Whole-genome metagenomic analysis of the oral microbiota in patients with obstructive sleep apnea.
The oral microbiota is closely associated with systemic health, but few studies have investigated the oral microbiota in patients with obstructive sleep apnea (OSA). This study aimed to identify the variation of oral microbiota among patients with severe OSA, and the change of oral microbiota after treatment with continuous positive airway pressure (CPAP). Participants were enrolled in the study from November 2020 to August 2021. Sleep parameters using full nocturnal polysomnography (PSG) were collected on healthy controls, patients with severe OSA, and patients with severe OSA after CPAP treatment for 3 months. Oral samples were also collected by rubbing disposable medical sterile swabs on the buccal mucosa. Routine blood tests and biochemical indicators were measured using the fully automated biochemical analyzer. Oral microbial composition of oral samples were determined using whole-genome metagenomic analysis in all participants. Correlations were analyzed between the oral microbiota and blood lipids. Study enrollment included 14 participants, 7 healthy controls and 7 patients with severe OSA. At the species level, the relative abundances of Prevotella, Alloprevotella, Bacteroides, Veillonella_tobetsuensis, Candidatus saccharimonas, and Leptotrichia in the groups with severe OSA were significantly lower than those in the healthy controls (P both < 0.05). The abundances of Capnocytophaga, Veillonella, Bacillus_anthracis, Eikenella, and Kingella were significantly higher whereas the abundances of Gordonia and Streptococcus were significantly lower in the group with severe OSA compared to the severe OSA-CPAP group (P < 0.05 for both). According to the Kyoto Encyclopedia of Genes and Genomes (KEGG), 4 pathways changed in the group with severe OSA compared with healthy controls (P both < 0.05). Pathways related to Novobiocin biosynthesis, 2-Oxocarboxylic acid metabolism, and Histidine metabolism were enriched in the patients with severe OSA. Nine pathways showed significant differences with regard to the relative abundances of phenylalanine metabolism; alanine, aspartate, and glutamate metabolism; one carbon pool by folate; monobactam biosynthesis; 2-oxocarboxylic acid metabolism; arginine biosynthesis and vitamin B6 metabolism; novobiocin biosynthesis; and arginine and proline metabolism, which were significantly higher in the group with severe OSA compared to the severe OSA-CPAP group (P both < 0.05). The Spearman correlation analysis between blood lipid parameters and oral microbiota components showed that negative correlations were observed between total cholesterol and Streptomyces (r = - 0.893, P = 0.007), and high-density lipoprotein cholesterol (HDL-C) and Gordonia (r = - 0.821, P = 0.023); positive correlations were observed between HDL-C and Candidatus saccharimonas (r = 0.929, P = 0.003), and low-density lipoprotein cholesterol (LDL-C) and Capnocytophaga (r = 0.893, P = 0.007). There was an apparent discrepancy of the oral microbiota and metabolic pathways between the group with severe OSA and controls, and CPAP significantly changed oral microbial abundance and metabolic pathways in patients with severe OSA. Correlation analysis showed that these oral bacteria were strongly correlated with the blood lipids level.


Targetable elements in SARS-CoV-2 S2 subunit for the design of pan-coronavirus fusion inhibitors and vaccines.
The ongoing global pandemic of coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has caused devastating impacts on the public health and the global economy. Rapid viral antigenic evolution has led to the continual generation of new variants. Of special note is the recently expanding Omicron subvariants that are capable of immune evasion from most of the existing neutralizing antibodies (nAbs). This has posed new challenges for the prevention and treatment of COVID-19. Therefore, exploring broad-spectrum antiviral agents to combat the emerging variants is imperative. In sharp contrast to the massive accumulation of mutations within the SARS-CoV-2 receptor-binding domain (RBD), the S2 fusion subunit has remained highly conserved among variants. Hence, S2-based therapeutics may provide effective cross-protection against new SARS-CoV-2 variants. Here, we summarize the most recently developed broad-spectrum fusion inhibitors (e.g., nAbs, peptides, proteins, and small-molecule compounds) and candidate vaccines targeting the conserved elements in SARS-CoV-2 S2 subunit. The main focus includes all the targetable S2 elements, namely, the fusion peptide, stem helix, and heptad repeats 1 and 2 (HR1-HR2) bundle. Moreover, we provide a detailed summary of the characteristics and action-mechanisms for each class of cross-reactive fusion inhibitors, which should guide and promote future design of S2-based inhibitors and vaccines against new coronaviruses.


A newly-identified lineage of Schistosoma.
Because of their role in causing schistosomiasis, flukes of the genus Schistosoma are the best known of all digeneans. The genus has traditionally been divided into four familiar species groups. Here we report on three poorly known species of Schistosoma, one of which, Schistosoma hippopotami, is known from the hippopotamus, one of which is provisionally identified as Schistosoma edwardiense, another hippo parasite, and a third that has not previously been described. All were collected from freshwater snails obtained from Lake Edward, western Uganda, the type locality for both known hippo schistosomes. The three different kinds of schistosome cercariae differ from one another in size, and all are readily differentiated by their long tail stems from the cercariae of human-infecting species. Furthermore, each was recovered from a different genus of snail host, Biomphalaria sudanica, Bulinus truncatus or Ceratophallus natalensis. Molecular analysis, based on 8350 bases of combined nuclear and mitochondrial DNA, groups these three long tail-stem cercariae into a well supported clade that does not associate with any of the recognised species groups. The placement of this clade, basal to all African species plus several Asian species, suggests that there has been an ancient association between Schistosoma and hippos. This new African Schistosoma clade advocates the need for further modification of the traditional species group-based classification. Two of the four species groups are paraphyletic. It also suggests that Schistosoma has been remarkably plastic with respect to adapting to snail hosts-three distantly related genera of planorbid snails have been exploited by worms within a single clade. Finally, it adds a new layer of complexity to deciphering the origins of Schistosoma, often considered to be African but recently challenged as being Asian. In the late Cenozoic the distribution of hippo species straddled both Africa and Asia and they may have provided a means for the introduction of blood flukes to Africa.


The interaction of a polymeric persimmon proanthocyanidin fraction with Chinese cobra PLA2 and BSA.
To elucidate the anti-venom mechanism of persimmon tannin, the interaction between a polymeric persimmon proanthocyanidin fraction (PT40) and phospholipase A2 (PLA2) or bovine serum albumin (BSA) were studied using a competitive binding assay and spectroscopic methods including Fourier transform infrared spectroscopy (FT-IR), circular dichroism (CD), and resonance light scattering (RLS) spectroscopy. The results revealed that PT40 has a higher affinity for PLA2 than for BSA at physiological pH and induced greater conformational changes in PLA2 than in BSA. PT40 covalently bound to PLA2 in a reaction probably involving Lys residues. We propose that the high affinity of PT40 for PLA2 and the covalent modification of PLA2 by PT40 may be responsible for the ability of the tannin to irreversibly inhibit PLA2 catalytic activity, to prevent edema, and to neutralize the lethality of Chinese cobra PLA2 in vivo.


The right hemisphere and the dark side of consciousness.
Self-awareness and Theory of Mind comprise the main elements of higher-order consciousness. In attempting to localize these abilities, it appears that regions of the right hemisphere including fronto-temporal regions are capable of sustaining a sense of self-awareness. The right hemisphere appears dominant for tasks involving Theory of Mind as well as deception, which may be important for understanding the brain and laterality in terms of evolution. These findings are critical for our understanding of hemispheric differences in terms of higher-order consciousness.


Combination propranolol and bepridil therapy in stable angina pectoris.
The safety and efficacy of bepridil plus propranolol therapy were investigated in a placebo-controlled, parallel-design, double-blind trial in 56 patients who were not responding to propranolol alone. Patients entering the study were receiving an average propranolol dosage of 131 mg/day (range 20 to 240). For the first 2 weeks of the study they were given placebo in addition to their propranolol dose, and then were randomized to receive continued placebo plus propranolol or bepridil plus propranolol therapy. The bepridil dosage was adjusted over the 8 weeks of active treatment to an average of 273 mg/day (range 200 to 400). The double-blind treatment period was followed by a 3-week washout period during which all patients received propranolol and placebo. The effects of treatment on the frequency of angina attacks, nitroglycerin consumption, exercise performance (treadmill-modified Bruce protocol) and Holter electrocardiogram (ECG) were assessed. Propranolol and bepridil plasma levels also were obtained. Improved antianginal efficacy and reduced nitroglycerin consumption were noted when bepridil was added to propranolol (p less than 0.01). During 8 weeks of combination treatment, exercise tolerance increased 1.0 +/- 1.2 minutes from a baseline of 7.3 +/- 2.2 with bepridil plus propranolol compared with an increase of 0.02 +/- 1.3 minutes from a baseline of 7.6 +/- 2.9 with placebo plus propranolol (p less than 0.01). With bepridil plus propranolol, there were also increases in exercise time to onset of angina (p less than 0.04), exercise time to 1-mm electrocardiographic ST-segment depression (p less than 0.06) and total work (p less than 0.03) compared with placebo plus propranolol therapy.(ABSTRACT TRUNCATED AT 250 WORDS)


A rotary nano ion pump: a molecular dynamics study.
The dynamics of a rotary nano ion pump, inspired by the F (0) part of the F(0)F(1)-ATP synthase biomolecular motor, were investigated. This nanopump is composed of a rotor, which is constructed of two carbon nanotubes with benzene rings, and a stator, which is made of six graphene sheets. The molecular dynamics (MD) method was used to simulate the dynamics of the ion nanopump. When the rotor of the nanopump rotates mechanically, an ion gradient will be generated between the two sides of the nanopump. It is shown that the ion gradient generated by the nanopump is dependant on parameters such as the rotary frequency of the rotor, temperature and the amounts and locations of the positive and negative charges of the stator part of the nanopump. Also, an electrical potential difference is generated between the two sides of the pump as a result of its operation.


Bovine abortion associated with mixed Movar 33/63 type herpesvirus and bovine viral diarrhea virus infection.
A herpesvirus and cytopathogenic bovine viral diarrhea virus were isolated from a bovine fetus aborted in the 6th month of gestation. The herpesvirus was serologically indistinguishable from bovine herpesviruses DN-599 and Movar 33/63.


Herpesviruses and the microbiome.
The focus of this article will be to examine the role of common herpesviruses as a component of the microbiome of atopic patients and to review clinical observations suggesting that atopic patients might be predisposed to more severe and atypical herpes-related illness because their immune response is biased toward a TH2 cytokine profile. Human populations are infected with 8 herpesviruses, including herpes simplex virus HSV1 and HSV2 (also termed HHV1 and HHV2), varicella zoster virus (VZV or HHV3), EBV (HHV4), cytomegalovirus (HHV5), HHV6, HHV7, and Kaposi sarcoma-associated herpesvirus (termed KSV or HHV8). Herpesviruses are highly adapted to lifelong infection of their human hosts and thus can be considered a component of the human "microbiome" in addition to their role in illness triggered by primary infection. HSV1 and HSV2 infection and reactivation can present with more severe cutaneous symptoms termed eczema herpeticum in the atopic population, similar to the more severe eczema vaccinatum, and drug reaction with eosinophilia and systemic symptoms syndrome (DRESS) is associated with reactivation of HSV6 and possibly other herpesviruses in both atopic and nonatopic patients. In this review evidence is reviewed that primary infection with herpesviruses may have an atypical presentation in the atopic patient and conversely that childhood infection might alter the atopic phenotype. Reactivation of latent herpesviruses can directly alter host cytokine profiles through viral expression of cytokine-like proteins, such as IL-10 (EBV) or IL-6 (cytomegalovirus and HHV8), viral encoded and secreted siRNA and microRNAs, and modulation of expression of host transcription pathways, such as nuclear factor κB. Physicians caring for allergic and atopic populations should be aware of common and uncommon presentations of herpes-related disease in atopic patients to provide accurate diagnosis and avoid unnecessary laboratory testing or incorrect diagnosis of other conditions, such as drug allergy or autoimmune disease. Antiviral therapy and vaccines should be administered promptly when indicated clinically.


The association between psychosocial factors, protective factors, and its associated triggers with psychological distress among Bolivian adolescents.
The purpose of this study is to investigate the association between psychosocial factors, protective factors, and its associated triggers with psychological distress among Bolivian adolescents. This cross-sectional study was conducted by investigating the 2018 Bolivia global school-based student health survey (GSHS). The total number of students who participated in this survey was 7931, and the final sample was 7377. The mean age of the participants was 15.3 years (SD = 1.4). Psychological distress was assessed with a 2-item screener (loneliness and worry induced sleep disturbance). In all 22.3% of participants reported experiencing psychological distress, with 18.1% among adolescent males and 26.2% among adolescent females. In adjusted logistic regression analysis (AOR, 95% CI), there are two significant directions of association. One is the negative association, such as parental involvement as a protective factor. School adolescents who had more parental involvement were less likely to experience psychological distress. Parents understand problems or worries (0.64, 0.54-0.75, p < .001) and parents disregard privacy (0.69, 0.58-0.82, p < .001). On the other hand, many psycho-social factors are significantly positively associated with psychological distress. School adolescents who experience more psychosocial factors are more likely to experience psychological distress. Physical assault in the previous year (1.83, 1.59-2.11, p < .001), being bullied at school (1.27, 1.07-1.52, p < .01), being bullied outside of school (1.36, 1.15-1.61, p < .001), and being cyberbullied (1.60, 1.37-1.88, p < .001), were all significantly associated with psychological distress. Healthy relationships in a family, and interventions to reduce violence and bullying, should be encouraged and promoted.


Coexistence of TSH-secreting pituitary adenoma and autoimmune hypothyroidism.
TSH-secreting pituitary adenomas account for about 1-2% of all pituitary adenomas. Their diagnosis may be very difficult when coexistence of other diseases masquerades the clinical and biochemical manifestations of TSH-hypersecretion. A 41-yr-old female patient, weighing 56 kg, was referred for evaluation of an intra- and suprasellar mass causing menstrual irregularities. Eight yr before, the patient had been given a diagnosis of subclinical autoimmune hypothyroidism because of slightly elevated TSH levels and low-normal free T4 (FT4). Menses were normal. Despite increasing doses of levo-T4 (L-T4; up to 125 microg/day), TSH levels remained elevated and the patient developed mild symptoms of hyperthyroidism. After 7 yr, the menstrual cycle ceased. Gonadotropins were normal, whereas PRL level was elevated at 70 microg/l and magnetic resonance imaging (MRI) of the hypothalamic- pituitary region revealed a pituitary lesion with slight suprasellar extension. The tumor was surgically removed and histological examinations revealed a pituitary adenoma strongly positive for TSH. Three months after surgery the patient was well while receiving L-T4 75 microg/day and normal menses had resumed. MRI of the hypothalamic-pituitary region showed no evidence of residual tumor. At the last follow-up, 16 months after surgery, serum TSH, free T3 (FT3), and FT4 levels were normal. Coexistence of autoimmune hypothyroidism and TSH-secreting pituitary adenoma may cause further delays in the diagnosis of the latter. In patients with autoimmune hypothyroidism, one should be aware of the possible presence of a TSH-secreting pituitary adenoma when TSH levels do not adequately suppress in the face of high doses of L-T4 replacement therapy and elevated serum thyroid hormone levels.


Second primary germ cell tumors in patients with seminoma of the testis.
In a review of our experience with seminoma 9 cases of bilateral primary testis germ cell tumors were encountered, including 2 simultaneous and 7 successive. Of the 9 cases 6 were bilateral seminomas and 7 were stage I, contributing to the good survival experience. Treatment policy is specified and discussed.


Cerebral blood flow with the continuous infusion of oxygen-15-labeled water.
This work describes the determination of CBF in eight normal human subjects with positron emission tomographic (PET) imaging using the continuous intravenous infusion of H2(15)O. A whole-brain CBF model is described that permits the comparison of the CBF values determined using PET with those obtained using other methods. This model includes a correction for whole-brain recovery coefficient, a correction for the underestimation of flow due to the nonlinearity of the CBF model when considering tissue that includes both gray and white matter, the use of in vitro-determined brain-blood partition coefficients for gray and white matter, and a variation of the equilibrium model that permits the arterial concentration to vary. CBF values using this method compare well with values determined previously. Regional determinations using a brain overlay atlas are presented. Radiation dosimetry for the continuous infusion of H2(15)O is also included.


Monitoring versus interim analysis of clinical trials: a perspective from the pharmaceutical industry.
The definitions of "interim analysis" and "monitoring" of clinical trials are often ambiguous in the current literature. The resulting confusion can lead to erroneous conclusions and misguided decisions, especially when activities that are operational or observational are evaluated in a probabilistic sense as inferential. The authors seek to define "interim analysis" and "monitoring" in a mutually exclusive fashion. These definitions will then provide the opportunity to review and categorize existing clinical trial practices and procedures. This will clarify such issues as "when to look" and "when to pay a price" (e.g., test size and power) and characterize such issues in the context of pharmaceutical industry drug development.


Subjective cognitive complaints and cardiovascular risk factors in older Mexican Americans: A cross-sectional study.
Subjective cognitive complaints (SCC) are associated with higher risk of mild cognitive impairment (MCI) and dementia. Cardiovascular risk factors (CVRF) have been also associated with cognitive decline, MCI, and dementia. Few studies have examined the associated of CVRF and SCC. Participants were cognitively normal Mexican Americans from the HABLE study. Participants were categorized as with and without SCC, and SCC was also measured as a continuous variable. CVRF diagnosis were ascertained during consensus review. Cognitive measures used were MMSE, Trails B, SEVLT, and digit span. Logistic regression and linear regression were used to asses the association of SCC with CVRF and cognitive scores. A total of 673 participants [mean age 63.3 (SD=7.71), 69.2% female] were included. SCC was present in 323 participants (47.99%). Dyslipidemia and depression were associated with SCC. Individuals with dyslipidemia had 1.72 times the odds (95% CI = 1.20 to 2.47) of SCC, and those with depression had 3.15 times the odds (95% CI = 2.16 to 4.59) of self-reporting SCC. Higher SCC scores, were significantly associated with MMSE (B = 0.07; SE = 0.03; p = 0.02), and SEVLT immediate and delayed (B= -0.03; SE = 0.00; p = 0.000 and B = -0.03; SE = 0.00; p = 0.000, respectively). In a cognitively normal Mexican Americans sample of older adults, depression and dyslipidemia were correlated with self-reported SCC. A greater self-perception of cognitive decline correlated with lower scores on the MMSE and SEVLT.


Correction to: Reoperative Bariatric Surgery: a Systematic Review of the Reasons for Surgery, Medical and Weight Loss Outcomes, and Relevant Behavioral Factors.
Eating and lifestyle behaviors, difficulty in embracing the required lifestyle changes, and reappearance of depressive and anxious symptoms have been associated with failure of weight loss or weight regain after primary surgeries. These aspects are further developed in the original manuscript.


The impact of nurse staffing levels and nurse's education on patient mortality in medical and surgical wards: an observational multicentre study.
Growing evidence indicates that improved nurse staffing in acute hospitals is associated with lower hospital mortality. Current research is limited to studies using hospital level data or without proper adjustment for confounders which makes the translation to practice difficult. In this observational study we analysed retrospectively the control group of a stepped wedge randomised controlled trial concerning 14 medical and 14 surgical wards in seven Belgian hospitals. All patients admitted to these wards during the control period were included in this study. Pregnant patients or children below 17 years of age were excluded. In all patients, we collected age, crude ward mortality, unexpected death, cardiac arrest with Cardiopulmonary Resuscitation (CPR), and unplanned admission to the Intensive Care Unit (ICU). A composite mortality measure was constructed including unexpected death and death up to 72 h after cardiac arrest with CPR or unplanned ICU admission. Every 4 months we obtained, from 30 consecutive patient admissions across all wards, the Charlson comorbidity index. The amount of nursing hours per patient days (NHPPD) were calculated every day for 15 days, once every 4 months. Data were aggregated to the ward level resulting in 68 estimates across wards and time. Linear mixed models were used since they are most appropriate in case of clustered and repeated measures data. The unexpected death rate was 1.80 per 1000 patients. Up to 0.76 per 1000 patients died after CPR and 0.62 per 1000 patients died after unplanned admission to the ICU. The mean composite mortality was 3.18 per 1000 patients. The mean NHPPD and proportion of nurse Bachelor hours were respectively 2.48 and 0.59. We found a negative association between the nursing hours per patient day and the composite mortality rate adjusted for possible confounders (B = - 2.771, p = 0.002). The proportion of nurse Bachelor hours was negatively correlated with the composite mortality rate in the same analysis (B = - 8.845, p = 0.023). Using the regression equation, we calculated theoretically optimal NHPPDs. This study confirms the association between higher nurse staffing levels and lower patient mortality controlled for relevant confounders.


Treatment patterns of targeted and nontargeted therapies and survival effects in patients with locally advanced head and neck cancer in Taiwan.
Taiwan's National Health Insurance has covered targeted therapy, namely cetuximab, for locally advanced head and neck cancers (LAHNC) since July 2009. This study examines treatment trends and survival effects of locally advanced head and neck cancer patients before and after Taiwan's National Health Insurance covered cetuximab. We examined treatment trends and survival effects for patients with LAHNC using Taiwan's National Health Insurance Research Database. Patients who received treatment within 6 months were categorized as either nontargeted or targeted therapy groups. We analyzed treatment trends with the Cochran-Armitage trend test and explored factors associated with treatment selection and survival effects using multivariable logistic regression and Cox proportional hazards models. Of the 20,900 LAHNC patients included in the study, 19,696 received nontargeted therapy, while 1,204 received targeted therapy. Older patients with more comorbid conditions, advanced stages and patients with hypopharynx and oropharynx cancers were more likely to receive targeted therapy with concomitant cetuximab treatment. Patients who received targeted therapy in addition to other treatment modalities had a greater risk of one-year and long-term all-cause mortality or cancer-specific mortality than those without receiving targeted therapy (P < 0.001). Our study found an increasing trend in cetuximab utilization among LAHNC after reimbursement in Taiwan, but overall usage rates were low. LAHNC patients receiving cetuximab with other treatments had higher mortality risk than those receiving cisplatin, suggesting cisplatin may be preferred. Further research is needed to identify subgroups that could benefit from concomitant cetuximab treatment.


Reusable carboxylesterase immobilized in ZIF for efficient degradation of chlorpyrifos in enviromental water.
The past few decades have witnessed biodegradation of pesticides as a significant method in remediation of the environment for its specificity, efficiency and biocompatibility. However, the tolerability and recyclability of the enzymes in pesticide degradation and the development of enzymes that biodegrad pesticides are still urgent problems to be solved so far. Herein, a novel hyper-thermostable and chlorpyrifos-hydrolyzing carboxylesterase EstC was immobilized by biomineralization using zeolitic imidazolate framework (ZIF), one of the metal-organic frameworks (MOFs) with highly diverse structure and porosity. Compared with free enzyme, EstC@ZIF with a cruciate flower-like morphology presented scarcely variation in catalytic efficiency and generally improved the tolerance to organic solvents or detergents. Furthermore, there was scarcely decrease in the catalytic efficiency of EstC@ZIF and it also showed good reusability with about 50% residual activity after 12 continuous uses. Notably, EstC@ZIF could be used in actual water environment with an excellent value of degradation rate of 90.27% in 120 min, and the degradation efficiency remained about 50% after 9 repetitions. The present strategy of immobilizing carboxylesterase to treat pesticide-contaminated water broadens the method of immobilized enzymes on MOFs, and envisions its recyclable applicability in globe environmental remediation.


Clinical characteristics in unilateral vestibular atelectasis.
Unilateral vestibular atelectasis (UVA), an entity first described by Merchant and Schuknecht in 1988, has rarely been reported in vivo as of yet. We specify here the clinical characteristics of 22 patients diagnosed with UVA. Patients with a radiological diagnosis of UVA who underwent delayed inner ear MRI were included between April 2017 and January 2020. Full clinical testing including ocular infrared video-oculography, oVEMPs, cVEMPs, vHIT, bithermal caloric testing and auditory testing was performed. There were 13 men and 9 women, of mean age 58.6 ± 13.7 years. Onset was more frequently sudden (73%) than insidious (27%) though both clinical presentations were reported, and positional vertigo was described in 41% of cases. There were only two (9%) patients reporting Tullio's phenomenon. Vestibular testing showed that in 90% of cases, there was utricular dysfunction on oVEMP, while in 77% of cases, saccular function was preserved on cVEMP. vHIT showed high-velocity canal function impairment in all 22 patients: 8 patients (36%) had one impaired canal, 5 (23%) had two and 9 (41%) had all three canals affected. Caloric tests found complete unilateral areflexia, in 65% of tested cases, and partial deficiency in 35% of cases. Nine patients (40%) displayed asymmetrical hearing. We described in this study the various clinical presentations of a disease rarely reported in vivo, UVA. Initial clinical presentation can appear similar to an acute vestibular deficit, a recurrent positional vertigo, or fluctuating dizziness.


Reduction of epithelial secretion in male rat distal colonic mucosa by bile acid receptor TGR5 agonist, INT-777: role of submucosal neurons.
Recent evidence from rat neuron-free mucosa study suggests that the membrane bile acid receptor TGR5 decreases colonic secretion under basal and stimulated conditions. As submucosal neurons are key players in secretory processes and highly express TGR5, we investigated their role in TGR5 agonist-induced inhibition of secretion and the pathways recruited. TGR5 expression and localization were assessed in rat proximal (pC) and distal (dC) colon by qPCR and immunohistochemistry with double labeling for cholinergic neurons in whole-mount preparations. The influence of a selective (INT-777) or weak (ursodeoxycholic acid, UDCA) TGR5 agonist on colonic secretion was assessed in Ussing chambers, in dC preparation removing seromuscular ± submucosal tissues, in the presence of different inhibitors of secretion pathways. TGR5 mRNA is expressed in full thickness dC and pC and immunoreactivity is located in colonocytes and pChAT-positive neurons. Addition of INT-777, and less potently UDCA, decreased colonic secretion in seromuscular stripped dC by -58.17± 2.6%. INT-777 effect on basal secretion was reduced in neuron-free and TTX-treated mucosal-submucosal preparations. Atropine, hexamethonium, indomethacin, and L-NAME all reduced significantly INT-777's inhibitory effect while the 5-HT4 antagonist, RS-39604, and lidocaine abolished it. INT-777 inhibited stimulated colonic secretion induced by nicotine, but not cisapride, carbachol or PGE2. TGR5 activation inhibits basal and stimulated distal colonic secretion in rats by acting directly on epithelial cells and also inhibiting submucosal neurons. This could represent a counter-regulatory mechanism, at the submucosal level, of the known prosecretory effect of bile acids in the colon.


A new type of gap junction in the phylum Brachiopoda.
Gap junctions in the phylum Brachiopoda are shown to be of the P type as has been previously reported in invertebrates only in molluscs and possibly bryozoans. However, the mollusc gap junctions appear identical to vertebrae gap junctions in the fracture face on which particles remain, in particle spacing and in particle arrangement. The brachiopod gap junction, on the other hand, has general appearance traits which are more commonly seen in other invertebrate gap junction types. The freeze-fracture appearance of the brachiopod gap junction is described and discussed in relation to other types of gap junction which have been reported.


Fistula of the fourth branchial pouch.
Most of the congenital anomalies of the branchial apparatus are from the second arch, pouch, and cleft; some are from the first and third arches. Fourth branchial pouch remnants are very rare. We present a rare case of fourth branchial fistula, with characteristic clinical features of recurrent left-sided neck abscess that burst spontaneously, forming a fistula. Diagnosis of fourth branchial pouch fistula was confirmed by contrast radiography (ie, fistulogram and barium swallow) revealing the internal opening in the apex of the left pyriform fossa.


Thymoquinone accelerates osteoblast differentiation and activates bone morphogenetic protein-2 and ERK pathway.
Thymoquinone (TQ), the active component of Nigella sativa L. is well known for its various beneficial effects against several diseases. However, its detailed effect on bone metabolism has not been studied before. Therefore, the aim of the present study is to evaluate the effect of TQ on the proliferation, differentiation, and mineralization of MC3T3-E1 osteoblast cells. Our data shows that TQ induced the proliferation of MC3T3-E1 cells and proved to be non-toxic for up to 72 h of incubation. TQ induced the mineralization of MC3T3-E1 cells as evidenced by an increase in bone nodule formation 14 days post TQ treatment. qRT-PCR analysis shows that TQ induced the expression levels of differentiation related genes including alkaline phosphatase, osteocalcin, and osteopontin, while no effect was seen on collagen 1a1. TQ also induced the expression levels of bone morphogenetic protein-2 (BMP-2) and upregulated the phosphorylation of ERK signaling pathway. In summary, the present study shows for the first time that TQ has anabolic effects on MC3T3-E1 cells and that this effect is mediated by an increase in the expression of BMP-2 along with the involvement of the ERK signaling pathway. This study also reveals that TQ may be beneficial in inducing osteogenesis.


Type 1 diabetes: current concepts in epidemiology, pathophysiology, clinical care, and research.
Type 1 diabetes (T1D) is an autoimmune disease mediated by a combination of genetic and environmental triggers resulting in lymphocytic infiltration of pancreatic islets, destruction of beta cells, and lifelong dependency on exogenous insulin. Although T1D is prevalent (1 in 300) and its incidence is steadily increasing worldwide (3% per year), the exact gene-environment interactions precipitating the disease remain unknown. Living with T1D is challenging for patients, families, and caregivers. Because of the relative paucity of pediatric endocrinologists, general pediatricians and other subspecialists may occasionally be faced with the task of managing diabetes-related complaints. Herein, we provide a comprehensive review of the natural history, pathophysiology, and contemporary management of T1D. In addition, recent advances in T1D research are discussed.


Bacterial contamination of bone marrow grafts intended for autologous and allogeneic bone marrow transplantation. Incidence and clinical significance.
In a series of 100 bone marrow harvests, the incidence of bacterial contamination of the bone marrow graft was 17 percent. Ex vivo manipulation of some of the grafts prior to infusion may have caused additional bacterial contamination. All isolated bacteria were common skin flora, and no serious sequelae were observed in the patients receiving the culture-positive bone marrow grafts. Samples of harvested bone marrows purposely contaminated with an isolate of Staphylococcus epidermidis demonstrated a bactericidal property that was maximal early after bone marrow collection. Bone marrow collection and ex vivo manipulation may result in considerable bacterial contamination. Procedures must be developed to assure that marrow collection and processing do not result in clinically significant contamination.


Traumatic Injury Reduces Amyloid Plaque Burden in the Transgenic 5xFAD Alzheimer's Mouse Spinal Cord.
Axonal injury has been implicated in the development of amyloid-β in experimental brain injuries and clinical cases. The anatomy of the spinal cord provides a tractable model for examining the effects of trauma on amyloid deposition. Our goal was to examine the effects of axonal injury on plaque formation and clearance using wild type and 5xFAD transgenic Alzheimer's disease mice. We contused the spinal cord at the T12 spinal level at 10 weeks, an age at which no amyloid plaques spontaneously accumulate in 5xFAD mice. We then explored plaque clearance by impacting spinal cords in 27-week-old 5xFAD mice where amyloid deposition is already well established. We also examined the cellular expression of one of the most prominent amyloid-β degradation enzymes, neprilysin, at the lesion site. No plaques were found in wild type animals at any time points examined. Injury in 5xFAD prevented plaque deposition rostral and caudal to the lesion when the cords were examined at 2 and 4 months after the impact, whereas age-matched naïve 5xFAD mice showed extensive amyloid plaque deposition. A massive reduction in the number of plaques around the lesion was found as early as 7 days after the impact, preceded by neprilysin upregulation in astrocytes at 3 days after injury. At 7 days after injury, the majority of amyloid was found inside microglia/macrophages. These observations suggest that the efficient amyloid clearance after injury in the cord may be driven by the orchestrated efforts of astroglial and immune cells.


Count on trust: the indirect effect of trust in government on policy compliance with health behavior instructions.
Trust in government is considered a prominent factor for enhancing public compliance with government policies and instructions. The Coronavirus pandemic demonstrates the crucial role public compliance with governmentally issued health guidelines has in mitigating the pandemic. However, the mechanism explaining the trust-compliance association, particularly in regard to health-behavior compliance, is unclear. This article develops a new theoretical model, the Mediated Trust Model (MTM), for explaining the relationship between trust in government and public compliance with health instructions. The model extends the classic Health Belief Model for predicting health behavior by claiming that the perceptions regarding the instructions' costs, benefits and one's ability to perform them are affected by trust in government and mediate the trust-compliance association. The MTM was tested in four cross-sectional studies performed during the first 20 months of the Coronavirus pandemic in Israel on 3732 participants, for various health instructions. Implications for public health literature, policy compliance theory and policy makers are discussed.


Real clinical outcomes of nivolumab plus ipilimumab for renal cell carcinoma in patients over 75 years old.
Although nivolumab plus ipilimumab is the standard treatment for metastatic renal cell carcinoma (RCC), its efficacy and safety in older patients remain unclear. Therefore, this study aimed to assess the clinical outcomes of nivolumab plus ipilimumab for metastatic RCC in patients aged ≥ 75 years. We enrolled 120 patients with metastatic RCC treated with nivolumab plus ipilimumab from August 2015 to January 2023. Objective response rates (ORRs) were compared between patients aged < 75 and ≥ 75 years. Progression-free survival (PFS), overall survival (OS), and adverse events were compared between the groups. Adverse events were evaluated according to the Response Evaluation Criteria in Solid Tumors 1.1. Among the patients, 57 and 63 were classified as intermediate and poor risk, respectively, and one could not be classified. The median follow-up duration after the initiation of treatment was 16 months. The patient characteristics between the groups, except for age, were not significantly different. Intergroup differences in ORR (42% vs. 40%; p = 0.818), PFS (HR: 0.820, 95% CI 0.455-1.479; p = 0.510), and median OS (HR: 1.492, 95% CI 0.737-3.020; p = 0.267) were not significant. The incidence of adverse events (50% vs. 67%; p = 0.111) and nivolumab plus ipilimumab discontinuation due to adverse events was not significantly different between the groups (14% vs. 13%; p = 0.877). The effectiveness of nivolumab plus ipilimumab was comparable between patients with metastatic RCC aged < 75 and those ≥ 75 years with respect to their ORRs, PFS, OS, and adverse event rates.


Quantification of antigen-reactive T cells by a modified ELISPOT assay based on freshly isolated blood dendritic cells.
The enzyme-linked immunospot (ELISPOT) assay has become a widely employed method for quantification of antigen-reactive T lymphocytes. In recent years, various types of antigen-presenting cells (APCs) have been tested as stimulator cells in ELISPOT protocols to achieve a highly sensitive and rapid assay which is not impaired by a marked nonspecific cytokine release. However, the currently available APCs still have disadvantages, such as significant background reactivities, limited sensitivity, and time-consuming preparation procedures. Recently, we succeeded in defining a novel subpopulation of circulating dendritic cells (DCs) that can easily be prepared from human blood. These M-DC8+ DCs proved to be very effective in the induction of antigen-specific T cell responses. In the present study we provide evidence that M-DC8+ DCs are particularly well suited as APCs for the detection of antigen-specific CD8+ T cells after challenge with viral or tumor peptides in ELISPOT assays. In addition, protein-loaded M-DC8+ DCs proved to be quite efficient in the presentation of MHC class II-bound peptides, thus allowing the determination of frequencies of antigen-reactive CD4+ T cells. The use of M-DC8+ DCs as stimulator cells can improve the ELISPOT assay by combining high sensitivity, rapidity, and low background reactivity.


Engineering a novel immunogenic chimera protein utilizing bacterial infections associated with atherosclerosis to induce a deviation in adaptive immune responses via Immunoinformatics approaches.
Recent studies have established the role of bacteria including Streptococcus pneumoniae, Helicobacter pylori, Chlamydia pneumonia, Mycobacterium tuberculosis, and Porphyromonas gingivalis in the development of atherosclerosis. These bacteria contribute to plaque formation via promoting Th1 immune responses and speeding up ox-LDL formation. Hence, we employed computational reverse vaccinology (RV) approaches to deviate immune response toward Th2 via engineering a novel immunogenic chimera protein. Prominent atherogenic antigens from related bacteria were identified. Then, machine learning-based servers were employed for predicting CTL and HTL epitopes. We selected epitopes from a wide variety of HLAs. Then, a chimeric protein sequence containing TAT peptide, adjuvant, IL-10 inducer, and linker-separated epitopes was designed. The conformational structure of the vaccine was built via multiple-template homology modelling using MODELLER. The initial structure was refined and validated by Ramachandran plot. The vaccine was also docked with TLR4. After that, molecular dynamics (MD) simulation of the docked vaccine-TLR4 was conducted. Finally, the immune simulation of the vaccine was conducted via the C-ImmSim server. A chimera protein with 629 amino acids was built and, classified as a non-allergenic probable antigen. An improved ERRAT score of 80.95 for the refined structure verified its stability. Additionally, validation via the Ramachandran plot showed 98.09% of the residues were located in the most favorable and permitted regions. MD simulations showed the vaccine-TLR4 complex reached a stable conformation. Also, RMS fluctuations analysis revealed no sign of protein denaturation or unfolding. Finally, immune response simulations indicated a promising response by innate and adaptive immunity. In summary, we built an immunogenic vaccine against atherosclerosis and demonstrated its favorable properties via advanced Immunoinformatics analyses. This study may pave the path for combat against atherosclerosis.


Food insecurity, sleep, and cardiometabolic risks in urban American Indian/Alaska Native youth.
Food insecurity contributes to racial/ethnic disparities in health. This is the first study to examine associations among food insecurity, sleep, and cardiometabolic outcomes in urban American Indian/Alaska Native (AI/AN) youth. Participants were 142 urban AI/AN youth (mean age = 14 years, 58% female). Food insecurity and self-reported sleep disturbance were measured using validated surveys. A multi-dimensional sleep health composite was derived using questionnaires (ie, satisfaction, alertness) and actigraphy-derived indices (ie, duration, efficiency, regularity, timing). Cardiometabolic measures included body mass index, blood pressure, glycosylated hemoglobin, waist circumference, cholesterol, and triglycerides. Covariates were sex, age, and single-parent household. Greater food insecurity was significantly associated with greater body mass index (b = 0.12, p = .015), higher systolic blood pressure (b = 0.93, p = .03), and greater sleep disturbance (b = 1.49, p < .001), and marginally associated with lower sleep health composite scores (b = -0.09, p = .08). There was a significant indirect path from greater food insecurity to greater waist circumference through poorer sleep health (0.11, 95% bootstrapping CI: [0.01, 0.30]). Food insecurity is an important correlate of sleep and cardiometabolic health among urban AI/AN youth and should be addressed to reduce emerging health risks during this important developmental period. Policies to reduce food insecurity and increase access to healthy foods as well as sleep interventions for these youth could help, as preliminary findings suggest that sleep health may mediate the negative impact of food insecurity on cardiometabolic risks.


Profiling the Binding Sites of RNA-Binding Proteins with Nucleotide Resolution Using iCLIP.
The importance of posttranscriptional regulation in cellular metabolism has recently gone beyond what was previously appreciated. The regulatory mechanisms are controlled by RNA-binding proteins (RBPs), which form complexes with RNA and regulate RNA processing, stability, and localization, among others. Consistently, mutations in RBPs result in defects in developmental processes, diseases, and cancer. Gaining deeper insights into the biology of RNA-RBP interactions will lead to a better understanding of regulatory processes and disease development. Several techniques have been developed to capture the properties of RNA-RBP interactions. Furthermore, the development of high-throughput sequencing has broadened the capability of these methods. Here, we summarize individual-nucleotide resolution UV cross-linking and immunoprecipitation (iCLIP), a powerful technique that provides genome-wide information on RNA-RBP interactions at nucleotide resolution. In this chapter, we outline the iCLIP protocol and list possible controls that allow a targeted and cost-minimizing optimization of the protocol for an RBP-of-interest. Moreover, we provide notes on experimental design and a troubleshooting guideline for common problems that can occur during iCLIP library preparation.


Effect of Lavage Solution Type on Bronchoalveolar Lavage Fluid Cytology in Clinically Healthy Horses.
Equine bronchoalveolar lavage (BAL) is usually performed with 250-500 mL of isotonic saline at pH 5.5. The acidic pH of saline may cause an increase in airway neutrophil count 48 h after BAL. Other isotonic solutions such as Ringer's solution, phosphate-buffered saline (PBS) or Plasma-Lyte 148® have a neutral pH of 7.4 and might be a better choice for BAL by not provoking inflammation and the influx of neutrophils into airways. BAL was performed in four healthy horses in four different lung lobes using four different solutions in a randomized crossover design. In each lobe, BAL was performed twice with a 48 h interval using 250 mL of solution. Automated total nucleated cell counts (TNCs) were recorded, and differential cell counts in lavage fluid were determined by two investigators blinded to treatments. The mean volume of BAL fluid retrieved was 51 ± 14%. The mean neutrophil percentage (%N) increased from 1.5 ± 0.9% to 14.7 ± 9.6% at 48 h (p < 0.001) but was not significantly affected by the solution used or the lung lobe sampled. In conclusion, in this study, the influx of neutrophils into airways after BAL was independent of the type of isotonic solution used and the lung lobe sampled. Saline remains an appropriate solution for BAL in horses.


The cytotoxicity of malathion and essential oil of Nepeta crispa (lamiales: lamiaceae) against vertebrate and invertebrate cell lines.
Pesticides are used as essential tools to control vector-borne diseases and agricultural pests and maintain quality and quantity crop production. Scientists attempt to use derived plant natural products due to environmental safety and low mammalian toxicity. Therefore, the cytotoxicity of malathion and Nepeta crispa essential oil against vertebrate L929 and invertebrate Sf9 cell lines were investigated. About 2×103 cells were placed into the wells of a 96-well plate experiments. Then appropriate concentrations of malathion and N. crispa essential oil added to the wells. The cells were allowed to grow for 3-5 days and estimated the cell numbers. Control cell wells contained only cells with DMSO. All treatments and controls repeated at least four replicates. About 2×103 cells were placed into the wells of a 96-well plate experiments. Then appropriate concentrations of malathion and N. crispa essential oil added to the wells. The cells were allowed to grow for 3-5 days and estimated the cell numbers. Control cell wells contained only cells with DMSO. All treatments and controls repeated at least four replicates. Plant essential oil not only had no negative effects but also had boosting effects on the L929 cell viability. Nepeta crispa essential oil had negative effects on the Sf9 cell viability with the differences that derived plant natural products containing environmentally friendly and readily biodegradable derivatives, hydrolyzing rapidly in nature and nearly having no destructive effects on mammals and environment.


Management of medicinally useful plants by European migrants in South America.
Using the example of Polish migrants living in the subtropics of Argentina, we attempt to expand knowledge about migrant strategies for retaining their agency in medicinal plant procurement. Is to state which environments play a pivotal role as a source of medicinal plants for the study community, and if a gradient of relevance exists in the exploitation of medicinally useful species between the most proximate and the most distant habitats. We particularly aim to answer the following questions: (1) if Polish migrants have changed their patterns of obtaining medicinal plants during the migratory process; and (2) if the choice of strategies for medicinal plants depends on: (a) the degree of floristic and environmental similarity between the home and host country; (b) the perception and usefulness of certain environments as a source of medicinal plants; (c) the degree of contact with the local population in the host country, and/or (d) the degree of contact between migrants and their homeland. The analysis is grounded in data from different types of interviews and a homegarden inventory addressed to 72 study participants. Voucher specimens of species mentioned were gathered and identified. Two indices were used as proxy measures: (1) the number of species obtained from each habitat, and (2) the number of citations for both modes and places of obtaining medicinal plants. Due to different flora found in Argentina, Polish migrants could reconstruct only bits and pieces of their native pharmacopoeia. They could not acquire medicinal plants either from relatives in Poland or via importation. Therefore they had to develop new strategies for securing medicinal resources. During the migratory process, Poles in Misiones changed forms and places of obtaining medicinal plants. Cultivated species from homegardens play the most important role, while in the native country homegarden species were used sparsely. The second most important environment for medicinal plant procurement is the forest, whose exploitation was increased by contact with local mestizos and indigenous groups. This study clearly shows that traditional knowledge can be adaptive and resilient. New species have been selected and incorporated from mestizos, and indigenous people, and at the same time the use of some legacy plants has been preserved. The importance of home gardens as a venue for medicinal plants is another facet of this adaptive process. Traditional knowledge is resilient too, because despite the many changes that have occurred, Polish people have maintained phytotherapy as their preferred form of treating ailments, and managed to retain certain species brought from Europe as the most relevant.


Quality of life in cancer patients receiving palliative care.
The main focus of palliative care services is to improve the patient's quality of life (QOL), which is defined as the subjective evaluation of life as a whole or the patient's appraisal and satisfaction with their current level of functioning compared with what they perceive to be possible or ideal. In this prospective study we attempt to validate the Hindi version of a questionnaire designed by the functional assessment of chronic illness therapy (FACIT) measurement system; to measure the subjective QOL of cancer patients receiving home-based palliative care, determine ease of use of the questionnaire and correlate the QOL of these patients with the objective assessment of their Karnofsky's performance status and their numerical pain score. One hundred cancer patients receiving free home-based palliative care in New Delhi, India. A multidisciplinary palliative home care team using the Functional Assessment of Cancer Therapy-General (FACT-G(©)) questionnaire in Hindi. Microsoft Excel Correlation. The FACT-G(©) questionnaire in Hindi is a useful tool in measuring QOL and can be used to monitor the patient's progress and symptom control during the course of the disease. It is simple to use and does not take too much time to complete. The results are tabulated in English and can be used for comparison purposes globally; the scoring process is very simple. Increasing QOL and KPS showed a positive correlation whereas increasing pain and better QOL show negative correlation, as do better performance status and increasing pain score.


Evaluation of an I-box wind tunnel model for assessment of behavioral responses of blow flies.
The behavioral response of flies to olfactory cues remains the focus of many investigations, and wind tunnels have sometimes been employed for assessment of this variable in the laboratory. In this study, our aim was to design, construct, and operate a new model of I-box wind tunnel with improved efficacy, highlighting the use of a new wind tunnel model to investigate the behavioral response of the medically important blow fly, Chrysomya megacephala (Fabricius). The I-box dual-choice wind tunnel designed for this study consists of seven conjoined compartments that resulted in a linear apparatus with clear glass tunnel of 30 × 30 × 190 cm ended both sides with wooden "fan compartments" which are equipped with adjustable fans as wind source. The clear glass tunnel consisted of two "stimulus compartments" with either presence or absence (control) of bait; two "trap compartments" where flies were attracted and allowed to reside; and one central "release compartment" where flies were introduced. Wind tunnel experiments were carried out in a temperature-controlled room, with a room light as a light source and a room-ventilated fan as odor-remover from tunnel out. Evaluation of testing parameters revealed that the highest attractive index was achieved with the use of 300 g of 1-day tainted pork scrap (pork meat mixed with offal) as bait in wind tunnel settings wind speed of 0.58 m/s, during 1.00-5.00 PM with light intensity of 341.33 lux from vertical light and 135.93 lux from horizontal light for testing a group of 60 flies. In addition, no significant response of well-fed and 24 h staved flies to this bait under these conditions was found. Results of this study supported this new wind tunnel model as a suitable apparatus for investigation of behavioral response of blow flies to bait chemical cues in the laboratory.


Autologous peripheral stem cell transplantation for aggressive hemophagocytic syndrome associated with T-cell lymphoma: case study and review.
Hemophagocytic syndrome (HPS) is a rare clinical presentation infrequently associated with lymphoproliferative disorders. We describe a 29-year-old male with aggressive HPS and T-cell lymphoma managed successfully with high-dose chemotherapy and autologous peripheral stem cell transplantation (APSCT), in remission at 41 months of follow-up. In reviewing the literature, this case illustrates the 2nd longest surviving individual post stem cell transplant for aggressive HPS.


Editorial: Heavy Adolescent Alcohol Use: An Accelerant of Impulsivity?
It has been difficult to disentangle factors conferring vulnerability to substance use disorders (SUDs) from the consequences of substance use. Reward sensitivity and impulsivity have been identified as adolescent risk factors that confer vulnerability for later problematic substance use.1,2 Studies also suggest, however, that substance use itself affects brain development and behavior and that some of the same risk factors that predispose youth to SUD (eg, reward sensitivity and impulsivity) may be brought on or worsened by the neurotoxicity of drugs of abuse.3,4 Studies examining neural and behavioral correlates of SUDs commonly include youth with varying degrees of substance exposure; thus development of vulnerabilities to substance abuse are difficult to separate from the effects of substance use. In this issue of JACC, Ivanov et al.5 advance our field's knowledge in this area by leveraging longitudinal data from the European IMAGEN dataset (n = 2,200)6 in order to characterize predictors of alcohol use at age 16 as well as trajectories of impulsivity. This design allows investigation into whether alcohol drinking in adolescence may actually be related to worsening impulsivity. The authors followed a subset of the IMAGEN sample, 304 substance-naïve 14-year-olds over 2 years. Reward system function in the brain was assessed at baseline by collecting functional magnetic resonance imaging scans during a Monetary Incentive Delay (MID)7 task which assessed neural response to reward anticipation and outcome. Impulsivity and delay discounting, the propensity to select smaller immediate rewards versus larger, delayed rewards, were also assessed at baseline and follow-up. Linear regression was used to evaluate longitudinal associations among the frequency of alcohol use at age 16 and impulsivity, delay discounting, and reward system function at age 14. Reward system function was measured by activation in medial orbitofrontal cortex (mOFC) and ventral striatum regions of interest during the highest versus the lowest reward levels on the MID task. These regions are involved in key aspects of reward processing, including valuation of rewarding stimuli and outcomes.8,9.


Connexin43 deficiency causes dysregulation of coronary vasculogenesis.
The connexin43 knockout (Cx43alpha1 KO) mouse dies at birth from outflow obstruction associated with infundibular pouches. To elucidate the origin of the infundibular pouches, we used microarray analysis to investigate gene expression changes in the pouch tissue. We found elevated expression of many genes encoding markers for vascular smooth muscle (VSM), endothelial cells, and fibroblasts, cell types that are epicardially derived and essential for coronary vasculogenesis. This was accompanied by increased expression of VEGF and genes in the TGFbeta and VEGF/Notch/Eph cell-signaling pathways known to regulate vasculogenesis/angiogenesis. Using immunohistochemistry and a VSM lacZ reporter gene, we confirmed an abundance of ectopic VSM and endothelial cells in the infundibular pouch and in some regions of the right ventricle forming secondary pouches. This was associated with distinct thinning of the compact myocardium. TUNEL labeling showed increased apoptosis in the pouch tissue, in agreement with the finding of altered expression of many apoptotic genes. Defects in vascular remodeling were indicated by a marked reduction in the branching complexity of the distal coronary arteries. In the near term KO mouse, we also observed a profusion of large coronary vascular plexuses subepicardially. This was associated with elevated epicardial expression of VEGF and abnormal epicardial cell morphology. Together, these observations indicate that dysregulated coronary vasculogenesis plays a pivotal role in formation of the infundibular pouches and suggests an essential role for Cx43alpha1 gap junctions in coronary vasculogenesis and vascular remodeling.


The geography of COVID-19 misinformation: using geospatial maps for targeted messaging to combat misinformation on COVID-19, South Africa.
The proliferation of false information on COVID-19 mostly through social media is adversely affecting control efforts. The objective of this study was to identify areas where targeted effective messaging can be useful in demystifying misinformation against COVID-19. The study showed high levels of misinformation on COVID-19 in the study area [mean score 2.71; standard deviation (SD) 1.5]. The highest levels of misinformation were observed in Dr. Ruth Segomotsi Mompati district, North West province (mean score: 3.84; SD: 2.1) and Sedibeng district, Gauteng province (mean score: 3.56; SD 1.7). Higher levels of misinformation were reported by those aged 18-24 years (mean score: 3.48; SD: 1.8), and men (mean score: 2.73; SD: 1.8). Across the two provinces, we identified geospatial hot and coldspots of misinformation highlighting the need to implement point of care strategies such as targeted messaging. Findings showed the need for targeted interventions to young people, students, those with low levels of education and the self-employed in the two districts more importantly, as South Africa expands its nationwide vaccination roll-out.


Implications for patient follow-up of changes in inferior vena cava filter removal plan practices.
To assess the improvements during patient follow-up after implementing a proactive plan for retrieval of inferior vena cava (IVC) filters. A retrospective study was undertaken including all patients who had a temporary IVC filter inserted over a 10-year period. Patient demographics, insertion dates, retrieval dates, retrieval rates, and complications were recorded. Patients were categorised into three groups as follows: group A, no intervention where retrieval was not suggested; group B, passive intervention where retrieval was suggested but no appointment made; and group C, proactive intervention where a retrieval date was booked by the radiologist. Five hundred and nine patients had a temporary IVC filter inserted at Royal Liverpool University Hospital over the study period. One per cent of patients in group C were lost to follow-up compared to 41% and 29% in groups A and B respectively (p<0.001). The number of patients who had an IVC retrieval attempt also increased to 80% in group C compared to 53% and 64% in groups A and B (p<0.001). Incorporation of a proactive retrieval booking system has significantly reduced the number of patients lost to follow-up to 1%, leading to an increased number of IVC filter retrieval attempts.


Isolated limb perfusion in the management of locally advanced extremity soft tissue sarcoma.
In conclusion, ILP is an interesting and important treatment option in the management of locally advanced extremity soft tissue sarcomas. Large medical centers, dealing with referrals and an important caseload of STS patients, should develop this treatment option and have it readily available to offer patients the best chances for limb salvage. In Europe, the success of TNF-based ILP has lead to the training, accreditation, and activation of TNF-based ILP programs in over 30 cancer centers since the approval of TNF for this indication in 1999. Thus, country by country centers for referral programs are established to deal with those categories of patients that can greatly benefit from the availability and integration of this treatment option in the STS treatment programs.


Two-step discriminant analysis based multi-view polarimetric SAR image classification with high confidence.
Polarimetric synthetic aperture radar (PolSAR) image classification is a hot topic in remote sensing field. Although recently many deep learning methods such as convolutional based networks have provided great success in PolSAR image classification, but they need a high volume of labeled samples, which are not usually available in practice, or they cause a high computational burden for implementation. In this work, instead of spending cost for network training, the inherent nature of PolSAR image is used for generation of convolutional kernels for extraction of deep and robust features. Moreover, extraction of diverse scattering characteristics contained in the coherency matrix of PolSAR and fusion of their output classification results with a high confidence have high impact in providing a reliable classification map. The introduced method called discriminative features based high confidence classification (DFC) utilizes several approaches to deal with difficulties of PolSAR image classification. It uses a multi-view analysis to generate diverse classification maps with different information. It extracts deep polarimetric-spatial features, consistent and robust with respect to the original PolSAR data, by applying several pre-determined convolutional filters selected from the important regions of image. Convolutional kernels are fixed without requirement to be learned. The important regions are determined with selecting the key points of image. In addition, a two-step discriminant analysis method is proposed to reduce dimensionality and result in a feature space with minimum overlapping and maximum class separability. Eventually, a high confidence decision fusion is implemented to find the final classification map. Impact of multi-view analysis, selection of important regions as fixed convolutional kernels, two-step discriminant analysis and high confidence decision fusion are individually assessed on three real PolSAR images in different sizes of training sets. For example, the proposed method achieves 96.40% and 98.72% overall classification accuracy by using 10 and 100 training samples per class, respectively in L-band Flevoland image acquired by AIRSAR. Generally, the experiments show high efficiency of DFC compared to several state-of-the-art methods especially for small sample size situations.


Intrinsically disordered protein PID-2 modulates Z granules and is required for heritable piRNA-induced silencing in the Caenorhabditis elegans embryo.
In Caenorhabditis elegans, the piRNA (21U RNA) pathway is required to establish proper gene regulation and an immortal germline. To achieve this, PRG-1-bound 21U RNAs trigger silencing mechanisms mediated by RNA-dependent RNA polymerase (RdRP)-synthetized 22G RNAs. This silencing can become PRG-1-independent and heritable over many generations, a state termed RNA-induced epigenetic gene silencing (RNAe). How and when RNAe is established, and how it is maintained, is not known. We show that maternally provided 21U RNAs can be sufficient for triggering RNAe in embryos. Additionally, we identify PID-2, a protein containing intrinsically disordered regions (IDRs), as a factor required for establishing and maintaining RNAe. PID-2 interacts with two newly identified and partially redundant eTudor domain-containing proteins, PID-4 and PID-5. PID-5 has an additional domain related to the X-prolyl aminopeptidase APP-1, and binds APP-1, implicating potential N-terminal proteolysis in RNAe. All three proteins are required for germline immortality, localize to perinuclear foci, affect size and appearance of RNA inheritance-linked Z granules, and are required for balancing of 22G RNA populations. Overall, our study identifies three new proteins with crucial functions in C. elegans small RNA silencing.


Low-dose Adriamycin remission maintenance therapy for pediatric solid tumors.
A low-dose maintenance schedule of Adriamycin was evaluated in six children whose metastatic solid tumors regressed following toxic induction therapy with Adriamycin. Three of these children are now disease-free more than one year following discontinuation of therapy. Adriamycin can be given on a low-dose maintenance schedule free from alopecia, fever, stomatitis, myelosuppression and recognizable cardiomyopathy. Further studies of similar schedules are warranted.


Localization of somatostatin-like immunoreactivity in the area postrema of the rat and cat.
The distribution of somatostatin-like immunoreactive (SOM-LI) fibers within the area postrema (AP) of rat and cat, using the peroxidase-anti-peroxidase (PAP) technique, is described. In rat, SOM-LI fibers were scattered throughout the AP while moderate accumulations of SOM-LI fibers occurred near the ventrolateral border. In cat, scattered SOM-LI fibers occurred throughout the caudal AP, with slightly higher numbers present along the AP border with the nucleus tractus solitarius. In rostral AP levels only small patches of scattered SOM-LI fibers were present. AP SOM-LI cell bodies were absent in both species. The immunohistochemical localization of SOM in the AP supports the concept that the gastric and cardiovascular effects of SOM are mediated in part through the AP.


Psychometric Properties of the Persian Version of the Spiritual Well-Being Scale Among Iranian Potential Organ Donors.
The Spiritual Well-Being Scale (SWBS) is a widely used scale that requires evaluation for the Iranian population. This study aimed to determine the psychometric properties of the SWBS among a sample of potential Iranian organ donors. The research was conducted in 2022 in Iran using cross-sectional methodological design. The sample consisted of 316 individuals from the Iranian community who expressed willingness to donate organs. Convenience sampling was used to select the participants. The construct validity and reliability of the Persian version of the SWBS were assessed. The results from maximum likelihood (ML) and confirmatory factor analysis (CFA) revealed three factors labeled as higher power, the meaning of life, and the purpose of life, which explained 50.2% of the total variance in the concept of Spiritual Well-Being (SWB). The results indicated that the model was appropriate. Cronbach's Alpha and McDonald's Omega demonstrated an acceptable internal consistency of the scale. All the statistical analyses were performed using SPSS 27 and AMOS 27 software. These findings suggest that the Persian version of the SWBS is a reliable and valid scale for assessing the spiritual well-being of individuals who are willing to donate organs. The scale comprises 16 items related to higher power, meaning of life, and sense of life.


Antipsychotic dose, dopamine D2 receptor occupancy and extrapyramidal side-effects: a systematic review and dose-response meta-analysis.
Antipsychotic drugs differ in their propensity to cause extrapyramidal side-effects (EPS), but their dose-effects are unclear. Therefore, we conducted a systematic review and dose-response meta-analysis. We searched multiple electronic databases up to 20.02.2023 for fixed-dose studies investigating 16 second-generation antipsychotics and haloperidol (all formulations and administration routes) in adults with acute exacerbations of schizophrenia. The primary outcome was the number of participants receiving antiparkinsonian medication, and if not available, the number of participants with extrapyramidal side-effects (EPS) and the mean scores of EPS rating scales were used as proxies. The effect-size was odds ratio (ORs) compared with placebo. One-stage random-effects dose-response meta-analyses with restricted cubic splines were conducted to estimate the dose-response curves. We also examined the relationship between dopamine D2 receptor (D2R) occupancy and ORs by estimating occupancies from administrated doses. We included data from 110 studies with 382 dose arms (37193 participants). Most studies were short-term with median duration of 6 weeks (range 3-26 weeks). Almost all antipsychotics were associated with dose-dependent EPS with varied degrees and the maximum ORs ranged from OR = 1.57 95%CI [0.97, 2.56] for aripiprazole to OR = 7.56 95%CI [3.16, 18.08] for haloperidol at 30 mg/d. Exceptions were quetiapine and sertindole with negligible risks across all doses. There was very low quality of findings for cariprazine, iloperidone, and zotepine, and no data for clozapine. The D2R occupancy curves showed that the risk increased substantially when D2R occupancy exceeded 75-85%, except for D2R partial agonists that had smaller ORs albeit high D2R occupancies. In conclusion, we found that the risk of EPS increases with rising doses and differs substantially in magnitude among antipsychotics, yet exceptions were quetiapine and sertindole with negligible risks. Our data provided additional insights into the current D2R therapeutic window for EPS.


Effect of NO synthesis blocker on electrical activity of rat stomach and small intestine in the early postoperation period.
The effect of NO on electrical activity of the stomach and small intestine was studied in chronic experiments on rats during the early postoperation period. Generation of migrating myoelectric complex in the stomach and small intestine was impaired after abdominal surgery. Administration of NO synthesis blocker suppressed the inhibitory influences, which potentiated generation of the myoelectric complex in the small intestine. Our results suggest that NO is involved in the regulation of the migrating myoelectric complex in the small intestine during postoperation motor disturbances in the gastrointestinal tract.


Subantral recovery technique (T.R.O.A.S.).
Extensive pneumatization of the maxillary sinus can limit rehabilitation with maxillary implants. During the last 30 years, several graft addition techniques have been proposed to solve this problem. A technique without graft has been developed by the authors and the results of this technique are analyzed.


Oxime effects on the rate constants of carbamylation and decarbamylation of acetylcholinesterase for pyridostigmine, physostigmine and insecticidal carbamates.
The effects of the oximes 2-pyridine aldoxime methiodide (PAM), HI-6, HS-6, toxogonin and TMB-4 on the rate of carbamylation of membrane-bound bovine erythrocyte acetylcholinesterase were studied. The second-order rate constant of carbamylation (ki) and the first-order rate constant of decarbamylation (k3) were calculated from the proportion of free acetylcholinesterase at equilibrium and the rate of approach to equilibrium. Twenty insecticidal carbamates plus physostigmine and pyridostigmine were studied. The oximes increased ki for several carbamates, with HI-6 causing an increase in the most number of cases (12) and PAM the least (3). HI-6 was also a potent accelerator of decarbamylation (increase in k3) in all cases, whereas PAM caused a significant decrease in k3 in 15 cases and a nonsignificant decrease in the other 7. Toxogonin and TMB-4 increased k3 or had no significant effect. The results were generally consistent with a proposal in the literature that there is a correlation between increased ki and increased toxicity of the carbamate in the presence of an oxime.


Outcomes of lensectomy with glued intraocular lens and factors associated with failure in eyes with spherophakia and glaucoma.
To evaluate the outcomes of lensectomy with a glued intraocular lens (IOL) in spherophakic eyes with secondary glaucoma and assess factors associated with failure. We prospectively evaluated outcomes of lensectomy with glued IOL in 19 eyes with spherophakia and secondary glaucoma (intraocular pressure (IOP) ≥22 mm Hg and/or glaucomatous optic disc damage) between 2016 and 2018. The vision, refractive error, IOP, antiglaucoma medications (AGMs), optic disc changes, need for glaucoma surgery, and complications were assessed. Success was defined as complete when IOP was ≥5 and ≤21 mmHg without AGMs; qualified success as similar IOP with up to 3 AGM; the need for >3AGM/additional surgery for IOP control was considered a failure. Preoperatively, the median (interquartile range: IQR) age was 18 (13.5-30) years. IOP was 16 (14-22.5) mmHg on a median of 3 (2,3) AGMs. Median postoperative follow up was 27.7 months (11.9, 39.7). Postsurgery, most patients achieved emmetropia, with significantly decreased refractive error from a median spherical equivalent of -12.5D to + 0.5D, P < 0.0002. The complete success probability was 47% (95% confidence intervals (CIs): 29-76%) at 3 months and was 21% (8 - 50%) at 1 year and 3 years. The qualified success probability was 93% (82-100%) at 1 year, which reduced to 79% (60-100%) in 3 years. None of the eyes had any retinal complications. The higher number of preoperative AGM was found to be a significant risk factor (p < 0.02) for the failure of complete success. One-third of the eyes had IOP control without the need for AGM postlensectomy with glued IOL. Surgery resulted in significant improvement in visual acuity. The higher number of preoperative AGM was associated with poor glaucoma control after glued IOL surgery.


Contribution of inflammasome genetics in Plasmodium vivax malaria.
Recent reports showed that, in mice, symptomatic Plasmodium infection triggers NLRP3/NLRP12-dependent inflammasome formation and caspase-1 activation in monocytes. In humans, few works demonstrated that inflammasome is activated in malaria. As Plasmodiumvivax is a potent inducer of inflammatory response we hypothesised that inflammasome genetics might affect P. vivax malaria clinical presentation. For this purpose, selected SNPs in inflammasome genes were analysed among patients with symptomatic P. vivax malaria. 157 Brazilian Amazon patients with P. vivax malaria were genotyped for 10 single nucleotide polymorphisms (SNPs) in inflammasome genes NLRP1, NLRP3, AIM2, CARD8, IL1B, IL18 and MEFV. Effect of SNPs on hematologic and clinical parameters was analysed by multivariate analysis. Our data suggested an important role of NLRP1 inflammasome receptor in shaping the clinical presentation of P. vivax malaria, in term of presence of fever, anaemia and thrombocytopenia. Moreover IL1B rs1143634 resulted significantly associated to patients' parasitaemia, while IL18 rs5744256 plays a protective role against the development of anaemia. Polymorphisms in inflammasome genes could affect one or other aspects of malaria pathogenesis. Moreover, these data reveal novel aspects of P.vivax/host interaction that involved NLRP1-inflammasome.


Whey Protein-Enriched and Carbohydrate-Rich Breakfasts Attenuate Insulinemic Responses to an ad libitum Lunch Relative to Extended Morning Fasting: A Randomized Crossover Trial.
Typical breakfast foods are rich in carbohydrate, so they not only elevate blood glucose during the morning, but also elicit a second-meal effect that can attenuate blood glucose responses in the afternoon. To determine whether a reduced-carbohydrate protein-enriched breakfast can elicit similar effects on glucose control later in the day but without hyperglycemia in the morning. In a randomized crossover design, 12 healthy men and women (age 22 ± 2 y, BMI 24.1 ± 3.6 kg·m-2; Mean ± SD) completed 3 experimental conditions. In all conditions, participants consumed an ad libitum lunch at 1200 ± 1 h but differed in terms of whether they had fasted all morning (control) or had consumed a standardized porridge breakfast at 0900 ± 1 h (320 ± 50 kcal; prescribed relative to resting metabolic rate) that was either carbohydrate-rich (50 ± 10 g CHO) or protein-enriched (that is, isoenergetic substitution of carbohydrate for 15 g whey protein isolate). The protein-enriched breakfast reduced the morning glycemic response (iAUC 87 ± 36 mmol·L-1·180 min) relative to the carbohydrate-rich breakfast (119 ± 37 mmol·L-1·180 min; P = 0.03). Despite similar energy intake at lunch in all 3 conditions (protein-enriched 769 ± 278 kcal; carbohydrate-rich 753 ± 223 kcal; fasting 790 ± 227 kcal), postlunch insulinemic responses were markedly attenuated when breakfasts had been consumed that were either protein-enriched (18.0 ± 8.0 nmol·L-1·120 min; P = 0.05) or carbohydrate-rich (16.0 ± 7.7 nmol·L-1·120 min; P = 0.005), relative to when lunch was consumed in an overnight fasted state (26.9 ± 13.5 nmol·L-1·120 min). Breakfast consumption attenuates insulinemic responses to a subsequent meal, achieved with consumption of energy-matched breakfasts typically high in carbohydrates or enriched with whey protein isolate relative to extended morning fasting. NCT03866720 (clinicaltrials.gov).


[What diagnosis can be made by the pathologist in a case of sclerosing thyroiditis?].
The pathology of thyroiditis seems well-established with a recognized classification based on clinical and pathological features. However, problems of differential diagnosis remain between both Riedel's and Quervain's thyroiditis and sclerosing Hashimoto's thyroiditis: these entities sometimes lack the characteristic histological pattern, and the clinico-biological data are not always available to the pathologist. We re-examined 18 cases of thyroiditis with sclerosis, retrieved from our files, diagnosed as Riedel's thyroiditis in five cases, Quervain's thyroiditis in five other cases and sclerosing Hashimoto thyroiditis in eight cases. Only two diagnosed cases of Riedel's thyroiditis were pathognomic. Three cases of Quervain's thyroiditis and four cases of sclerosing Hashimoto's thyroiditis presented a slight or moderate extension of the fibrosis in perithyroidal soft-tissues, raising the differential diagnosis of an incipient Riedel's thyroiditis. A definite diagnosis of the type of thyroiditis with sclerosis remains difficult, because all three pathologies present common points. In cases with a characteristic pattern, the diagnosis is straightforward. However, it appears in our study that half of the diagnoses remain ambiguous, because of the existence of histological features common to different entities. In these cases, we think the diagnosis of sclerosing thyroiditis NOS would be more appropriate, the histology not being sufficiently characteristic to make a more specific diagnosis.


LC-MS/MS method development for quantitative analysis of acetaminophen uptake by the aquatic fungus Mucor hiemalis.
Acetaminophen is a pharmaceutical, frequently found in surface water as a contaminant. Bioremediation, in particular, mycoremediation of acetaminophen is a method to remove this compound from waters. Owing to the lack of quantitative analytical method for acetaminophen in aquatic organisms, the present study aimed to develop a method for the determination of acetaminophen using LC-MS/MS in the aquatic fungus Mucor hiemalis. The method was then applied to evaluate the uptake of acetaminophen by M. hiemalis, cultured in pellet morphology. The method was robust, sensitive and reproducible with a lower limit of quantification of 5 pg acetaminophen on column. It was found that M. hiemalis internalize the pharmaceutical, and bioaccumulate it with time. Therefore, M. hiemalis was deemed a suitable candidate for further studies to elucidate its pharmaceutical tolerance and the longevity in mycoremediation applications.


Watching a video together creates social closeness between children and adults.
Human social relationships are often formed through shared social activities in which individuals share mental states about external stimuli. Previous work on joint attention has shown that even minimal shared experiences such as watching something together facilitates social closeness between individuals. Here, we examined whether young children already connect with others through joint attention. In the current studies, children sat next to a novel adult who either watched a film with them or was not able to see the film and read a book instead. After the video, we measured children's willingness (i.e., latency) to approach the experimenter holding out a toy. In both studies, the 2.5-year-olds who watched the film together approached more quickly than the other children. These results show that both minimally interactive shared experiences and noninteractive shared experiences lead children to feel more comfortable with a novel adult. This suggests that joint attention interactions, and shared experiences in general, play an important role not only in children's cognitive development but also in their social development and the formation of their social relationships.


Determination of seven preservatives in cosmetic products by micellar electrokinetic chromatography.
A micellar electrokinetic chromatography method using cetyltrimethylammonium bromide (CTAB) as a cationic surfactant, coupled with UV-Vis detection, was developed for the simultaneous determination of seven preservatives, including methyl-, ethyl-, propyl- and butyl-paraben and phenol, phenoxyethanol and resorcinol. The method involved optimizing the pH of the phosphate buffer and concentrations of CTAB, ethanol and 2-hydroxypropyl-β-cyclodextrin (HP-β-CD). The preservatives were well separated using optimum conditions and separated within 10 min at a separation voltage of -12.5 kV with the 1.0 mM phosphate buffer (pH 7.0) containing 90 mM CTAB, 25 mM HP-β-CD and 10% (v/v) ethanol. Satisfactory recoveries (84.1-103.0%), migration time (RSD < 3.1%) and peak area (RSD < 4.5%) repeatabilities were achieved. Detection limits of the preservatives were between 0.31 and 1.52 μg mL(-1) (S/N = 3, n = 5). The optimized method was successfully applied to the simultaneous determination of these preservatives in 10 commercial cosmetic products.


A modular molecular photovoltaic system based on phospholipid/alkanethiol hybrid bilayers: photocurrent generation and modulation.
Monolayer quantities of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), incorporated with either fullerenes or ruthenium tris(bipyridyl) (Ru(bpy)(3)(2+)) complexes, were formed on ferrocene-terminated C11-alkanethiol self-assembled monolayers (SAMs) through lipid fusion. Thus formed hybrid structures are characterized by quartz crystal microbalance, UV-vis spectroscopy, cyclic voltammetry and impedance analysis. In comparison to lipid monolayers deposited on C12-alkanethiol SAMs, photocurrent generation from these ferrocene-based structures is significantly modulated, displaying attenuated anodic photocurrents and enhanced cathodic photocurrents. While a similar trend was observed for the two photoagents studied, the degree of such modulations was always found to be greater in fullerene-incorporated bilayers. These findings are evaluated in the context of the film structure, energetics of the involved photo(electrochemical) species and cross-membrane electron-transfer processes.


Contrast-enhanced US of ampullary carcinoma: correlations with pathological findings.
To investigate the utility of contrast-enhanced transabdominal ultrasonography (CEUS) in the diagnosis of ampullary cancer (AC). In 12 patients with AC, the presence of tumor enhancement, histological type, amount of connective tissue in cancer, and tumor growth pattern were evaluated. The correlation between the tumor growth pattern and enhancement shape, that between the enhancement pattern and surrounding tissue, and the presence of pancreatic infiltration were evaluated. The depth of cancer invasion was mucosa in five patients, within Oddi's sphincter in four, beyond the duodenum but not to the pancreas in two, and pancreatic invasion less than 5 mm in one. The tumor was enhanced in 11 patients but not in the patient with much connective tissue in the tumor. Enhancement was observed in all patients with the intraluminal papillary or mixed type, but not in those with the periductal invasive type. The enhancement shape was predominantly round or irregular in the intraluminal papillary type and predominantly serrated in the mixed type. The enhancement shape differed among the growth patterns. The normal pancreatic parenchyma around the tumor was enhanced in all patients, and pancreatic infiltration was regarded as absent when there was a continuous unenhanced area between the tumor and pancreatic parenchyma. CEUS correlates with pathological findings of ampullary cancer.


Effect of GT-Peptide 10 and Triethyl Citrate on P. acnes Biofilm Formation, Viability, and Dispersion.
P. acnes biofilms are emerging topics in acne vulgaris pathogenesis and may be responsible for antibiotic tolerance.
 To investigate the efficacy of GT peptide 10 either alone or in combination with triethyl citrate (TEC) in in vitro model of P. acnes biofilm.
 Six-day-old P. acnes biofilms were treated with various concentrations of these substances and biofilm dispersion and cell viability were monitored.
 A 24-hour exposure of preformed biofilms to a combination of GT peptide 10/TEC led to killing of up to 92% of bacterial cells inside the biofilm. Neither the single substance nor the combination of both substances affected the biofilm integrity or resulted in biofilm dispersal.
 A combination of GT peptide 10/TEC shows antibacterial effects in in vitro model of P. acnes biofilm.

 J Drugs Dermatol. 2016;15(6):778-781.


SMAD7 and SMAD4 expression in colorectal cancer progression and therapy response.
Inhibitory SMAD7 and common mediator SMAD4 play crucial roles in SMAD-dependent TGF-β signaling that is often disrupted in colorectal cancer (CRC). This study aimed to profile the expression of SMAD7 and SMAD4 in primary and metastatic CRC and to evaluate their significance in disease progression and therapy response. The expression of SMAD7 and SMAD4 genes was analyzed by quantitative real-time PCR in tissues from 35 primary and metastatic CRC patients and in vitro in 7 human cell lines originating from colon tissue. Expression levels of SMAD7 and SMAD4, as well as their ratio, were determined and their association with tumor characteristics and response to therapy were evaluated. SMAD4 level was significantly lower in tumors compared to non-tumor tissues in both primary (p = 0.001) and metastatic (p = 0.001) CRC patients, while tumor expression of SMAD7 was significantly lower from non-tumor tissue only in metastatic patients (p = 0.017). SMAD7/SMAD4 ratio was elevated in CRC primary tumor tissues and cell lines compared to corresponding non-tumor tissues and cell line, respectively (p = 0.003). SMAD7 expression was significantly elevated in primary tumor tissues obtained from responders to neoadjuvant chemoradiotherapy (nCRT) compared to non-responders (p = 0.014). Alterations of expression and ratio of SMAD7 and SMAD4 in CRC cell lines, primary rectal cancer, and liver metastasis emphasize the importance of these genes in different stages of disease progression. Differential expression of SMAD7 in responders versus non-responders to nCRT should be further investigated for its potential predictive value.


Allyl-isothiocyanate against colorectal cancer via the mutual dependent regulation of p21 and Nrf2.
Allyl-isothiocyanate (AITC) is a common Isothiocyanates (ITC) and its chemo-preventive and anti-tumor effects are believed to be related to the activation of NF-E2 p45-related Factor 2 (Nrf2). However, its anti-tumor effects on colorectal cancer (CRC) are not well elucidated. Here, we investigated the therapeutic in vitro and/or in vivo effects and mechanisms of action (MOA) for AITC on CRC cell line HCT116 (human) and MC38 (mouse). AITC treatment in a low concentration range (1 mg/kg in vivo) significantly inhibited the tumor cell growth and increased the expression of p21 and Nrf2. The AITC-mediated induction of p21 was dependent on Nrf2 but independent on p53 in vitro and in vivo at low dose. In contrast, the high dose of AITC (5 mg/kg in vivo) failed to increase substantial levels of p21/MdmX, and impaired the total antioxidant capacity of tumors and subsequent anti-tumor effect in vivo. These results suggest that an optimal dose of AITC is important and required for the proper Nrf2 activation and its anti-CRC effects and thus, providing insights into the potential applications of AITC for the prevention and treatment of CRC.


Supramolecular organization of the yeast F1Fo-ATP synthase.
The yeast mitochondrial F(1)F(o)-ATP synthase is a large complex of 600 kDa that uses the proton electrochemical gradient generated by the respiratory chain to catalyse ATP synthesis from ADP and P(i). For a large range of organisms, it has been shown that mitochondrial ATP synthase adopts oligomeric structures. Moreover, several studies have suggested that a link exists between ATP synthase and mitochondrial morphology. In order to understand the link between ATP synthase oligomerization and mitochondrial morphology, more information is needed on the supramolecular organization of this enzyme within the inner mitochondrial membrane. We have conducted an electron microscopy study on wild-type yeast mitochondria at different levels of organization from spheroplast to isolated ATP synthase complex. Using electron tomography, freeze-fracture, negative staining and image processing, we show that cristae form a network of lamellae, on which ATP synthase dimers assemble in linear and regular arrays of oligomers. Our results shed new light on the supramolecular organization of the F(1)F(o)-ATP synthase and its potential role in mitochondrial morphology.


Atherosclerotic cardiovascular disease in women with endometriosis: a systematic review of risk factors and prospects for early surveillance.
Endometriosis and atherosclerotic cardiovascular disease (ASCVD) share similar pathogenic mechanisms. Hence, this systematic review evaluates the association between endometriosis and lifetime ASCVD risk including co-prevalence with dyslipidaemia, atherosclerosis and non-invasive markers of endothelial dysfunction. The electronic databases Embase, PubMed, MEDLINE, Cochrane Register of Trials and ClinicalTrials.gov were systematically searched for relevant articles. Two prospective cohort studies demonstrated an increased lifetime ASCVD risk after controlling for demographic and lifestyle confounders in women with endometriosis, as measured by higher incidence of myocardial infarction (relative risk [RR] 1.52), angiography-confirmed angina (RR 1.91), or requiring coronary artery bypass graft surgery (RR 1.35). Among 10 studies that included 407 patients with surgically proven endometriosis and 557 controls, RR of developing hypercholesterolemia and hypertension were 1.25 and 1.14, respectively, while higher serum lipoprotein a and lower paraoxonase 1 levels were found in women with endometriosis that was negatively correlated with stage of disease (r = -0.74, P < 0.0001). Hence, currently available evidence suggests that women with endometriosis are at higher lifetime risk of developing ASCVD than women without endometriosis. However, robust causal evidence is still lacking and future studies are needed to determine whether women with endometriosis represent a high-risk population for lifelong ASCVD risk.


Mcl-1 as a Therapeutic Target in Acute Myelogenous Leukemia (AML).
The B-cell lymphoma-2 (Bcl-2) family of proteins regulates the intrinsic, or mitochondrial pathway of apoptosis, the final common mechanism of cell death in response to a variety of physiologic and pharmacologic signals, and plays a central role in AML pathogenesis, prognosis and responsiveness to chemotherapy. Traditionally thought to be an important survival factor for multiple myeloma cells, the anti-apoptotic Bcl-2 family protein myeloid cell leukemia-1 (Mcl-1) has recently been shown in preclinical studies to be critical to the development and maintenance of AML, making it an attractive therapeutic target in this disease. Several characteristics, such as its very short half-life, distinguish Mcl-1 from other anti-apoptotic Bcl-2 family members. Additionally, Mcl-1 levels are regulated by a large number of pathways affecting its transcription, translation and degradation. A variety of approaches exploiting these features has been developed to inhibit directly or indirectly the anti-apoptotic function of Mcl-1. Many of these lend themselves well to combination therapies, leading to striking synergism, at least in preclinical models. In this brief review, we highlight some of the more promising strategies targeting Mcl-1 in AML, with a particular emphasis on rational combinations of novel agents.


Preparation of large numbers of plasmid DNA samples in microtiter plates by the alkaline lysis method.
A protocol is described for the growth and preparation of plasmid DNAs from small culture volumes (250 microliters) and utilizing standard 96-well plates. Several hundred plasmids can be prepared simultaneously, yielding sufficient DNA for subsequent analysis by restriction digestion and gel electrophoresis. This protocol may be useful for rapid screening of clones arising in recombinant DNA work such as site-directed mutagenesis, oligonucleotide cassette cloning, deletion analysis, etc. The technique was initially developed to meet our requirement to provide large numbers of cosmid DNAs for restriction enzyme fingerprint analyses in genome mapping projects.