Combining clinical and molecular characterization of CDH1: a multidisciplinary approach to reclassification of a splicing variant.
Pathogenic germline variants (PGVs) in the CDH1 gene are associated with diffuse gastric and lobular breast cancer syndrome (DGLBC) and can increase the lifetime risk for both diffuse gastric cancer and lobular breast cancer. Given the risk for diffuse gastric cancer among individuals with CDH1 PGVs is up to 30-40%, prophylactic total gastrectomy is often recommended to affected individuals. Therefore, accurate interpretation of CDH1 variants is of the utmost importance for proper clinical decision-making. Herein we present a 45-year-old female, with lobular breast cancer and a father with gastric cancer of unknown pathology at age 48, who was identified to have an intronic variant of uncertain significance in the CDH1 gene, specifically c.833-9 C > G. Although the proband did not meet the International Gastric Cancer Linkage Consortium (IGCLC) criteria for gastric surveillance, she elected to pursue an upper endoscopy where non-targeted gastric biopsies identified a focus of signet ring cell carcinoma (SRCC). The proband then underwent a total gastrectomy, revealing numerous SRCC foci, but no invasive diffuse gastric cancer. Simultaneously, a genetic testing laboratory performed RNA sequencing to further analyze the CDH1 intronic variant, identifying an abnormal transcript from a novel acceptor splice site. The RNA analysis in conjunction with the patient's gastric foci of SRCC and family history was sufficient evidence for reclassification of the variant from uncertain significance to likely pathogenic. In conclusion, we report the first case of the CDH1 c.833-9 C > G intronic variant being associated with DGLBC and illustrate how collaboration among clinicians, laboratory personnel, and patients is crucial for variant resolution.


Herbal polysaccharides and cough reflex.
In the last decades plant substances have become a leading form of treatment of many respiratory symptoms, including cough. It has been shown that compounds purified form polysaccharides from Adhatoda vasica, Withania somnifera, and Glycyrrhiza glabra have various biological activities, such as antioxidant, anti-inflammatory, immunomodulating, antispasmodic action, or antiallergic properties, and they often act as cough suppressants. This work demonstrates new natural substitutes for synthetic antitussives whose application is associated with numerous adverse effects. We investigated pharmacodynamic characteristics of arabinogalacatan samples extracted from A. vasica, W. somnifera, and G. glabra. These extracts showed the ability to reduce citric acid-induced cough in awake guinea pigs after oral administration in a dose of 50mg/kg. The strongest antitussive effect (81%) was found after application of the extract from G. glabra. There was a 67% cough suppression with A. vasica and 61% with W. somnifera, which was comparable with the antitussive activity of codeine (62%).


Characterization of non-hemodynamic functional signal measured by spin-lock fMRI.
Current functional MRI techniques measure hemodynamic changes induced by neural activity. Alternative measurement of signals originated from tissue is desirable and may be achieved using T1ρ, the spin-lattice relaxation time in the rotating-frame, which is measured by spin-lock MRI. Functional T1ρ changes in the brain can have contributions from vascular dilation, tissue acidosis, and potentially other contributions. When the blood contributions were suppressed with a contrast agent at 9.4 T, a small tissue-originated T1ρ change was consistently observed at the middle cortical layers of cat visual cortex during visual stimulation, which had different dynamic characteristics compared to hemodynamic fMRI such as a faster response and no post-stimulus undershoot. Functional tissue T1ρ is highly dependent on the magnetic field strength and experimental parameters such as the power of the spin-locking pulse. With a 500Hz spin-locking pulse, the tissue T1ρ without the blood contribution increased during visual stimulation, but decreased during acidosis-inducing hypercapnia and global ischemia, indicating different signal origins. Phantom studies suggest that it may have contribution from concentration decrease in metabolites. Even though the sensitivity is much weaker than BOLD and its exact interpretation needs further investigation, our results show that non-hemodynamic functional signal can be consistently observed by spin-lock fMRI.


The Janus-faced nature of the C(2)B domain is fundamental for synaptotagmin-1 function.
Synaptotagmin-1 functions as a Ca2+ sensor in neurotransmitter release and was proposed to act on both the synaptic vesicle and plasma membranes through interactions involving the Ca2+ binding top loops of its C(2) domains and the Ca2+-independent bottom face of the C(2)B domain. However, the functional importance of the C(2)B domain bottom face is unclear. We now show that mutating two conserved arginine residues at the C(2)B domain bottom face practically abolishes synchronous release in hippocampal neurons. Reconstitution experiments reveal that Ca2+-synaptotagmin-1 can dramatically stimulate the rate of SNARE-dependent lipid mixing, and that the two-arginine mutation strongly impairs this activity. These results demonstrate that synaptotagmin-1 function depends crucially on the bottom face of the C(2)B domain and strongly support the notion that synaptotagmin-1 triggers membrane fusion and neurotransmitter release by bringing the vesicle and plasma membranes together, much like the SNAREs do but in a Ca2+-dependent manner.


Clinicopathological features of acute undifferentiated leukaemia with a stem cell phenotype.
Over a 4 1/2-year period, 141 patients with acute leukaemia had morphologic, immunophenotypic and cytochemical studies performed at King's College Hospital. Seven cases were noted to have blast cells which did not express myeloid or lymphoid antigens or cytochemical staining indicative of differentiation but were HLA DR and CD 34 positive. Based on these criteria we have used the term stem cell acute leukaemia to denote these patients. There were five women and two men with a median age of 61 years (16-86). Presentation marrows were heavily infiltrated with blasts (greater than 95% in 6/7) which were usually pleomorphic. Type 2 blasts. Auer rods and dysplastic features were absent. Two of six cases studied showed clonal karyotypic abnormalities. Four patients were treated with high dose chemotherapy. Three of these achieved a complete remission but relapsed at 3, 6 and 7 months respectively. The median survival of the group was 7 months (2-12). We conclude that the stem cell acute leukaemias are a distinct clinicomorphological group which appear to have a poor prognosis with conventional chemotherapy.


Outside-in transscleral fixation for ciliary sulcus intraocular lens placement.
We describe a new technique of transscleral intraocular lens fixation in the absence of adequate posterior capsular support. This technique of suture placement within the ciliary sulcus employs routinely used instruments, requires minimum manipulation, and has good visual outcome and negligible complications. We performed five intraocular lens implantations using the technique with good visual outcomes in each. No significant intraoperative or postoperative complications were noted.


Distal stenting technique for coil embolization of early branch aneurysms of middle cerebral artery.
Stent-assisted coil embolization of early branch aneurysms of the middle cerebral artery (MCA) can sometimes be challenging due to the small diameter and acute angle of the branches. This study aimed to analyze the treatment results and report the feasibility and outcomes of the distal stenting technique for these aneurysms. The distal stenting technique was used for 15 wide-neck MCA aneurysms (females, 10; males, 5; mean age, 65.1 years) originating from the early branch between December 2018 and October 2021. The average sizes of the dome, depth, and neck of the aneurysms were 4.17 mm (range: 2.99-6.21 mm), 2.86 mm (range: 1.82-3.72 mm), and 3.42 mm (range: 2.44-4.32 mm), respectively. The average diameter of the stents was 4.0 mm (3.0 mm, 3; 4.0 mm, 6; 4.5 mm, 6). Stents were successfully deployed in all 15 aneurysms (100%). The average length of the procedure was 69 min (range: 45-117 min). On postoperative angiography, nine (60.0%) aneurysms were completely occluded, four (26.7%) had neck remnants, and two (13.3%) had contrast flow in the sac. During the procedure, vasospasm and thrombus formation occurred in one case each, but no neurological sequelae were observed. On follow-up digital subtraction angiography of the 10 aneurysms after an average of 13.7 months (range: 12-18 months), the branches were well preserved, and neck remnants were noted in four aneurysms (40.0%). No thromboembolic events occurred during the clinical follow-up. The distal stenting technique using an open-cell stent can be a good option for coil embolization of early branch MCA aneurysms.


Roux-en-Y Gastric Bypass for the Treatment of Leak Following Sleeve Gastrectomy.
Laparoscopic sleeve gastrectomy (LSG) is estimated to be its most severe complication. An aggressive management with surgical reconstructive procedures can be proposed in patients in whom all the conservative endoscopic techniques fail. The purpose of the present study was to report our experience with Roux-en-Y gastric bypass (RYGBP) as treatment for the chronic leak after LSG. Between January 2013 and July 2019, 17 consecutive patients underwent RYGBP for the treatment of chronic leak after LSG. The initial intervention, the endoscopic approach and the definitive surgical repair were carefully reviewed. Seventeen patients (13 women) with a median age of 39 years (24-67) with a median body mass index (BMI) of 40 kg/m2 (30-52) underwent RYGBP for persistent fistula. Sixteen patients had their early LSG performed in another hospital. Eleven patients had an initial endoscopic treatment by pigtail drains following laparoscopic drainage and 6 other patients had the endoscopic stent as the first-choice line treatment. The overall average fistula diagnosis was done at 7.7 months (2-49 months) for 12 patients. For the rest of five patients, the procedure was performed almost in the acute setting (< 30 days). All procedures were performed by laparotomy but one. Five patients had a gastrojejunal anastomosis leak diagnosed by salivary flow in the drainage, but all patients were treated conservatively. No post-operative mortality was recorded. Surgery should be considered in case of failure of the endoscopic treatment of chronic leak after LSG. Further research is needed to clearly identify the appropriate treatment, but in our experience, RYGBP approach including the leak site offers a low morbidity rate.


Continuous cytomegalovirus seroconversion in a large group of healthy blood donors.
Transmission of cytomegalovirus (CMV) to seronegative, immunocompromised recipients can cause serious and fatal complications. Although the seroprevalence of CMV is high, the risk of primary CMV infection among healthy blood donors has not yet been analysed in a large population. We developed an algorithm to determine the rate of CMV seroconversion in an overall cohort of 24,260 subjects who donated 176,474 blood units during an 11-year observation period. We detected CMV seroconversion in all relevant age groups (18-60 years) with an overall seroconversion rate of 0.55% per year. Both CMV seroconversion and seroprevalence occurred more frequently in female donors (P = 0.02 and P < 0.001, respectively). We identified 30-35-year-old blood donors as the group with the highest rate of CMV seroconversion per year (1.33% vs. 0.46%; P < 0.0001). We conclude that the risk of primary CMV infection is a continuous lifelong event and correlates with age and female gender.


Prevalence of Demodex spp. in patients with chronic blepharitis.
Blepharitis is a very common disease in ophthalmology, dermatology and allergy practice. It generally follows a chronic course and is frequently associated with objective and/or subjective symptoms such as epiphora, red eye, dandruff, gritty sensation, itching, burning, photophobia, and blurred vision. The purpose of this study is to analyze the prevalence of Demodex spp. in patients with symptoms of chronic blepharitis. An analytical cross-sectional study was conducted in the period between 2016 and 2020. All patients with symptoms of chronic blepharitis who underwent a parasitological test of eyelashes (Rapitest) in the Dermatology Department of the Hospital Italiano de Buenos Aires were included. Those with previously established blepharitis due to another infectious cause were excluded. We analyzed 972 patients. Sixty percent (n=585) underwent a positive Rapitest for the presence of Demodex spp. Seventy five percent (n=728) were women. There were no significant differences in the prevalence associated with sex (p=0.38). Among the patients positive for Demodex spp., 65% (n=628) were older than 60 years old. The most frequently associated symptom was itching, present in 35% (n=342). A statistically significant decrease in the number of consultations was observed during the cold months of the year (May-June-July-August). Our results show a high prevalence of Demodex spp. in patients with chronic blepharitis. As its presence reveals a direct association with age, we recommend looking for this parasite in this age group.


Injecting Opioid Use Disorder Treatment in Jails and Prisons: The Potential of Extended-release Buprenorphine in the Carceral Setting.
As the opioid overdose cases rise, policy-makers and researchers should target interventions to populations at highest risk. Incarceration serves as a risk factor for opioid overdose (Gan et al. Addiction 2021) and a large portion of recent overdose deaths have had encounters in the criminal justice system.Medications for opioid use disorder in the criminal justice system can save lives, though unique administrative barriers in jails and prisons hinder access. As facilities expand medications for opioid use disorder access (due to new legislation and court rulings across states), extended-release buprenorphine offers an opportunity to overcome these barriers including logistics of administration, diversion concern, patient stigma, and an increased bridge of treatment during re-entry to the community.As extended-release buprenorphine has practical advantages in correctional health delivery, future research and policy discussions should investigate its optimal role in treating opiate addiction in a carceral setting.


Hepatocellular carcinoma in Japanese patients with nonalcoholic fatty liver disease and alcoholic liver disease: multicenter survey.
In Japan, the prevalence of hepatocellular carcinoma (HCC) associated with nonviral liver disease, especially with nonalcoholic fatty liver disease (NAFLD-HCC) and alcoholic liver disease (ALD-HCC), has been increasing. Clarification of the clinical features of NAFLD-HCC and ALD-HCC is needed. We performed a large retrospective multicenter survey to clarify the clinical course of these two types of HCC. Clinical characteristics, survival, and recurrence were examined in 532 patients with ALD-HCC and 209 patients with NAFLD-HCC who were diagnosed between January 2000 and December 2013. The ALD-HCC patients were predominantly male and were younger than the patients with NAFLD-HCC. Lifestyle-related diseases were significantly more common in the NAFLD-HCC group, but the prevalence of cirrhosis was significantly higher in the ALD-HCC group. The histological diagnosis of NAFLD-HCC showed a gender difference (F4; 72.7 % in the females vs. 37.6 % in the males). The characteristic features of HCC including histology, survival rate, and recurrence rate were quite similar in the NAFLD-HCC and ALD-HCC groups: 5-year survival rates 49.1 vs. 43.7 %; 5-year recurrence rates 69.6 vs. 65.4 %, respectively. However, the risk factors for recurrence differed between the two groups: des-gamma-carboxy prothrombin was a risk factor in NAFLD-HCC and α-fetoprotein was a risk factor in ALD-HCC. Although the characteristic features underlying these two diseases are different, the two HCC groups showed a similar clinical course. The recurrence rates of the two HCC groups were relatively high. We found that critical tumor markers for recurrence differed between the two diseases.


Replicative fitness of historical and recent HIV-1 isolates suggests HIV-1 attenuation over time.
Changes in virulence during an epidemic are common among pathogens, but still unexplored in the case of HIV-1. Here we used primary human cells to study the replicative fitness of primary HIV-1 isolates from untreated patients, comparing historical (1986-1989) and recent samples (2002-2003). Head-to-head dual virus infection/competition assays were performed in both peripheral blood mononuclear cells and human dendritic cell/T-cell co-cultures with pairs of 12 carefully matched historical and recent HIV-1 isolates from untreated patients. Sensitivity to inhibition by lamivudine (3TC) and TAK-779 of historical and recent R5 HIV-1 isolates was measured in a subset of samples. Overall, the historical HIV-1 out-competed the recent HIV-1 isolates in 176 of 238 competitions and in 9 of 12 competitions carefully matched for CD4 cell count. The mean relative replicative fitness (W) of all historical HIV-1 strains was significantly greater than that of recent HIV-1 isolates (W(1986-1989) = 1.395 and W(2002-2003) = 0.545, P < 0.001 (t test)). The more fit viruses (mean W > 1) from 1986-1989 appeared less sensitive to TAK-779 and 3TC than did the less fit (mean W < 1) 2002-2003 viruses. These findings suggest that HIV-1 replicative fitness may have decreased in the human population since the start of the pandemic. This 'attenuation' could be the consequence of serial bottlenecks during transmission and result in adaptation of HIV-1 to the human host.


FMNL formins boost lamellipodial force generation.
Migration frequently involves Rac-mediated protrusion of lamellipodia, formed by Arp2/3 complex-dependent branching thought to be crucial for force generation and stability of these networks. The formins FMNL2 and FMNL3 are Cdc42 effectors targeting to the lamellipodium tip and shown here to nucleate and elongate actin filaments with complementary activities in vitro. In migrating B16-F1 melanoma cells, both formins contribute to the velocity of lamellipodium protrusion. Loss of FMNL2/3 function in melanoma cells and fibroblasts reduces lamellipodial width, actin filament density and -bundling, without changing patterns of Arp2/3 complex incorporation. Strikingly, in melanoma cells, FMNL2/3 gene inactivation almost completely abolishes protrusion forces exerted by lamellipodia and modifies their ultrastructural organization. Consistently, CRISPR/Cas-mediated depletion of FMNL2/3 in fibroblasts reduces both migration and capability of cells to move against viscous media. Together, we conclude that force generation in lamellipodia strongly depends on FMNL formin activity, operating in addition to Arp2/3 complex-dependent filament branching.


Disorders of pyruvate metabolism.
Pyruvate dehydrogenase and pyruvate carboxylase deficiency are the most common disorders in pyruvate metabolism. Diagnosis is made by enzymatic and DNA analysis after basic biochemical tests in plasma, urine, and CSF. Pyruvate dehydrogenase has three main subunits, an additional E3-binding protein and two complex regulatory enzymes. Most frequent are deficiencies in PDH-E1α. There is a spectrum of clinical presentations in E1α deficiency, ranging in boys from severe neonatal lactic acidosis, Leigh encephalopathy, to later onset of neurological disease such as intermittent ataxia or dystonia. Females tend to have a more uniform presentation resembling nonprogressive cerebral palsy. Neuroradiological abnormalities such as corpus callosum agenesis are seen more frequently in girls, basal ganglia and midbrain disturbances in boys. Deficiencies in the other subunits have also been described, but in a smaller number of patients. Pyruvate carboxylase deficiency has three clinical phenotypes. The infantile type is characterized mainly by severe developmental delay, failure to thrive, and seizures. The second type is characterized by neonatal onset of severe lactic acidosis with rigidity and hypokinesia. A third form is rarer with intermittent episodes of lactic acidosis and ketoacidosis. Neuroradiological findings such as cystic periventricular leukomalacia have been described.


New chlorinated amphetamine-type-stimulants disinfection-by-products formed during drinking water treatment.
Previous studies have demonstrated high removal rates of amphetamine-type-stimulants (ATSs) through conventional drinking water treatments; however the behaviour of these compounds through disinfection steps and their transformation into disinfection-by-products (DBPs) is still unknown. In this work, for the first time, the reactivity of some ATSs such as amphetamine, methamphetamine, 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxymethamphetamine (MDMA) and 3,4-methylenedioxyethylamphetamine (MDEA) with chlorine has been investigated under simulated and real drinking water treatment conditions in order to evaluate their ability to give rise to transformation products. Two new DBPs from these illicit drugs have been found. A common chlorinated-by-product (3-chlorobenzo)-1,3-dioxole, was identified for both MDA and MDEA while for MDMA, 3-chlorocatechol was found. The presence of these DBPs in water samples collected through drinking water treatment was studied in order to evaluate their formation under real conditions. Both compounds were generated through treatment from raw river water samples containing ATSs at concentration levels ranging from 1 to 15 ng/L for MDA and from 2.3 to 78 ng/L for MDMA. One of them, (3-chlorobenzo)-1,3-dioxole, found after the first chlorination step, was eliminated after ozone and GAC treatment while the MDMA DBP mainly generated after the postchlorination step, showed to be recalcitrant and it was found in final treated waters at concentrations ranging from 0.5 to 5.8 ng/L.


Alterations in vitamin A metabolism by polyhalogenated aromatic hydrocarbons.
Adequate stores and adequate tissue levels of vitamin A are maintained by a balance of tissue demands and dietary intake of the vitamin and are modified by many factors, including xenobiotics. It is well established that exposure to polyhalogenated aromatic hydrocarbons (PHAH) decreases hepatic content of vitamin A. Recent findings indicate that hepatic depletion of vitamin A is accompanied by an increase in serum and renal vitamin A content and enhanced excretion of vitamin A metabolites in urine and feces. Examination of tissue retinoid profiles reveals that PHAH exposure causes the generation of increased amounts of polar retinoids. It is very likely that PHAH affect enzymes crucial for regulation of vitamin A storage as well as enhance activities of specific enzymes in vitamin A metabolic pathway.


Gasless laparoendoscopic single-site assisted extracorporeal ovarian cystectomy through the umbilicus for the management of ovarian torsion at 33 weeks of gestation: A case report with literature review.
Ovarian torsion in the third trimester of pregnancy is a rare but potentially dangerous non-obstetric complication for both mother and fetus and presents a diagnostic and therapeutic challenge for the treating physicians. A 39-year-old woman (gravida 2, para 1) presented at 7 weeks of gestation. Asymptomatic bilateral small ovarian cysts were diagnosed at the initial presentation. Progesterone was intramuscularly administered every 2 weeks after 28 weeks of gestation because of uterine cervical length shortening. Sudden onset of right lateral abdominal pain was reported at 33 weeks and 2 days of gestation. Emergency laparoendoscopic single-site (LESS) surgery was performed through the umbilicus under strong suspicion of right adnexal torsion with ovarian cyst, as indicated by magnetic resonance imaging a day after admission. Isolated right ovarian torsion without fallopian tube involvement was identified under laparoscopic view. The contents of the right ovarian cyst were aspirated after confirming resumption of color tone of the right ovary after detorsion. Then, the right adnexal tissue was grasped through the umbilicus, followed by a successful ovarian cystectomy under direct vision. Tocolysis was postoperatively attempted by intravenous ritodorine hydrochloride and magnesium sulfate administration, which was continued through to 36 weeks and 4 days of gestation, because of increased uterine contraction frequency. The next day, spontaneous labor occurred, followed by the vaginal delivery of a healthy 2108-g female infant. The postnatal course was uneventful. Transumbilical LESS-assisted extracorporeal ovarian cystectomy is a feasible and minimally invasive option for ovarian torsion management in the third trimester of pregnancy.


Combination therapy: A New Tool for the Management of Obesity.
Obesity is a long-term lifestyle problem that can lead to disastrous consequences. Lifestyle interventions are one of the initial lines of management strategies for obesity but in long term, it is not very effective in the management of obesity. Some people also manage their weight with bariatric surgery, which is now the most effective obesity treatment. Synthetic pharmaceuticals make a bridge between lifestyle modification and bariatric surgery-based obesity management. The major problem associated with monotherapy without side effects is that these are moderately effective and also need in higher doses. The combination therapy is already used for many serious and complicated disease treatments and management and has shown efficacy as well. Generally, we use two or more medicines with different of actions for a better effect. The commonly used combination therapy for obesity management includes low-dose phentermine and prolonged and slow-releasing mechanism topiramate; naltrexone along with bupropion. Phentermine with inhibitors of Na-glucose cotransporter-2 or glucagon-like peptide-1 (GLP-1) agonist with gastric hormone or Na-glucose cotransporter-2 is two more viable combo therapy. This combination strategy aims to achieve success in the bariatric surgery and the scientific community is working in this direction.


Combined radiotherapy and immunotherapy in urothelial bladder cancer: harnessing the full potential of the anti-tumor immune response.
Radiotherapy (RT), as part of trimodal therapy, is an attractive alternative treatment in patients with urothelial muscle-invasive bladder cancer (MIBC). There is accumulating evidence suggesting the immunomodulatory effects of RT and its potential synergy when combined with immunotherapy. The aim of this review was to report on the most recent advances on this combination, including the mechanisms of RT immunomodulation, practical approach to combining RT and immunotherapy, and ongoing clinical trials in bladder cancer. Using the PubMed database, we identified articles published between March 2004 and April 2020 on the combination of RT with immunotherapy in localized or metastatic MIBC. A search of the Clinicaltrials.gov and Clinicaltrialsregister.eu/ retrieved ongoing clinical trials on the topic as well. Combination of RT with immunotherapy leads to immunogenic cell death and an increase in immune markers thus leading to improved tumor control. For localized MIBC, there are safety concerns related to the use of concurrent immunotherapy with hypofractionated RT, thus neoadjuvant or adjuvant immunotherapy is preferred. In the metastatic setting, the combination of multi-site RT with SBRT-like doses (≥ 6 Gy per fraction) and concurrent immunotherapy seems most efficacious at harnessing the abscopal effect. At least 25 clinical trials combining immunotherapy and RT in MIBC are currently ongoing and will answer pending questions on safety, efficacy, and practical considerations on RT scheduling, fractionation, and targets volumes. RT has the potential to synergize with immunotherapy to improve oncological outcomes in patient with localized or metastatic MIBC. Clinical trials results are eagerly awaited.


A statistical theory of amino acid mutation.
Assuming that the observed mutation frequency of an amino acid depends on two factors. The first is mutation coefficient which describes the rate of the nucleotide substitution stochastically and the second is the similarity of amino acids which represents the fitness of a mutant under the selective pressure. A statistical theory is proposed and 380 mutation frequencies are calculated, only 10 of which disagree obviously with the observed data.


Fetal factors disrupt placental and maternal iron homeostasis in murine β-thalassemia.
Pregnancy rates in β-thalassemia are increasing but the risk of complications is higher; thus, better understanding of maternal and fetal iron homeostasis in this disorder is needed. HbbTh3/+ (Th3/+) mice model human β-thalassemia. Both the murine and human diseases are characterized by low hepcidin, high iron absorption, and tissue iron overload, with concurrent anemia. We hypothesized that disordered iron metabolism in pregnant Th3/+ mice would negatively affect their unborn offspring. The experimental design included these groups: wild-type (WT) dams carrying WT fetuses (WT1); WT dams carrying WT and Th3/+ fetuses (WT2); Th3/+ dams carrying WT and Th3/+ fetuses (Th3/+); and age-matched, nonpregnant adult females. Serum hepcidin was low, and mobilization of splenic and hepatic storage iron was enhanced in all 3 groups of experimental dams. Intestinal 59Fe absorption was lower in Th3/+ dams (as compared with WT1/2 dams) but splenic 59Fe uptake was higher. Th3/+ dams had hyperferremia, which led to fetal and placenta iron loading, fetal growth restriction, and placentomegaly. Notably, Th3/+ dams loaded Th3/+ and WT fetuses, with the latter situation more closely mirroring human circumstances when mothers with thalassemia have relatively unaffected (thalassemia trait) offspring. Iron-related oxidative stress likely contributed to fetal growth impairment; enhanced placental erythropoiesis is a probable cause of placental enlargement. Moreover, high fetal liver iron transactivated Hamp; fetal hepcidin downregulated placental ferroportin expression, limiting placental iron flux and thus mitigating fetal iron loading. Whether gestational iron loading occurs in human thalassemic pregnancy, when blood transfusion can further elevate serum iron, is worth consideration.


Demolishing the great wall of biofilms in Gram-negative bacteria: To disrupt or disperse?
Bacterial infections lead to high morbidity and mortality globally. While current therapies against bacteria often employ antibiotics, most bacterial pathogens can form biofilms and prevent effective treatment of infections. Biofilm cells can aggregate and encased themselves in a self-secreted protective exopolymeric matrix, to reduce the penetration by antibiotics. Biofilm formation is mediated by c-di-GMP signaling, the ubiquitous secondary messenger in bacteria. Synthesis of c-di-GMP by diguanylate cyclases leads to biofilm formation via the loss of motility, increased surface attachment, and production of biofilm matrix, whereas c-di-GMP degradation by phosphodiesterases causes biofilm dispersal to new sites via increased bacterial motility and matrix breakdown. The highly variable nature of biofilm development and antimicrobial tolerance imposes tremendous challenges in conventional antimicrobial therapies, indicating an imperative need to develop anti-biofilm drugs against biofilm infections. In this review, we focus on two main emergent approaches-active dispersal and disruption. While both approaches aim to demolish biofilms, we will discuss their fundamental differences and associated methods. Active dispersal of biofilms involves signaling the bacterial cells to leave the biofilm, where resident cells ditch their sessile lifestyle, gain motility and self-degrade their matrix. Biofilm disruption leads to direct matrix degradation that forcibly releases embedded biofilm cells. Without the protection of biofilm matrix, released bacterial cells are highly exposed to antimicrobials, leading to their eradication in biofilm infections. Understanding the advantages and disadvantages of both approaches will allow optimized utility with antimicrobials in clinical settings.


Fast Lane to Slow Science.
Fast Science is bad for scientists and bad for science. Slow Science may actually help us to make faster progress, but how can we slow down? Here, I offer preliminary suggestions for how we can transition to a healthier and more sustainable research culture.


Effects of local administration of vascular endothelial growth factor on properties of the in situ frozen-thawed anterior cruciate ligament in rabbits.
In the autogenous tendon for anterior cruciate ligament reconstruction, intrinsic fibroblasts are necrotized immediately after surgery, and repopulation and revascularization occur. Vascular endothelial growth factor is considered to be a potent mediator of angiogenesis. An application of vascular endothelial growth factor significantly enhances angiogenesis in the in situ frozen anterior cruciate ligament, and the application significantly affects mechanical properties of the in situ frozen anterior cruciate ligament. Controlled laboratory study. Right anterior cruciate ligaments from 66 rabbits underwent the freeze-thaw treatment, and animals were then divided into 3 groups. Group I served as a freeze-thaw but otherwise untreated control. In group II, 0.2 mL phosphate-buffered saline alone was applied. In group III, 30 mug vascular endothelial growth factor was applied. The groups were compared on the basis of histologic revascularization examinations using the Chalkley score, an indicator of the microvessel density, and mechanical evaluations, which included the anterior-posterior translation of the tibia relative to the femur during +/- 10 N of anterior-posterior load and the mechanical properties of the anteromedial bundle of the anterior cruciate ligament. Group III's Chalkley score was significantly greater than that of groups I and II. The tensile strength and the tangent modulus of anterior cruciate ligaments in groups I, II, and III were significantly lower than those of a normal anterior cruciate ligament, although there were no significant differences among groups I, II, and III. Vascular endothelial growth factor, as administered in this study, significantly promoted angiogenesis in the devitalized anterior cruciate ligament with in situ freeze-thaw treatment, but it did not affect the mechanical properties of the in situ frozen-thawed anterior cruciate ligament in the rabbit model. An application of the recombinant anterior cruciate ligament is a potential future strategy to enhance revascularization of the autograft in anterior cruciate ligament reconstruction.


The Influence of Pretherapeutic and Preoperative Sarcopenia on Short-Term Outcome after Esophagectomy.
By introducing minimally invasive surgery the rate of postoperative morbidity in esophageal cancer patients could be reduced. But esophagectomy is still associated with a relevant risk of postoperative morbidity and mortality. Patients often present with nutritional deficiency and sarcopenia even at time of diagnosis. This study focuses on the influence of skeletal muscle index (SMI) on postoperative morbidity and mortality. Fifty-two patients were included in this study. SMI was measured using computer tomographic images at the time of diagnosis and before surgery. Then, SMI and different clinicopathological and demographic features were correlated with postoperative morbidity. There was no correlation between SMI before neoadjuvant therapy (p = 0.5365) nor before surgery (p = 0.3530) with the short-term postoperative outcome. Regarding cholesterol level before surgery there was a trend for a higher risk of complications with lower cholesterol levels (p = 0.0846). Our findings suggest that a low preoperative SMI does not necessarily predict a poor postoperative outcome in esophageal cancer patients after esophagectomy but that there are many factors that influence the nutritional status of cancer patients. To improve nutritional status, cancer patients at our clinic receive specialized nutritional counselling during neoadjuvant treatment as well as after surgery.


Successful treatment of aortic root aneurysm after orthotopic heart transplantation: case report.
Aortic root aneurysm after orthotopic heart transplantation (HTx) is rare. It may originate from cystic medial necrosis of the donor heart aorta. Herein we have reported a 64-year-old man who received an orthotopic HTx due to dilated cardiomyopathy. Although asymptomatic, follow-up echocardiography revealed dilatation of the aortic root and severe aortic regurgitation at 3 years after the transplantation. He underwent a Bentall procedure with a prosthetic valved conduit. The post-operative course was uneventful. This case demonstrated that a heart-transplant recipient with a late aortic root aneurysm can be successfully treated with an excellent outcome.


A simple method for percutaneous cannulation of the internal jugular vein.
A safe and rapid technic for internal jugular vein cannulation is described. The success rate is almost as high and the serious complication rate is much lower than subclavian vein cannulation.


Prognostic significance of tumor deposits in gastric cancer patients who underwent radical surgery.
To investigate the prognostic significance of tumor deposits (TDs) in gastric cancers patients who underwent radical surgery. Clinicopathologic and prognostic data from 2998 gastric cancer patients who underwent R0 surgery with D2/D3 lymphadenectomy were retrospectively reviewed. A TD was defined as discrete foci of tumor found in the perigastric fat or in adjacent ligament away from the leading edge of the tumor and showing no evidence of residual lymph node tissue, but within the lymph drainage area of the primary carcinoma. TDs were detected in 17.8% of patients. TDs were more frequently observed in cancers of larger size, of Borrmann type 4, with lymphovascular invasion, deeper in depth of invasion, and with extended lymph node metastasis. Multivariate analysis confirmed the presence of TDs as 1 of independent factors predicting a poorer outcome. When stratified by pN category, significant differences in survival were observed between patients with and without TDs for those in pN0/pT1-3, pN1/pT3, pN2/pT1-3 and pN3/pT2-3 category, but not for those in pT4a and pT4b category. Moreover, for cancers in each pN category, the prognosis for patients with TDs in pT1-4a category was similar with that of those without TDs in pT4a category, but significantly better than that of those with or without TDs in pT4b category. A revised pT category and a revised pTNM system were proposed, in which all the cancers with TDs in pT1-4a category were incorporated into those without TDs in pT4a category according to the pN category. Further analysis revealed the revised pT category and the revised pTNM system had better homogeneity, discriminatory ability, and monotonicity of gradients than the American Joint Committee on Cancer (AJCC) pT category and the AJCC pTNM system, respectively, representing optimum prognostic stratification. TDs significantly correlated with gastric cancer patients' survival. It might be more suitable for TDs to be treated as a form of serosal invasion. Consequently, en bloc resection of the primary carcinoma is crucially important, and adjuvant chemotherapy should always be considered if TDs have been detected.


Humor-based interventions for patients undergoing hemodialysis: A scoping review.
There is evidence that humor interventions can impact chronic kidney disease (CKD) patients. Synthesize relevant literature to understand the elements and mechanisms of implementation of humor interventions. Relevant peer-reviewed articles were searched. Data were extracted according to the Template for Intervention Description and Replication (TIDieR) checklist. The initial search yielded 261 articles. Nine were included in the scoping review. The studies included laughter (n = 5) and humorous video interventions (n = 4) in a face-to-face group format. The tailoring and fidelity process needs to be more detailed in most studies. None of the studies mentioned the theoretical basis. The humor videos were provided by nurses and required various types of equipment. The video interventions were conducted during dialysis. The laughter intervention consisted of 3-4 themes guided by qualified therapists. The timing of the laughter intervention is chosen based on the patient's dialysis schedule and is maintained for 30 min. The characteristics of the current study somewhat limit the understanding, replication, and implementation of evidence-based humor interventions. Future studies need to clarify the theoretical basis, tailoring, fidelity, and control groups. Ongoing reporting and evaluation of the implementation of humor interventions may help better understand their underlying mechanisms.


Purification, crystallization, and properties of D-ribose isomerase from Mycobacterium smegmatis.
D-Ribose isomerase, which catalyzes the conversion of D-ribose to D-ribulose, was purified from extracts of Mycobacterium smegmatis grown on D-ribose. The purified enzyme crystalized as hexagonal plates from a 44% solution of ammonium sulfate. The enzyme was homogenous by disc gel electrophoresis and ultracentrifugal analysis. The molecular weight of the enzyme was between 145,000 and 174,000 by sedimentation equilibrium analysis. Its sedimentation constant of 8.7 S indicates it is globular. On the basis of sodium dodecyl sulfate gel electrophoresis in the presence of Mn2+, the enzyme is probably composed of 4 identical subunits of molecular weight about 42,000 to 44,000. The enzyme was specific for sugars having the same configuration as D-ribose at carbon atoms 1 to 3. Thus, the enzyme could also utilize L-lyxose, D-allose, and L-rhamnose as substrates. The Km for D-ribose was 4 mM and for L-lyxose it was 5.3 mM. The enzyme required a divalent cation for activity with optimum activity being shown with Mn2+. the Km for the various cations was as follows: Mn2+, 1 times 10(-7) M, Co2+, 4 times 10(-7) M, and Mg2+, 1.8 times 10(-5) M. The pH optimum for the enzyme was 7.5 to 8.5. Polyols did not inhibit the enzyme to any great extent. The product of the reaction was identified as D-ribulose by thin layer chromatography and by preparation of the O-nitrophenylhydrazone derivative.


Innovative technique to reconstruct two branches of the right hepatic artery in living donor liver transplantation.
Hepatic arterial thrombosis is a critical complication in living donor liver transplantation (LDLT). Two separate branches of the right hepatic artery (RHA) are sometimes observed and addressed by anastomosis of the larger branch first, then checking backflow from the smaller branch. If not good, the smaller branch must be reconstructed. We used the cystic artery as a conduit for the reconstruction. Meticulous dissection was performed to identify all branches of the hepatic artery in the donor operation. The length of cystic artery preserved was as long as possible. The cystic arterial stump was anastomosed to the stump of the posterior branch the of RHA under microscopic guidance on the back table. Patency was checked through the stump of the anterior branch of the RHA. With this technique, only one orifice, the stump of right anterior hepatic artery, was used for hepatic artery reconstruction. We have performed this technique in two patients. Both had good arterial flow after living donor liver transplantation. This innovative technique is easy and safe, and requires only one anastomosis, which, in theory, decreases the adds of developing hepatic arterial thrombosis.


Influence of catecholamines on cytokine production and expression of adhesion molecules of human neutrophils in vitro.
The impact of catecholamines on cytokine production and expression of adhesion molecules by human neutrophils was evaluated in vitro. Neutrophils were separated from venous blood of healthy subjects. The generation of intracellular cyclic adenosine monophosphate (cAMP) and Ca2+ was determined after incubation with catecholamines. Resting and lipopolysaccharide (LPS)-activated neutrophils were tested for synthesis of interleukins (IL-6, IL-8) and tumor necrosis factor alpha (TNF-alpha). In addition, the expression of the adhesion molecules CD15, CD44, and CD54 was evaluated in resting and activated neutrophils. Increasing concentrations (1 nM-1 mM) of epinephrine (EPI) were used to study the influence of activation of beta2-adrenergic receptors (beta2R) on cytokine production and adhesion molecule expression. Incubation with catecholamines induced an increase in cAMP but not Ca2+ in neutrophils. Only IL-8 was detected following stimulation with LPS and was unchanged upon co-incubation with EPI. The expression of CD15 and CD44 decreased spontaneously in vitro. The density of CD44 increased in the presence of very high doses of EPI (1 mM). Expression of CD54 on resting neutrophils increased upon activation. The density of CD54 on activated neutrophils was reduced upon co-incubation with 1 mM EPI for 6 h. However, 1 mM EPI for 12 and 18 h decreased the spontaneous loss of CD54 on resting neutrophils. Beta2R are functionally coupled to signalling cascades in human neutrophils. Nevertheless, the impact of catecholamines on IL-8 synthesis and expression of CD15, CD44, and CD54 is limited.


Pre-clinical evaluation of an innovative oral nano-formulation of baicalein for modulation of radiation responses.
There is an unmet medical need for non-toxic and effective radiation countermeasures for prevention of radiation toxicity during planned exposures. We have earlier shown that intraperitoneal administration of baicalein (BCL) offers significant survival benefit in animal model. Safety, tolerability, pharmacokinetics (PK) and pharmacodynamics of baicalein has been reported in pre-clinical model systems and also in healthy human volunteers. However, clinical translation of baicalein is hindered owing to poor bioavailability due to lipophilicity. In view of this, we fabricated and characterized in-situ solid lipid nanoparticles of baicalein (SLNB) with effective drug entrapment and release kinetics. SLNB offered significant protection to murine splenic lymphocytes against 4 Gy ionizing radiation (IR) induced apoptosis. Oral administration of SLNB exhibited ~70% protection to mice against whole body irradiation (WBI 7.5 Gy) induced mortality. Oral relative bioavailability of BCL was enhanced by over ~300% after entrapment in the SLNB as compared to BCL. Oral dosing of SLNB resulted in transient increase in neutrophil abundance in peripheral blood. Interestingly, we observed that treatment of human lung cancer cells (A549) with radioprotective dose of SLNB exhibited radio-sensitization as evinced by decrease in survival and clonogenic potential. Contrary to antioxidant nature of baicalein in normal cells, SLNB treatment induced significant increase in cellular ROS levels in A549 cells probably due to higher uptake and inhibition of TrxR. Thus, a pharmaceutically acceptable SLNB exhibited improved bioavailability, better radioprotection to normal cells and sensitized cancer cells to radiation induced killing as compared to BCL suggesting its possible utility as an adjuvant during cancer radiotherapy.


Longitudinal association between grip strength and the risk of heart diseases among European middle-aged and older adults.
Few multi-country European studies have investigated the association between grip strength and heart diseases incidence. Thus, the aim of this study is to analyse the longitudinal relationship between grip strength and the diagnosis of heart diseases in European middle-aged and older adults. A prospective cohort study was conducted using data from the Survey of Health, Aging and Retirement in Europe (2004-2017). Participants were 20,829 middle-aged and older adults from 12 countries. GS was objectively measured by a dynamometer and heart diseases diagnosis was self-reported. Incidence rate of heart diseases was calculated and a Cox proportional hazard regression was performed. The heart diseases incidence rate decreased from 930 per 100,000 person-years in the lowest quartile to 380 per 100,000 person-years in the highest grip strength quartile. During the 13 years of follow-up, compared to being in the lowest grip strength quartile, being in the highest quartile decreased the hazard of being diagnosed with a heart disease in 36 % (95 % confidence interval [CI]: 0.53, 0.78) for the whole sample, 35 % (95 % CI: 0.51, 0.84) for men and 46 % (95 % CI: 0.40, 0.73) for women. Grip strength seems to be inversely associated with the incidence of heart diseases among European middle-aged and older adults. Scientific evidence has highlighted the potential role of grip strength as a risk stratifying measure for heart diseases, suggesting its potential to be included in the cardiovascular risk scores used in primary care. However, further research is still needed to clarify it.


Rapidly stopping hemorrhage by enhancing blood clotting at an opened wound using chitosan/polylactic acid/polycaprolactone wound dressing device.
Doxycycline and monosodium glutamate (MSG) loaded chitosan (CHI)/polylactic acid (PLA)/polycaprolactone (PCL) blend film was studied as a model device to deliver drug to targeted human organ which in this case was the skin with opened wound. The CHI/PLA/PCL blend film containing 60 % CHI, 28 % PLA, and 12 % PCL exhibited the good properties for making the dressing device. It was observed that doxycycline/MSG loaded CHI/PLA/PCL blend film could rapidly deliver both doxycycline and MSG at the high release percentage approaching 100 % loaded. MSG accelerated blood clotting and fibrin formation; thus, it exhibited the good hemostatic activity. The antibacterial activity of doxycycline loaded CHI/PLA/PCL blend film against Staphylococcus aureus and Escherichia coli as model bacteria was investigated. Doxycycline release played the crucial role in bacterial inhibition as observed from the lowest bacterial cell dry weight observed when compared with the control bacterial culture or the bacterial cultures with the presence of other films studied.


A repressor (MAT alpha 2 Product) and its operator control expression of a set of cell type specific genes in yeast.
The product of the yeast MAT alpha 2 gene, alpha 2 protein, negatively regulates expression of a class of yeast cell type specific genes, the a-specific genes. In this paper we show that alpha 2 is a sequence-specific DNA binding protein. It recognizes a 32 base pair DNA sequence (operator) located upstream of an a-specific gene, STE6. A strongly homologous sequence is present upstream of each of the other four known a-specific genes. A synthetic STE6 operator, when placed in a test promoter (CYC1), brings the promoter under negative control by alpha 2 in vivo. This operator brings about repression when it is placed between the CYC1 upstream activation sequence (UAS) and the transcription start and when it is placed upstream of the UAS, outside the promoter. Thus, the operator need not overlap with essential promoter sequences to permit repression by alpha 2.


Reprogramming astrocytic NDRG2/NF-κB/C3 signaling restores the diabetes-associated cognitive dysfunction.
Dementia is a serious complication in patients with diabetes-associated cognitive dysfunction (DACD). In this study, we aim to explore the protective effect of exercise on DACD in diabetic mice, and the role of NDRG2 as a potential guarder for reversing the pathological structure of neuronal synapses. Seven weeks of standardized exercise at moderate intensity was carried out using an animal treadmill in the vehicle + Run and STZ + Run groups. Based on quantitative transcriptome and tandem mass tag (TMT) proteome sequencing, weighted gene co-expression analysis (WGCNA) and gene set enrichment analysis (GSEA) were used to investigate the activation of complement cascades to injury neuronal synaptic plasticity. Golgi staining, Western blotting, immunofluorescence staining, and electrophysiology were used to verify the reliability of sequencing data. The role of NDRG2 was assessed by overexpressing or inhibiting the NDRG2 gene in vivo. Moreover, we estimated the cognitive function in diabetic or normal patients using DSST scores. Exercise reversed the injury of neuronal synaptic plasticity and the downregulation of astrocytic NDRG2 in diabetic mice, which succeeded in attenuating DACD. The deficiency of NDRG2 aggravated the activation of complement C3 by accelerating the phosphorylation of NF-κB, ultimately leading to synaptic injury and cognitive dysfunction. Conversely, the overexpression of NDRG2 promoted astrocytic remodeling by inhibiting complement C3, thus attenuating synaptic injury and cognitive dysfunction. Meanwhile, C3aR blockade rescued dendritic spines loss and cognitive deficits in diabetic mice. Moreover, the average DSST score of diabetic patients was significantly lower than that of non-diabetic peers. Levels of complement C3 in human serum were elevated in diabetic patients compared to those in non-diabetic patients. Our findings illustrate the effectiveness and integrative mechanism of NDRG2-induced improvement of cognition from a multi-omics perspective. Additionally, they confirm that the expression of NDRG2 is closely related to cognitive function in diabetic mice and the activation of complement cascades accelerated impairment of neuronal synaptic plasticity. NDRG2 acts as a regulator of astrocytic-neuronal interaction via NF-κB/C3/C3aR signaling to restore synaptic function in diabetic mice. This study was supported by the National Natural Science Foundation of China (No. 81974540, 81801899, 81971290), the Key Research and Development Program of Shaanxi (Program No. 2022ZDLSF02-09) and Fundamental Research Funds for the Central Universities (Grant No. xzy022019020).


[Traumatic brain injury: impact on timing and modality of fracture care].
Traumatic brain injury (TBI) represents the major "killing factor" after trauma in young individuals. Those patients who survive the initial injury are highly susceptible to secondary insults to the injured brain which are mainly caused by hypotension and/or hypoxia in the early resuscitative period. Furthermore, a potent inflammatory cascade is initiated within the injured brain which leads to the development of brain edema and delayed neuronal cell death. This profound endogenous neuroinflammatory response after TBI, which is phylogenetically aimed at repairing lesioned tissue and defending the brain from invading pathogens, is in large part responsible for the extent of secondary brain damage and adverse outcome. Thus, the optimal management of the multiply injured patient, based on a thorough understanding of the pathophysiological alterations after TBI, should avoid an iatrogenic "second hit" which may be devastating to the injured brain. The standard approach of "early total care" for isolated fractures should be strictly avoided in brain-injured patients in favor of an "orthopedic damage control" concept with temporary external fixation of long bone fractures and priority given to early transfer to intensive care. The present review provides an up-to-date overview on the neuroinflammatory pathophysiology of brain injury and its implications for an optimized concept of fracture care in TBI patients.


An ontology for developmental processes and toxicities of neural tube closure.
In recent years, the development and implementation of animal-free approaches to chemical and pharmaceutical hazard and risk assessment has taken off. Alternative approaches are being developed starting from the perspective of human biology and physiology. Neural tube closure is a vital step that occurs early in human development. Correct closure of the neural tube depends on a complex interplay between proteins along a number of protein concentration gradients. The sensitivity of neural tube closure to chemical disturbance of signalling pathways such as the retinoid pathway, is well known. To map the pathways underlying neural tube closure, literature data on the molecular regulation of neural tube closure were collected. As the process of neural tube closure is highly conserved in vertebrates, the extensive literature available for the mouse was used whilst considering its relevance for humans. Thus, important cell compartments, regulatory pathways, and protein interactions essential for neural tube closure under physiological circumstances were identified and mapped. An understanding of aberrant processes leading to neural tube defects (NTDs) requires detailed maps of neural tube embryology, including the complex genetic signals and responses underlying critical cellular dynamical and biomechanical processes. The retinoid signaling pathway serves as a case study for this ontology because of well-defined crosstalk with the genetic control of neural tube patterning and morphogenesis. It is a known target for mechanistically-diverse chemical structures that disrupt neural tube closure The data presented in this manuscript will set the stage for constructing mathematical models and computer simulation of neural tube closure for human-relevant AOPs and predictive toxicology.


Infection control: the case for horizontal rather than vertical interventional programs.
The authors define two types of infection control interventions: horizontal, in which all infections at any site are reduced; and vertical, in which only specific organisms are targeted. We suggest that horizontal programs should form the platform of all infection control programs and the key question should be, what is the incremental value of a new vertical program?


Effects of intravenous acetaldehyde, acrolein, formaldehyde and propionaldehyde on arterial blood pressure following acute guanethidine treatment.
Guanethidine (G) is currently used in the treatment of essential hypertension. Acetaldehyde (A), acrolein (AR), formaldehyde (F) and propionaldehyde (P) are constituents of cigarette smoke and (A) is also an intermediate oxidative metabolite of ethanol. These aldehydes are known to produce sympathomimetic effects by the release of NE from adrenergic neurons and to exert cardioinhibitory effects. The type of predominant effect is dose-dependent. This study was undertaken to determine if these aldehydes result in sympathomimetic effects in the presence of G (15 mg/kg iv) in anesthetized rats. G enhanced the pressor responses to A and P in dose ranges of 5-20 and 5-10 mg/kg respectively. However, AR and F at dose ranges of 0.05-5 and 0.1-10 mg/kg, respectively, elicited only depressor responses after G. Thus, A and P exerted greater sympathomimetic effects through the release of intraneuronal NE in the presence of G. The adrenergic neuronal blocking action of G changes the blood pressure effects of AR and F. When these two compounds are administered in the absence of G, they cause predominant pressor effects, whereas in the presence of G a depressor response is noted. This depressor response is possibly by vagal stimulation and/or direct vasodilation.


Influence of a selective histamine H3 receptor antagonist on hypothalamic neural activity, food intake and body weight.
This study was conducted to elucidate whether antagonistic targeting of the histamine H3 receptor increases hypothalamic histamine levels, in parallel with decreases in food intake and body weight. The competitive antagonist potency of a recently synthesized histamine H3 receptor antagonist, NNC 38-1049, was studied in intact HEK293 cells expressing human or rat histamine H3 receptor, in which NNC 38-1049 was allowed to antagonize the effect of the H3 receptor agonist R-alpha-methylhistamine on isoprenaline-induced accumulation of cAMP. The affinity of NNC 38-1049 for a number of variants of the histamine receptor was also determined. Following single dosing of normal rats with NNC 38-1049, hypothalamic histamine levels were assessed by means of microdialysis. Plasma and brain levels of NNC 38-1049 and acute effects on food intake and energy expenditure were followed after oral doses of 3-60 mg/kg. Potential side effects were examined with rat models of behaviour satiety sequence (BSS), pica behaviour and conditioned taste aversion (CTA). Intakes of food and water together with body weight were recorded for 15 days during daily dosing of dietary obese rats. NNC 38-1049 was found to be a highly specific and competitive antagonist towards both human and rat histamine H3 receptors, and measurable amounts of NNC 38-1049 were found in the plasma of rats following single oral doses of 3-60 mg/kg and in the brain after 15-60 mg/kg. Following single intraperitoneal injections of NNC 38-1049 (20 mg/kg), significant increases in extracellular histamine concentrations were observed. The same dose did not change BSS or pica behaviour acutely, nor did it induce CTA following repeated administration for 7 days. Reductions in food intake were seen very soon after administration, and occurred in a dose-dependent fashion. Energy expenditure was unchanged, but the respiratory quotient (RQ) tended to decrease at higher doses, indicating an increase in lipid oxidation. Twice daily administration of 20 mg/kg of NNC 38-1049 in old and dietary obese rats resulted in sustained reduction of food intake throughout a 2-week study, and was associated with a highly significant (P<0.01) decrease in body weight compared with controls (-18.4+/-3.4 vs +0.4+/-2.7 g). The same dose of NNC 38-1049 produced an acute decrease of water intake, but 24 h intakes were not significantly changed. The results of this study strongly support the idea that an increase in the hypothalamic concentration of histamine produces a specific reduction of food intake and that this effect can be translated into a decrease in body weight.


[Prevention of bacterial risk: pathogen inactivation/detection of bacteria].
Bacterial contamination of blood products remains the most important infectious risk of blood transfusion in 2013. Platelet concentrates (PC) are in cause in the majority of the transfusion reaction due to bacterial contaminations. A lot of prevention methods have been developed over the last 10 years (pre-donation interview, skin decontamination, diversion of the first 30 mL of the donation, leuko-reduction...), they have focused on limiting the contamination of the donations and prevent the bacterial growth in donations and/or in the blood products. These measures were effective and led to significantly reducing the risk of adverse effects associated with bacterial growth. However, every year there are about six accidents (with a high level of imputability) and one death. The reduction of the bacterial risk remains a priority for the French Blood Establishment (EFS). The procedure for skin disinfection is going to be improved in order to further strengthen this crucial step to avoid the contamination of donation. Methods of pathogen inactivation applied to plasma and PC are available in France and their effectiveness is demonstrated on the bacterial risk. Methods for bacterial detection of PC are used in many countries now. Automated culture is the most common. Alternatives are now available in the form of rapid tests able to analyze the PC just before the delivery and avoid false negatives observed with automated culture. Assessments are under way to confirm these benefits in 2013.


Circ_0000218 plays a carcinogenic role in laryngeal cancer through regulating microRNA-139-3p/Smad3 axis.
Laryngeal squamous cell carcinoma (LSCC) accounts for about 85%-90% of all cases of laryngeal cancer. So far, the role and molecular mechanism of circular RNA 0,000,218 (circ_0000218)/microRNA (miR)-139-3p in laryngeal cancer are not clear. The present study aimed to investigate the role and regulatory mechanism of circ_0000218/miR-139-3p in laryngeal cancerin vitro and in vivo. quantitative real time polymerase chain reaction (qRT-PCR) was used to detect the expression of circ_0000218/miR-139-3p in LSCC cells. Dual luciferase reporter assay and RNA immunoprecipitation (RIP) assay were used to confirm binding sites between miR-139-3p and smad family member 3 (Smad3), and circ_0000218 and miR-139-3p. Cell Counting Kit-8 (CCK-8) and cell apoptosis analysis were used to detect cell viability and apoptosis. Xenograft experiment was performed to show in vivo effect of circ_0000218/miR-139-3p on the growth of LSCC. Circ_0000218 was highly expressed in LSCC cells. miR-139-3p, lower expressed in LSCC cells, was negatively regulated by circ_0000218 in LSCC cells. Besides, the findings suggested that circ_0000218 silencing inhibited the LSCC cell viability and promoted apoptosis by negatively regulating miR-139-3p expression. Furthermore, the data indicated that miR-139-3p inhibited the viability of LSCC cells and promoted apoptosis, and these effects were reversed by Smad3 over-expression. In addition, the in vivo effects of circ_0000218/miR-139-3p on LSCC were consistent with the in vitro study. circ_0000218 inhibition inhibited the growth of LSCC by targeting miR-139-3p/Smad3 axis. Our present study provided a new target for laryngeal cancer treatment.


Gene therapy applications for intervertebral disc degeneration.
Review of current approaches to gene therapy as a potential treatment modality for regeneration of the degenerating intervertebral disc. Describe recent advances in the field of gene therapy pertaining to treatment of the degenerating intervertebral disc. The potential application of gene therapy to the treatment of degenerative disc disease has been an area of exciting and active research. Several in vitro studies have documented the capacity of gene therapy to favorably modify the biologic functions of the intervertebral disc cells with the delivery of the cDNA for various growth factors, whereas in vivo studies have reinforced this potential in animal models. Currently, investigators are exploring the therapeutic potential of gene delivery of the cDNA for several promising factors. Literature review. The application of gene therapy to the treatment of the degenerating intervertebral disc holds much clinical promise and continues to expand as an area of active research. The therapeutic potential of multiple agents, as well as possible vectors, are currently being investigated with encouraging results.


Impact of the 2018 Society of Critical Care Medicine Pain, Agitation/Sedation, Delirium, Immobility, and Sleep Guidelines on Nonopioid Analgesic Use and Related Outcomes in Critically Ill Adults After Major Surgery.
We compared ICU nonopioid analgesic use, opioid use, and pain before and after Pain, Agitation/Sedation, Delirium, Immobility, and Sleep guideline publication at one academic center among critically ill adults receiving an opioid infusion and greater than or equal to 24 hours of mechanical ventilation after major surgery. The 2017 (n = 77) and 2019 (n = 57) groups were similar at baseline. The 2019 (vs 2017) patients were more likely to receive scheduled IV/oral acetaminophen (84% vs 69%; p = 0.05), less likely to receive a lidocaine patch (33% vs 50%; p = 0.05), and just as likely to receive ketamine (4% vs 3%; p = 1.0), an nonsteroidal anti-inflammatory drug (7% vs 3%; p = 0.26), or gabapentin/pregabalin (16% vs 9%; p = 0.23). Daily average opioid exposure (in IV morphine milligram equivalent) was not different (70 [42-99] [2017] vs 78 mg [49-109 mg]; p = 0.94). The 2019 (vs 2017) group spent more ICU days with severe pain (p = 0.04). At our center, Pain, Agitation/Sedation, Delirium, Immobility, and Sleep guideline publication had little effect on nonopioid analgesic or opioid prescribing practices in critically ill surgical adults.


Feasibility of intraoperative ultrasound of the small bowel during Crohn's disease surgery.
Intraoperative assessment of the extent and location of Crohn's disease is not standardised and relies on a mixture of surgeons' experience, tactile feedback and macroscopic appearance. To overcome this variability, we developed a protocol for full intraoperative ultrasound scan of the small bowel and we here report the results of "Assessing the Feasibility and Safety of Using Intraoperative Ultrasound in Ileocolic Crohn's Disease-The IUSS CROHN Study". This is a prospective single centre observational study with enrolment of all patients undergoing elective surgery for terminal ileal Crohn's disease from January 2019 to March 2020. Patients underwent laparoscopic ileocolic resection, according to a standardised technique. Ultrasound intraoperative quantitative assessment was performed according to the METRIC (MREnterography or ulTRasound in Crohn's disease) scoring guide. Intraoperative ultrasound was successfully performed in 6 patients from the ileocaecal valve to the proximal jejunum. The median time required was 23.5 min (range 17-37 min) as compared to 6.5 min (5-12 min) required for the macroscopic evaluation performed by the surgeon. In 3 patients, intraoperative ultrasound identified more disease than surgical evaluation. This feasibility study demonstrated the safety of intraoperative ultrasound and allowed the development of a standardised protocol for intraoperative ultrasound and the data collection required to inform a randomised multicentre study.


A proposal for new definition (s) and management approach to paediatric refractory ITP: Reflections from the Intercontinental ITP Study Group.
Immune thrombocytopenia (ITP) in children is a relatively mild and self-limited disorder with the majority of children demonstrating normalization of platelet count by 12 months from diagnosis. Because of this, many children with ITP can be observed without the need for treatment. When needed, treatment with either intravenous immunoglobulin (IVIG) or corticosteroids is highly effective (>80% IVIG and >95% corticosteroids). For those children who require second-line therapies, response rates of >60% are seen with both the thrombopoietin-receptor agonists and rituximab. Despite this, some children will have 'refractory' ITP (rITP) with poor or transient responses to platelet-raising therapies. Here, we review the clinical features of rITP in children, outline proposed classifications and explore potential predictors for children with rITP.


Engineered biosynthesis of alkyne-tagged polyketides.
Polyketides have demonstrated their significance as therapeutics, industrial products, pesticides, and biological probes following intense study over the past decades. Tagging polyketides with a bioorthogonal functionality enables various applications such as diversification, quantification, visualization and mode-of-action elucidation. The terminal alkyne moiety, as a small, stable and highly selective clickable functionality, is widely adopted in tagging natural products. De novo biosynthesis of alkyne-tagged polyketides offers the unique advantage of reducing the background from feeding the biorthogonal moiety itself, leading to the accomplishment of in situ generation of a clickable functionality for bioorthogonal reactions. Here, we introduce several engineering strategies to apply terminal alkyne biosynthetic machinery, represented by JamABC, which produces a short terminal alkyne-bearing fatty acyl chain on a carrier protein, to functions with different downstream polyketide synthases (PKSs). Successful results in engineering type III and type I PKSs provide engineering guidelines and strategies that are applicable to additional PKSs to produce targeted alkyne-tagged metabolites for chemical and biological applications.


Induction of Bax protein and degradation of lamin A during p53-dependent apoptosis induced by chemotherapeutic agents in human cancer cell lines.
In this study, subcellular fractionation analysis was performed to investigate the intracellular localization of Bax protein. We demonstrated that Bax protein is localized primarily in the nuclear and heavy membrane fractions. The expression of Bax protein in the nuclear membrane was induced in wild-type p53 human cancer cells (COLO 205 and Hep G2) by a wide variety of cancer chemotherapeutic agents in order to scrutinize further the biologic function of the Bax protein in the nuclear membrane. We found that lamin A and poly-(ADP ribose) polymerase (PARP) protein degradation coincided when the Bax protein level was elevated in the nuclear membrane of cells affected by drug stimuli. By using anti-sense oligodeoxynucleotides specific to human Bax mRNA, we further demonstrated that inhibition of Bax expression could specifically block lamin A but not PARP cleavage in apoptotic cancer cells.


Urgent, resective surgery for medically refractory, convulsive status epilepticus.
Refractory, convulsive status epilepticus has significant mortality and morbidity. Urgent resective surgery may be of benefit in selected cases where medical therapies have failed. At our institution, from January 1992 until December 2005, urgent resective surgery was performed in three children with medically refractory convulsive status epilepticus. The etiologies were microdysgenesis, focal cortical dysplasia, and bilateral Rasmussen's syndrome and cortical dysplasia. In two cases, surgery resulted in termination of status epilepticus. In all three cases, surgery permitted discontinuation of high-dose suppression therapy and allowed the patient to leave the intensive care unit, either by terminating status epilepticus or by providing important histopathological information about the nature of the underlying disease and prognosis.


The Rikers Island Hot Spotters: Defining the Needs of the Most Frequently Incarcerated.
We used "hot spotting" to characterize the persons most frequently admitted to the New York City jail system in 2013. We used our Correctional Health Services electronic health record to identify 800 patients admitted in 2013 who returned most since November 2008. We compared them to a randomly selected control group of 800 others admitted in 2013, by using descriptive statistics and cross-tabulations, including data through December 2014. The frequently incarcerated individuals had a median of 21 incarcerations (median duration 11 days), representing 18 713 admissions and $129 million in custody and health costs versus $38 million for the controls. The frequently incarcerated were significantly older (42 vs 35 years), and more likely to have serious mental illness (19% vs 8.5%) and homelessness (51.5% vs 14.7%) in their record. Significant substance use was highly prevalent (96.9% vs 55.6%). Most top criminal charges (88.7%) for the frequently incarcerated were misdemeanors; assault charges were less common (2.8% vs 10.4%). Frequently incarcerated persons have chronic mental health and substance use problems, their charges are generally minor, and incarceration is costly. Tailored supportive housing is likely to be less costly and improve outcomes.


The lack of active bile acid transport in AS-30 D ascites hepatoma cells.
The uptake of cholic acid as well as taurocholic acid into AS-30 D ascites hepatoma cells showed linearity with respect to incubation concentrations. It has been suggested that these processes can be described as simple diffusion. In further experiments it could be shown that ascites hepatoma cells were unable to conjugate cholic acid. These results may have significance in the phalloidin action on hepatocytes.


New chemosensory component in the U.S. National Health and Nutrition Examination Survey (NHANES): first-year results for measured olfactory dysfunction.
The U.S. NHANES included chemosensory assessments in the 2011-2014 protocol. We provide an overview of this protocol and 2012 olfactory exam findings. Of the 1818 NHANES participants aged ≥40 years, 1281 (70.5 %) completed the exam; non-participation mostly was due to time constraints. Health technicians administered an 8-item, forced-choice, odor identification task scored as normosmic (6-8 odors identified correctly) versus olfactory dysfunction, including hyposmic (4-5 correct) and anosmic/severe hyposmic (0-3 correct). Interviewers recorded self-reported smell alterations (during past year, since age 25, phantosmia), histories of sinonasal problems, xerostomia, dental extractions, head or facial trauma, and chemosensory-related treatment and changes in quality of life. Olfactory dysfunction was found in 12.4 % (13.3 million adults; 55 % males/45 % females) including 3.2 % anosmic/severe hyposmic (3.4 million; 74 % males/26 % females). Selected age-specific prevalences were 4.2 % (40-49 years), 12.7 % (60-69 years), and 39.4 % (80+ years). Among adults ≥70 years, misidentification rates for warning odors were 20.3 % for smoke and 31.3 % for natural gas. The highest sensitivity (correctly identifying dysfunction) and specificity (correctly identifying normosmia) of self-reported olfactory alteration was among anosmics/severe hyposmics (54.4 % and 78.1 %, respectively). In age- and sex-adjusted logistic regression analysis, risk factors of olfactory dysfunction were racial/ethnic minority, income-to-poverty ratio ≤ 1.1, education <high school, and heavy drinking. Moderate-to-vigorous physical activity reduced risk of impairment. Olfactory dysfunction is prevalent, particularly among older adults. Inexpensive, brief odor identification tests coupled with questions (smell problems past year, since age 25, phantosmia) could screen for marked dysfunction. Healthcare providers should be prepared to offer education on non-olfactory avoidance of hazardous events.


Safety and effectiveness of sling fiber preservation POEM to reduce severe post-procedural erosive esophagitis.
Peroral endoscopic myotomy (POEM) is standard treatment for achalasia. Gastroesophageal reflux disease (GERD) after POEM has been an important challenge since the early days of POEM implementation. The esophagogastric junction anti-reflux barrier consists of internal lower esophageal sphincter (LES) (i.e., intrinsic muscles of distal esophagus with sling fibers) and external LES (crural diaphragm and phrenoesophageal ligament anchors the distal esophagus to the crural diaphragm). During conventional POEM, the entire internal LES is unintentionally dissected. Preservation of the sling fiber may reduce post-POEM GERD, but its safety and effectiveness have been unclear. In this study, we investigated the safety and effectiveness of sling fiber preservation POEM (SP-POEM) for reducing severe post-procedural erosive esophagitis. We analyzed data of 236 patients who underwent POEM; of these, 203 patients underwent posterior myotomy without (Group 1) or with attempted (Group 2) sling fiber preservation. Group 1 (N = 79; sling fiber excision, N = 68) and Group 2 (N = 90; sling fiber preservation, N = 81) were compared. Post-procedural erosive esophagitis (Los Angeles classification) were assessed and the area (direction and length) of mucosal break was also investigated. The gastroesophageal flap valve was evaluated by Hill's classification. Severe erosive esophagitis exceeding grade C (Los Angeles classification) occurred in 44.1% of patients (30/68) in Group 1 and in 18.5% of patients (15/81) in Group 2. In mapping of erosive esophagitis, mucosal breaks appeared widely in all directions in Group 1; they tended to be limited in Group 2 (especially in the 2 o'clock direction). Assessment using the Hill's classification showed that the gastroesophageal flap valve was preserved after SP-POEM. SP-POEM is safe and effective, with a success rate of 90%. The rate of severe erosive esophagitis can be decreased by preserving sling fibers.


Role of human 3α-hydroxysteroid dehydrogenase isoforms (AKR1C1-AKR1C3) in the extrahepatic metabolism of the steroidal aromatase inactivator Formestane.
The clinical use of the steroidal aromatase inhibitor Formestane (4-hydroxandrostenedione, 4-OHA) in the treatment of advanced ER+ breast cancer has been discontinued, and therefore, interest in this remarkable drug has vanished. As a C-19 sterol, 4-OHA can undergo extensive intracellular metabolism depending on the expression of specific enzymes in the corresponding cells. We used the metabolites 4β-hydroxyandrosterone, 4β-hydroxyepiandrosterone and its 17β-reduced derivative as standards for the proof of catalytic activity present in the cell culture medium and expressed by the isolated enzymes. All of the aldo-keto reductases AKR1C1, AKR1C2, AKR1C3 and AKR1C4 catalysed the reduction of the 3-keto-group and the Δ4,5 double bond of 4-OHA at the same time. Molecular docking experiments using microscale thermophoresis and the examination of the kinetic behaviour of the isolated enzymes with the substrate 4-OHA proved that AKR1C3 had the highest affinity for the substrate, whereas AKR1C1 was the most efficient enzyme. Both enzymes (AKR1C1and AKR1C3) are highly expressed in adipose tissue and lungs, exhibiting 3β-HSD activity. The possibility that 4-OHA generates biologically active derivatives such as the androgen 4-hydroxytestosterone or some 17β-hydroxy derivatives of the 5α-reduced metabolites may reawaken interest in Formestane, provided that a suitable method of administration can be developed, avoiding oral or intramuscular depot-injection administration.


Accounting for urban biogenic fluxes in regional carbon budgets.
Many ecosystem models incorrectly treat urban areas as devoid of vegetation and biogenic carbon (C) fluxes. We sought to improve estimates of urban biomass and biogenic C fluxes using existing, nationally available data products. We characterized biogenic influence on urban C cycling throughout Massachusetts, USA using an ecosystem model that integrates improved representation of urban vegetation, growing conditions associated with urban heat island (UHI), and altered urban phenology. Boston's biomass density is 1/4 that of rural forests, however 87% of Massachusetts' urban landscape is vegetated. Model results suggest that, kilogram-for-kilogram, urban vegetation cycles C twice as fast as rural forests. Urban vegetation releases (RE) and absorbs (GEE) the equivalent of 11 and 14%, respectively, of anthropogenic emissions in the most urban portions of the state. While urban vegetation in Massachusetts fully sequesters anthropogenic emissions from smaller cities in the region, Boston's UHI reduces annual C storage by >20% such that vegetation offsets only 2% of anthropogenic emissions. Asynchrony between temporal patterns of biogenic and anthropogenic C fluxes further constrains the emissions mitigation potential of urban vegetation. However, neglecting to account for biogenic C fluxes in cities can impair efforts to accurately monitor, report, verify, and reduce anthropogenic emissions.


14q+ marker chromosome in an EBV-genome-negative lymph node without signs of malignancy in a patient with EBV-genome-positive nasopharyngeal carcinoma.
In a patient with an EBV-genome-positive nasopharyngeal carcinoma, an EBV-genome-negative inguinal lymph node without histological evidence of malignant lymphoma or metastatic carcinoma growth was found to contain a 14q+ marker chromosome, identified as an 8;14 translocation, in all cells analyzed. This observation indicates that chromosome aberrations may precede histological signs of malignancy. The possible implication of this finding in relation to the postulated role of the 14q+ marker and lymphoma development is discussed.


The Mesothelioma epidemic in Western Europe: an update.
The number of male deaths from pleural cancer in France, Germany and Italy increased from about 8750 in 1990-1994 to 9550 in 1995-1999, suggesting that mesothelioma deaths in males may be levelling off in most of Western Europe.


Superhydrophobic lignin-based multifunctional polyurethane foam with SiO2 nanoparticles for efficient oil adsorption and separation.
Superhydrophobic polyurethane foam is one of the most promising materials for oil-water separation. However, there are only limited studies prepared matrix superhydrophobic foams as adsorbents. In this paper, SiO2 modified by 1H, 1H, 2H, 2H-perfluorododecyl trichlorosilane (F-SiO2) was added into the lignin-based foam matrix by a one-step foaming technique. The average diameter of F-SiO2 was about 480 nm with an water contact angle (WCA) of 160.3°. The lignin-based polyurethane foam with F-SiO2 had a superhydrophobic water contact angle of 151.3°. There is no obvious change in contact angle after 100 cycles of compression or after cutting and abrasion. Scanning electron microscopy (SEM) analysis showed that F-SiO2 was distributed both on the surface and inside of the foam. The efficiency for oil-water separation reached 99 %. Under the light intensity of 1 kW/m2, the surface temperature of the lignin-based foam rose to 77.6 °C. In addition, the foam exhibited self-cleaning properties and degraded within 2 h in an alcoholic alkali solution. Thus, in this study, we developed a novel matrix superhydrophobic lignin-based polyurethane foam with an excellent promise to be used as oil water separation adsorbents in industrial wastewater treatment and oil spill clean-up processes.


The sharp structural switch of covalent cages mediated by subtle variation of directing groups.
It is considered a more formidable task to precisely control the self-assembled products containing purely covalent components, due to a lack of intrinsic templates such as transition metals to suppress entropy loss during self-assembly. Here, we attempt to tackle this challenge by using directing groups. That is, the self-assembly products of condensing a 1:2 mixture of a tetraformyl and a biamine can be precisely controlled by slightly changing the substituent groups in the aldehyde precursor. This is because different directing groups provide hydrogen bonds with different modes to the adjacent imine units, so that the building blocks are endowed with totally different conformations. Each conformation favors the formation of a specific product that is thus produced selectively, including chiral and achiral cages. These results of using a specific directing group to favor a target product pave the way for accomplishing atom economy in synthesizing purely covalent molecules without relying on toxic transition metal templates.


Recombinant interferon-alpha therapy of Philadelphia chromosome-negative myeloproliferative disorders with thrombocytosis.
The clinical course of patients with myeloproliferative disorders and excessive thrombocytosis may be complicated by serious hemorrhagic or thrombotic events. We have previously reported that interferon-alpha can control severe refractory thrombocytosis in patients with advanced chronic myelogenous leukemia. Therefore, we treated a group of thrombocythemic patients with Ph-negative myeloproliferative disorders, including polycythemia vera and essential thrombocythemia, with recombinant interferon-alpha (rIFN-alpha 2a). Eight patients with profound elevations in platelet counts received a median induction dose of 5.4 X 10(6) U/day (range, 5.0 to 10.0 X 10(6) U/day) of rIFN-alpha 2a administered intramuscularly or subcutaneously. We observed a significant decline in platelet counts from a median baseline value of 1,929 X 10(9)/L (range, 960 to 2,960 X 10(9)/L) to a median posttreatment value of 431 X 10(9)/L (range, 71 to 1,150 X 10(9)/L) (p less than 0.01). Concomitantly, white blood cell counts declined from a median baseline value of 20.8 X 10(9)/L (range, 10.5 to 40.8 X 10(9)/L) to a median posttreatment value of 6.1 X 10(9)/L (range, 2.9 to 29.0 X 10(9)/L) (p less than 0.02). Correction of thrombocytosis was rapid, with a median of only eight days from the start of therapy to the achievement of a platelet count less than 1,000 X 10(9)/L. Six of eight patients have shown an ongoing response with a median follow-up period of 11 months (range, one to 30 months). There have been no bleeding or thrombotic events during the study. Side effects of rIFN-alpha 2a therapy consisted of fever and flu-like symptoms, with tachyphylaxis developing after one to two weeks of therapy. Our observations suggest that alpha interferon may be a promising therapeutic agent for myeloproliferative disorders characterized by thrombocytosis.


Therapeutic efficacy of new botulinum toxin identified in CCUG 7968 strain.
Botulinum neurotoxin type A (BoNT/A) induces muscle atrophy by cleaving synaptosomal-associated protein 25. Thus, BoNT/A has been actively utilized for the treatment of masseter and gastrocnemius hypertrophy. In this study, INI101 toxin was newly identified from the CCUG 7968 strain, and its therapeutic efficacy was evaluated both in vitro and in vivo. The INI101 toxin showed identical genetic sequence, amino acid sequence, and protein subunit composition to BoNT/A produced from strain Hall A. Electromyography (EMG), and immunofluorescence staining demonstrated that INI101 (at 2 ~ 8 U/rat) effectively blocked the neuromuscular junction with no toxicity in a rat model. The EMG results showed INI101 toxin-induced weight loss and volume reduction of the gastrocnemius, similar to the effects of Botox® (BTX). Histological and immunofluorescence staining was consistent with this EMG result, showing that INI101 toxin caused muscle fiber reduction in the gastrocnemius. Notably, INI101 toxin diffused less into adjacent muscle tissue than BTX, indicating that INI101 toxin may reduce potential side effects due to diffusion into normal tissues. INI101 toxin isolated from the novel strain CCUG 7968 is a newly identified meaningful biopharmaceutical comparable to the conventional BoNT/A in the medical field. KEY POINTS: • Botulinum neurotoxin type A (BoNT/A, INI101) was identified from the CCUG 7968 strain. • INI101 toxin showed similar safety and therapeutic efficacy comparable to conventional BoNT/A both in vitro and in vivo. • INI101 toxin is a meaningful biopharmaceutical comparable to the conventional BoNT/A in the medical field.


Management of cytomegalovirus infection in inflammatory bowel diseases.
Cytomegalovirus is a deoxyribonucleic acid virus that infects a large part of the human population; after primary infection, it develops a latent state and can be reactivated, notably after a decrease in host immune defences. In patients with inflammatory bowel diseases, cytomegalovirus is frequently involved, either as an agent of colitis or through local asymptomatic reactivation. Due to the immune context of inflammatory bowel diseases and to the immunosuppressive therapies that are used to treat them, cytomegalovirus entertains complex relationships with these diseases. Whereas Crohn's disease seems little impacted by cytomegalovirus, this agent interferes strongly with the natural progression of ulcerative colitis. While immune treatments have a clear influence on the occurrence of cytomegalovirus colitis in ulcerative colitis (favourable for steroids and cyclosporine and rather inhibitory for infliximab), the role of cytomegalovirus infection on ulcerative colitis is more debated with roles ranging from innocent bystander to key pathogen suggested. There is however growing evidence for a participation of intestinal cytomegalovirus infection in the resistance of ulcerative colitis to steroids and the investigation of cytomegalovirus infection in intestinal biopsies by immunohistochemistry or quantitative polymerase chain reaction assay is strongly recommended. In several studies, treatment of cytomegalovirus infection by ganciclovir was shown to restore the response to immunomodulatory therapies and even to prevent the need for colectomy. All of these recently acquired data need to be validated by randomised clinical trials conducted on a large panel of ulcerative colitis patients.


Influence of positive end-expiratory pressure ventilation on survival during severe hemorrhagic shock.
Although a moderate positive end-expiratory pressure (PEEP) level is widely recommended, it is unknown whether moderate PEEP during mechanical ventilation has adverse effects during severe hemorrhagic shock. Therefore, the purpose of our study was to evaluate the effects of 0 cm H2O PEEP versus 5 cm H2O PEEP versus 10 cm H2O PEEP on short-term survival in a porcine model of severe hemorrhagic shock. Secondary study endpoints were hemodynamic variables and blood gases. Twenty-four anesthetized pigs were bled approximately 45 mL/kg, randomized into 3 groups, and then ventilated with 0, 5, or 10 cm H2O PEEP. Survival rates were compared using Kaplan-Meier methods with log rank (Mantel Cox) comparison of cumulative survival by treatment group. Seven of 8 0 cm H2O PEEP animals survived the 120-minute study period, but 8 of 8 5 cm H2O PEEP animals died within 30 minutes, and 8 of 8 10 cm H2O PEEP animals were dead within 20 minutes (P<.0001). Ventilation with 0 cm H2O PEEP prevented a further reduction of mean arterial blood pressure and cardiac output. When compared with the 0 cm H2O PEEP group, end-tidal CO2 declined in the 5 cm H2O PEEP and 10 cm H2O PEEP animals. Compared with the 0 cm H2O PEEP animals, those ventilated with 5 or 10 cm H2O PEEP had higher lactate levels after 10 minutes. When compared with pigs ventilated with either 5 or 10 cm H2O PEEP, those ventilated with 0 cm H2O PEEP during untreated, severe hemorrhagic shock had significantly improved short-term survival.


Neoadjuvant chemoradiotherapy versus neoadjuvant chemotherapy followed by minimally invasive esophagectomy for locally advanced esophageal squamous cell carcinoma: a prospective multicenter randomized clinical trial.
Neoadjuvant therapy is recommended for locally advanced esophageal cancer, but the optimal strategy remains unclear. We aimed to evaluate the safety and efficacy of neoadjuvant chemoradiotherapy (nCRT) versus neoadjuvant chemotherapy (nCT) followed by minimally invasive esophagectomy (MIE) for locally advanced esophageal squamous cell carcinoma (ESCC). Eligible patients staged as cT3-4aN0-1M0 ESCC were randomly assigned (1 : 1) to the nCRT or nCT group stratified by age, cN stage, and centers. The chemotherapy, based on paclitaxel and cisplatin, was administered to both groups, while concurrent radiotherapy was added for the nCRT group; then MIE was carried out. The primary endpoint was 3-year overall survival. This study is registered with ClinicalTrials.gov (NCT03001596). A total of 264 patients were eligible for the intention-to-treat analysis. By 30 November 2021, 121 deaths had occurred. The median follow-up was 43.9 months (interquartile range 36.6-49.3 months). The overall survival in the intention-to-treat population was comparable between the nCRT and nCT strategies [hazard ratio (HR) 0.82, 95% confidence interval (CI) 0.58-1.18; P = 0.28], with a 3-year survival rate of 64.1% (95% CI 56.4% to 72.9%) versus 54.9% (95% CI 47.0% to 64.2%), respectively. There were also no differences in progression-free survival (HR 0.83, 95% CI 0.59-1.16; P = 0.27) and recurrence-free survival (HR 1.07, 95% CI 0.71-1.60; P = 0.75), although the pathological complete response in the nCRT group (31/112, 27.7%) was significantly higher than that in the nCT group (3/104, 2.9%; P < 0.001). Besides, a trend of lower risk of recurrence was observed in the nCRT group (P = 0.063), while the recurrence pattern was similar (P = 0.802). NCRT followed by MIE was not associated with significantly better overall survival than nCT among patients with cT3-4aN0-1M0 ESCC. The results underscore the pending issue of the best strategy of neoadjuvant therapy for locally advanced bulky ESCC.


Development of the National Marine Mammal Tissue Bank.
The National Marine Mammal Tissue Bank (NMMTB) has been established as part of a comprehensive effort to obtain reliable information on contaminant levels in marine mammal tissues. A four component program consisting of the NMMTB, stranding networks, monitoring and quality assurance has been developed. The development and current status of the NMMTB and its relationship to the other three components will be described.


CES2 sustains HNF4α expression to promote pancreatic adenocarcinoma progression through an epoxide hydrolase-dependent regulatory loop.
Intra-tumoral expression of the serine hydrolase carboxylesterase 2 (CES2) contributes to the activation of the pro-drug irinotecan in pancreatic ductal adenocarcinoma (PDAC). Given other potential roles of CES2, we assessed its regulation, downstream effects, and contribution to tumor development in PDAC. Association between the mRNA expression of CES2 in pancreatic tumors and overall survival was assessed using The Cancer Genome Atlas. Cell viability, clonogenic, and anchorage-independent growth assays as well as an orthotopic mouse model of PDAC were used to evaluate the biological relevance of CES2 in pancreatic cancer. CES2-driven metabolic changes were determined by untargeted and targeted metabolomic analyses. Elevated tumoral CES2 mRNA expression was a statistically significant predictor of poor overall survival in PDAC patients. Knockdown of CES2 in PDAC cells reduced cell viability, clonogenic capacity, and anchorage-independent growth in vitro and attenuated tumor growth in an orthotopic mouse model of PDAC. Mechanistically, CES2 was found to promote the catabolism of phospholipids resulting in HNF4α activation through a soluble epoxide hydrolase (sEH)-dependent pathway. Targeting of CES2 via siRNA or small molecule inhibitors attenuated HNF4α protein expression and reduced gene expression of classical/progenitor markers and increased basal-like markers. Targeting of the CES2-sEH-HNF4α axis using small molecule inhibitors of CES2 or sEH reduced cell viability. We establish a novel regulatory loop between CES2 and HNF4α to sustain the progenitor subtype and promote PDAC progression and highlight the potential utility of CES2 or sEH inhibitors for the treatment of PDAC as part of non-irinotecan-containing regimens.