Functional analyses of melanocortin-4 receptor mutations identified from patients with binge eating disorder and nonobese or obese subjects.
Whether mutations in the melanocortin-4 receptor (MC4R) are the cause of binge eating disorder was controversial. In addition, the penetrance of mutations in the MC4R in causing obesity was debated. We investigated whether MC4R variants identified from obese patients with binge eating disorder (T11A, F51L, T112M, and M200V) and variants identified in nonobese (I102T, F202L, and N240S) or obese (I102S, A154D, and S295P) subjects cause loss-of-function and what are the defects. Variant or wild-type MC4Rs were expressed in HEK293 cells and examined for their pharmacological characteristics. The study setting was in vitro bench-top laboratory experiments. Ligand binding, signaling, and cell surface expression of the variant MC4Rs were compared with wild-type MC4R. Our data clearly show a loss-of-function phenotype in vitro for I102T and N240S variants identified in nonobese individuals. Furthermore, not all MC4R variants identified in obese subjects exhibit a loss-of-function phenotype in vitro. Finally, the MC4R variants T11A, F51L, T112M, and M200V identified from patients with binge eating disorder displayed normal function with regards to the parameters measured in our study. Patients harboring loss-of-function MC4R mutations do not always exhibit obesity. Novel MC4R variant identified from an obese patient cannot be assumed to be the cause of obesity without demonstrating a loss-of-function phenotype in vitro for the variant MC4R. Whether MC4R mutations are involved in the pathogenesis of binge eating disorder needs additional investigation.


Erythropoietin-coated ZP-microneedle transdermal system: preclinical formulation, stability, and delivery.
To evaluate the feasibility of coating formulated recombinant human erythropoietin alfa (EPO) on a titanium microneedle transdermal delivery system, ZP-EPO, and assess preclinical patch delivery performance. Formulation rheology and surface activity were assessed by viscometry and contact angle measurement. EPO liquid formulation was coated onto titanium microneedles by dip-coating and drying. Stability of coated EPO was assessed by SEC-HPLC, CZE and potency assay. Preclinical in vivo delivery and pharmacokinetic studies were conducted in rats with EPO-coated microneedle patches and compared to subcutaneous EPO injection. Studies demonstrated successful EPO formulation development and coating on microneedle arrays. ZP-EPO patch was stable at 25°C for at least 3 months with no significant change in % aggregates, isoforms, or potency. Preclinical studies in rats showed the ZP-EPO microneedle patches, coated with 750 IU to 22,000 IU, delivered with high efficiency (75-90%) with a linear dose response. PK profile was similar to subcutaneous injection of commercial EPO. Results suggest transdermal microneedle patch delivery of EPO is feasible and may offer an efficient, dose-adjustable, patient-friendly alternative to current intravenous or subcutaneous routes of administration.


Fasting state and episodes of vomiting in children receiving nitrous oxide for dental treatment.
The purpose of this controlled-crossover study was to determine the frequency of vomiting during nitrous oxide / oxygen analgesia (NOA) and assess the relationship between fasting status and vomiting. One hundred and thirteen children (64 male, 49 female), ranging in age from 24-160 months (mean = 74) and a mean weight of 23 kg (range 11-60 kg), participated in the study. At the initial examination, subjects were randomly assigned to be either fasting on the first appointment and non-fasting during the second appointment or alternatively be non-fasting for the first appointment and fasting for the second. The average time interval between eating and treatment in the fasting sessions was 6 hours and in the non-fasting group, 1 hour before treatment. Vomiting occurred in only one subject, immediately after cessation of treatment resulting in a frequency of 1% of subjects or 0.5% of sessions. No other differences were found between fasting and non-fasting subjects. During dental treatment with NOA using the rapid induction method, constant nonfluctuating concentration/ flow, and treatment time of under 35 minutes, the frequency of vomiting during NOA was found to be 0.5%.


Long-term persistence of anti-rods and rings antibodies in patients with chronic hepatitis C after antiviral treatment.
Anti-rods and rings (anti-RR) antibodies are related to hepatitis C virus (HCV) in patients treated with pegylated interferon (PEG-IFN) and ribavirin (RBV). Only RBV induces rods/rings structures in vitro; but in vivo, the antibody appearance is related to the combination of these drugs, because data about patients using just one of these drugs alone is missing. Some studies suggest disappearance of these antibodies over time. The aim of this study was to describe the occurrence of anti-RR in patients with chronic hepatitis C treatment-naïve or previously PEG-IFN/RBV-experienced, evaluating the persistence of anti-RR antibodies long after PEG-IFN/RBV treatment. From 2016 to 2017, 70 HCV-infected patients were screened for anti-RR using indirect immunofluorescence. Demographic and clinical data about previous treatments against HCV were assessed. Thirty-four patients (49%) had been previously treated with PEG-IFN/RBV and the average time since they had received the last antiviral treatment was 85.4 months. Anti-RR seropositivity was detected in 16 patients (23%), and all of these had used PEG-IFN/RBV (corresponding to 47% of experienced patients). Previous antiviral treatment and previous exposure time to RBV were associated with anti-RR positivity. Median time elapsed since last treatment was similar between anti-RR-positive and anti-RR-negative patients. Anti-RR seropositivity was not observed in treatment-naïve patients, but was detected in almost half of patients previously treated with PEG-IFN and RBV, even after a long period without exposure to these drugs. This antibody was related to extended prior exposure to ribavirin.


Morphology and morphometry of the human embryonic brain: A three-dimensional analysis.
The three-dimensional dynamics and morphology of the human embryonic brain have not been previously analyzed using modern imaging techniques. The morphogenesis of the cerebral vesicles and ventricles was analyzed using images derived from human embryo specimens from the Kyoto Collection, which were acquired with a magnetic resonance microscope equipped with a 2.35-T superconducting magnet. A total of 101 embryos between Carnegie stages (CS) 13 and 23, without apparent morphological damage or torsion in the brain ventricles and axes, were studied. To estimate the uneven development of the cerebral vesicles, the volumes of the whole embryo and brain, prosencephalon, mesencephalon, and rhombencephalon with their respective ventricles were measured using image analyzing Amira™ software. The brain volume, excluding the ventricles (brain tissue), was 1.15 ± 0.43 mm(3) (mean ± SD) at CS13 and increased exponentially to 189.10 ± 36.91 mm(3) at CS23, a 164.4-fold increase, which is consistent with the observed morphological changes. The mean volume of the prosencephalon was 0.26 ± 0.15 mm(3) at CS13. The volume increased exponentially until CS23, when it reached 110.99 ± 27.58 mm(3). The mean volumes of the mesencephalon and rhombencephalon were 0.20 ± 0.07 mm(3) and 0.69 ± 0.23 mm(3) at CS13, respectively; the volumes reached 21.86 ± 3.30 mm(3) and 56.45 ± 7.64 mm(3) at CS23, respectively. The ratio of the cerebellum to the rhombencephalon was approximately 7.2% at CS20, and increased to 12.8% at CS23. The ratio of the volume of the cerebral vesicles to that of the whole embryo remained nearly constant between CS15 and CS23 (11.6-15.5%). The non-uniform thickness of the brain tissue during development, which may indicate the differentiation of the brain, was visualized with surface color mapping by thickness. At CS23, the basal regions of the prosencephalon and rhombencephalon were thicker than the corresponding dorsal regions. The brain was further studied by the serial digital subtraction of layers of tissue from both the external and internal surfaces to visualize the core region (COR) of the thickening brain tissue. The COR, associated with the development of nuclei, became apparent after CS16; this was particularly visible in the prosencephalon. The anatomical positions of the COR were mostly consistent with the formation of the basal ganglia, thalamus, and pyramidal tract. This was confirmed through comparisons with serial histological sections of the human embryonic brain. The approach used in this study may be suitable as a convenient alternative method for estimating the development and differentiation of the neural ganglia and tracts. These findings contribute to a better understanding of brain and cerebral ventricle development.


HCV NS4B induces apoptosis through the mitochondrial death pathway.
The hepatitis C virus (HCV) NS4B protein is known to induce the formation of a membranous web that is thought to be the site of viral RNA replication. However, the exact functions of NS4B remain poorly characterized. In this study, we found that NS4B induced apoptosis in 293T cells and Huh7 cells, as confirmed by Hoechst staining, DNA fragmentation, and annexin V/PI assays. Furthermore, protein immunoblot analysis demonstrated that NS4B triggered the cleavage of caspase 3, caspase 7, and poly(ADP-ribose) polymerase (PARP). Further studies revealed that NS4B induced the activation of caspase 9, the reduction of mitochondrial membrane potential and the release of cytochrome c from the mitochondria. However, NS4B expression did not trigger XBP1 mRNA splicing and increase the expression of binding immunoglobulin protein (BiP, or GRP78) and C/EBP homologous protein (CHOP), which serves as the indicators of ER stress. Taken together, our results suggest that HCV NS4B induces apoptosis through the mitochondrial death pathway.


Mandarin Chinese vowel-plus-tone identification in noise: Effects of language experience.
Several studies found better English vowel identification in English multi-talker babble (MTB) and temporally-modulated (TM) noise, but not in quiet condition for native Chinese listeners in the US (CNU) with the US residency of 1-3 years than native Chinese listeners in China (CNC) with no residency history in English speaking countries. Two possible explanations were proposed: (1) CNU listeners used temporal dips of noise more efficiently than CNC listeners; and (2) CNU listeners had less informational masking of MTB than their CNC peers. The current study explored whether the difference in noise processing between CNU and CNC listeners was also presented for their native speech perception. Chinese vowel-plus-tone identifications were measured for CNU and CNC in quiet, stationary and TM noise, babble-modulated noise, and MTB. The identification scores of CNU listeners were significantly higher than CNC listeners in most noisy backgrounds, whereas both groups had the same performance in quiet. Moreover, compared with CNC listeners, CNU listeners gained greater masking releases from the temporal modulation in TM noise at low SNRs, whereas no significant difference was found in informational masking between the two groups. In conclusion, the native English experience may improve native Chinese listeners' capacity to use temporal glimpses in TM noise, possibly depending on the modulation frequency and depth, while it may not improve their ability to resist the informational masking of babble when perceiving their native speech.


Eating disorders.
Anorexia nervosa and bulimia nervosa present special challenges for physicians. Considerable skill and flexibility, as well as a collaborative spirit, are required to address the important physical, psychological, and social dimensions of these eating disorders. The ideal treatment is an individualized program designed to (1) eliminate the multifaceted effects of a body weight driven well below the biologic set point, (2) develop self-esteem, including a healthy attitude toward the body, and (3) strengthen those aspects of the individual and the family that orient the patient to health instead of illness.


Clinical presentation of infective endocarditis caused by different groups of non-beta haemolytic streptococci.
Streptococci are common causes of infective endocarditis (IE) and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has provided a practical tool for their species determination. We aimed to investigate if particular groups of non-beta heamolytic streptococci were associated with IE or to specific presentations thereof. The Swedish Registry of Infective Endocarditis was used to identify cases of IE caused by streptococci and a local database to identify cases of streptococcal bacteremia. The bacteria were grouped using MALDI-TOF MS and the clinical characteristics of IE caused by different groups were compared. We identified a group of 201 streptococcal IE isolates: 18 isolates belonged to the anginosus, 19 to the bovis, 140 to the mitis, 17 to the mutans, and seven to the salivarius groups. The mitis and mutans groups were significantly more common and the anginosus group less common among IE cases as compared to all cause bacteremia. Patients infected with the bovis group isolates were older, had more cardiac devices, and had more commonly prosthetic valve IE compared to IE caused by streptococci of the other groups. Twenty-one percent of patients needed surgery, and in-hospital mortality was 8% with no significant differences between the groups. Grouping of non-beta haemolytic streptococci using MALDI-TOF MS can provide a basis for decision-making in streptococcal bacteremia. IE caused by bovis group isolates have clinical characteristics distinguishing them from IE caused by other groups of Streptococcus.


Mental Health Screening of Medically-Admitted Patients With Cystic Fibrosis.
Multi-national studies have identified an increased risk for depression and anxiety among the cystic fibrosis population. People with cystic fibrosis and depression have decreased lung function, adherence, and quality of life, and increased health care utilization. This is a pilot study of mental health screening and referral of patients with cystic fibrosis in a large tertiary medical center. Patients with a diagnosis of cystic fibrosis aged 8 and older, medically admitted to a tertiary hospital, were screened for eligibility and offered mental health screening for depression and anxiety. Patients indicating elevated rates of anxiety, depression, or suicidal ideation were offered a psychiatric consultation, and all participants were offered mental health referrals. Health-related outcomes were gathered via medical record review. The pediatric population showed elevated rates/at risk of depression (17%), anxiety (22%) and clinically-elevated depression (5%), and anxiety (11%). Twenty-two percent of the youth reported suicidal ideation. The adult population reported mild rates of depression (11%), anxiety (28%), and suicidality (11%). The mental health screening process resulted in 1 mental health referral, 16 patients eligible for psychiatric consultation, and 4 completed psychiatric consultations. This study represents a pilot mental health screening in the inpatient medical setting. The results indicate an elevated rate of depression, anxiety, and suicidal ideation, and a protocol for responding to elevated responses via psychiatric consultation. This study indicates the need for further exploration of implementation of mental health screening, rapid response to suicidal ideation, referral process, and treatment interventions.


The association between nonsteroidal anti-inflammatory drugs and skin cancer: Different responses in American and European populations.
To conduct a comprehensive systematic meta-analysis investigating the association of nonsteroidal anti-inflammatory drugs (NSAIDs) and their subtypes with skin cancer (SC) and its subclasses (basal cell carcinoma BCC; squamous cell carcinoma SCC; melanoma; nonmelanoma skin cancer NMSC) in general, American and European populations. PubMed, Embase, the Cochrane Library, the China National Knowledge Infrastructure and ClinicalTrials.gov were searched up to 24 February 2019. Pooled effect sizes and 95% confidence intervals were used to estimate associations. Results based on 26 original studies including 223,619 cases and 1,398,507 controls showed both NSAIDs and nonselective Cyclooxygenase (COX) inhibitors to be statistically significantly associated with a reduced risk of SC, BCC, SCC and NMSC but not with melanoma. Conversely, no association was observed between selective Cyclooxygenase 2 (COX-2) inhibitors and SC or its subclasses. Further subgroup analysis showed that the results analyzed for American populations were almost the same as those for the general population. For European populations, neither NSAIDs nor its subtypes correlated significantly with susceptibility to SC or its subclasses. The use of NSAIDs might reduce the risk of SC, but many factors including study population, drug subtype, and disease subclass affect the significance of the association.


Associations between childhood trauma and childhood psychiatric disorders in Brazil: a population-based, prospective birth cohort study.
Childhood trauma is a proposed transdiagnostic risk factor for psychopathology, but epidemiological evidence from low-income and middle-income countries (LMICs) is scarce. We investigated associations between trauma and child psychiatric disorders in a birth cohort in Brazil. The 2004 Pelotas Birth Cohort is an ongoing, population-based, prospective birth cohort, including all hospital births occurring between Jan 1 and Dec 31, 2004, in the city of Pelotas, Brazil. When the children were aged 6 and 11 years, trained psychologists administered the Development and Well-Being Assessment clinical interview to caregivers to assess current child psychiatric disorders (anxiety disorders, mood disorders, ADHD and hyperactivity disorders, and conduct and oppositional disorders), and lifetime trauma exposure (ie, experiencing or witnessing life-threatening events) including interpersonal and non-interpersonal events. Analyses used multiple imputation and logistic regression models. Of 4263 live births, 4231 children were included in the study sample, and 4229 (2195 [51·9%] boys and 2034 [48·1%] girls; 2581 [61·7%] with White mothers and 1600 [38·3%] with Black or mixed race mothers) were included in the imputed analyses. 1154 (34·3%) of 3367 children with complete data at age 11 years had been exposed to trauma by that age. After adjusting for confounders, at age 6 years, trauma was associated with increased odds of anxiety disorders (adjusted odds ratio 1·79 [95% CI 1·33-2·42]) and any psychiatric disorder (1·59 [1·22-2·06]), and at age 11 years, with any psychiatric disorder (1·45 [1·17-1·79]) and all four specific diagnostic classes of anxiety disorders (1·47 [1·04-2·09]), mood disorders (1·66 [1·08-2·55]), ADHD and hyperactivity disorders (1·47 [1·01-2·13]), and conduct and oppositional disorders (1·76 [1·19-2·61]). Interpersonal trauma and non-interpersonal trauma were each associated with increased odds of multiple psychiatric disorders, even when adjusting for their co-occurrence. A considerable mental health burden associated with childhood trauma is already evident by middle childhood in this sample from Brazil. Evidence-based efforts to reduce the incidence of childhood trauma in Brazil and address its consequences are urgently needed. Children's Pastorate, WHO, National Support Program for Centres of Excellence, Brazilian National Research Council, Brazilian Ministry of Health, São Paulo Research Foundation, University of Bath, Wellcome Trust. For the Portuguese translation of the abstract see Supplementary Materials section.


An early loaded implant-supported mandibular complete arch fixed prosthesis in a young completely edentulous patient: a case report.
The restoration of the mandibular arch up to the first molars with 5 one-piece implants presents a viable and cost-effective treatment plan in patients with adequate bone volume and favorable ridge relationships. The early loading of the implants with a provisional restoration reduces the period of edentulousness and restores the patient's ability to eat, talk, and smile effectively. Implants in younger patients prevent resorption of the residual alveolar ridge. This article presents a case report of the restoration of an edentulous mandibular arch with an early loaded implant-supported fixed restoration in a young patient.


[The value of erythrocyte sedimentation rate and C-reactive protein in evaluating disease activity in ankylosing spondylitis].
To determine whether erythrocyte sedimentation rate (ESR) or C-reactive protein (CRP) is more appropriate in measuring the disease activity in ankylosing spondylitis (AS). We studied 126 consecutive patients with AS. The external criteria for disease activity were Cowling clinical assessment of disease activity and the Bath AS disease activity index (BASDAI). In each measure we defined 3 levels of disease activity i.e. no activity, ambiguous activity and definite activity. The patients with AS were divided into 2 groups: those with spinal involvement only and those with peripheral arthritis as well. For each criterion of disease activity, the patients without activity and with definite activity were included in receiver operating characteristic curve, which was used to determine cutoff values with the highest sensitivity and specificity. We also calculated Spearman correlation. The median ESR and CRP were 25.3 mm/1 h and 11.1 mg/L in the spinal group and 30.0 mm/1 h and 15.0 mg/L in the peripheral group. In both groups the Spearman correlation coefficients between ESR and CRP were around 0.30. There was no correlation between ESR, CRP, and the 2 disease activity variables (0.027-0.282). Sensitivity for both ESR and CRP was between 39.4 %and 81.3% for Cowling assessment of disease activity and the BASDAI, while specificity was between 40.0% and 86.7% for all disease activity measures. It is concluded that neither ESR nor CRP is superior for assessing disease activity in ankylosing spondylitis.


Effect of breed and management practices on reproductive and milking performance of rangeland goats.
The traditional goat production systems on arid and semi-arid rangelands of northern Mexico are characterized by minimum external inputs; therefore, little is known about goat response to technological improvements. This observational study aimed to ascertain the effect of using improved goat production technologies and breed on milk yield and reproductive performance of goats on rangeland. The study was conducted from January 2017 to June 2018. Thirty-seven goat herds comprising 6393 animals were used. Goats supplemented with 250 g of concentrate (S-goats; 14% crude protein) 30 days before breeding produced significantly more daily milk yield (218 ± 61 vs. 200 ± 60 g) than the unsupplemented (UNS-goats) goats. Milk production per lactation was 12 percentage points greater in goats with access to plain salt throughout the year than those without salt access. The absence of deworming significantly depressed daily milk yield (189 ± 55 vs. 221 ± 61 for non-treated and treated goats). In 5 months, the lowest total milk yield was for Boer goats (26.9 ± 8.3 kg) and the highest for Nubian goats (36.1 ± 9.2 kg). The kidding rate was significantly higher in the S-goats than in the UNS-goats group (66.9 vs. 62.8%). Salt-supplemented goats had a significantly greater kidding rate than goats not receiving salt (68.5 vs. 61.2%). The lowest kidding rate was for Saanen goats and the highest for Nubian goats (66.2%). These findings indicate that milk yield and reproductive performance were primarily influenced by supplementing grazing goats with concentrates and salt in this semi-arid rangeland.


SARS-CoV-2: a new dimension to our understanding of coronaviruses.
Coronaviruses have marked their significant emergence since the twenty-first century with the outbreaks of three out of the seven existing human coronaviruses, including the severe acute respiratory syndrome coronavirus (SARS-CoV) in 2003, Middle East respiratory syndrome coronavirus (MERS-CoV) in 2012, and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in 2019. These viruses have not only acquired large-scale transmission during their specified outbreak period, but cases of MERS-CoV still remain active, although there is only limited transmission. While, on the other hand, SARS-CoV-2 continues to remain a rising threat to global public health. The recent novel coronavirus, SARS-CoV-2, responsible for the ongoing coronavirus disease 2019 (COVID-19), emerged during December 2019 in Wuhan, China, and has repeatedly raised questions about its characteristic variability. Despite belonging to the same family, SARS-CoV-2 has proven to be quite difficult to control and contain in terms of transmissibility, leading to around 19.8 million reported cases and more than 730,000 deaths of individuals worldwide. Here, we discuss how SARS-CoV-2 differs from its two other related human coronaviruses in terms of genome composition, site of infection, and transmissibility, among several other notable aspects-all indicating to the possibility that it is these variations in addition to other unknowns that are contributing to this virus' differing deadly pattern.


Low-dose every-second-day LHRH treatment following bilateral orchidopexy in children with bilateral cryptorchidism may improve their fertility outcome.
Currently the standard treatment for bilateral cryptorchidism is bilateral surgical orchidopexy. Whether a hormonal treatment should be routinely administered postoperatively to increase fertility is debatable. Low-dose postoperative luteinizing hormone releasing hormone (LHRH) can increase spermatogonial numbers, but the effect of native LHRH (Kryptocur®) on adult fertility is unclear. To determine if low-dose every-second-day postoperative LHRH administration in children with bilateral cryptorchidism improves fertility in adulthood and if Nistal testicular histological grading could guide the decision to administer LHRH. All patients, actually at least 16yr of age, that underwent a bilateral orchidolysis and orchidopexy for bilateral cryptorchidism (surgery between 1997 and 2018) were contacted and offered a clinical exam, hormone levels, sperm analysis, and a scrotal ultrasound. At the original surgery, testicular biopsy was performed (if 60% of the tubuli contain >1 spermatogonia, this is normal = Nistal-1, if 30-60% filled = Nistal-2, if <30% = Nistal-3 and if Sertoli only = Nistal-4) and if in at least one testis impaired. A low dose native LHRH treatment was offered to the patients, as this treatment is known to increase the number of spermatogonia in a short term. Kryptocur® (LHRH, Gonadorelin, Hoechst®) was prescribed and dosed at 200 μg (one spray in one nostril) every other day for 6-8 months. Forty-two men were eligible for this study. 20/42 accepted the invitation for a clinical and hormonal evaluation. 16/20 men accepted the invitation for an additional sperm analysis. Fourteen of 20 men received low-dose LHRH postoperatively in a nonrandomized manner. Three men had Nistal grade 1, eight grade 2, seven grade 3, and two had grade 4. Inhibin B levels were higher in men with Nistal 1 and 2 compared with Nistal 3 and 4 P ≤ 0.037). Severe oligospermia/azoospermia (<1 × 106/ejaculate) was observed in 33% of the treated group vs 67% of the untreated group (P ≤ 0.036.) DISCUSSION AND CONCLUSIONS: Low-dose every-second-day postoperative LHRH treatment improves fertility outcome in bilateral cryptorchidism. Histological analysis of prepubertal testes according to Nistal grading cannot be used as a predictive diagnostic test for LHRH treatment.


Autogenous breast reconstruction with the deep inferior epigastric perforator flap.
Muscle-sparing autogenous breast reconstruction has enhanced the multidisciplinary care that is available to patients who have breast cancer. The DIEP flap has proven reliability, a low complication rate, and is applicable to many clinical scenarios (Figs. 8-12). Avoidance of muscle sacrifice in the abdomen ultimately translates into greater patient satisfaction. The increased demands, in terms of surgical expertise, are more than offset by decreased postoperative pain and decreased donor site morbidity. The methods that were used to innovate the DIEP flap have been applied to other donor sites and the available options for patients have been expanded.


LU 73068, a new non-NMDA and glycine/NMDA receptor antagonist: pharmacological characterization and comparison with NBQX and L-701,324 in the kindling model of epilepsy.
The aim of this study was to assess whether a drug which combines an antagonistic action at both NMDA and non-NMDA receptors offers advantages for treatment of epileptic seizures compared to drugs which antagonize only one of these ionotropic glutamate receptors. The novel glutamate receptor antagonist LU 73068 (4,5-dihydro-1-methyl-4-oxo-7-trifluoromethylimidazo[1,2a]quinoxal ine-2-carbonic acid) binds with high affinity to both the glycine site of the NMDA receptor (Ki 185 nM) and to the AMPA receptor (Ki 158 nM). Furthermore, binding experiments with recombinant kainate receptor subunits showed that LU 73068 binds to several of these subunits, particularly to rGluR7 (Ki 104 nM) and rGluR5 (Ki 271 nM). In comparison, the prototype non-NMDA receptor antagonist NBQX (2,3-dihydroxy-6-nitro-7-sulphamoyl-benzo[f]quinoxaline) binds with high affinity to AMPA receptors only. Both NBQX and LU 73068 were about equieffective after i.p. injection in mice to block lethal convulsions induced by AMPA or NMDA. In the rat amygdala kindling model of temporal lobe epilepsy, LU 73068 dose-dependently increased the focal seizure threshold (afterdischarge threshold, ADT). When rats were stimulated with a current 20% above the individual control ADT, LU 73068 completely blocked seizures with an ED50 of 4.9 mg kg(-1). Up to 20 mg kg(-1), only moderate adverse effects, e.g. slight ataxia, were observed. NBQX, 10 mg kg(-1), and the glycine/NMDA site antagonist L-701,324 (7-chloro-4-hydroxy-3-(3-phenoxy)phenyl-quinoline-2(1H)one), 2.5 or 5 mg kg(-1), exerted no anticonvulsant effects in kindled rats when administered alone, but combined treatment with both drugs resulted in a significant ADT increase. The data indicate that combination of glycine/NMDA and non-NMDA receptor antagonism in a single drug is an effective means of developing a potent and effective anticonvulsant agent.


Evaluation of crossed cerebellar diaschisis in 30 patients with major cerebral artery occlusion by means of quantitative I-123 IMP SPECT.
Quantitative crossed cerebellar diaschisis (CCD) and the correlation with a reduction in supratentorial regional cerebral blood flow (rCBF) and cerebrovascular reserve capacity (CVR) were investigated in clinically stable patients with major cerebral artery occlusion by the iodine-123-N-isopropyl-p-iodoamphetamine (I-123 IMP) single photon emission computed tomography (SPECT) method. Thirty patients with major cerebral artery occlusion underwent SPECT by the I-123 IMP autoradiographic method. Regional CBF was measured in the cerebral hemisphere, frontal and parietal lobes, temporo-parietal lobe, and cerebellum both at rest and after administration of acetazolamide. Eighteen of 30 patients (60%) had CCD. CCD was significantly related to magnetic resonance imaging evidence of infarction. Quantitative CCD was 17% and the CVR in the cerebellum was preserved in patients with CCD. There was a significant difference in CBF and CVR between the affected and normal sides in all regions of interest in the patients without CCD [CBF (ml/100 g/min): hemisphere (H), normal side (N): 31.4 +/- 6.8, affected side (A): 27.5 +/- 7.4; p < 0.05. CVR: H, N: 0.56 +/- 0.38, A: 0.42 +/- 0.18; p < 0.01]. CCD is common in patients with major cerebral artery occlusion, and quantitative I-123 IMP SPECT is helpful in detecting CCD in clinically stable patients with occlusion of major cerebral arteries.


Colocalization of neutrophils, extracellular DNA and coagulation factors during NETosis: Development and utility of an immunofluorescence-based microscopy platform.
Neutrophils, the most populous innate immune cell type, are the first responders to sites of infection and inflammation. Neutrophils can release their DNA to form extracellular traps (NETs), webs of DNA and granular proteases that contribute to pathogen clearance and promote thrombus formation. At present, the study of NETs is in part limited to the qualitative analysis of fluorescence microscopy-based images, thus quantification of the interactions between NETs and coagulation factors remains ill-defined. Develop a quantitative method to measure the spatial distribution of DNA and colocalization of coagulation factor binding to neutrophils and NETs utilizing fluorescence-based microscopy. Human neutrophils were purified from peripheral blood, bound to fibronectin and treated with the PKC-activator phorbol myristate acetate (PMA) to induce neutrophil activation and NETs formation. Samples were incubated with purified coagulation factors or plasma before staining with a DNA-binding dye and coagulation factor-specific antibodies. The spatial distribution of DNA and coagulation factors was imaged via fluorescence microscopy and quantified via a custom-built MATLAB-based image analysis algorithm. The algorithm first established global thresholding parameters on a training set of fluorescence image data and then systematically quantified intensity profiles across treatment conditions. Quantitative comparison of treatment conditions was enabled through the normalization of fluorescent intensities using the number of cells per image to determine the percent and area of DNA and coagulation factor binding per cell. Upon stimulation with PMA, NETs formation resulted in an increase in the area of DNA per cell. The coagulation factor fibrinogen bound to both the neutrophil cell body as well as NETs, while prothrombin, FX and FVIIa binding was restricted to the neutrophil cell body. The Gla domain of FX was required to mediate FX-neutrophil binding. Activated protein C (APC), but not Gla-less APC, bound to neutrophil cell bodies and NETs in a punctate manner. Neither FXIIa nor FXIa were found to bind either neutrophil cell bodies or NETs. Fibrinogen binding was dependent on extracellular DNA, while FX and APC required phosphatidylserine exposure for binding to activated neutrophils. We have developed a quantitative measurement platform to define the spatial localization of fluorescently-labeled coagulation factor binding to neutrophils and extracellular DNA during NETosis.


FAK is required for the assembly of podosome rosettes.
Podosomes are dynamic actin-enriched membrane structures that play an important role in invasive cell motility and extracellular matrix degradation. They are often found to assemble into large rosettelike structures in highly invasive cells. However, the mechanism of this assembly remains obscure. In this study, we identified focal adhesion kinase (FAK) as a key molecule necessary for assembly. Moreover, phosphorylation of p130Cas and suppression of Rho signaling by FAK were found to be important for FAK to induce the assembly of podosome rosettes. Finally, we found that suppression of vimentin intermediate filaments by FAK facilitates the assembly of podosome rosettes. Collectively, our results strongly suggest a link between FAK, podosome rosettes, and tumor invasion and unveil a negative role for Rho signaling and vimentin filaments in podosome rosette assembly.


Psychotherapy and the Mormon faith.
The Church of Jesus Christ of Latter-day Saints, also known as the Mormon Church, is a Christian faith with a large presence across the globe. Although Mormon doctrine suggests that faith in Jesus allows people to overcome weakness and heal from pain, Mormon people are not immune from experiencing periods of mental and emotional suffering. The deeply held religious beliefs of Mormons can influence the nature of the psychological difficulties a Mormon individual is prone to experiencing, how and when they choose to seek treatment, as well as the types of treatment that may be most beneficial.


Nutritional intake from food sources of high school football athletes.
Male high school football athletes served as subjects (no. = 134; age = 15.0 +/- 1.2 years, range = 12 to 18 years; weight = 67.7 +/- 13.9 kg) (mean +/- standard deviation) in a dietary survey project to characterize nutritional intake from food sources. Dietary intake data were collected using the recall method for 1 weekday during the summer when athletes participated in unsupervised, informal conditioning. Subjects were grouped by age as junior high (JR-HI: 12 to 14 years) or senior high (SR-HI: 15 to 18 years) students. Absolute mean energy and nutrient intakes, except for vitamin A, were statistically greater for SR-HI relative to JR-HI (p less than .02). These findings are consistent with age-related growth on nutritional intake. Mean nutritional intakes from food sources for SR-HI met or exceeded the RDAs. For JR-HI, mean intakes met or exceeded the RDAs except for energy (94% RDA) and zinc (87% RDA). Mean intakes exceeded those of a representative sample of same-age boys in the larger American population.


[Electron microscope observations on the microfibrillar arrangement of collagen fibers in the tail tendon of the rat].
We have examined the rat tail tendon microfibrillar arrangement by electron microscopy and freeze-etching technique. Collagen fibers have been examined in unstretched and stretched conditions. Our data indicate the freeze-etching technique as a particularly useful to interpretate the microfibrillar arrangement. We are able to observe that the microfibrils are arranged in straight lines along the axis of the fibrils. The most suitable condition for the interpretation of this datum is obtainable when: 1) the fibers are stretched and therefore the crimping are reduced; 2) the fracture movement follows a direction parallel to the bulk fiber axis.


Electrochemically switchable cucurbit[7]uril-based pseudorotaxanes.
[Structure: see text] The binding location of cucurbit[7]uril can be controlled via redox conversions in these novel, structurally simple pseudorotaxanes.


COVID-19 control accelerates the decline in illegal drug use in China.
This work illustrates the accelerated decline in illegal drug use during the COVID-19 pandemic in China. We first reviewed the global effects of the COVID-19 pandemic on the situation of illegal drugs. We then compared the data of the pre-pandemic (2016-2019) and pandemic period (2020-2022) for drug seizures, individuals identified as using drugs, registered and newly discovered illegal drug users and the number of community-treated drug users to demonstrate the fast decline in the use of illegal drugs in China. We then discussed the possible reasons and additional considerations for these changes. Overall, the COVID-19 controls in China, such as all-staff nucleic acid testing and ID-based dynamic monitoring, substantially reduced illegal drug use. Being wary of a possible rebound in drug use and preventing new types of drug crimes are still essential in post-COVID China.


Effects of different implant-abutment connections on micromotion and stress distribution: prediction of microgap formation.
The aim of this study was to analyse micromotion and stress distribution at the connections of implants and four types of abutments: internal hexagonal, internal octagonal, internal conical and trilobe. A three dimensional (3D) model of the left posterior mandible was reconstructed from medical datasets. Four dental implant systems were designed and analysed independently in a virtual simulation of a first molar replacement. Material properties, contact properties, physiological loading and boundary conditions were assigned to the 3D model. Statistical analysis was performed using one-way analysis of variance (ANOVA) with a 95% confidence interval and Tukey's Honestly Significant Difference (HSD) multiple comparison test. The internal hexagonal and octagonal abutments produced similar patterns of micromotion and stress distribution due to their regular polygonal design. The internal conical abutment produced the highest magnitude of micromotion, whereas the trilobe connection showed the lowest magnitude of micromotion due to its polygonal profile. Non-cylindrical abutments provided a stable locking mechanism that reduced micromotion, and therefore reduced the occurrence of microgaps. However, stress tends to concentrate at the vertices of abutments, which could lead to microfractures and subsequent microgap formation.


Entamoeba histolytica cysteine proteinases with interleukin-1 beta converting enzyme (ICE) activity cause intestinal inflammation and tissue damage in amoebiasis.
The protozoan parasite Entamoeba histolytica causes intestinal inflammation and ulceration. Amoebic trophozoites activate the transcription factor NF-kappa B in human intestinal epithelial cells, initiating an inflammatory response programme with resultant damage to the intestinal tissue. Amoebic cysteine proteinases have been proposed as important virulence factors for amoebiasis. To test the role of amoebic cysteine proteinases in the pathogenesis of amoebic colitis, human intestinal xenografts in SCID mice were infected with E. histolytica trophozoites expressing an antisense message to ehcp5. The cysteine proteinase-deficient amoeba failed to induce intestinal epithelial cell production of the inflammatory cytokines interleukin (IL)-1B and IL-8, and caused significantly less gut inflammation and damage to the intestinal permeability barrier. The critical role of amoebic cysteine proteinases in human gut inflammation and tissue damage may be explained by our discovery that amoebic cysteine proteinases possess IL-1B converting enzyme (ICE) activity. This ICE activity could contribute to intestinal inflammation by activating human pIL-1B released by damaged intestinal cells. These results demonstrate for the first time that amoebic cysteine proteinases are a key virulence factor in amoebic colitis, and provide a novel mechanism for their activity.


Implementing a Food is Medicine Program to Address Food Insecurity in an Academic Medical Center.
Food insecurity is a public, social, and health concern. A Food is Medicine Program was developed to address food insecurity. A quality improvement initiative was piloted on 3 acute care units. Patients were screened for Social Determinant of Health (SDoH) needs and if identified as food insecure, linked to community resources and provided with a bag of food on discharge. Education was offered to nursing staff and a pre- and postsurvey was administered to assess SDoH knowledge and confidence. Over a 3-month period, 2354 patients were admitted; 2063 (88%) were screened for SDoH and 220 (10%) were positive for food insecurity. Patients (n = 1525, 74%) were linked to community resources. Nearly all (97%) nurses participated in education and demonstrated increased knowledge and confidence (P < .001). These data provide preliminary outcomes from the Food is Medicine Program.


Effects of BPA and BPS exposure limited to early embryogenesis persist to impair non-associative learning in adults.
Bisphenol-A (BPA) is a polymerizing agent used in plastic bottles and several routinely used consumer items. It is classified among endocrine disrupting chemicals suspected to cause adverse health effects in mammals ranging from infertility and cancer to behavioral disorders. Work with the invertebrate lab model Caenorhabditis elegans has shown that BPA affects germ cells by disrupting double-stranded DNA break repair mechanisms. The current study utilizes this model organism to provide insight into low-dose and long-term behavioral effects of BPA and bisphenol-S (BPS), a supposed safer replacement for BPA. Experiments presented in our report demonstrate that the effects of embryonic exposure to considerably low levels of BPA persist into adulthood, affecting neural functionality as assayed by measuring habituation to mechano-sensory stimuli in C. elegans. These results are noteworthy in that they are based on low-dose exposures, following the rationale that subtler effects that may not be morphologically apparent are likely to be discernible through behavioral changes. In addition, we report that embryonic exposure to BPS follows a pattern similar to BPA. Building upon previous observations using the C. elegans model, we have shown that exposure of embryos to BPA and BPS affects their behavior as adults. These long-term effects are in line with recommended alternate low-dose chemical safety testing approaches. Our observation that the effects of BPS are similar to BPA is not unexpected, considering their structural similarity. This, to our knowledge, is the first reported behavioral study on low-dose toxicity of any endocrine disrupting chemical in C. elegans.


Opportunities and Challenges for Antibodies against Intracellular Antigens.
Therapeutic antibodies are one most significant advances in immunotherapy, the development of antibodies against disease-associated MHC-peptide complexes led to the introduction of TCR-like antibodies. TCR-like antibodies combine the recognition of intracellular proteins with the therapeutic potency and versatility of monoclonal antibodies (mAb), offering an unparalleled opportunity to expand the repertoire of therapeutic antibodies available to treat diseases like cancer. This review details the current state of TCR-like antibodies and describes their production, mechanisms as well as their applications. In addition, it presents an insight on the challenges that they must overcome in order to become commercially and clinically validated.


Design and evaluation of an innovative floating and bioadhesive multiparticulate drug delivery system based on hollow structure.
In this study a gastric-retentive delivery system was prepared by a novel method which is reported here for the first time. An innovative floating and bioadhesive drug delivery system with a hollow structure was designed and prepared. The floating and bioadhesive drug delivery system was composed of a hollow spherical shell, a waterproof layer (Stearic acid), a drug layer (Ofloxacin), a release retarding film (the novel blended coating materials) and a bioadhesive layer (Carbomer 934P) prepared by using a liquid multi-layering process. A novel blended coating material was designed and investigated to solve the problem of the initial burst release of the formulation and the release mechanism of the novel material was analyzed in this study. The optimized formulation provided the sustained release characteristic and was able to float for 24h. The SEM cross-section images showed that the particulates were hollow with a spherical shell. X-ray images and pharmacokinetic studies (Frel = 124.1 ± 28.9%) in vivo showed that the gastric-retentive delivery system can be retained in the stomach for more than 6h. The floating and bioadhesive particulate drug delivery system based on a hollow structure with a dual function presented here is a viable alternative to other for gastroretentive drug delivery system.


A liquid chromatography-tandem mass spectrometry-based investigation of the lamellar interstitial metabolome in healthy horses and during experimental laminitis induction.
Lamellar bioenergetic failure is thought to contribute to laminitis pathogenesis but current knowledge of lamellar bioenergetic physiology is limited. Metabolomic analysis (MA) can systematically profile multiple metabolites. Applied to lamellar microdialysis samples (dialysate), lamellar bioenergetic changes during laminitis (the laminitis metabolome) can be characterised. The objectives of this study were to develop a technique for targeted MA of lamellar and skin dialysates in normal horses, and to compare the lamellar and plasma metabolomic profiles of normal horses with those from horses developing experimentally induced laminitis. Archived lamellar and skin dialysates (n = 7) and tissues (n = 6) from normal horses, and lamellar dialysate and plasma from horses given either 10 g/kg oligofructose (treatment group, OFT; n = 4) or sham (control group, CON; n = 4) were analysed. The concentrations of 44 intermediates of central carbon metabolism (CCM) were determined using liquid chromatography-tandem mass spectrometry. Data were analysed using multivariate (MVA) and univariate (UVA) analysis methods. The plasma metabolome appeared to be more variable than the lamellar metabolome by MVA, driven by malate, pyruvate, aconitate and glycolate. In lamellar dialysate, these metabolites decreased in OFT horses at the later time points. Plasma malate was markedly increased after 6 h in OFT horses. Plasma malate concentrations between OFT and CON at this time point were significantly different by UVA. MA of lamellar CCM was capable of differentiating horses developing experimental laminitis from controls. Lamellar malate, pyruvate, aconitate and glycolate, and plasma malate alone were identified as the source of differentiation between OFT and CON groups. These results highlighted clear discriminators between OFT and CON horses, suggesting that changes in energy metabolism occur locally in the lamellar tissue during laminitis development. The biological significance of these alterations requires further investigation.


Sinonasal small cell neoplasm developing after radiation therapy for retinoblastoma: an immunohistologic, ultrastructural, and cytogenetic study.
Patients with retinoblastoma have an increased risk of developing second primary tumors. Only a few examples of sinonasal small cell neoplasms developing after radiation therapy for retinoblastoma have been reported. We report one such case that developed 18 years after treatment for retinoblastoma. Histologic examination revealed a small, blue, round cell tumor without rosettes or cytoplasmic glycogen. Immunohistochemically, the tumor cells were positive for neuron-specific enolase, synaptophysin, and S-100 protein, but negative for epithelial and mesenchymal markers, suggesting that this was a primitive neuroectodermal tumor. Cytogenetic studies of this tumor failed to reveal the chromosome 13 abnormality typical of retinoblastoma and the t(11:22) translocation typical of the group of peripheral neuroepitheliomas.


The immunotherapeutic effect of anti Lyt-1 antibodies on local and metastatic tumor growth.
Monoclonal antibodies against the T cell differentiation antigen Lyt-1 were effective in the therapy of the murine Lewis lung carcinoma (3LL). A possible mechanism whereby anti Lyt-1 antibodies directly effect the tumor cells was unlikely because 3LL cells lack the Lyt-1 antigen. In addition, the curative effect of anti Lyt-1 antibodies was abrogated in mice depleted of T cells, supporting a mechanism whereby host Lyt-1+ cells were involved in tumor therapy. Treatment with anti Lyt-1 antibodies was not accompanied by depletion of Lyt-1+ cells from lymphoid organs, indicating that the administered antibodies altered Lyt-1+ cell functions without effecting their frequency. In view of the in vitro enhancing effects of anti Lyt-1 antibodies on responses, it is suggested that the potentiation of Lyt-1+ cell activity by passively administered anti Lyt-1 antibodies results in tumor rejection.


Calcium supplementation and bone mineral density in postmenopausal women using estrogen replacement therapy.
Both estrogen and calcium have been shown to be of benefit in reducing the incidence of osteoporosis in postmenopausal women. It has been suggested that estrogen promotes the intestinal absorption of calcium, and this effect may especially benefit postmenopausal women with a low dietary intake of calcium. The purpose of this study was to examine the effect of calcium supplementation on the bone mineral density (BMD) of postmenopausal women using estrogen replacement therapy. BMD measurements were made over 12 months in two groups of postmenopausal women, one having treatment with estrogen alone, and the second having estrogen and 1000 mg of supplemental calcium each day. There was no significant reduction in BMD between the two groups over the 12-month study period. Those in the group having supplemental calcium had a significant increase in BMD at the femoral neck (p = 0.023), but not in other areas of the femur nor in the lumbar spine. The results of this study suggest that the addition of supplemental calcium may improve the bone mass of postmenopausal women using estrogen who have a low dietary calcium intake. This benefit appears to be more pronounced in corticocancellous than in trabecular bone, and may therefore have a greater effect on the femoral neck than the lumbar spine.


Modulators of TRPM7 and its potential as a drug target for brain tumours.
TRPM7 is a non-selective divalent cation channel with an alpha-kinase domain. Corresponding with its broad expression, TRPM7 has a role in a wide range of cell functions, including proliferation, migration, and survival. Growing evidence shows that TRPM7 is also aberrantly expressed in various cancers, including brain cancers. Because ion channels have widespread tissue distribution and result in extensive physiological consequences when dysfunctional, these proteins can be compelling drug targets. In fact, ion channels comprise the third-largest drug target type, following enzymes and receptors. Literature has shown that suppression of TRPM7 results in inhibition of migration, invasion, and proliferation in several human brain tumours. Therefore, TRPM7 presents a potential target for therapeutic brain tumour interventions. This article reviews current literature on TRPM7 as a potential drug target in the context of brain tumours and provides an overview of various selective and non-selective modulators of the channel relevant to pharmacology, oncology, and ion channel function.


Induced chromosomal aberrations in somatic cells of Nigella sativa L. by mitomycin C.
A cytological study was carried out on root tips of Nigella sativa L. by treatment with Mitomycin C at 0.001% for six time intervals (10, 15, 20, 30, 40, and 50 min). The chromosomal abnormalities were increasingly proportionate to the increase in time of treatment. The seedlings treated with a 0.001% concentration of Mitomycin C for 10 min. did not show any significant effect. At other time intervals, the effect was observed to be quite significant. Beyond 40 min. treatment almost all the cells would become sticky. Thirty minutes' treatment showed significant effect, inducing various types of chromosomal aberrations in the anaphase, such as bridges and fragments of 34.13% and 48.07%, respectively.


Reporting of uncertainty at the 2013 annual meeting of the American Society for Radiation Oncology.
The annual meeting of the American Society for Radiation Oncology (ASTRO) is designed to disseminate new scientific findings and technical advances to professionals. Best practices of scientific dissemination require that some level of uncertainty (or imprecision) is provided. A total of 279 scientific abstracts were selected for oral presentation in a clinical session at the 2013 ASTRO Annual Meeting. A random sample of these abstracts was reviewed to determine whether a 95% confidence interval (95% CI) or analogous measure of precision was provided for time-to-event analyses. A sample of 140 abstracts was reviewed. Of the 65 abstracts with Kaplan-Meier or cumulative incidence analyses, 6 included some measure of precision (6 of 65 = 9%; 95% CI, 2-16). Of the 43 abstracts reporting ratios for time-to-event analyses (eg, hazard ratio, risk ratio), 22 included some measure of precision (22 of 43 = 51%; 95% CI, 36-66). Measures of precision are not provided in a significant percentage of abstracts selected for oral presentation at the Annual Meeting of ASTRO.


Accelerated water removal from frozen thin films containing bacteria.
Freeze-drying, or lyophilization, is widely used to produce pharmaceutical solids that contain temperature-sensitive materials. Herein, using Escherichia coli as a model live organism, whose viability in dry powders is highly sensitive to the water content in the powders, we demonstrated that the drying rate from the frozen thin films generated by thin-film freezing (TFF) is significantly faster than from the bulk frozen solids in conventional shelf freeze-drying. This is likely because the loosely stacked frozen thin films provided a larger solid-air interface and the low thickness of the thin films provided a low mass transfer resistance. The highly porous microstructure and high specific surface area of the thin-film freeze-dried powders may also be related to the faster drying observed. Moreover, we demonstrated that TFF can be applied to produce dry powders of E. coli, a Gram-negative bacterium, or Lactobacillus acidophilus, a Gram-positive bacterium, with minimum bacterial viability loss (i.e., within one log reduction). It is concluded that the TFF technology is promising in accelerating water removal from frozen samples.


Oropharyngeal Geometry of Maxilectomized Patients Rehabilitated with Palatal Obturators in the Trans-surgical Period: Repercussions on the Voice.
Patients undergoing maxillectomies may present alterations in the stomatognathic functions involved in oral communication. Rehabilitative treatment should favor the rescue of these functions, through surgical flaps, obturator prostheses or both. The present study aims to present the impact of the use of the palatal obturator on the oropharyngeal geometry and on the voice of patients undergoing maxillectomies, after adaptation to trans-surgical palatine obturators (TPO). Twelve patients treated at a Cancer Hospital, submitted to maxillectomy and rehabilitated during surgery were evaluated. The oropharyngeal geometry was measured by acoustic pharyngometry and the vocal parameters were evaluated through auditory-perceptual and acoustic analyses. The comparison between the results with and without TPO was analyzed using the Wilcoxon test and the correlation between oropharyngeal measurements and acoustic parameters using Spearman's correlation coefficient, all with a significance level of 5%. There was a decrease in the following oropharyngeal measurements with the use of TPO: length of the pharyngeal cavity and vocal tract, volume of the oral cavity, pharyngeal and vocal tract and area of the oropharyngeal junction. There was no difference in the length of the oral cavity and in the glottic area between situations with and without TPO. In the vocal evaluation, changes in intelligibility and resonance were observed in the situation without TPO and, in only one case, mild hypernasality was detected in the situation with TPO. It is concluded that the use of TPO brought the oropharyngeal measurements closer to normal values and provided an improvement in speech intelligibility and vocal resonance in maxilectomized individuals.


Results of operative treatment of stress urinary incontinence with special reference to preoperative clinical and radiological evaluation.
Operative results of stress urinary incontinence are discussed on the basis of a follow-up study made 3-5 years after the operation. The series consisted of 301 patients operated on during 1969-1971. The most significant anamnestic feature was the great number of multiparas: 67% of the patients had given birth to at least 4 children. The choice of the operative method was based on Green's urethrocystographic classification into type I and type II. The healing was complete or satisfactory in 85% of the total material. In the group operated on according to type I the outcome was good in 88%, while in the type II group it was good in 84%. Incontinence of degree I was cured in 89% of the cases, while patients of degree III in only 72%. If there was a significant urgency component observable in the incontinence symptoms preoperatively, a good outcome was only attained in 64%. In pure or dominant stress incontinence the corresponding outcome was 88%. Hysterectomy combined to the operation seemed to improve the results. Age, parity, duration of the incontinence symptoms, weight and changes of weight after the operation had no significant effect on the healing. The authors emphasize the importance of careful differential diagnosis based on the anamnesis and examinations in choosing the therapeutic method for incontinence patients.


Gender differences in the surgical management and early clinical outcome of coronary artery disease: Single centre experience.
To investigate the gender disparity in the distribution of patient-related risk factors and their effect on the surgical management and clinical outcome of coronary artery disease in Saudi population. We carried out a retrospective analysis of prospectively collected data of 971 patients undergoing isolated coronary artery bypass grafting (CABG) at our institution between January 2005 and December 2008. Seven hundred and eighty seven patients (81%) were males and 184 patients (19%) were females. We analyzed gender-based difference in clinical presentation and patient-related pre-operative risk factors and studied their impact on surgical management and clinical outcome. The mean age was 59.5 years in males and 63.4 years in females (p = <0.0001). Associated co-morbidities were higher in females. Prevalence of diabetes mellitus was 61.2% in males and 78.8% in females (p-value = <0.0001); hypertension 61.9% in males and 79.9% in females (p-value <0.0001); hyperlipidemia 66.7% in males and 77.7% in females (p-value 0.0035); morbid obesity 24.7% in males and 45.1% in females (p-value <0.0001); and Hypothyroidism 2.5% in males and 13.6% in females (p-value <0.0001). Smoking was the only risk factor with higher prevalence in males compared to females (44.2% v/s 2.2%; p-value <0.0001). The mean logistic euroSCORE was 3.94 in males and 5.51 in females (p < 0.0003). On-pump and off-pump CABG was carried out in equal numbers in two groups. Females required urgent surgery and less than 3 grafts more frequently while males underwent elective surgery and more than 3 grafts in greater numbers. No significant difference was present between the two gender groups in aortic occlusion times and bypass times. Univariant analysis revealed females gender as an independent risk factor for higher in-hospital mortality (1.1% versus 4.9% p = 0.0026) and higher incidence of post-operative complications like surgical wound infection, need for prolonged ventilation, low cardiac output state and multi-organ failure (p-values 0.01 or less). Female gender is an independent predictor of adverse outcome after isolated CABG due to significantly higher co-morbidities and acute presentation and independent of their peri-operative management. Therefore, major socioeconomic education and preventive measures are needed to reduce the burden of major co-morbidities in females and to seek early cardiac advice and care.


Glycosyl transfers from UDP-sugars to lipids of plant membranes: identification and specificity of the transferases.
Microsome preparations extracted from wheat roots or sycamore cell suspensions catalyzed the transfer of sugar from nucleotide-sugars to endogenous lipidic acceptors. The nature of the products biosynthesized from UDP-Glc, GDP-Glc, UDP-Gal, UDP-Xyl or UDP-Arab was examined. Sterylglycosides were obtained from UDP-Gglc, GDP-Glc or UDP-Xyl. Galactosyldiglycerides were synthesized from UDP-Gal. When UDP-Glc or UDP-Gal was used as a substrate, a membrane-bound 4-epimerase interconverted the epimeric nucleotide-sugars, thereby allowing the simultaneous biosynthesis of galactosyldiglycerides and sterylglucosides. The biosynthesis of free and acylated sterylglucosides from UDP-Glc, without interference of other glycosyl transfer reactions, was obtained by the omission of Mg++ ions from the incubation medium. The biosynthesis of galactosyldiglycerides from UDP-Gal without interference of other transfer reactions was obtained when digitonin was added to the incubation medium of sycamore microsomes.


The radiation inactivation method provides evidence that membrane-bound mitochondrial creatine kinase is an oligomer.
Lyophilized suspensions of rabbit heart mitochondria have been irradiated with varying doses of gamma rays. Mitochondrial creatine kinase activity was inactivated exponentially with a radiation inactivation size of 352 or 377 kDa depending upon the initial medium. These values are in good agreement with the molecular mass previously deduced from by permeation experiments: 357 kDa. This is the first direct evidence showing that the native form of mitochondrial creatine kinase is associated to the inner membrane as an oligomer, very likely an octamer.


Presentation of bilateral thalamic infarction on CT, MRI and PET.
Paramedian thalamic structures and part of the upper midbrain are frequently supplied by posterior thalamoperforating arteries originating from one common trunk. Local impairment of flow entails a bilateral more or less symmetric thalamic infarction with varying involvement of the midbrain. Diagnosis usually can neither be firmly established on clinical grounds nor by angiography alone. In the present series of four patients the two cases observed before the CT era were diagnosed correctly only at autopsy. Only one patient presented the classical syndrome of hypersomnia, thalamic dementia, and oculomotor nerve paralysis, while in the others clinical signs were probably masked by serious impairment of consciousness. In two cases X-ray computed tomography and magnetic resonance tomography (one case) afforded precise definition of infarct localization and size. Infarction in the described terminal vascular supply territory may be detected more often by these modern diagnostic techniques than anticipated from previous clinico-pathological experience as the underlying cause of coma in the elderly-a group of patients at particular risk for low-flow states. Positron emission tomography repeat studies with 18F-2-fluorodeoxyglucose (one case) revealed complex disturbances of brain energy metabolism; correlative analysis of clinical function and metabolic patterns during the course of the disease may not only advance individual prognostication but also contribute to the understanding and localization of brain function.


Corrigendum: Immune tolerance induced by hematopoietic stem cell infusion after HLA identical sibling kidney transplantation.
[This corrects the article DOI: 10.3389/fimmu.2022.995243.].


Emergency department visits after pancreatoduodenectomy: examining a novel quality metric.
Postoperative emergency department (ED) visits represent fragmented care, are costly, and often evolve into readmission. Readmission rates after pancreatoduodenectomy (PD) are defined, while ED visits following PD are not. We examined the pattern of 30-day post-discharge ED visits for PD patients. A quaternary institutional database analysis of adult patients who underwent PD between 2010-2017 was reviewed for ED utilization within 30 days from discharge. Of the 1,004 patients who underwent PD, 12% (N = 117) patients sought care in the ED within 30 days from postoperative discharge. The median time to ED presentation was 5 days post-discharge (IQR 3-9). Half of ED visits occurred during nights and weekends (N = 59, 50%). Of ED-utilizing patients, 64% (N = 76) were admitted to the hospital and 29% (N = 34) were discharged from the ED. ED visits were associated with a Clavien-Dindo Classification of 0 in 10.2% (N = 13) of patients, I-II in 62.4% (N = 73), and III-V in 26.5% (N = 31). Post-discharge ED utilization is a novel quality metric and represents a potential target population for reducing hospital readmissions. Over two-thirds (72%) of ED visits were associated with low acuity complications, and promoting institutional strategies addressing postoperative ED visits may improve patient care and efficient utilization of healthcare resources.


Serum concentrations of ursodeoxycholic acid in portal venous and systemic venous blood of fasting humans as determined by isotope dilution-mass spectrometry.
The fasting concentrations of ursodeoxycholic acid were determined in peripheral and portal venous serum of untreated (n = 12) and ursodeoxycholic acid-treated (n = 7) patients undergoing cholecystectomy. The levels of ursodeoxycholic acid were also determined in peripheral venous serum of 9 healthy subjects before and during treatment with ursodeoxycholic acid. Ursodeoxycholic acid, as well as cholic, chenodeoxycholic, and deoxycholic acids, were analyzed by a highly specific method based on isotope dilution-mass spectrometry. The fasting peripheral venous serum concentration of total (unconjugated plus conjugated) ursodeoxycholic acid averaged 0.14 mumol/L in the untreated gallstone patients and 0.19 mumol/L in the healthy subjects. The corresponding value in portal venous serum was 0.44 mumol/L. Treatment with ursodeoxycholic acid raised the level of this bile acid about 25-fold in portal as well as in peripheral venous serum. The proportion of unconjugated ursodeoxycholic acid was 34% in portal and 49% in peripheral venous serum of treated subjects. The mean hepatic uptake of ursodeoxycholic acid was calculated to be about 60% both in untreated and treated subjects. This uptake was significantly lower than that of cholic acid (83%). The hepatic uptake of ursodeoxycholic acid also tended to be lower than that of chenodeoxycholic acid (68%). This was mainly due to a lower hepatic uptake of unconjugated ursodeoxycholic acid (34%) compared with unconjugated chenodeoxycholic acid (49%). The relatively low hepatic uptake of unconjugated ursodeoxycholic acid explains why serum levels of the administered bile acid are higher during treatment with ursodeoxycholic acid than during treatment with chenodeoxycholic acid. Our results also give evidence that the hepatic uptake of ursodeoxycholic acid cannot be saturated under physiologic conditions.


Protection against tabun toxicity in mice by prophylaxis with an enzyme hydrolyzing organophosphate esters.
We demonstrate here the correlation between protection afforded by pretreatment alone with parathion hydrolase purified from Pseudomonas sp. against tabun toxicity in mice and the kinetic parameters which are assumed to determine the in vivo detoxification of tabun by the same enzyme. Results show that 15 and 22 micrograms of parathion hydrolase per animal conferred a protective ratio of 3.94 and 5.65 respectively, against tabun toxicity, without post-exposure treatment.


Preoperative MRI of pure intraductal breast carcinoma--a valuable adjunct to mammography in assessing cancer extent.
The aim of the study was to evaluate the contribution of preoperative breast magnetic resonance imaging (MRM) as an adjunct to mammography in assessing extent of pure ductal carcinoma in situ (DCIS) and to relate magnetic resonance imaging (MRI) findings to histopathological features. A retrospective analysis was conducted of 86 histologically proven cases of pure DCIS of the breast. Two experienced radiologists with knowledge of clinical and histopathological findings at the time of the review evaluated mammographic and preoperative MRI results by consensus. Compared to histopathology, mammography or MRM alone underestimated DCIS extent in 18.6% and 31.4% of cases, respectively. When both imaging modalities were considered, DCIS extent was underestimated in 8% of cases. Combined use of mammography and MRM revealed good agreement with histopathology to assess DCIS extent (kappa=0.439; P<0.001). MR enhancement of DCIS was related to histologic size (P=0.011). Mammography is more accurate than MRM in assessing cancer extent of pure DCIS, but combined use of both imaging techniques leads to improved accuracy.


Studies on the distribution and metabolism of N-[14C]nitrosopyrrolidine in mice.
Pretreatments with pyrazole, ethanol, nialamide or diethyldithiocarbamate were found to strongly depress the exhalation of 14CO2 and the incorporation of radioactivity in the acid-insoluble fraction of the liver in mice injected with N-[14C]nitrosopyrrolidine. Whole-body autoradiography performed with hemisections of mice at -80 degrees C (to prevent evaporation of the volatile N-nitrosopyrrolidine) and with dry tape-sections (to localize the non-volatile metabolites), using pretreated and non-pretreated mice, indicated a uniform distribution of the non-metabolized N-nitrosopyrrolidine in the tissues. At the shortest survival intervals (1 and 5 min), a high level of metabolites were found in the liver, the tracheo-bronchial and nasal mucosa and Harder's gland, indicating a local formation of metabolits in these tissues. At later survival intervals (0.5--24 h) metabolites were in addition found in tissues with a rapid cell turnover and a high rate of protein synthesis and in brown fat, which probably reflects incorporation of metabolites via normal biosynthetic pathways. Autoradiography of N-[14C]nitrosopyrrolidine in mice given the substance orally resulted in distribution pictures similar to those obtained after i.v. injections.


Neurodevelopmental outcome following whole body cooling for perinatal asphyxia.
This follow-up study conducted on children who underwent therapeutic cooling for hypoxic ischemic encephalopathy, showed normal neurodevelopmental outcome with normal milestones and normal developmental quotient in a minimum of 60% of children at 18-24 months of age. This study shows comparable neurodevelopmental outcome in infants who underwent cooling in a resource poor setting, when compared with existing literature.


Long-term survival and cause-specific mortality of patients newly diagnosed with tuberculosis in São Paulo state, Brazil, 2010-15: a population-based, longitudinal study.
Long-term survival and cause-specific mortality of patients who start tuberculosis treatment is rarely described. We aimed to assess the long-term survival of these patients and evaluate the association between vulnerable conditions (social, health behaviours, and comorbidities) and cause-specific mortality in a country with a high burden of tuberculosis. In this population-based, longitudinal study in São Paulo state, Brazil, we described the 5-year survival of patients who were newly diagnosed with tuberculosis in 2010. We included patients with newly-diagnosed tuberculosis, aged 15 years or older, and notified to the São Paulo State Tuberculosis Program in 2010. We excluded patients whose diagnosis had changed during follow-up (ie, they did not have tuberculosis) and patients who had multidrug-resistant (MDR) tuberculosis. We selected our population with tuberculosis from the dedicated electronic system TBweb. Our primary objective was to estimate the excess mortality over 5 years and within the group who survived the first year, compared with the general São Paulo state population. We also estimated the association between social vulnerability (imprisonment and homelessness), health behaviours (alcohol and drug use), and comorbidities (diabetes and mental disorders) with all-cause and cause-specific mortality. We used the competing risk analysis framework, estimating cause-specific hazard ratios (HRs) adjusted for potential confounding factors. In 2010, there were 19 252 notifications of tuberculosis cases. We excluded 550 cases as patients were younger than 15 years, 556 cases that were not tuberculosis, 2597 retreatments, and 48 cases of MDR tuberculosis, resulting in a final cohort of 15 501 patients with tuberculosis. Over a period of 5 years from tuberculosis diagnosis, 2660 (17%) of 15 501 patients died. Compared with the source population, matched by age, sex, and calendar year, the standardised mortality ratio was 6·47 (95% CI 6·22-6·73) over 5 years and 3·93 (3·71-4·17) among those who survived the first year. 1197 (45%) of 2660 deaths were due to infection. Homelessness and alcohol and drug use were associated with death from infection (adjusted cause-specific HR 1·60, 95% CI 1·39-1·85), cardiovascular (1·43, 1·06-1·95), and external or ill-defined causes of death (1·80, 1·37-2·36). Diabetes was associated with deaths from cardiovascular causes (1·70, 1·23-2·35). Patients newly diagnosed with tuberculosis were at a higher risk of death than were the source population, even after tuberculosis treatment. Post-tuberculosis sequelae and vulnerability are associated with excess mortality and must be addressed to mitigate the tuberculosis burden worldwide. Wellcome Trust.


The conceptual grouping effect: categories matter (and named categories matter more).
Do conceptual categories affect basic visual processing? A conceptual grouping effect for familiar stimuli is reported using a visual search paradigm. Search through conceptually-homogeneous non-targets was faster and more efficient than search through conceptually-heterogeneous non-targets. This effect cannot be attributed to perceptual factors and is not explained by a long-term representational reorganization due to perceptual-learning. Rather, conceptual categories seem to modulate visual representations dynamically, and are sensitive to task-demands. Verbally labeling a visual target further exaggerates the degree to which conceptual categories penetrate visual processing.


FR183998, a Na+/H+ exchange inhibitor, suppresses both IL-8 content and myocardial infarct size in a cardiac ischaemia-reperfusion model in rats.
The aim of this study was to determine the effect of FR183998 (5-(2,5-dichlorothiophen-3-yl)-3-[(2-dimethylaminoethyl)carbamoyl]benzoylguanidine dihydrochloride), an Na+/H+ exchange inhibitor, on myocardial interleukin-8 (IL-8) content and myocardial infarct size in a rat ischaemia and reperfusion model. Rats underwent 30 min of ischaemia followed by 1 to 24 h of reperfusion. IL-8 content rapidly increased in reperfused rat hearts. The maximum increase in IL-8 was obtained after 3 h of reperfusion. Intravenous administration of FR183998 at 1 and 3.2 mg kg(-1), 5 min before ischaemia, significantly reduced the IL-8 level after 3 h of reperfusion (122 +/- 16 and 149 +/- 23 pg mg(-1) protein, respectively), compared with that of the saline-treated group (258 +/- 27 pg mg(-1) protein). Myeloperoxidase activity after 3 h of reperfusion was also reduced by FR183998 (from 0.83+0.19 unit g(-1) weight of tissue in the saline-treated group to 0.36 +/- 0.09 and 0.33 +/- 0.06 unit g(-1) weight of tissue in FR183998-treated groups at 1.0 and 3.2 mg kg(-1), respectively). Myocardial infarction induced by 30 min of ischaemia and 24 h of reperfusion was significantly suppressed by the same doses of FR183998 (14.0 +/- 1.5,13.5 +/- 1.9% at 1.0 and 3.2 mg kg(-1)), compared with 22.2+2.7% in the saline-treated group. These results suggestthat IL-8 may contribute to the generation of myocardial infarction in an ischaemia and reperfusion model in rats.


Safety and feasibility of robotic major hepatectomy for novice surgeons in robotic liver surgery: A prospective multicenter pilot study.
Robotic liver resection has not yet been widely implemented. We aimed to evaluate the feasibility and safety of robotic major liver resection by performing a prospective multicenter study. From July 2017 to December 2018, five surgeons from five tertiary hospitals who were novices in robotic liver resection but experienced in open and laparoscopic liver resection performed 46 cases of robotic major anatomical liver resections. Perioperative clinical data and surgical data, including detailed procedure times were prospectively collected. All operations were performed according to a protocol for unify surgical techniques and instruments. Twenty-two cases of left hemihepatectomy, one case of extended left hemihepatectomy, 14 cases of right hemihepatectomy, two cases of right anterior sectionectomy, six cases of right posterior sectionectomy, and one case of central bisectionectomy were performed. The most common indications were hepatocellular carcinoma (21 cases) followed by intrahepatic duct stones (10 cases), intrahepatic cholangiocellular carcinoma (7 cases), liver metastases (3 cases), intraductal papillary neoplasms (2 cases), sarcoma (1 case), mucinous cystic neoplasm (1 case), and hemangioma (1 case). Surgical resection margins for all tumor cases were negative. The mean operation time was 378.58 ± 124.31 (190-696) minutes and the estimated intraoperative blood loss was 276.67 ± 397.41 mL (range, 10-2600 mL). Overall complications developed in 16 cases (34.8%). There were three cases of severe surgical complications (Clavien-Dindo classification of III or more). Only one of 46 cases was converted to conventional open left hemihepatectomy because of bleeding. The mean hospital stay was 7.3 ± 2.5 (4-18) days. The results of this study indicate that robotic anatomic major liver resection can be safely performed by robotic beginners who are advanced open and laparoscopic liver surgeons.


The gene for pycnodysostosis maps to human chromosome 1cen-q21.
Pycnodysostosis (OMIM 265800) is an autosomal recessive skeletal disorder first described by Maroteaux and Lamy that is characterized by short stature, increased bone density, delayed closure of cranial sutures, loss of the mandibular angle, dysplastic clavicles, dissolution of the terminal phalanges of the hands and feet, dental abnormalities and increased bone fragility. Patients have a typical appearance secondary to prominence of the calvarium, smallness of the facial features, prominent nose and micrognathia. The French painter, Henri de Toulouse Lautrec (1864-1901), is believed to have had the disorder. Although more than 100 cases have been reported, we are aware of only two large consanguinous pedigrees in which the pycnodysostosis disorder segregates. We have studied the segregation of the pycnodysostosis phenotype in a large consanguinous Mexican pedigree, the clinical features of which are very similar to those described in the Arab pedigree studied by Edelson et al. Here, we report linkage for the pycnodysostosis phenotype in the 1cen-q21 region of human chromosome 1, and discuss candidate genes for this skeletal disorder.


Analysis of UBQLN1 variants in a Polish Alzheimer's disease patient: control series.
Late-onset Alzheimer's disease (LOAD) is the most common neurodegenerative disorder, and has a complex etiology. Recently an intronic polymorphism in the ubiquilin 1 gene (UBQLN1) and a particular haplotype was reported to be associated with LOAD. We investigated whether variants in UBQLN1 confer a risk for the disease in 407 Polish LOAD patients and 407 controls. We observed a weak association with the rs2781002 polymorphism, however, contrary to the initial reports, in our group the association was with the A allele. Risk estimation for AA versus GG genotypes showed that the AA genotype is a weak risk factor for AD (OR = 1.8, 95% CI = 1.1-3.1, p = 0.025). This effect was stronger in a group of LOAD patients without APOE4 allele. Haplotype analyses indicate that there is an increase of haplotypes with an A allele in the case group. Also, the specific haplotypes with the A allele that increase AD risk differ between the APOE4-positive and APOE4-negative pools. However, the association observed seems to be driven mostly by rare (<5%) haplotypes. Results suggest a need for additional association studies and in silico analysis of the UBQLN1 locus.


Scabietic vasculitis: Report of 2 cases.
The infectious causes of cutaneous vasculitis are well known and include streptococcal infections among others. Cases resulting from parasitic infection are less frequent. Scabies, which is currently on the increase, has only been reported in a few isolated cases. Herein, we report two noteworthy cases of profuse scabies complicated by cutaneous vasculitis. Case 1: a 90-year-old woman, residing in a nursing home, was admitted to our dermatology department complaining of pruritus, present for one month, predominantly on the inside of the thighs and on the buttocks, associated with purpuric lesions on the lower limbs. A skin biopsy revealed leukocytoclastic vasculitis. A diagnosis of scabies was based on severe pruritus and hypereosinophilia and was confirmed by microscopic examination of the parasitology sample and the skin biopsy sample. Despite thorough investigation, no other cause of vasculitis could be found. Complete regression of the skin lesions was achieved with scabies treatment only, without any specific treatment for the vasculitis. Case 2: a 74-year-old man, living in a nursing home, was hospitalized for purpuric papules on the lower limbs, present for one month. Physical examination revealed linear patterns in the interdigital spaces associated with scabies evident on dermoscopic examination. The skin biopsy revealed signs of vasculitis. As in our first case, no aetiology of vasculitis was found and a favorable outcome was achieved by means of scabies treatment alone with no specific treatment for vasculitis. Both of our patients presented scabies and vasculitis. In view of the absence of other causes of vasculitis and of the complete regression of lesions due to vasculitis without recurrence achieved with the scabies treatment alone, a diagnosis was made of scabietic vasculitis, probably as a result of cutaneous hypersensitivity reaction to humeral mediators.


ECM stiffness-regulated exosomal thrombospondin-1 promotes tunneling nanotubes-based cellular networking in breast cancer cells.
Intercellular communication is pivotal in various stages of cancer progression. For smart and effective communication, cancer cells employ diverse modes of messaging that may be further fine-tuned by the microenvironmental changes. Extracellular matrix (ECM) stiffening due to excess deposition and crosslinking of collagen is one of the crucial tumor-microenvironmental changes that influence a plethora of cellular processes, including cell-cell communication. We herein studied the crosstalk between exosomes and tunneling nanotubes (TNT), the two distinct means of cell-cell communication under varying ECM-stiffness conditions. We show that exosomes promote the formation of tunneling nanotubes in breast cancer cells, which results in cellular internet. Interestingly, exosomes drastically increased the fraction of cells connected by TNT; however, they elicited no effect on the number of TNTs per pair of connected cells or the length of TNT. The observed pro-TNT effects of exosomes were found to be ECM-stiffness dependent. ECM-stiffness tuned exosomes were found to promote TNT formation predominantly via the 'cell dislodgment model'. At the molecular level, exosomal thrombospondin-1 was identified as a critical pro-TNT factor. These findings underline the influence of ECM stiffening on two diverse modes of cell communication and their interdependence, which may have significant implications in cancer biomedical research.


Application of a Novel Two-Dimensional Echocardiographic Calcium Quantification Method to Assess All Severities of Aortic Stenosis.
Aortic valve (AV) calcification (AVC) is a strong predictor of aortic stenosis (AS) severity. The two-dimensional AVC (2D-AVC) ratio, a gain-independent ratio composed of the average pixel density of the AV and the aortic annulus, has previously shown strong correlations with two-dimensional (2D) echocardiographic hemodynamic parameters for severe AS and AVC by cardiac computed tomography. We hypothesize that the 2D-AVC ratio correlates with hemodynamic parameters in all severities of AS. A total of 285 patients with a normal AV (n = 49), aortic sclerosis (n = 75), or mild (n = 38), moderate (n = 72), or severe (n = 51) AS undergoing 2D echocardiography were retrospectively evaluated, and the 2D-AVC ratios were correlated to mean AV gradient, peak AV velocity, AV area, and dimensionless index. The 2D-AVC ratios of various AS severities were compared against each other via area under the curve (AUC) analysis. The 2D-AVC ratio is strongly correlated with mean AV gradient (r = 0.79, P < .0001) and peak AV velocity (r = 0.78, P < .0001). There was moderate correlation with the AV area (r = -0.58, P < .0001) and dimensionless index (r = -0.67, P < .0001) across all AS severities. The 2D-AVC ratio also distinguished nonmoderate AS (mild AS + normal AV) from moderate or greater (moderate + severe) AS (AUC = 0.93) and moderate versus severe AS (AUC = 0.88). The 2D-AVC ratio exhibits moderate to strong correlation with 2D echocardiographic hemodynamic parameters across all severities of AS.


Enhancement of fibroblastic proliferation on chitosan surfaces by immobilized epidermal growth factor.
Chitosan membranes were modified with mouse epidermal growth factor (EGF) by a photochemical technique. Photochemical immobilization was performed via a two-step process, in which EGF was first reacted with a heterobifunctional cross-linker sulfo-SANPAH (sulfosuccinimidyl 6(4'-azido-2'-nitrophenyl-amino)hexanoate) and then immobilized on the chitosan membrane by UV irradiation. The success of immobilization process was checked by Fourier transform infrared attenuated total reflection spectroscopy and X-ray photoelectron spectroscopy. Atomic force microscopy was used to evaluate the surface topography. The mitogenic effect of the EGF-modified chitosan membrane was investigated using mouse fibroblasts (L929 cell line), and cell proliferation was investigated by MTT and crystal violet assays. The results obtained from cell culture experiments showed that immobilized EGF stimulated fibroblast growth on chitosan membranes, and a considerable difference in cell proliferation was detected on EGF-modified chitosan membranes.


NF-κB as a target for oncogenic viruses.
NF-κB is a pivotal transcription factor that controls cell survival and proliferation in diverse physiological processes. The activity of NF-κB is tightly controlled through its cytoplasmic sequestration by specific inhibitors, IκBs. Various cellular stimuli induce the activation of an IκB kinase, which phosphorylates IκBs and triggers their proteasomal degradation, causing nuclear translocation of activated NF-κB. Under normal conditions, the activation of NF-κB occurs transiently, thus ensuring rapid but temporary induction of target genes. Deregulated NF-κB activation contributes to the development of various diseases, including cancers and immunological disorders. Accumulated studies demonstrate that the NF-κB signaling pathway is a target of several human oncogenic viruses, including the human T cell leukemia virus type 1, the Kaposi sarcoma-associated herpesvirus, and the Epstein-Bar virus. These viruses encode specific oncoproteins that target different signaling components of the NF-κB pathway, leading to persistent activation of NF-κB. This chapter will discuss the molecular mechanisms by which NF-κB is activated by the viral oncoproteins.


Elucidation of resistance signaling and identification of powdery mildew resistant mapping loci (ClaPMR2) during watermelon-Podosphaera xanthii interaction using RNA-Seq and whole-genome resequencing approach.
Watermelon is an important vegetable crop and is widely cultivated in USA with an approximate global production of > 100 million tons. Powdery mildew (PM) caused by Podosphaera xanthii is a major production-limiting factor on watermelon and other cucurbits. Numerous PM and multiple disease resistant (MDR) watermelon germplasm lines have been developed by the USDA in Charleston, SC. To gain a better understanding of the innate and activated molecular defense mechanisms involved during compatible and incompatible PM-watermelon interactions, we inoculated PM susceptible (USVL677-PMS) and resistant (USVL531-MDR) watermelon plants with 105 conidia ml-1 of P. xanthii. RNA-seq profiling was done on leaf samples collected at 0, 1, 3, and 8 days post inoculation (DPI). A total of 2,566 unique differentially expressed genes (DEGs) were identified between compatible and incompatible interactions with P. xanthii. The compatible interactions resulted in distinct plant gene activation (> twofold unique transcripts, 335:191:1762 :: 1:3:8 DPI) as compared to incompatible interaction (> twofold unique transcripts, 314:681:487 :: 1:3:8 DPI). Further, comparative whole-genome resequencing analysis of USVL531-PMR, USVL677-PMS and four introgressed PM resistant recombinant inbred lines (RIL, USVL531-PMR × USVL677-PMS) were performed to identify the region of PM resistance introgressed break points along with other traits inherent by USVL531-PMR by comparing the SNPs and InDels. Based on SNPs identification and CAPS markers, the resistance gene was identified as ClaPMR2, Citrullus lanatus PM Resistance gene 2 {Chr2 : 26750001 .. 26753327 (-)}, a NBS-LRR resistance protein (R) with homology to the Arabidopsis thaliana PM resistance protein, RPW8. The transcriptome data also revealed a complex regulatory network associated with the introgressed junctions mediated by PM resistance R proteins (R genes) that may involve multiple signal regulators and transducers, carbohydrate metabolism, cell wall modifications and the hormone-signaling pathway.


Multilevel D-loop PCR identification of hunting game.
The control region of mtDNA (D-loop) was used for hair samples of the five hunting game species identification: red deer (Cervus elaphus), roe deer (Capreolus capreolus), fallow deer (Dama dama), mouflon (Ovis aries musimon), and wild boar (Sus scrofa). For D-loop multilevel PCR detection scheme was applied in six primers (CE CVZV 1 = 5'-GATCACGAGCTTGATCACCA-3'; CE CVZV 2 = 5'-AGGAGTGGGCGATTTTAGGT-3'; DD CVZV 3 = 5'-CGCGTGAAACCAACAACCCGC-3'; DD CVZV 4 = 5'-CCGGGTCGGGGCCTTAGACG-3'; SSW CVZV 5 = 5'-ACACGTGCGTACACGCGCATA-3'; SSW CVZV 6 = 5'-GGTGCCTGCT T TCGTAGCACG-3') designed to identify unknown biological samples of the hunting game animals. The PCR reaction volume was 25 μl at conditions 95 °C for 2 min, 94 °C for 30 s, 60 °C for 30 s, 72 °C for 30 s, 35 cycles, with last extension at 72 °C for 10 min. D-loop mtDNA amplicons of the game animals are characterized with specific PCR product sizes depending on species: red deer = 163 bp and 140 bp, fallow deer = 280 bp and 138 bp, roe deer = 303 bp, 280 bp, 160 bp and 138 bp, mouflon = 299 bp and 178 bp, wild boar = 137 bp and 229 bp.


Differential diagnosis of negated avian notifiable disease cases in Great Britain.
This focus article has been prepared by David Welchman, veterinary lead of the APHA Avian Expert Group, Rowena Hansen, veterinary lead for avian virology at APHA Weybridge, and Alex Schock, team leader of the diagnostic and consultant avian pathology services at APHA Lasswade.


Roles of p53 and p27(Kip1) in the regulation of neurogenesis in the murine adult subventricular zone.
The tumor suppressor protein p53 (Trp53) and the cell cycle inhibitor p27(Kip1) (Cdknb1) have both been implicated in regulating proliferation of adult subventricular zone (aSVZ) cells. We previously reported that genetic ablation of Trp53 (Trp53-/-) or Cdknb1 (p27(Kip1-/-) ) increased proliferation of cells in the aSVZ, but differentially affected the number of adult born neuroblasts. We therefore hypothesized that these molecules might play non-redundant roles. To test this hypothesis we generated mice lacking both genes (Trp53-/- ;p27(Kip1-/-) ) and analysed the consequences on aSVZ cells and adult neuroblasts. Proliferation and self-renewal of cultured aSVZ cells were increased in the double mutants compared with control, but the mice did not develop spontaneous brain tumors. In contrast, the number of adult-born neuroblasts in the double mutants was similar to wild-type animals and suggested a complementation of the p27(Kip1-/-) phenotype due to loss of Trp53. Cellular differences detected in the aSVZ correlated with cellular changes in the olfactory bulb and behavioral data on novel odor recognition. The exploration time for new odors was reduced in p27(Kip1-/-) mice, increased in Trp53-/- mice and normalized in the double Trp53-/- ;p27(Kip1-/-) mutants. At the molecular level, Trp53-/- aSVZ cells were characterized by higher levels of NeuroD and Math3 and by the ability to generate neurons more readily. In contrast, p27(Kip1-/-) cells generated fewer neurons, due to enhanced proteasomal degradation of pro-neural transcription factors. Together, these results suggest that p27(Kip1) and p53 function non-redundantly to modulate proliferation and self-renewal of aSVZ cells and antagonistically in regulating adult neurogenesis.


Assignment of a human diaphorase (DIA4) to chromosome 16.
A human FAD-dependent diaphorase, DIA4, has been studied in 29 independent human-rodent hybrids and in 17 subclones. The results suggest that the locus DIA4 is on chromosome 16.