, as well as itch. Moreover, chronic inflammatory pain and neurogenic inflammation were unaffected by loss of the two peptides. Finally, neuropathic pain evoked by nerve injury or chemotherapy treatment was also preserved in peptide-deficient mice. Thus, our results demonstrate that even in combination, Substance P and CGRPα are not required for the transmission of acute and chronic pain.


Dietary modification of Parkinson's disease.
To investigate the efficacy of a low-protein diet suitable for long-term use, and to observe the effect in a double-blind manner of re-introducing large neutral amino acids or an inactive placebo amino acid, eight patients with 'on-off' phenomenon were given a low-protein diet suitable for long-term use, and then the diet was supplemented with large neutral amino acids (LNAA) or placebo in a double-blind crossover trial. Five patients had improved Parkinsonian disability on a low-protein diet compared to normal diet. On supplementing the diet with LNAA or placebo, three subjects' Parkinson's disease was significantly better on placebo and there was a trend towards improvement on placebo noticed subjectively by seven patients; however, there was no overall significant difference in disability measured by mobility diaries. Thus, we have been unable to show that the improvement on a low-protein diet is due to a reduction in LNAA, rather than a placebo effect, possibly due to the paucity of subjects; however, the patients' identification of the LNAA supplement suggests that it is a genuine effect.


Maintenance therapy in patients following the surgical treatment of peri-implantitis: a 5-year follow-up study.
To evaluate the outcomes of conventional periodontal maintenance therapy on patients surgically treated for peri-implantitis. 27 patients with 149 dental implants were monitored during 5 years every 6 months. At each recall visit, the prostheses were removed to have proper access for implant examination and supra- and sub-gingival instrumentation. Sub-gingival instrumentation was performed using an ultrasonic instrument with under 0.12% chlorhexidine irrigation. At baseline (6 months following peri-implant surgery), 149 implants (78 not treated and 71 treated) were available for analysis. Of the 71 treated implants, 43 presented healthy peri-implant condition, while 28 had residual peri-implant pockets either of 4-5 mm or ≥ 6 mm associated with bleeding on probing/suppuration. The longitudinal evaluation revealed that the plaque and the bleeding index scores were low during the entire follow-up period, and healthy peri-implant conditions were maintained for both the 78 non-treated and the 43 treated "healthy" implants. Of the 28 implants with residual pockets, nine showed clinical attachment loss during the 5-year follow-up. Thus, of 71 treated implants, probing attachment loss occurred in only in 9 (13%) of the implants in four patients during the 5-year period. The presence of residual pockets at three or four sites of the implants (circumferential type of pockets) was frequently associated with increased probing pocket depth (PPD) and attachment loss, while this was not the case for implants with the presence of pockets at one or two sites only (site specific). In patients with a high standard of oral hygiene and enrolled in a recall system every 6 months, the peri-implant conditions obtained following peri-implant surgery were maintained stable for the majority of subjects and implants during a 5-year period. Presence of residual pockets around the circumference of the implants seemed to be a high predictor for disease progression.


Functional specificity of a long-range horizontal connection in cat visual cortex: a cross-correlation study.
Anatomical investigations of the visual cortex revealed a regular set of interlaminar connections and long-range horizontal connections. An important component of the intrinsic cortical circuit is the projection from layer 5 to layer 6 over long horizontal distances. Previous work has shown that when layer 5 is locally inactivated, layer 6 cells lose their response in a segment of their receptive fields corresponding to the blocked area in layer 5. It has therefore been proposed that the long receptive fields characteristic of layer 6 cells might be generated by concatenating layer 5 cells with similar orientation preferences. In the present study, we used cross-correlation analysis to examine both source and target cells of the interlaminar connection from layer 5 to layer 6. We found correlated firing between cells separated by up to 4.2 mm, the longest horizontal distance studied. The occurrence of correlated firing depended on the functional properties and on the topographic position of the cells in layer 5 and layer 6. Interactions were only observed if the cells had matching orientation tuning and similar eye preference and if the layer 5 cells lay within the summation area of the layer 6 cells. Both simple and complex cells in layer 6 fired in synchrony with layer 5 cells. In layer 5, mainly standard complex cells, but very few special complex cells, participated in correlated firing with layer 6 cells. These results suggest that cells in layer 6 receive their input from cooriented, coaxially aligned standard complex cells in layer 5. This topographic arrangement and the length summation properties of standard complex cells in layer 5 are well suited to generate long receptive fields in layer 6. Thus, our experiments provide evidence for the functional specialization of a particular component of cortical circuitry.


A novel dengue virus detection method that couples DNAzyme and gold nanoparticle approaches.
Recent epidemics of dengue viruses (DENV) coupled with new outbreaks on the horizon have renewed the demand for novel detection methods that have the ability to identify this viral pathogen prior to the manifestation of symptoms. The ability to detect DENV in a timely manner is essential for rapid recovery from disease symptoms. A modified lab-derived 10-23 DNAzyme tethered to gold nanoparticles provides a powerful tool for the detection of viruses, such as DENV. We examined the effectiveness of coupling DNAzyme (DDZ) activation to the salt-induced aggregation of gold nanoparticles (AuNP) to detect dengue virus (DENV) progeny in mosquito cells. A DNAzyme was designed to recognize the 5' cyclization sequence (5' CS) that is conserved among all DENV, and conjugated to AuNPs. DDZ-AuNP has demonstrated the ability to detect the genomic RNA of our model dengue strain, DENV-2 NGC, isolated from infected Aedes albopictus C6/36 cells. These targeting events lead to the rapid aggregation of AuNPs, resulting in a red to clear color transition of the reaction mixes, and thus positive detection of the DENV RNA genome. The inclusion of SDS in the reaction mixture permitted the detection of DENV directly from cell culture supernatants without additional sample processing. Specificity assays demonstrated detection is DENV-specific, while sensitivity assays confirm detection at levels of 1 × 10(1) TCID50 units. These results demonstrate DDZ-AuNP effectively detects DENV genomes in a sequence specific manner and at concentrations that are practical for field use. We have developed an effective detection assay using DNAzyme catalysis coupled with AuNP aggregation for the detection of DENV genomes in a sequence specific manner. Full development of our novel DDZ-AuNP detection method will provide a practical, rapid, and low cost alternative for the detection of DENV in mosquito cells and tissues, and possibly infected patient serum, in a matter of minutes with little to no specialized training required.


Na+/Ca2+ Exchanger a Druggable Target to Promote β-Cell Proliferation and Function.
An important feature of type 2 diabetes is a decrease in β-cell mass. Therefore, it is essential to find new approaches to stimulate β-cell proliferation. We have previously shown that heterozygous inactivation of the Na+/Ca2+ exchanger (isoform 1; NCX1), a protein responsible for Ca2+ extrusion from cells, increases β-cell proliferation, mass, and function in mice. Here, we show that Ncx1 inactivation also increases β-cell proliferation in 2-year-old mice and that NCX1 inhibition in adult mice by four small molecules of the benzoxyphenyl family stimulates β-cell proliferation both in vitro and in vivo. NCX1 inhibition by small interfering RNA or small molecules activates the calcineurin/nuclear factor of activated T cells (NFAT) pathway and inhibits apoptosis induced by the immunosuppressors cyclosporine A (CsA) and tacrolimus in insulin-producing cell. Moreover, NCX1 inhibition increases the expression of β-cell-specific genes, such as Ins1, Ins2, and Pdx1, and inactivates/downregulates the tumor suppressors retinoblastoma protein (pRb) and miR-193a and the cell cycle inhibitor p53. Our data show that Na+/Ca2+ exchange is a druggable target to stimulate β-cell function and proliferation. Specific β-cell inhibition of Na+/Ca2+ exchange by phenoxybenzamyl derivatives may represent an innovative approach to promote β-cell regeneration in diabetes and improve the efficiency of pancreatic islet transplantation for the treatment of the disease.


Allocation of Treatment Responsibility in Adolescents With Epilepsy: Associations With Cognitive Skills and Medication Adherence.
To describe allocation of treatment responsibility (ATR) in adolescents with epilepsy, investigate associations between cognitive skills and ATR, and examine whether ATR for antiepileptic drugs (AEDs) predicted electronically monitored adherence. Sixty adolescents with epilepsy and their caregivers completed the Allocation of Treatment Responsibility Scale and a battery of self-report measures. Medical chart review data and electronically monitored AED adherence were collected for 1 year. Descriptive data assessed ATR for caregivers and adolescents; multivariate hierarchical regressions tested associations between variables. ATR for labs and clinic appointments was greatest for caregivers, while ATR for AEDs was more likely to be shared between caregiver and adolescent. Poorer attention was associated with greater caregiver responsibility for AEDs. Greater caregiver responsibility for AEDs was associated with higher electronically monitored adherence over 12 months. In adolescents with epilepsy, caregivers are responsible for most treatment tasks, although responsibility for taking medication was shared with the adolescent. Greater caregiver responsibility for medication results in better long-term AED adherence. ATR is an important construct that warrants further attention in research and clinical practice, especially in the context of transition and health outcomes in pediatric epilepsy.


Seeking help because of others' drinking.
Many individuals contact and are assisted by community and emergency services because of someone else's drinking. Previous studies have focused on family members accessing services, such as Alcoholics Anonymous due to significant others' drinking; however, little is known about service use in the broad community. This paper aims to estimate the prevalence of contacting the police and seeking help from health services because of others' drinking and to compare the profiles of individuals seeking services with those who did not contact a service. A total of 2649 adult Australians were surveyed about their experience of harm from others' drinking, including use of emergency and community services. In the 12 months prior to the survey, 13% of respondents had contacted the police and 5% had sought help from a health-related service. Using logistic regression, being older and having a secondary school education were associated with a decreased likelihood of contacting police because of others' drinking, whereas residing in a non-metropolitan location was positively associated with using health-related services. Having a partner was negatively associated with use of health-related services. The extent of self-reported harm from others' drinking was the only factor associated with use of both police and health-related services. Results suggest differences in the profile of respondents who call the police and those who seek health-related services due to others' drinking. This supports the need for tailored services to support and address the needs of people experiencing harm from others' drinking.


Oxygen-linked S-nitrosation in fish myoglobins: a cysteine-specific tertiary allosteric effect.
The discovery that cysteine (Cys) S-nitrosation of trout myoglobin (Mb) increases heme O2 affinity has revealed a novel allosteric effect that may promote hypoxia-induced nitric oxide (NO) delivery in the trout heart and improve myocardial efficiency. To better understand this allosteric effect, we investigated the functional effects and structural origin of S-nitrosation in selected fish Mbs differing by content and position of reactive cysteine (Cys) residues. The Mbs from the Atlantic salmon and the yellowfin tuna, containing two and one reactive Cys, respectively, were S-nitrosated in vitro by reaction with Cys-NO to generate Mb-SNO to a similar yield (∼0.50 SH/heme), suggesting reaction at a specific Cys residue. As found for trout, salmon Mb showed a low O2 affinity (P50 = 2.7 torr) that was increased by S-nitrosation (P50 = 1.7 torr), whereas in tuna Mb, O2 affinity (P50 = 0.9 torr) was independent of S-nitrosation. O2 dissociation rates (koff) of trout and salmon Mbs were not altered when Cys were in the SNO or N-ethylmaleimide (NEM) forms, suggesting that S-nitrosation should affect O2 affinity by raising the O2 association rate (kon). Taken together, these results indicate that O2-linked S-nitrosation may occur specifically at Cys107, present in salmon and trout Mb but not in tuna Mb, and that it may relieve protein constraints that limit O2 entry to the heme pocket of the unmodified Mb by a yet unknown mechanism. UV-Vis and resonance Raman spectra of the NEM-derivative of trout Mb (functionally equivalent to Mb-SNO and not photolabile) were identical to those of the unmodified Mb, indicating that S-nitrosation does not affect the extent or nature of heme-ligand stabilization of the fully ligated protein. The importance of S-nitrosation of Mb in vivo is confirmed by the observation that Mb-SNO is present in trout hearts and that its level can be significantly reduced by anoxic conditions.


[Epilepsies with midline projected foci: study with quantitative and topographic EEG (brain mapping)].
We report a topographic and quantitative EEG (EEGQT) study of four patients with analogic EEG diagnostics of midline foci. The new study with EEGQT offered an increased definition of the electrical source with advantages in foci localization. These findings should be confirmed with studies including a greater number of patients.


MAA3 (MAGATAMA3) helicase gene is required for female gametophyte development and pollen tube guidance in Arabidopsis thaliana.
The female gametophyte plays a central role in the sexual reproduction of angiosperms. We previously isolated the maa3 (magatama3) mutant of Arabidopsis thaliana, defective in development of the female gametophyte, micropylar pollen tube guidance, and preventing the attraction of multiple pollen tubes. We here observed that the nucleolus of polar nuclei is small, and that the fusion of polar nuclei often did not occur at the time of pollination. The MAA3 gene encodes a homolog of yeast Sen1 helicase, required for RNA metabolism. It is suggested that MAA3 may regulate RNA molecules responsible for nucleolar organization and pollen tube guidance.


A Comparative Biochemical Study Between L-Carnosine and β-Alanine in Amelioration of Hypobaric Hypoxia-Induced Skeletal Muscle Protein Loss.
Rathor, Richa, Sukanya Srivastava, and Geetha Suryakumar. A comparative biochemical study between L-carnosine and β-alanine in amelioration of hypobaric hypoxia-induced skeletal muscle protein loss. High Alt Med Biol 00:000-000, 2023. Background: Carnosine (CAR; β-alanyl-L-histidine), a biologically active dipeptide is known for its unique pH-buffering capacity, metal chelating activity, and antioxidant and antiglycation property. β-Alanine (ALA) is a nonessential amino acid and used to enhance performance and cognitive functions. Hypobaric hypoxia (HH)-induced muscle protein loss is regulated by multifaceted signaling pathways. The present study investigated the beneficial effects of CAR and ALA against HH-associated muscle loss. Methodology: Simulated HH exposure was performed in an animal decompression chamber. Gastric oral administration of CAR (50 mg·kg-1) and ALA (450 mg·kg-1) were given daily for 3 days and at the end of the treatment, hindlimb skeletal muscle tissue was excised for western blot and biochemical assays. Results: Cosupplementation of CAR and ALA alone was able to ameliorate the hypoxia-induced inflammation, oxidative stress (FOXO), ER stress (GRP-78), and atrophic signaling (MuRF-1) in the skeletal muscles. Creatinine phospho kinase activity and apoptosis were also decreased in CAR- and ALA-supplemented rats. However, CAR showed enhanced protection in HH-induced muscle loss as CAR supplementation was able to enhance protein concentration, body weight, and decreased the protein oxidation and ALA administration was not able to restore the same. Conclusions: Hence, the present comprehensive study supports the fact that CAR (50 mg·kg-1) is more beneficial as compared with ALA (450 mg·kg-1) in ameliorating the hypoxia-induced skeletal muscle loss.


Phylogenomics of the narrowly endemic Eurycheilichthys (Siluriformes: Loricariidae): Sympatric species with non-sister relationships suggest mainly allopatric speciation.
Armored catfishes of the genus Eurycheilichthys are endemic to Southern Brazil and Misiones (Argentina) comprising nine species of small size, with a high degree of sympatry and species diversity distributed in two river basins. Here we use new genome-wide data to infer a species phylogeny and test species boundaries for this poorly known group. We estimate 1) the phylogenetic relationships of the species of Eurycheilichthys based on 29,350 loci in 65 individuals of nine species plus outgroups, and 2) the population structure and differentiation based on 43,712 loci and 62 individuals to estimate how geography may have acted on speciation and formation of the sympatric species groups. Analyses support the monophyly of the genus and suggest two species-inclusive clades (East and West) with high support and very recently diverged species. Western clade contains E. limulus (from upper Jacuí River basin) that is sister to Western species of the Taquari-Antas basin plus E. paucidens. The Eastern clade contains E. pantherinus (from Uruguay River basin) sister to the Eastern species of the Taquari-Antas basin E. coryphaenus, plus the central-distributed species E. planus and E. vacariensis, and the more widely-distributed species E. luisae. Eurycheilichthys luisae is not monophyletic and may contain one or more cryptic species or hybrid individuals. A stronger diversity on structure of lineages on the Taquari-Antas, when compared to upper Uruguay and Jacuí River basins, and the fact that most of the sympatrically distributed taxa have non-sister relationships suggest a scenario of mainly allopatric speciation and may indicate a more dynamic landscape with headwater capture events among these tributaries.


Temporal changes in MMP mRNA expression in the lens epithelium during anterior subcapsular cataract formation.
Transforming growth factor beta (TGFbeta) has been known to play a role in anterior subcapsular cataract (ASC) formation and posterior capsule opacification (PCO), both of which are fibrotic pathologies of the lens. Several models have been utilized to study ASC formation, including the TGFbeta1 transgenic mouse model and the ex-vivo rat lens model. A distinct characteristic of ASC development within these models includes the formation of isolated fibrotic plaques or opacities which form beneath the lens capsule. A hallmark feature of ASC formation is the epithelial to mesenchymal transition (EMT) of lens epithelial cells (LECs) into myofibroblasts. Recently, the matrix metalloproteinases (MMPs) have been implicated in the formation of these cataracts through their involvement in EMT. In the present study, we sought to further investigate the role of MMPs in subcapsular cataract formation in a time course manner, through the examination of gene expression and morphological changes which occur during this process. RT-QPCR and immunohistochemical analysis was carried out on lenses treated with TGFbeta for a period of 2, 4 and 6 days. Laser capture microdissection (LCM) was utilized to specifically isolate cells within the plaque region and cells from the adjacent epithelium in lenses treated for a 6 day period. Multilayering of LECs was observed as early as day 2, which preceded the presence of alpha smooth muscle actin (alpha-SMA) immunoreactivity that was evident following 4 days of treatment with TGFbeta. A slight reduction in E-cadherin mRNA was detected at day 2, although this was not significant until the day 4 time point. Importantly, our results also indicate an early induction of MMP-9 mRNA following 2 days of TGFbeta treatment, whereas MMP-2 was found to be upregulated at the later 4 day time point. Further experiments using FHL 124 cells show an induction in MMP-2 protein levels following treatment with recombinant MMP-9. Together these findings suggest an upstream role for MMP-9 in ASC formation.


Medialization thyroplasty with a customized silicone implant: clinical experience.
The authors implemented medialization thyroplasty with a customized silicone implant in a total of 43 operations (36 patients) in 1999-2003. In 5 of these patients, the medialization thyroplasty was combined with cricothyroid subluxation (3 cases) or adduction of arytenoid cartilage (3 cases). One patient received medialization thyroplasty, cricothyroid subluxation and adduction of arytenoid cartilage. Postoperatively 36 patients reported substantial reduction of their complaints, 5 patients found their voice improved and only 2 patients (5.6%) stated that their voice had not changed. The subjective evaluation was consistent with the findings of laryngoscopy and the preoperative and postoperative phonation parameters (maximum phonation time, maximum sound pressure level, jitter and shimmer). Average maximum phonation time was 6.5 s before surgery and 12.5 s after surgery. Maximum vocal sound pressure level was, on average, about 4 dB higher after surgery. Jitter was reduced from 5.3 to 3.7% and shimmer from 32.3 to 18.6%. The differences between presurgical and postsurgical parameters in our study were all statistically significant, indicating voice improvement. Medialization thyroplasty with a silicone implant was proven to be a successful and safe surgical method for the treatment of vocal fold paralysis.


Depression in nonalcoholic fatty liver disease and all-cause/cause-specific mortality.
Depression has been associated with nonalcoholic fatty liver disease (NAFLD). Data addressing the impact of depression on NAFLD-related mortality are evolving. We aim to study the association of depression in NAFLD and all-cause/cause-specific mortality in the United States. A total of 11,877 individuals with NAFLD in the 2007-2016 National Health and Nutrition Examination Survey with the availability of linked mortality through 2019 were analysed. NAFLD was defined by utilizing the hepatic steatosis index in the absence of known causes of chronic liver disease. Depression and functional impairment due to depression were assessed using the Patient Health Questionnaire. During the median follow-up of 7.6 years, individuals with depression among individuals with NAFLD had a 35% higher all-cause mortality than those without depression (hazard ratio [HR]: 1.35, 95% confidence interval [CI]: 1.03-1.75) after adjusting for demographic, lifestyle and clinical risk factors. NAFLD with functional impairment due to depression had a 62% higher all-cause mortality than NAFLD without functional impairment (HR: 1.62, 95% CI: 1.10-2.39). Depression in NAFLD was associated with an approximately 50% increase in the risk for cardiovascular mortality, with a 2-fold higher cardiovascular mortality in those with functional impairment compared to those without (HR: 2.07, 95% CI: 1.30-3.30). However, there was no significant difference in cancer- and accident-related mortalities in NAFLD with or without depression. Depression among individuals with NAFLD was associated with a higher risk for all-cause and cardiovascular mortality in the United States.


[End-stage cardiac manifestations and autopsy findings in patients with cardiac fabry disease].
Fabry disease is an X-linked recessive disorder resulting from a deficiency of alpha-galactosidase A with multi-organ dysfunction. Patients with manifestations limited to the heart have been reported recently as a disease variant. We have previously reported a 3% prevalence of this cardiac variant in men with left ventricular hypertrophy, which we designated cardiac Fabry disease. The purposes of the current study were to evaluate the end-stage cardiac manifestations and autopsy findings in patients with cardiac Fabry disease. We evaluated five autopsied male patients with cardiac Fabry disease. One died of ventricular fibrillation and four of heart failure. Electrocardiograms obtained at hospitalization revealed the presence of conduction abnormalities and nonsustained ventricular tachycardia. Echocardiograms and autopsy findings showed the presence of left ventricular hypertrophy in all patients. Localized thinning of the basal posterior wall of the left ventricle was detected in four patients who died of heart failure. All patients had severe left ventricular dysfunction. Histologically, myocardial cells showed glycosphingolipid accumulation in all of the patients but no accumulation was observed in other organs or in systemic vascular endothelial cells. Severe left ventricular dysfunction, conduction disturbances and ventricular arrhythmias occur in end-stage cardiac Fabry patients. Furthermore, left ventricular hypertrophy commonly associated with thinning of the base of the left ventricular posterior wall is present. The accumulation of glycosphingolipids can be observed in myocardial cells but not in other organs.


Combined effects of packaging atmosphere and lactic acid on growth and survival of Listeria monocytogenes in crayfish tail meat 4 degrees C.
The effect of lactic acid on growth and survival of Listeria monocytogenes in crayfish tail meat stored under refrigeration and various gas environments was investigated. Frozen crayfish tail meat was thawed overnight, autoclaved, cooled, and inoculated with approximately 4 log colony-forming units (CFU) of a mixed-strain (Scott A and F5027) L. monocytogenes culture per gram of meat. Inoculated samples were blended with 0, 0.5, 1.0, 1.5, or 2.0% lactic acid and packaged under air, vacuum, or modified atmosphere (74.8% CO2, 10.4% O2, and 14.8% N2) and stored at 4 degrees C for 20 days. Results demonstrated that modified atmosphere packaging inhibited the growth of L. monocytogenes more than air and vacuum packaging at 0 and 1% lactic acid. Microbial counts declined steadily in crayfish tail meat treated with 2% lactic acid, with no differences among the packaging atmospheres. The lag phase was extended by 8 days in samples treated with 1% lactic acid and modified atmosphere compared to that in air or vacuum packaging. Overall, the combination of lactic acid and modified atmosphere had the greatest potential to prevent growth of L. monocytogeines.


Chronic pain and pattern of health care utilization among Malaysian elderly population: National Health and Morbidity Survey III (NHMS III, 2006).
The aims of this study were to report prevalence of chronic pain and to examine whether chronic pain influence healthcare usage among elderly Malaysian population. This was a sub-population analysis of the elderly sample in the Malaysia's Third National Health and Morbidity Survey (NHMS III) 2006, a nation-wide population based survey. A subset of 4954 elderly aged 60 years and above was used in the analysis. Chronic pain, pain's interference and outcome variables of healthcare utilization (hospital admission and ambulatory care service) were all measured and determined by self-report. Prevalence of chronic pain among elderly Malaysian was 15.2% (95% CI: 14.5, 16.8). Prevalence of chronic pain increased with advancing age, and the highest prevalence was seen among the old-old group category (21.5%). Across young-old and old-old groups, chronic pain was more prevalent among females, Indian ethnicity, widows/widowers, rural residency and those with no educational background. Our study showed that chronic pain alone increased hospitalization but not visits to ambulatory facilities. Presence of chronic pain was significantly associated with the frequency of hospitalization (aIRR 1.11; 95% CI 1.02, 1.38) but not ambulatory care service. Chronic pain is a prevalent health problem among the elderly in Malaysia and is associated with higher hospitalization rate among the elderly population. This study provides insight into the distribution of chronic pain among the elderly and its relationship with the patterns of healthcare utilization.


Recent key advances in human immunodeficiency virus medicine and implications for China.
In this article we summarize several recent major developments in human immunodeficiency virus treatment, prevention, outcome, and social policy change. Updated international guidelines endorse more aggressive treatment strategies and safer antiretroviral drugs. New antiretroviral options are being tested. Important lessons were learned in the areas of human immunodeficiency virus vaccines and microbicide gels from clinical studies, and additional trials in prevention, especially pre-exposure prophylaxis, are nearing completion. Insight into the role of the virus in the pathogenesis of diseases traditionally thought to be unrelated to acquired immunodeficiency syndrome has become a driving force for earlier and universal therapy. Lastly, we review important achievements of and future challenges facing China as she steps into her eighth year of the National Free Antiretroviral Treatment Program.


The development of the human brain, the closure of the caudal neuropore, and the beginning of secondary neurulation at stage 12.
Twenty-four embryos of stage 12 (26 days) were studied in detail and graphic reconstructions of five of them were prepared. The characteristic features of this stage are 21-29 pairs of somites, incipient or complete closure of the caudal neuropore, and the appearance of upper limb buds. The caudal neuropore closes during stage 12, generally when 25 somitic pairs are present. The site of final closure is at the level of future somite 31, which corresponds to the second sacral vertebral level. Non-closure of the neuropore may be important in the genesis of spina bifida aperta at low levels. The primitive streak probably persists until the caudal neuropore closes, when it is replaced by the caudal eminence or end-bud (Endwulst oder Rumpfknospe). The caudal eminence, which appears at stage 9, gives rise inter alia to hindgut, notochord, caudal somites, and the neural cord. The material for somites 30-34 (which appear in stage 13) is laid down during stage 12, and its absence would be expected to result in sacral agenesis. Aplasia of the caudal eminence results in cloacal deficiency and various degrees of symmelia. The junction of primary and secondary development (primäre und sekundäre Körperentwicklung) is probably at the site of final closure of the caudal neuropore. Secondary neurulation begins during stage 12. The cavity of the already formed spinal cord extends into the neural cord, and isolated spaces are not found within the neural cord. Primary and secondary neurulation are probably coextensive with primary and secondary development of the body, respectively. The telencephalon medium has enlarged, two mesencephalic segments (M1 and M2) are distinguishable, and rhombomere 4 is reduced. The sulcus limitans is detectable in the spinal cord and hindbrain (RhD), and in the mesencephalon and diencephalon, where it extends as far rostrally as the optic sulcus in D1. A marginal layer is appearing in the rhombencephalon and mesencephalon. The first nerve fibres are differentiating, chiefly within the hindbrain (from the nucleus of the lateral longitudinal tract). Optic neural crest is at its maximum, and the otic vesicle is giving crest cells to ganglion 7/8. Neural crest continues to develop in the brain and contributes to cranial ganglia 5, 7/8, and 10/11. The spinal crest extends as far caudally as somites 18-19 but shows no subdivision into ganglia yet. Placodal contribution to the trigeminal ganglion is not certain at stage 12. Such a contribution to ganglion 7/8 is not unlikely.(ABSTRACT TRUNCATED AT 400 WORDS)


GDNF mediates glioblastoma-induced microglia attraction but not astrogliosis.
High-grade gliomas are the most common primary brain tumors. Their malignancy is promoted by the complex crosstalk between different cell types in the central nervous system. Microglia/brain macrophages infiltrate high-grade gliomas and contribute to their progression. To identify factors that mediate the attraction of microglia/macrophages to malignant brain tumors, we established a glioma cell encapsulation model that was applied in vivo. Mouse GL261 glioma cell line and human high-grade glioma cells were seeded into hollow fibers (HF) that allow the passage of soluble molecules but not cells. The glioma cell containing HF were implanted into one brain hemisphere and simultaneously HF with non-transformed fibroblasts (controls) were introduced into the contralateral hemisphere. Implanted mouse and human glioma- but not fibroblast-containing HF attracted microglia and up-regulated immunoreactivity for GFAP, which is a marker of astrogliosis. In this study, we identified GDNF as an important factor for microglial attraction: (1) GL261 and human glioma cells secret GDNF, (2) reduced GDNF production by siRNA in GL261 in mouse glioma cells diminished attraction of microglia, (3) over-expression of GDNF in fibroblasts promoted microglia attraction in our HF assay. In vitro migration assays also showed that GDNF is a strong chemoattractant for microglia. While GDNF release from human or mouse glioma had a profound effect on microglial attraction, the glioma-induced astrogliosis was not affected. Finally, we could show that injection of GL261 mouse glioma cells with GDNF knockdown by shRNA into mouse brains resulted in reduced tumor expansion and improved survival as compared to injection of control cells.


Activity of quinupristin/dalfopristin and its components against Haemophilus influenzae.
Quinupristin/dalfopristin is an injectable streptogramin antibiotic that is constituted in a 30:70 (w/w) ratio of the two components. Quinupristin and dalfopristin are thought to act synergically by binding to two separate sites on the bacterial 50S ribosomal subunit. The in-vitro activities of the two components separately and together in different ratios were determined for a collection of 100 Haemophilus influenzae strains representing various antimicrobial resistance phenotypes. The NCCLS microdilution susceptibility testing procedure incorporating Haemophilus test medium (HTM) broth was used to determine MICs of quinupristin, dalfopristin and seven other antimicrobial agents. The MIC50 and MIC90 values were 4 and 8, 4 and 16, and 64 and 128 mg/L for quinupristin/dalfopristin (30:70), dalfopristin and quinupristin, respectively. MICs of quinupristin and dalfopristin were also determined in Mueller-Hinton lysed horse blood broth and by HTM agar dilution testing. Compared with HTM broth-derived results, the MICs of quinupristin/dalfopristin and its components were the same or one dilution higher in lysed horse blood and HTM agar incubated in air, and were equivalent or one dilution lower in HTM agar incubated in a CO2 atmosphere. The MICs of quinupristin and dalfopristin separately or together were directly proportional to erythromycin MICs, but were otherwise unaffected by any of the resistance mechanisms represented in these strains. MICs of quinupristin and dalfopristin combined in ratios of 10:90, 70:30 and 90:10 did not differ significantly from those of the 30:70 ratio. Thus, unlike the synergic activity noted against Gram-positive bacteria, the activity of quinupristin/dalfopristin against H. influenzae appears to be due almost entirely to the dalfopristin component of the combination.


Follicular GH and IGF1 Levels Are Associated With Oocyte Cohort Quality: A Pilot Study.
Oocyte quality contributes to the development of an optimal embryo and thus a successful pregnancy. The objective of this study was to analyse the association between oocyte cohort quality and the follicular levels of growth hormone (GH), insulin-like growth factor 1 (IGF1), 25-hydroxy vitamin D (25OHD), thyroid-stimulating hormone (TSH), free triiodothyronine (fT3), free thyroxine (fT4) and antithyroid antibodies, as a function of intracytoplasmic sperm injection (ICSI) outcomes. We conducted a prospective comparative pilot study from January 2013 to December 2017. 59 ICSI cycles constituted an abnormal oocyte cohort (n=34 cycles, in which more than 50% of oocytes presented at least one morphological abnormality) and a normal oocyte cohort (n=25 cycles, in which 50% or less of the oocytes presented at least one morphological abnormality). GH, IGF1, 25OHD, TSH, fT3, fT4 and antithyroid antibodies were measured in follicular fluid. The fertilisation rate was lower in the abnormal oocyte cohort (65.5% vs. 80%, respectively, p=0.012). Oocytes' proportion with at least one abnormality was 79.4% in the abnormal oocyte cohort and 29.0% in the normal oocyte cohort. The mean number of morphological abnormalities per oocyte was significantly higher in the abnormal oocyte cohort. The follicular levels of GH (4.98 vs. 2.75 mIU/L, respectively; p <0.01) and IGF1 (72.1 vs. 54.2 ng/mL, respectively; p=0.05) were higher in the normal oocyte cohort. There was no association with follicular levels of TSH, fT3, fT4, antithyroid antibodies, or 25OHD. Oocyte cohort quality appears to be associated with follicular levels of GH and IGF1.


First Global-Scale Synoptic Imaging of Solar Eclipse Effects in the Thermosphere.
A total solar eclipse occurred in the Southern Hemisphere on 2 July 2019 from approximately 17 to 22 UT. Its effect in the thermosphere over South America was imaged from geostationary orbit by NASA's Global-scale Observation of Limb and Disk (GOLD) instrument. GOLD observed a large brightness reduction (>80% around totality) in OI 135.6 nm and N2 LBH band emissions compared to baseline measurements made 2 days prior. In addition, a significant enhancement (with respect to the baseline) in the ΣO/N2 column density ratio (~80%) was observed within the eclipse's totality. This enhancement suggests that the eclipse induced compositional changes in the thermosphere. After the eclipse passed, a slight enhancement in ΣO/N2 column density ratio (~7%) was also seen around the totality path when compared to measurements before the eclipse. These observations are the first synoptic imaging measurements of an eclipse's thermospheric effects with the potential to drastically improve and test our understanding of how the thermosphere responds to rapid, localized changes in solar short wavelength radiation.


Global distribution and genomic characteristics of carbapenemase-producing Escherichia coli among humans, 2005-2023.
Carbapenem-resistant Escherichia coli (CREC) has become a major public health problem worldwide. To date, there is a limited understanding of the global distribution of CREC. In this study, we performed a comprehensive genomic analysis of 7, 731 CRECs of human origin collected from different countries worldwide between 2005 and 2023. Our results showed that these CRECs were distributed in 75 countries, mainly from the United States (17.49%), China (14.88%), and the United Kingdom (14.73%). Eight carbapenemases were identified among the CRECs analyzed, including KPC, IMP, NDM, VIM, OXA, FRI, GES, and IMI. NDM was the most predominant carbapenemase (52.15%), followed by OXA (30.09%) and KPC (14.72%). Notably, all CRECs carried multiple antibiotic resistance genes (ARGs), with 178 isolates carrying mcr-1 and 9 isolates carrying tet(X). The CREC isolates were classified into 465 known sequence types (STs), with ST167 being the most common (11.5%). Correlation analysis demonstrated the significant role of mobile genetic elements in facilitating the transfer of carbapenem resistance genes. Furthermore, some CRECs from different countries showed high genetic similarity, suggesting clonal transmission exists. According to the GWAS results, the genetic difference of blaNDM-positive CRECs from China were mainly enriched in bacterial Type IV secretion system pathways compared with those from the United Kingdom and the United States. Therefore, continuous global surveillance of CRECs is imperative in the future.


Hepatic fat and macrophages are increased in livers of diabetic patients without NAFLD.
Diabetes mellitus (DM), especially type 2, is strongly associated with non-alcoholic fatty liver disease (NAFLD). Recent studies indicate that particularly in DM patients, "simple" liver steatosis can progress into more severe disease. However, little is known about putative hepatic histopathological changes in DM patients without NAFLD. In this study we therefore analysed fat content and inflammatory cell infiltration in the livers of deceased DM and non-DM patients without NAFLD, and analysed age/sex effects hereon. Hepatic fat and inflammatory cells were studied through (immuno)histochemical analysis in liver tissue from 24 DM patients and 66 non-diabetic controls, without histopathological characteristics of NAFLD. We observed a 2-fold increase in fat percentage/mm2 and a near 5-fold increase in the number of fat-containing cells/mm2 in DM patients compared to non-diabetic controls. Fat content was significantly higher in patients with type 2 DM, but not type 1 DM, compared to non-diabetic controls, while the number of CD68+ cells/mm2 was significantly elevated in both DM groups. Hepatic fat and number of macrophages are increased in patients with DM without NAFLD, which may reflect a higher risk on development of steatosis and steatohepatitis.


The management of postoperative nausea and vomiting: current thoughts and protocols.
The postoperative course of surgical patients can have a tremendous impact on the surgical outcome and on patient satisfaction. One of the most significant issues is postoperative nausea and vomiting (PONV) which, despite being a common side effect of general anesthesia, has received little attention in the plastic surgery literature. The incidence and potential consequences of PONV are frequently underestimated and consequently the need for prophylaxis is often overlooked. There are significant consequences to this seemingly minor morbidity that extend beyond patient discomfort and dissatisfaction. In addition to being considered a significant undesirable outcome by patients, severe cases of PONV may result in postoperative complications and unplanned hospital admissions. In this article we overview the mechanism, pathophysiology, and risk factors for PONV and provide a comprehensive algorithmic approach to its management. This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .


Climate change primer for respirologists.
Climate change is already affecting the cardiorespiratory health of populations around the world, and these impacts are expected to increase. The present overview serves as a primer for respirologists who are concerned about how these profound environmental changes may affect their patients. The authors consider recent peer-reviewed literature with a focus on climate interactions with air pollution. They do not discuss in detail cardiorespiratory health effects for which the potential link to climate change is poorly understood. For example, pneumonia and influenza, which affect >500 million people per year, are not addressed, although clear seasonal variation suggests climate-related effects. Additionally, large global health impacts in low-resource countries, including migration precipitated by environmental change, are omitted. The major cardiorespiratory health impacts addressed are due to heat, air pollution and wildfires, shifts in allergens and infectious diseases along with respiratory impacts from flooding. Personal and societal choices about carbon use and fossil energy infrastructure should be informed by their impacts on health, and respirologists can play an important role in this discussion.


Exploration of the epidemiological consequences of resistance to gastro-intestinal parasitism and grazing management of sheep through a mathematical model.
Predicting the impacts of selection for decreased faecal egg count (FEC) (i.e. host resistance) in grazing ruminants is difficult, due to complex interactions between parasite epidemiology, management and host responses. A mathematical model including heritable between lamb variation in host-parasite interactions, Teladorsagia circumcincta epidemiology and anthelmintic drenching, was developed and used to (i) address such interactions and their impact on outcomes including FEC, live weight (LW, kg) and pasture larval contamination (PC, larvae/kg DM), and (ii) investigate how grazing management strategies, aimed at reducing host exposure to infective larvae via pasture moves at 40 day intervals, affect these outcomes. A population of 10,000 lambs was simulated and resultant FEC predictions used to assign the 1,000 lambs with the highest and lowest predicted FEC to 'susceptible' (S) and 'resistant', (R) groups, respectively. The predicted average FEC of the S group was ∼8.5-fold higher than the R group across a grazing season. The R and S groups were then simulated to graze separate pastures (R(sep) and S(sep)); and repeated for 3 grazing seasons to allow predictions to diverge and stabilize. Further, different grazing strategies were superimposed on all groups. PC and average FEC were affected by whether lambs of different resistance genotype grazed together or separately, with differences increasing across grazing seasons. By the third grazing season the average PC of the R(sep) group was reduced by ∼83%, and the S(sep) group was increased by ∼240%, in comparison to the whole population average. Average FEC of the R(sep) group was reduced by ∼40%, and the S(sep) group increased by ∼46% in comparison to the R and S groups, respectively, whilst drenching had little impact on the proportional differences in FEC between groups. Predicted LW was similar for the R and R(sep) groups irrespective of anthelmintic treatment, whilst LW of the S(sep) group was reduced by ∼14% compared to the S group for un-drenched lambs, and by ∼4% for drenched lambs. The differing grazing strategies were predicted to have little impact on FEC or LW, with the exception of the S(sep) group which was predicted to have a 2 kg increase in LW when drenched and moved to a clean pasture. Together, these results suggest that host genotype has a substantial impact on parasite epidemiology, however the benefits of anthelmintic treatment and grazing management should only be expected for susceptible animals. This supports the use of targeted selective treatment, focussing on susceptible animals.


Evaluation of a multiple-antigen enzyme-linked immunosorbent assay for detection of Mycobacterium tuberculosis infection in captive elephants.
Mycobacterium tuberculosis has become an important agent of disease in the captive elephant population of the United States, although current detection methods appear to be inadequate for effective disease management. This investigation sought to validate a multiple-antigen enzyme-linked immunosorbent assay (ELISA) for screening of M. tuberculosis infection in captive elephants and to document the elephant's serologic response over time using a cross-sectional observational study design. Serum samples were collected from 51 Asian elephants (Elephas maximus) and 26 African elephants (Loxodonta africana) from 16 zoos and circuses throughout the United States. Infection status of each animal was determined by mycobacterial culture of trunk washes. Reactivity of each serum sample against six antigens was determined, and the linear combination of antigens that accurately predicted the infection status of the greatest number of animals was determined by discriminant analysis. The resulting classification functions were used to calculate the percentage of animals that were correctly classified (i.e., specificity and sensitivity). Of the 77 elephants sampled, 47 fit the criteria for inclusion in discriminant analysis. Of these, seven Asian elephants were considered infected; 25 Asian elephants and 15 African elephants were considered noninfected. The remaining elephants had been exposed to one or more infected animals. The specificity and sensitivity of the multiple-antigen ELISA were both 100% (91.9-100% and 54.4-100%, respectively) with 95% confidence intervals. Mycobacterium bovis culture filtrate showed the highest individual antigen specificity (95%; 83.0-100%) and sensitivity (100%; 54.4-100%). Serum samples from 34 elephants were analyzed over time by the response to the culture filtrate antigen; four of these elephants were culture positive and had been used to calculate the discriminant function. Limitations such as sample size, compromised ability to ascertain each animal's true infection status, and absence of known-infected African elephants suggest that much additional research needs to be conducted regarding the use of this ELISA. However, the results indicate that this multiple-antigen ELISA would be a valuable screening test for detecting M. tuberculosis infection in elephant herds.


Structure and anticoagulant activity of sulfated fucans. Comparison between the regular, repetitive, and linear fucans from echinoderms with the more heterogeneous and branched polymers from brown algae.
Sulfated fucans are among the most widely studied of all the sulfated polysaccharides of non-mammalian origin that exhibit biological activities in mammalian systems. Examples of these polysaccharides extracted from echinoderms have simple structures, composed of oligosaccharide repeating units within which the residues differ by specific patterns of sulfation among different species. In contrast the algal fucans may have some regular repeating structure but are clearly more heterogeneous when compared with the echinoderm fucans. The structures of the sulfated fucans from brown algae also vary from species to species. We compared the anticoagulant activity of the regular and repetitive fucans from echinoderms with that of the more heterogeneous fucans from three species of brown algae. Our results indicate that different structural features determine not only the anticoagulant potency of the sulfated fucans but also the mechanism by which they exert this activity. Thus, the branched fucans from brown algae are direct inhibitors of thrombin, whereas the linear fucans from echinoderms require the presence of antithrombin or heparin cofactor II for inhibition of thrombin, as reported for mammalian glycosaminoglycans. The linear sulfated fucans from echinoderms have an anticoagulant action resembling that of mammalian dermatan sulfate and a modest action through antithrombin. A single difference of one sulfate ester per tetrasaccharide repeating unit modifies the anticoagulant activity of the polysaccharide markedly. Possibly the spatial arrangements of sulfate esters in the repeating tetrasaccharide unit of the echinoderm fucan mimics the site in dermatan sulfate with high affinity for heparin cofactor II.


[Formation of gamma-aminobutyric acid from glutamate and putrescine in rat hypothalamic and hippocampal synaptosomes and regulation of these processes by glucocorticoids].
It has been shown that single or multiple hydrocortisone and ACTH administrations to intact rats increased GABA content and its synthesis from glutamate and putrescine in synaptosomes of hypothalamus. The letter content was increased by single hormonal administration while multiple hormonal administration and adrenalectomy decreased it. Ornithine decarboxylase activity was increased by single hydrocortisone administration to intact animals, following adrenalectomy, and it was decreased by single hormonal administration to adrenalectomized rats. GABA synthesis in synaptosomes of hippocampus from putrescine was increased by single hydrocortisone and multiple hormonal administrations. GABA content was increased by multiple administration of both hormones and was decreased by adrenalectomy. Putrescine level was decreased by multiple hydrocortisone administration to intact and single administration to adrenalectomized rats; ornithine decarboxylase activity was decreased by multiple administration of both hormones.


[Sequential grafts of cutaneous cells: First results of a new procedure and review of the literature].
A new choice for major burn wounds is presented. An alternative method for skin grafting and cutaneous substitute for bum victims and chronic non healing wounds. The new method of skin grafting is applied repetitively and non-invasively until healing is achieved. PATIENTS AND PROCEDURE: 39 patients including 33 burns victims and six with chronic wounds were treated between February and September 2012. One to four square centimeters of autologous skin graft were transformed with biodegradation of cellular peptic bonds with the goal of obtaining a rich solution of cutaneous cells (1.5 million skin cells/cm2). The process duration averaged 100 minutes. Part of the solution was sprayed immediately on the recipient skin zones. The remaining solution was divided up and kept in cryotubes inside a container of liquid nitrogen. Successive sprays were performed at each dressing. Good esthetic results on the whole as well as functional for patients with deep 2nd degree burns; healing was achieved in 10-12 days. Third degree burns and chronic wounds showed encouraging results for small surfaces. We present a new original technique without prior in vitro culture that gives autologous cellular grafts, including epidermal, junctional and stem cells that are necessary for skin regeneration. The contraindications to the technique are infected, oozing, hemorrhagic or granulomatous wounds. This is a technique that deserves to be practiced and should have its place in the therapeutic arsenal of burns and chronic wounds.


Musculoskeletal complaints in cardiac rehabilitation: Prevalence and impact on cardiovascular risk factor profile and functional and psychosocial status.
To assess the prevalence of musculoskeletal complaints and their association with risk factor profile and functional and psychosocial status in patients on a cardiac rehabilitation program. In this cross-sectional study of 449 patients admitted within three months of an acute coronary syndrome, patients were divided into those with (MSC+) and those without (MSC-) musculoskeletal complaints. The Hospital Anxiety and Depression Scale and the Short Form 36 Health Survey were used to assess psychosocial status and quality of life, and the International Physical Activity Questionnaire for physical activity. Functional capacity was estimated from exercise testing. Musculoskeletal pain was present in 119 patients (27%), mainly in the lower limbs (56%). MSC+ were older (mean 56.5±9.9 vs. 53.2±9.5 years; p<0.001) and more frequently women (20.2% vs. 9.1%; p<0.001). MSC+ had a higher prevalence of dyslipidemia (68.6% vs. 51.2%; p<0.001), hypertension (51.7% vs. 35.5%; p<0.001), obesity (29.4% vs. 17.9%; p<0.001) and metabolic syndrome (44.5% vs. 31.5%; p<0.001). MSC+ showed higher body mass index and waist circumference, and lower physical activity levels (p<0.05), as well as lower functional capacity (8.6±2.2 vs. 9.6±2.1 MET; p<0.05), higher scores for depression (6 [3-9] vs. 3 [1-7]; p<0.05) and anxiety (7 [3-10] vs. 5 [2-8]; p<0.05), and lower scores for physical (44.1±8.7 vs. 47.6±7.6; p<0.05) and mental (39.2±13.0 vs. 44.0±13.0; p<0.05) quality of life. Musculoskeletal complaints are common in cardiac rehabilitation and predict lower levels of physical activity, worse cardiovascular risk factor profile, and poorer functional capacity and psychosocial status, irrespective of age and gender.


[Nocardiosis in 2 immunocompetent patients].
Two diagnosed clinical cases of nocardiosis in immunocompetent patients are presented, different in their clinical aspects, evolution and treatment. We report a 55 years-old man with disseminated nocardiosis and a 32 years-old man with splenectomy diagnosed of pulmonary nocardiosis. The medical literature is reviewed and emphasis is placed on the different illness and treatment of the nocardiosis.


The application of nanoparticles in theranostic systems targeting breast cancer stem cells: current progress and future challenges.
Breast cancer (BC) is one of the diseases with the highest female mortality rates in the world and is closely related to breast cancer stem cells (BCSCs). Conventional breast cancer chemotherapy drugs target noncancer stem cells (non-CSCs), while cancer stem cells (CSCs) can still survive, which is an important reason for breast cancer drug resistance and local recurrence or distant metastasis. How to eradicate BCSCs while killing BCs is the key factor to improve the effect, and it is also an important scientific problem to be solved urgently. Therefore, targeted BCSC therapy has become a research hotspot. Interestingly, the emergence of nanotechnology provides a new idea for targeting BCSCs. This study summarizes the current application status of nanomaterials in targeting BCSCs, and attempts to construct a new type of lipid nanoparticle (LNP) that can target BCSCs through mRNA, providing a new idea for the treatment of BC.


Neuronal polarity: an evolutionary perspective.
Polarized distribution of signaling molecules to axons and dendrites facilitates directional information flow in complex vertebrate nervous systems. The topic we address here is when the key aspects of neuronal polarity evolved. All neurons have a central cell body with thin processes that extend from it to cover long distances, and they also all rely on voltage-gated ion channels to propagate signals along their length. The most familiar neurons, those in vertebrates, have additional cellular features that allow them to send directional signals efficiently. In these neurons, dendrites typically receive signals and axons send signals. It has been suggested that many of the distinct features of axons and dendrites, including the axon initial segment, are found only in vertebrates. However, it is now becoming clear that two key cytoskeletal features that underlie polarized sorting, a specialized region at the base of the axon and polarized microtubules, are found in invertebrate neurons as well. It thus seems likely that all bilaterians generate axons and dendrites in the same way. As a next step, it will be extremely interesting to determine whether the nerve nets of cnidarians and ctenophores also contain polarized neurons with true axons and dendrites, or whether polarity evolved in concert with the more centralized nervous systems found in bilaterians.


Baseline periodontal status and modifiable risk factors are associated with tooth loss over a 10-year period: Estimates of population attributable risk in a Japanese community.
This study aimed to examine whether modifiable risk factors can predict tooth loss over 10 years and estimate population attributable risk (PAR) for a combination of modifiable factors. This longitudinal study included 1466 participants who underwent dental examinations in 2007 and 2017 and were aged 40 to 79 years at baseline. Periodontal conditions were assessed using the 2018 periodontal classification. Incident tooth loss was defined as ≥4 teeth lost over a 10-year period. We calculated the partial PAR (pPAR%) for tooth loss to estimate the combined effect of modifiable risk factors. Incidence of tooth loss was 17.5%. Directed acyclic graphs were used to identify risk factors for tooth loss. A logistic regression model showed that baseline periodontitis, dental caries experience, no regular dental visit, periodontal treatment, smoking, and obesity were associated with tooth loss after adjusting for covariates; pPAR% was 55.5% (95% confidence interval: 31.1% to 73.0%) in periodontitis Stage III to IV and 87.6% (50.4% to 97.4%) in the combination of all factors, respectively. The sex-stratified analysis showed that smoking and no regular dental visit in men and obesity in women were identified as potential risk factors for tooth loss. Modifiable factors accounted for most cases of incident tooth loss. Risk factors for tooth loss might differ by sex, suggesting that the appropriate approach for preventing tooth loss base on sex.


Optimal savings and the value of population.
We study a model of economic growth in which an exogenously changing population enters in the objective function under total utilitarianism and into the state dynamics as the labor input to the production function. We consider an arbitrary population growth until it reaches a critical level (resp. saturation level) at which point it starts growing exponentially (resp. it stops growing altogether). This requires population as well as capital as state variables. By letting the population variable serve as the surrogate of time, we are still able to depict the optimal path and its convergence to the long-run equilibrium on a two-dimensional phase diagram. The phase diagram consists of a transient curve that reaches the classical curve associated with a positive exponential growth at the time the population reaches the critical level. In the case of an asymptotic population saturation, we expect the transient curve to approach the equilibrium as the population approaches its saturation level. Finally, we characterize the approaches to the classical curve and to the equilibrium.


Influence of bone marrow fat embolism on coagulation activation in an ovine model of vertebroplasty.
Intraoperative cardiovascular deterioration as a result of pulmonary embolization of bone marrow fat is a potentially serious complication during vertebroplasty. The release of fatty material and thromboplastin from the bone marrow cavity during vertebroplasty may activate the coagulation cascade resulting in thrombogenesis, and pharmacological prophylaxis may therefore prevent cardiovascular complications. Thus, the effects of bone marrow fat embolism on coagulation activation during vertebroplasty were investigated with use of an animal model. Polymethylmethacrylate was injected into three lumbar vertebrae of six sheep in order to force bone marrow fat into the circulation. Invasive blood pressures and heart rate were recorded continuously until sixty minutes after the last injection. Cardiac output, arterial and mixed venous blood gas parameters, and coagulation parameters were measured at selected time-points. Postmortem lung biopsy specimens were assessed for the presence of intravascular fat. Embolization of bone marrow fat resulted in a sudden and dramatic increase in mean pulmonary arterial pressure and a decrease in mean arterial blood pressure. There were no significant changes in any coagulation parameter from before the injection to after the injection. Intravascular fat and bone marrow cells were present in all lung lobes. Injection of polymethylmethacrylate into vertebral bodies caused embolization of bone marrow fat with subsequent transient cardiovascular deterioration, but no changes in coagulation parameters were observed. Thromboembolism did not contribute to the observed cardiovascular changes.


Modeling trajectory of depressive symptoms among psychiatric inpatients: a latent growth curve approach.
Changes in the parameters of inpatient psychiatric care have inspired a sizable literature exploring correlates of prolonged intervention as well as symptom change over varying lengths of hospitalization. However, existing data offer limited insight regarding the nature of symptom change over time. Objectives of this longitudinal research were to (1) model the trajectory of depressive symptoms within an inpatient psychiatric sample, (2) identify characteristics associated with unique patterns of change, and (3) evaluate the magnitude of expected gains using objective clinical benchmarks. Participants included 1,084 psychiatric inpatients treated between April 2008 and December 2010. Latent growth curve modeling was used to determine the trajectory of Beck Depression Inventory II depressive symptoms in response to treatment. Age, gender, trauma history, prior hospitalization, and DSM-IV diagnoses were examined as potential moderators of recovery. Results indicate a nonlinear model of recovery, with symptom reductions greatest following admission and slowing gradually over time. Female gender, probable trauma exposure, prior psychiatric hospitalization, and primary depressive diagnosis were associated with more severe trajectories. Diagnosis of alcohol/substance use, by contrast, was associated with more moderate trajectories. Objective benchmarks occurred relatively consistently across patient groups, with clinically significant change occurring between 2-4 weeks after admission. The nonlinear trajectory of recovery observed in these data provides insight regarding the dynamics of inpatient recovery. Across all patient groups, symptom reduction was most dramatic in the initial week of hospitalization. However, notable improvement continued for several weeks after admission. Results suggest that timelines for adequate inpatient care are largely contingent on program-specific goals.


Angle Kappa Influence on Multifocal IOL Outcomes.
To characterize angle kappa and study the relationship between preoperative angle kappa and postoperative refractive accuracy, visual outcomes, and patient satisfaction in a large population of eyes with multifocal intraocular lens (MIOL) implantation. A comprehensive electronic medical record chart review of 26,470 consecutive eyes that underwent immediate sequential bilateral cataract or refractive lens exchange with MIOLs was conducted. The primary outcome measures were postoperative monocular uncorrected distance visual acuity (UDVA), manifest refraction sphere and cylinder, spherical equivalent (SEQ), defocus equivalent (DEQ), subjective quality of vision at near, intermediate, and distance, and the likelihood of recommending the procedure. Relationships between preoperative angle kappa and postoperative outcomes were assessed with Pearson correlations. Angle kappa followed a right-skewed normal distribution (R2 = 0.99) with a mean ± standard deviation of 0.64 ± 0.27 mm. No clinically meaningful relationship was found between preoperative angle kappa and postoperative sphere, cylinder, SEQ, and DEQ, all with R2 ⩽ 0.0005. Similarly, there was no clinically meaningful relationship between preoperative angle kappa and postoperative UDVA (R2 = 0.001), postoperative satisfaction for near, intermediate, and distance vision (all R2 ⩽ 0.0023), or for recommending the MIOL surgery to friends and relatives (R2 = 0.0000). Preoperative angle kappa does not have a predictive clinical impact on postoperative MIOL visual outcomes, refractive accuracy, or subjective patient satisfaction. Angle kappa as a single variable cannot be used to determine MIOL candidacy. [J Refract Surg. 2023;39(12):840-849.].


An Explainable MRI-Radiomic Quantum Neural Network to Differentiate Between Large Brain Metastases and High-Grade Glioma Using Quantum Annealing for Feature Selection.
Solitary large brain metastases (LBM) and high-grade gliomas (HGG) are sometimes hard to differentiate on MRI. The management differs significantly between these two entities, and non-invasive methods that help differentiate between them are eagerly needed to avoid potentially morbid biopsies and surgical procedures. We explore herein the performance and interpretability of an MRI-radiomics variational quantum neural network (QNN) using a quantum-annealing mutual-information (MI) feature selection approach. We retrospectively included 423 patients with HGG and LBM (> 2 cm) who had a contrast-enhanced T1-weighted (CE-T1) MRI between 2012 and 2019. After exclusion, 72 HGG and 129 LBM were kept. Tumors were manually segmented, and a 5-mm peri-tumoral ring was created. MRI images were pre-processed, and 1813 radiomic features were extracted. A set of best features based on MI was selected. MI and conditional-MI were embedded into a quadratic unconstrained binary optimization (QUBO) formulation that was mapped to an Ising-model and submitted to D'Wave's quantum annealer to solve for the best combination of 10 features. The 10 selected features were embedded into a 2-qubits QNN using PennyLane library. The model was evaluated for balanced-accuracy (bACC) and area under the receiver operating characteristic curve (ROC-AUC) on the test set. The model performance was benchmarked against two classical models: dense neural networks (DNN) and extreme gradient boosting (XGB). Shapley values were calculated to interpret sample-wise predictions on the test set. The best 10-feature combination included 6 tumor and 4 ring features. For QNN, DNN, and XGB, respectively, training ROC-AUC was 0.86, 0.95, and 0.94; test ROC-AUC was 0.76, 0.75, and 0.79; and test bACC was 0.74, 0.73, and 0.72. The two most influential features were tumor Laplacian-of-Gaussian-GLRLM-Entropy and sphericity. We developed an accurate interpretable QNN model with quantum-informed feature selection to differentiate between LBM and HGG on CE-T1 brain MRI. The model performance is comparable to state-of-the-art classical models.


A Combination of Apple Vinegar Drink with Bacillus coagulans Ameliorates High Fat Diet-Induced Body Weight Gain, Insulin Resistance and Hepatic Steatosis.
Obesity is a worldwide epidemic characterized by excessive fat accumulation, associated with multiple comorbidities and complications. Emerging evidence points to gut microbiome as a driving force in the pathogenesis of obesity. Vinegar intake, a traditional remedy source of exogenous acetate, has been shown to improve glycemic control and to have anti-obesity effects. New functional foods may be developed by supplementing traditional food with probiotics. B. coagulans is a suitable choice because of its resistance to high temperatures. To analyze the possible synergic effect of Vinegar and B. coagulans against the metabolic alterations induced by a high fat diet (HFD), we fed twelve-week-old C57BL/6 mice with HFD for 5 weeks after 2 weeks of acclimation on a normal diet. Then, food intake, body weight, blood biochemical parameters, histology and liver inflammatory markers were analyzed. Although vinegar drink, either alone or supplemented with B. coagulans, reduced food intake, attenuated body weight gain and enhanced glucose tolerance, only the supplemented drink improved the lipid serum profile and prevented hepatic HFD-induced overexpression of CD36, IL-1β, IL-6, LXR and SREBP, thus reducing lipid deposition in the liver. The beneficial properties of the B. coagulans-supplemented vinegar appear to be mediated by a reduction in insulin and leptin circulating levels.


Microelectrode recording (MER) findings during sleep-awake anesthesia using dexmedetomidine in deep brain stimulation surgery for Parkinson's disease.
The preferred choice of anesthesia for deep brain stimulation (DBS) has been local anesthesia due to the need of patients' cooperation during the procedure, and concern on the interference of sedatives on microelectrode recording (MER) results. However, local anesthesia during the whole procedure may be impossible in some patients due to uncontrolled anxiety, fear, delirium or exhaustion. Therefore, sedative drugs have been used for DBS, but findings of MER during the procedures have not been reported in detail, especially in the globus pallidus internus (GPi). We introduce our experience using 'asleep-awake' technique by dexmedetomidine (DEX) anesthesia with MER findings during DBS in idiopathic Parkinson's disease (IPD) patients. Data from 14 different subcortical nuclei from 8 consecutive IPD patients whom had DBS at the GPi (6 patients) and subthalamic nucleus (STN) (2 patients) were retrospectively reviewed. We used continuous DEX and intermittent small boluses of propofol during the painful procedure ('asleep phase'), accompanied with continuous intraoperative monitorings of bispectral index (BIS) and modified observer's assessment of sedation (MOAA/S). Then sedatives were discontinued during MER recording ('awake phase'). Characteristic findings and firing rates of neurons were analyzed and compared to those from other 6 patients who underwent surgery under local anesthesia. All patients were satisfactorily sedated using this technique without any respiratory or hemodynamic complications. Characteristics of spike activities of each nucleus were inspected and analyzed quantitatively. We could inspect changes of spike activities according to level of patients' consciousness in some cases, but the localizing value was good to decide the target in all cases. Firing rates of group whom sedatives were given during asleep phase ('sedatives') were significantly lower than those of group under local anesthesia ('no sedative'). Intraoperative length of target nuclei, postoperative imaging and postoperative changes of UPDRS III score indicated satisfactory outcome. We concluded that though MER findings may change during DEX-based monitored 'sleep-awake' anesthesia, it did not affect the results of target localization for the clinical purpose. However, it should be considered that use of sedatives before MER could result in changes of firing rate and pattern depending on the patient's state of consciousness.


Specific environmental release categories--A tool for improving chemical safety assessment in the EC--report of a multi-stakeholder workshop.
In April 2011, experts from industry and authorities met for a workshop to discuss experience and future developments regarding the use of specific environmental release categories (SPERCs) in chemicals safety assessment (CSA) under the European Chemicals Regulation Registration, Evaluation and Authorization of Chemicals (REACH). This article provides a summary of the workshop. It briefly explains what a SPERC is, why SPERCs are needed, where the challenges of the concept are, and what improvements are needed to make SPERCs a useful tool for assessments under REACH.


Immunity to canine adenovirus respiratory disease: a comparison of attenuated CAV-1 and CAV-2 vaccines.
Four litters of puppies were divided into three groups. One group was vaccinated with a live CAV-1 vaccine and another with a live CAV-2 vaccine. Throat swabs were collected from two dogs in each of these groups to monitor the possible excretion of vaccine virus, but none was found. Both groups, together with the third group of unvaccinated controls, were challenged 17 days later with an aerosol of virulent CAV-2. One dog from each group was killed on the third, fourth, seventh, ninth, 11th and 14th days after challenge. The unvaccinated dogs developed a clinical disease characterised by anorexia, dullness, coughing and tachypnoea. The lungs were consolidated and histological examination revealed the main lesion to be a severe necrotising bronchiolitis. Large amounts of virus were present in the respiratory tissues of these dogs and high titres of virus were isolated from throat swabs. In contrast, both groups of vaccinated dogs remained clinically almost normal with minimal lesions, present for a much shorter period of time. Virus was found on day 4 in the respiratory tissues of one dog vaccinated with CAV-1 but the other vaccinated animals contained little or no virus. In general, the degree of protection afforded by CAV-1 vaccine seemed similar to that provided by CAV-2 vaccine.


Emergency Medicine Resident Orientation: How Training Programs Get Their Residents Started.
The first formal orientation program for incoming emergency medicine (EM) residents was started in 1976. The last attempt to describe the nature of orientation programs was by Brillman in 1995. Now almost all residencies offer orientation to incoming residents, but little is known about the curricular content or structure of these programs. The purpose of this project was to describe the current composition and purpose of EM resident orientation programs in the United States. In autumn of 2014, we surveyed all U.S. EM residency program directors (n=167). We adapted our survey instrument from one used by Brillman (1995). The survey was designed to assess the orientation program's purpose, structure, content, and teaching methods. The survey return rate was 63% (105 of 167). Most respondents (77%) directed three-year residencies, and all but one program offered intern orientation. Orientations lasted an average of nine clinical (Std. Dev.=7.3) and 13 non-clinical days (Std. Dev.=9.3). The prototypical breakdown of program activities was 27% lectures, 23% clinical work, 16% skills training, 10% administrative activities, 9% socialization and 15% other activities. Most orientations included activities to promote socialization among interns (98%) and with other members of the department (91%). Many programs (87%) included special certification courses (ACLS, ATLS, PALS, NRP). Course content included the following: use of electronic medical records (90%), physician wellness (75%), and chief complaint-based lectures (72%). Procedural skill sessions covered ultrasound (94%), airway management (91%), vascular access (90%), wound management (77%), splinting (67%), and trauma skills (62%). Compared to Brillman (1995), we found that more programs (99%) are offering formal orientation and allocating more time to them. Lectures remain the most common educational activity. We found increases in the use of skills labs and specialty certifications. We also observed increases in time dedicated to clinical work during orientation. Only a few programs reported engaging in baseline or milestone assessments, an activity that could offer significant benefits to the residency program.


Left ventricular dyssynchrony from right ventricular pacing depends on intraventricular conduction pattern in intrinsic rhythm.
Right ventricular pacing (RVP) prolongs ventricular activation and may induce mechanical dyssynchrony. We hypothesized that the severity of RVP-associated ventricular mechanical dyssynchrony may depend on the intrinsic intraventricular conduction pattern. Sixty-five patients with a single- or dual-chamber RV pacemaker were included. Forty-seven patients with ejection fraction (EF) <or= 35%, 17 with no bundle branch block (BBB), 16 with right bundle branch block (RBBB), and 14 with left bundle branch block (LBBB). Eighteen patients with EF > 35% and no BBB served as a control group. Echocardiographic dyssynchrony parameters [aortic pre-ejection delay (AoPEP), interventricular mechanical delay, delayed posterior left ventricular wall motion, and septal-to-posterior wall motion delay (SPWMD)] were evaluated in all patients with and without RVP. No dyssynchrony was found in patients with no BBB, RBBB, and in the control group, whereas LBBB patients showed significant dyssynchrony in AoPEP and SPWMD. RVP had a significant negative impact on all dyssynchrony parameters in patients with no BBB or LBBB. RVP induced significantly less severe dyssynchrony in RBBB patients. With RVP 100, 94, 56 and 16% of patients with LBBB, without BBB, RBBB, and from the control group, respectively, fulfilled the CARE-HF criteria for ventricular dyssynchrony. RVP worsens mechanical ventricular dyssynchrony in patients with reduced EF. These effects are most pronounced in patients with either normal QRS width or LBBB during intrinsic rhythm. In contrast, patients with an RBBB during intrinsic rhythm without RVP evidenced a better preserved haemodynamic function and mechanical synchrony with RVP, despite a comparable extent of pacing-induced QRS prolongation.


The myocardial and neuronal infectivity of SARS-CoV-2 and detrimental outcomes.
The epidemiological outbreak of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), alias COVID-19, began in Wuhan, Hubei, China, in late December and eventually turned into a pandemic that has led to over 3.71 million deaths and over 173 million infected cases worldwide. In addition to respiratory manifestations, COVID-19 patients with neurological and myocardial dysfunctions exhibit a higher risk of in-hospital mortality. The immune function tends to be affected by cardiovascular risk factors and is thus indirectly related to the prognosis of COVID-19 patients. Many neurological symptoms and manifestations have been reported in COVID-19 patients; however, detailed descriptions on the prevalence and characteristic features of these symptoms are restricted due to insufficient data. It is thus advisable for clinicians to be vigilant for both cardiovascular and neurological manifestations to detect them at an early stage to avoid inappropriate management of COVID-19 and to address the manifestations adequately. Patients with severe COVID-19 are notably more susceptible to developing cardiovascular and neurological complications than non-severe COVID-19 patients. This review focuses on the consequential outcomes of COVID-19 on cardiovascular and neuronal functions, including other influencing factors.


A community pharmacist delivered adherence support service for dyslipidaemia.
Dyslipidaemia is a major risk factor for coronary heart disease. A key issue in management is adherence to therapy. Pharmacists are ideally situated to provide a service to increase medication adherence and optimize health outcomes. This study aimed to evaluate the impact of a community pharmacist-delivered adherence support service on patients' adherence and total cholesterol levels. A repeated measures [baseline (t = 1), post-intervention at 3-monthly intervals (t = 2,3,4)], randomized-controlled study in community pharmacies, with patients on chronic lipid-lowering therapy was conducted. Measures used were Brief Medication Questionnaire, Medication Adherence Report Scale, total cholesterol and pharmacist intervention data sheets. A baseline t-test of cholesterol levels between groups was conducted followed by a repeated-measures analysis of variance. Seventeen pharmacists recruited 142 patients (97 completions: 49 control, 48 intervention). Most patients missed either the third or last visit, thus data at t = 3 and 4 were combined. Although the intervention group started with a higher cholesterol level (5.1 vs. 4.8 mmol/l), a difference was observed in the drop in cholesterol levels of the intervention group between t = 2 and 3 (F(1,95) = 8.85, P < 0.01), and between the two groups over the study period (F(2,190) = 4.89, P < 0.05). No changes in medicine adherence scores were observed though there was an improvement in participants' exercise and eating habits. Patients significantly lowered their cholesterol levels probably as a result of the service delivered by their pharmacists within the short study time frame of ~9 months.


Genetic and cellular analysis of male germ cell development.
The evolution of the germline has been studied for many decades. Although the pathway of germ cell primordial migration and the kinetic evolution of the gonocytes are well known, their genetic and cellular controls are poorly understood. Recently, a genetic approach using gene knockout and a cellular investigation using several germ cell culture models has allowed a better understanding of the involvement of several genes and factors in the development of germ cells during fetal and neonatal life. Because of the obvious importance of the development of primordial germ cells and gonocytes in adult fertility and its eventual alteration by toxins or xenobiotics, a better knowledge of these interactions appears as an important research area.


Long-term effect of shoyu (Japanese soy sauce) on the gastric mucosa of the rat.
The long-term effect of shoyu (Japanese soy sauce) on the gastric mucosa was assessed in intact rats and in those with a fundusectomy. One hundred female 4-month-old Wistar rats were divided into 4 groups:intact and operated rats on a control diet and intact and operated rats on the same diet saturated with shoyu. The animals given shoyu were smaller than the controls; 15 intact rats receiving the sauce were healthy at 33 months compared with 7 controls. Only 9 of 50 operated rats lived 33 months. Breast tumors developed in 10 control rats but in none given shoyu. The distribution of other tumors was the same. Two malignant neoplasms appeared to originate in the perigastric tissues of operated rats: a reticulum cell sarcoma in the shoyu group and a lymphosarcoma in the control group. None involved the gastric mucosa. Most of the operated rats (with or without shoyu) developed small benign gastric tumors. Mucosal histology was well preserved in 22 intact rats on shoyu: In 15 it was normal, in 7 there was mucus loss and nuclear changes in the cells of the gastric pits; 2 of the 7 had mild gastritis. One intact control showed patchy mucus loss. Shoyu did not appear to be a carcinogen in the rat; its prolonged use impaired neither health nor longevity. Long-term use was associated with gastric mucus loss and nuclear changes in a few rats, occasionally with a mild gastritis.


N,N'-Bis(2-amino-benz-yl)ethane-1,2-diaminium dinitrate.
In the title salt, C16H24N4 (2+)·2NO3 (-), both the cation and anion are placed in general positions, although the cation displays non-crystallographic inversion symmetry, with the aliphatic chain extended in an all-trans conformation. The benzene rings are almost parallel, with a dihedral angle between their mean planes of 3.3 (6)°. The nitrate ions are placed in the vicinity of the protonated amine groups, forming efficient N-H⋯O inter-ion hydrogen bonds. Each nitrate ion in the asymmetric unit bridges two symmetry-related cations, forming an R 4 (4)(18) ring, a common motif in organic ammonium nitrate salts. This results in the formation of chains along [010] with alternating cations and anions. The neutral amine groups are involved in slightly weaker N-H⋯O hydrogen bonds with the nitrate O atoms, and there are also a number of C-H⋯O hydrogen bonds present. The resulting supra-molecular structure is based on a two-dimensional network extending in the ab plane.


Urolithin A, a Novel Natural Compound to Target PI3K/AKT/mTOR Pathway in Pancreatic Cancer.
Pancreatic ductal adenocarcinoma (PDAC) is an aggressive malignancy and is highly resistant to standard treatment regimens. Targeted therapies against KRAS, a mutation present in an overwhelming majority of PDAC cases, have been largely ineffective. However, inhibition of downstream components in the KRAS signaling cascade provides promising therapeutic targets in the management of PDAC and warrants further exploration. Here, we investigated Urolithin A (Uro A), a novel natural compound derived from pomegranates, which targets numerous kinases downstream of KRAS, in particular the PI3K/AKT/mTOR signaling pathways. We showed that treatment of PDAC cells with Uro A blocked the phosphorylation of AKT and p70S6K in vitro, successfully inhibited the growth of tumor xenografts, and increased overall survival of Ptf1aCre/+;LSL-KrasG12D/+;Tgfbr2flox/flox (PKT) mice compared with vehicle or gemcitabine therapy alone. Histologic evaluation of these Uro A-treated tumor samples confirmed mechanistic actions of Uro A via decreased phosphorylation of AKT and p70S6K, reduced proliferation, and increased cellular apoptosis in both xenograft and PKT mouse models. In addition, Uro A treatment reprogrammed the tumor microenvironment, as evidenced by reduced levels of infiltrating immunosuppressive cell populations such as myeloid-derived suppressor cells, tumor-associated macrophages, and regulatory T cells. Overall, this work provides convincing preclinical evidence for the utility of Uro A as a therapeutic agent in PDAC through suppression of the PI3K/AKT/mTOR pathway.


cis-Thioindigo (TI) - a new ligand with accessible radical anion and dianion states. Strong magnetic coupling in the {[TI-(μ2-O),(μ-O)]Cp*Cr}2 dimers.
Reaction of decamethylchromocene (Cp*2Cr) with thioindigo (TI) yields a coordination complex {[TI-(μ2-O), (μ-O)]Cp*Cr}2·C6H14 (1) in which one Cp* ligand in Cp*2Cr is substituted by TI. TI adopts cis-conformation in 1 allowing the coordination of both carbonyl groups to chromium. Additionally, one oxygen atom of TI becomes a μ2-bridge for two chromium atoms to form {[TI-(μ2-O), (μ-O)]Cp*Cr}2 dimers with a CrCr distance of 3.12 Å. According to magnetic data, diamagnetic TI2- dianions and two Cr3+ atoms with a high S = 3/2 spin state are present in a dimer allowing strong antiferromagnetic coupling between two Cr3+ spins with an exchange interaction of -35.4 K and the decrease of molar magnetic susceptibility below 140 K. Paramagnetic TI˙- radical anions with the S = 1/2 spin state have also been obtained and studied in crystalline {cryptand[2,2,2](Na+)}(TI˙-) (2) salt showing that both radical anion and dianion states are accessible for TI.


Microwave-assisted aqueous two-phase extraction of polysaccharides from Hippophae rhamnoide L.: Modeling, characterization and hypoglycemic activity.
Natural polysaccharides are concerned for their high biological activity and low toxicity. Two kinds of polysaccharides were extracted from Hippophae rhamnoide L. by microwave-assisted aqueous two-phase system. Under the optimal conditions predicted by RSM model (K2HPO4/ethanol (18.93 %/28.29 %), liquid to material ratio 77 mL/g, power 625 W and temperature 61 °C), the yield of total polysaccharides reached 35.91 ± 0.76 %. Moreover, the polysaccharides extraction was well fitted to the Weibull model. After purification by Sepharose-6B, the polysaccharides from top phase (PHTP, purity of 81.44 ± 1.25 %) and bottom phase (PHBP, purity of 88.85 ± 1.40 %) were obtained. GC, FT-IR, methylation and NMR analyses confirmed the backbone of PHTP was composed of a repeated unit →4)-β-D-Glcp-(1 → 2)-α-L-Rhap-(1 → 4)-β-D-Galp-(1 → 4)-α-D-GalAp-(1 → 3)-α-L-Araf-(1 → 3)-α-D-Manp-(1→, while the repeated unit in PHBP was →3)-α-L-Araf-(1 → 2)-α-L-Rhap-(1 → 4)-β-D-Glcp-(1 → 3)-α-D-Manp-(1 → 4)-β-D-Galp-(1 → 4)-α-D-GalAp-(1→. Compared with PHTP (6.46 × 106 g/mol), PHBP with relatively low molecular weight (8.2 × 105 g/mol) exhibited the smaller particle size, better water-solubility, thermal and rheological property, stronger anti-glycosylation and α-amylase inhibitory effects. Moreover, PHTP and PHBP displayed a reversible inhibition on α-amylase in a competitive manner. This study provides a high-efficient and eco-friendly method for polysaccharides extraction, and lays a foundation for sea buckthorn polysaccharides as potential therapeutic agents in preventing and ameliorating diabetes.


Strain-Induced Metastable Phase Stabilization in Ga2O3 Thin Films.
It is well known that metastable and transient structures in bulk can be stabilized in thin films via epitaxial strain (heteroepitaxy) and appropriate growth conditions that are often far from equilibrium. However, the mechanism of heteroepitaxy, particularly how the nominally unstable or metastable phase gets stabilized, remains largely unclear. This is especially intriguing for thin-film Ga2O3, where multiple crystal phases may exist under varied growth conditions with spatial and dimensional constraints. Herein, the development and distribution of epitaxial strain at the Ga2O3/Al2O3 film-substrate interfaces is revealed down to the atomic resolution along different orientations, with an aberration-corrected scanning transmission electron microscope. Just a few layers of metastable α-Ga2O3 structure were found to accommodate the misfit strain in direct contact with the substrate. Following an epitaxial α-Ga2O3 structure of about couple unit cells, several layers (4-5) of transient phase appear as the intermediate structure to release the misfit strain. Subsequent to this transient crystal phase, the nominally unstable κ-Ga2O3 phase is stabilized as the major thin-film phase form. We show that the epitaxial strain is gracefully accommodated by rearrangement of the oxygen polyhedra. When the structure is under large compressive strain, Ga3+ ions occupy only the oxygen octahedral sites to form a dense structure. With gradual release of the compressive strain, more and more Ga3+ ions occupy the oxygen tetrahedral sites, leading to volumetric expansion and the phase transformation. The structure of the transition phase is identified by high-resolution electron microscopy observation, complemented by the density functional theory calculations. This study provides insights from the atomic scale and their implications for the design of functional thin-film materials using epitaxial engineering.


Sensitive biochemical aggregate detection reveals aggregation onset before symptom development in cellular and murine models of Huntington's disease.
A CAG-repeat gene expansion translated into a pathogenic polyglutamine stretch at the N-terminus of huntingtin triggers Huntington's Disease. Mutated huntingtin is predicted to adopt toxic properties mainly if aggregation-prone N-terminal fragments are released by proteolysis. Huntingtin-aggregates are indeed a major hallmark of this disorder and could represent useful markers of disease-onset or progression. We designed a simple method for qualitative and quantitative characterization of aggregates. For this, we analyzed samples from in vitro and in vivo Huntington's Disease models by agarose gel electrophoresis and showed that in the brain of transgenic mice huntingtin-aggregates became larger as a function of disease progression. This appears to be a property of cytoplasmic but not nuclear aggregates. In cell cultures, treatment with Congo Red inhibited aggregate growth but not total load. Finally, we showed that in primary striatal neurons and in brains of R6/2 and HdhQ150 mice, the presence of aggregates preceded initiation of any other functional deficits. This observation argues for a pathogenic role of huntingtin-aggregation in Huntington's Disease. Our results emphasize that thorough analysis of huntingtin metabolism and aggregation is now feasible, thus significantly improving the power of studies assessing therapies designed to lower huntingtin levels or to interfere with its aggregation.


Leveraging Molecular Properties to Tailor Mixed-Dimensional Heterostructures beyond Energy Level Alignment.
The surface sensitivity and lack of dielectric screening in two-dimensional (2D) materials provide numerous intriguing opportunities to tailor their properties using adsorbed π-electron organic molecules. These organic-2D mixed-dimensional heterojunctions are often considered solely in terms of their energy level alignment, i.e., the relative energies of the frontier molecular orbitals versus the 2D material conduction and valence band edges. While this simple model is frequently adequate to describe doping and photoinduced charge transfer, the tools of molecular chemistry enable additional manipulation of properties in organic-2D heterojunctions that are not accessible in other solid-state systems. Fully exploiting these possibilities requires consideration of the details of the organic adlayer beyond its energy level alignment, including hybridization and electrostatics, molecular orientation and thin-film morphology, nonfrontier orbitals and defects, excitonic states, spin, and chirality. This Perspective explores how these relatively overlooked molecular properties offer unique opportunities for tuning optical and electronic characteristics, thereby guiding the rational design of organic-2D mixed-dimensional heterojunctions with emergent properties.


Atrial fibrillation classification based on convolutional neural networks.
The global age-adjusted mortality rate related to atrial fibrillation (AF) registered a rapid growth in the last four decades, i.e., from 0.8 to 1.6 and 0.9 to 1.7 per 100,000 for men and women during 1990-2010, respectively. In this context, this study uses convolutional neural networks for classifying (diagnosing) AF, employing electrocardiogram data in a general hospital. Data came from Anam Hospital in Seoul, Korea, with 20,000 unique patients (10,000 normal sinus rhythm and 10,000 AF). 30 convolutional neural networks were applied and compared for the diagnosis of the normal sinus rhythm vs. AF condition: 6 Alex networks with 5 convolutional layers, 3 fully connected layers and the number of kernels changing from 3 to 256; and 24 residual networks with the number of residuals blocks (or kernels) varying from 8 to 2 (or 64 to 2). In terms of the accuracy, the best Alex network was one with 24 initial kernels (i.e., kernels in the first layer), 5,268,818 parameters and the training time of 89 s (0.997), while the best residual network was one with 6 residual blocks, 32 initial kernels, 248,418 parameters and the training time of 253 s (0.999). In general, the performance of the residual network improved as the number of its residual blocks (its depth) increased. For AF diagnosis, the residual network might be a good model with higher accuracy and fewer parameters than its Alex-network counterparts.