The serum vitamin C levels in Behçet's disease.
Behçet's disease (BD) is a chronic inflammatory disorder of unknown aetiology, and recognised as a multi-system vasculitis. It has been postulated that an imbalance of the oxidant and antioxidant systems related to the disease are important in its pathogenesis. Previous publications have reported increased levels of enzymatic antioxidant defence systems in patients with BD. The non-enzymatic antioxidant systems, including vitamin C and uric acid, were looked for in the present study. For this aim, the serum malondialdehyde (MDA), an end product of lipid peroxidation, and vitamin C and uric acid, as endogenous antioxidants, were determined in 20 patients with BD (11 in active and 9 in inactive periods) and 20 healthy subjects. The MDA level was significantly higher in both the active and inactive period patients compared with the control group (p < 0.001, p < 0.05, respectively). The MDA level was also significantly higher in the active period patients compared with the inactive period patients (p < 0.05). The vitamin C levels were significantly lower in both the active and inactive period patients compared with the control group (p < 0.001, p < 0.05, respectively). There was no significant difference in the vitamin C level between the active and inactive period patients (p > 0.05). There was also no significant difference in uric acid levels between the groups (p > 0.05). In the patients group, a negative correlation was found between the levels of serum MDA and vitamin C (r=-0.517; p < 0.05). Our results indicate that decreased vitamin C and increased MDA levels reflect the increased levels of oxidative stress in BD patients, and this situation may be important in relation with its pathogenesis.


Population-based study of central post-stroke pain in Rimini district, Italy.
Central post-stroke pain (CPSP) is still an underestimated complication of stroke, resulting in impaired quality of life and, in addition to the functional and cognitive consequences of stroke, the presence of CPSP may be associated with mood disorders, such as depression, anxiety, and sleep disturbances. This type of pain may also impair activities of daily living and further worsen quality of life, negatively influencing the rehabilitation process. The prevalence of CSPS in the literature is highly variable (1%-12%) according to different studies, and this variability could be influenced by selection criteria and the different ethnic populations being investigated. With this scenario in mind, we performed a population-based study to assess the prevalence of CPSP and its main features in a homogeneous health district (Rimini, Italy), including five hospitals for a total population of 329,970 inhabitants. From 2008 to 2010, we selected 1,494 post-stroke patients and were able to interview 660 patients, 66 (11%) of whom reported pain with related tactile and thermal hyperesthesia, accompanied by needle puncture, tingling, swelling, and pressure sensations. Patients reported motor impairment and disability, which influenced their working ability, rehabilitation, and social life. Despite this severe pain state, there was a high percentage of patients who did not receive adequate treatment for pain.


Monograph use at an academic health sciences library: the first three years of shelf life.
To study the circulation of monographs during the first three years of shelf life at an academic health sciences library. A record was kept of monographs added to the circulating collection from mid-1994 to mid-1995. After three years, each monograph was located and the number of times it circulated during the first, second, and third year of shelf life determined by counting checkout stamps on the circulation slip. Of the 1,958 monographs studied, 1,674 had complete data for the first three years of shelf life. Of those 1,674 titles, 81.48% circulated at least once. A total of 7,659 circulations were recorded; 38.69% occurred in the first year of shelf life, 32.37% in the second year, and 28.95% in the third year. The data did not fit the well-known 80/20 rule. Instead, approximately 38% of monographs accounted for 80% of circulation. A small percentage, 2.21%, of monographs accounted for a substantial percentage of circulation, 21.84%. A large percentage of the monographs circulated and use did not decline sharply with age within the first three years of shelf life, indicating a high demand for monographs at this academic health sciences library. These results, combined with the findings of earlier studies, suggested two possibilities. First, academic health sciences libraries might exhibit use of a higher percentage of monograph acquisitions than other types of libraries; or, second, a low monograph-to-user ratio might result in a higher percentage of monographs being used. Perhaps both factors contributed to the results found in this study. Further investigation would be needed to determine the extent to which library type and monograph-to-user ratio influenced monograph use.


Recent developments in the electrodiagnosis of radiculopathies.
The accuracy and reliability of electrodiagnosis in radiculopathy has improved in recent years. A better understanding of EMG findings coupled with the development of several tests that have recently become available in clinical practice (F wave studies, H reflex studies, and somatosensory evoked potential studies) have made it a valuable tool in the complete work-up for suspected radiculopathy. The use of this technique is reviewed and an illustrative case report is presented.


Mechanisms connecting square dance to sleep quality among middle-aged and older Chinese females: serial mediation roles of social support and depressive symptoms.
Square dance is gaining increasing popularity among middle-aged and older Chinese women who are also at high risk of sleep disturbance. Although previous studies have shown exercise could improve sleep quality, the association between square dance and sleep quality remains to be discussed, and even less is known about the potential mechanism underlying this association. This study aims to investigate the relationship between square dance and sleep quality and test if social support and depressive symptoms together play a serial mediating role in the influence of square dance on sleep quality. A cross-sectional study was conducted among 549 middle-aged and older Chinese females from September to December 2020 in Shao Yang City, Hunan Province of China, with ethics approval granted (SYU [2020]002). Square dance involvement was assessed by three questions about the time participants spent in square dance. Social support, depressive symptoms, and sleep quality were measured using the Pittsburgh Sleep Quality Index (PSQI), Social Support Self-Rating Scale (SSRS), and 9-item Patient Health Questionnaire (PHQ-9), respectively. The serial mediation model was analyzed by the bootstrapping method to assess whether social support and depressive symptoms mediate the relationship between square dance and sleep quality. Two-thirds of the participants had high involvement in square dance and most reported a moderate and high level of social support (98.54%). The prevalence of depressive symptoms and sleep disturbance was 19.49 and 26.78%, respectively. The serial mediation model showed a significant association between square dance and sleep quality, which was fully mediated by social support and depressive symptoms in a serial model (total effect c = -0.114, 95%CI = -0.227 to -0.001; direct effect c' = -0.036, 95% CI = -0.138 to 0.065; total indirect effect ab = -0.077, 95% CI = -0.139 to-0.016). Our study extends the understanding of how square dance is associated with sleep quality through the serial mediating roles of social support and depressive symptoms. It provides crucial implications for developing square dance interventions to improve sleep quality among middle-aged and older Chinese females.


Fluorescence properties of native and photooxidised proteinase K: the X-ray model in the region of the two tryptophans.
The fluorescence properties of proteinase K are described and related to the X-ray model refined at 1.48 A resolution. Upon excitation of proteinase K at 295 nm the fluorescence is determined by the two tryptophan residues, Trp-8 and Trp-212. The tryptophans are partly buried just below the surface of the molecule. Neither Trp is in a highly hydrophobic environment, suggesting that this cannot be the explanation for the fluorescence at 330 nm: formation of exiplexes with adjacent peptide bonds would seem to be the more likely cause. Trp-8 is located in a 'cavity', close to an internal cluster of water molecules. The contribution of Trp-8 to the total indole emission is 60% and that of Trp-212 is 40%. The tryptophan fluorescence quantum yield is constant in the pH range 3-9. The fluorescence spectrum resulting from the simultaneous excitation of the tyrosyl and tryptophyl residues at 280 nm is dominated by the indole fluorophores: 61% of the light absorbed by the tyrosyl side chains is transferred to the two indole rings. Iodide and caesium are not efficient quenchers of the proteinase K tryptophan fluorescence, which is explained by restricted access of the ions to the somewhat buried Trp side chains and by electrostatic repulsion of caesium ions. Acrylamide quenching proceeds via both a dynamic and a static process and the data show homogeneity of the indole fluorescence arising from fluorophores in similar environments. The activation energy for the thermal deactivation of the excited tryptophans is 54 kJ mol-1. This value is substantially higher than those found for other proteinases from microorganisms and arises from the thermostability of proteinase K. Photooxidation of proteinase K in the presence of proflavine follows the kinetics of a first order reaction. The two tryptophans differ in their photoreactivity, Trp-212 being considerably more reactive.


Predominantly Antibiotic-resistant Intestinal Microbiome Persists in Patients With Pouchitis Who Respond to Antibiotic Therapy.
Pouchitis that develops in patients with ulcerative colitis after total proctocolectomy and ileal pouch anal anastomosis is usually treated with antibiotics. Some patients have recurrence of flares, or become antibiotic-dependent, and require repeated courses or prolonged periods of antibiotic therapy. We investigated microbial factors associated with response to antibiotic treatment and development of antibiotic dependence in patients with pouchitis. We performed a prospective study of 49 patients who had undergone pouch surgery at a tertiary center. Disease activity was determined based on clinical, endoscopic, and histologic criteria. Pouch phenotype was defined as recurrent-acute pouchitis (n = 6), chronic pouchitis and Crohn's-like disease of the pouch (n = 27), normal pouch from patient with ulcerative colitis (n = 10), and normal pouch from patient with familial adenomatous polyposis (n = 6). Fecal samples (n = 234) were collected over time during or in the absence of antibiotic treatment (ciprofloxacin and/or metronidazole). Thirty-three patients were treated with antibiotics, for a median of 425 days of cumulative antibiotic therapy, during follow-up. Calprotectin was measured and fecal DNA was sequenced using shotgun metagenomics and analyzed with specifically designed bioinformatic pipelines. Bacterial strains were isolated from fecal samples. We assessed their ciprofloxacin resistance and ability to induce secretion of inflammatory cytokines by HT-29 intestinal epithelial cells. Most antibiotic-treated patients (79%) had a clinical response to each course of antibiotics; however, 89% of those who completed a 4-week course relapsed within 3 months. Median calprotectin levels decreased by 40% in response to antibiotics. Antibiotic treatment reduced disease-associated bacteria such as Clostridium perfringens, Ruminococcus gnavus, and Klebsiella pneumoniae, but also beneficial species, such as Faecalibacterium prausnitzii. The microbiomes of antibiotic-responsive patients were dominated by facultative anaerobic genera (Escherichia, Enterococcus, and Streptococcus), with multiple ciprofloxacin-resistance mutations in drug target genes and confirmed drug resistance. However, these strains had lower potential for virulence and did not induce secretion of inflammatory cytokines by epithelial cells. After antibiotic cessation, patients had an abrupt shift in microbiome composition, with blooms of oral and disease-associated bacteria. In addition, antibiotic treatment enriched for strains that acquired multidrug resistance loci, encoding enzymes that confer resistance to nonrelated antibiotics, including extended-spectrum beta-lactamases. The efficacy of antibiotic treatment of pouchitis might be attributed to the establishment of an antibiotic-resistant microbiome with low inflammatory potential. This microbiome might provide resistance against colonization by bacteria that promote inflammation. To avoid progression to antibiotic-dependent disease and its consequences, strategies such as short-term alternating antibiotics and nutrition- and microbiome-based interventions should be considered.


Polymyositis and dermatomyositis: novel insights into the pathogenesis and potential therapeutic targets.
Polymyositis (PM) and dermatomyositis (DM) are the two major forms of inflammatory muscle diseases. They are characterized clinically primarily by proximal muscle weakness. The disease mechanisms that cause muscle damage and dysfunction are not fully understood. However, because of the association with other autoimmune diseases, the presence of autoantibodies, and response to immunosuppressive medication, they are believed to be autoimmune in origin. Recent studies have highlighted the importance of the innate immune system and non-immune mechanisms and described novel adaptive immune-based pathways in the pathogenesis of polymyositis and dermatomyositis. Stimulation of Toll-like receptors (TLRs) by endogenous antigens, e.g., aminoacyl-tRNA synthetase enzyme, may trigger activation of signaling pathways and thereby induces expression of multiple genes involved in the inflammatory response. High-mobility group box-1 (HMGB1) might interact with the same receptors and cause skeletal muscle inflammation. In addition, this protein may be involved in T lymphocyte survival in muscle tissue. A newly described T cell subset, CD28(-) T cells, may have strong myotoxic properties and comprises the predominant muscle-infiltrating T cell phenotype. Future studies should focus more on understanding the relative contribution of each pathway to the pathogenesis of inflammatory myopathies. Given the connections between the pathways, targeting multiple pathways through combination therapies may be beneficial.


Initial non-compliance with antihypertensive monotherapy is followed by complete discontinuation of antihypertensive therapy.
Discontinuation with treatment is a major problem in the treatment of hypertension. The objective of our study was to assess the association between non-compliance and discontinuation in patients who started using antihypertensive monotherapy. A nested case-control study within a cohort of new users of antihypertensive drugs between 1st January 1999 and 31st December 2002 was performed. We used data from the PHARMO database, a record linkage system containing drug-dispensing records from community pharmacies and linked hospital discharge records of approximately 950,000 subjects. Cases discontinued their use of antihypertensive monotherapy and were not switched to other antihypertensive treatment, controls stayed on their initially prescribed monotherapy. Conditional logistic regression was used to calculate odds ratios (OR) and their 95% confidence intervals (CI). In a cohort of 39,714 new users of antihypertensive drugs, we identified 9111 cases and 9111 matched controls. The percentage of non-compliant patients (compliance < 80%) among cases and controls was 14.0% and 5.8%, respectively [OR 2.86 (95%CI: 2.52-3.24)]. Patients who used less than 90 days had a higher risk on discontinuation [OR 3.10 (95%CI: 2.67-3.59)] than patients who used more than 90 days [OR 2.28 (95%CI: 1.79-2.92)]. The association was generally similar among males and females, among the different types of antihypertensives and among the different age groups. In patients who start antihypertensive monotherapy, non-compliance is often followed by discontinuation of this antihypertensive treatment. The pharmacy medication history is a valuable tool for physicians to identify patients who have a high risk on discontinuation with antihypertensive treatment.


Implementation and spread of a simple and effective way to improve the accuracy of medicines reconciliation on discharge: a hospital-based quality improvement project and success story.
Changes are often made to medications at times of transitions in care. Inadequate reconciliation during admission, transfer and discharge causes medication errors and increases risks of patient harm. Despite well-established multidisciplinary medicines reconciliation (MR) processes at hospital admission, our MR process at discharge; however, was poor. The main errors included failure to recommence withheld medicines and lack of documentation explaining changes made to medications on discharge. Our objective was to develop an intervention that supports prescribers to follow a simple standardised MR process at discharge to reduce these errors. Working closely as a multidisciplinary team, we used improvement methodologies to design and test a process that reliably directs prescribers in surgery to use the inpatient prescribing chart as well as the MR on admission form as sources to create accurate discharge prescriptions. The project was segmented into testing, implementation, spread and sustainability. The tested intervention helped the accuracy of discharge prescriptions steadily and quickly improve from 45% to 96% in the pilot ward. Following the successful implementation and sustainability in two separate pilot wards, the intervention was spread to the remaining eight wards producing a similar improvement. To improve patient safety, it is crucial to ensure that information about medicines is effectively communicated when care is transferred between teams. Although this can be challenging, we've shown that it can be done effectively and reliably if this responsibility is equally shared by healthcare professionals from all disciplines while being supported by safe systems that make it easy to do the right thing. Successfully implementing a standardised multidisciplinary MR process at discharge can also reduce the reliance on pharmacists therefore freeing them to undertake other clinical roles.


Dynamic functional connectivity and brain metastability during altered states of consciousness.
The scientific study of human consciousness has greatly benefited from the development of non-invasive brain imaging methods. The quest to identify the neural correlates of consciousness combined psychophysical experimentation with neuroimaging tools such as functional magnetic resonance imaging (fMRI) to map the changes in neural activity associated with conscious vs. unconscious percepts. Different neuroimaging methods have also been applied to characterize spontaneous brain activity fluctuations during altered states of consciousness, and to develop quantitative metrics for the level of consciousness. Most of these studies, however, have not explored the dynamic nature of the whole-brain imaging data provided by fMRI. A series of empirical and computational studies strongly suggests that the temporal fluctuations observed in this data present a non-trivial structure, and that this structure is compatible with the exploration of a discrete repertoire of states. In this review we focus on how dynamic neuroimaging can be used to address theoretical accounts of consciousness based on the hypothesis of a dynamic core, i.e. a constantly evolving and transiently stable set of coordinated neurons that constitute an integrated and differentiated physical substrate for each conscious experience. We review work exploring the possibility that metastability in brain dynamics leads to a repertoire of dynamic core states, and discuss how it might be modified during altered states of consciousness. This discussion prompts us to review neuroimaging studies aimed to map the dynamic exploration of the repertoire of states as a function of consciousness. Complementary studies of the dynamic core hypothesis using perturbative methods are also discussed. Finally, we propose that a link between metastability in brain dynamics and the level of consciousness could pave the way towards a mechanistic understanding of altered states of consciousness using tools from dynamical systems theory and statistical physics.


Protein environment facilitates O2 binding in non-heme iron enzyme. An insight from ONIOM calculations on isopenicillin N synthase (IPNS).
Binding of dioxygen to a non-heme enzyme has been modeled using the ONIOM combined quantum mechanical/molecular mechanical (QM/MM) method. For the present system, isopenicillin N synthase (IPNS), binding of dioxygen is stabilized by 8-10 kcal/mol for a QM:MM (B3LYP:Amber) protein model compared to a quantum mechanical model of the active site only. In the protein system, the free energy change of O2 binding is close to zero. Two major factors consistently stabilize O2 binding. The first effect, evaluated at the QM level, originates from a change in coordination geometry of the iron center. The active-site model artificially favors the deoxy state (O2 not bound) because it allows too-large rearrangements of the five-coordinate iron site. This error is corrected when the protein is included. The corresponding effect on binding energies is 3-6 kcal/mol, depending on the coordination mode of O2 (side-on or end-on). The second major factor that stabilizes O2 binding is van der Waals interactions between dioxygen and the surrounding enzyme. These interactions, 3-4 kcal/mol at the MM level, are neglected in models that include only the active site. Polarization of the active site by surrounding amino acids does not have a significant effect on the binding energy in the present system.


Hydroxystreptomycin production and resistance in Streptomyces glaucescens.
The wild-type strain of Streptomyces glaucescens produces hydroxystreptomycin and shows an inherent natural resistance to streptomycin group aminoglycoside antibiotics. Cell free extracts of the wild-type strain were able to inactivate streptomycin, hydroxystreptomycin and dihydrostreptomycin in the presence of ATP. The phosphotransferase did not inactivate other aminoglycoside antibiotics, including spectinomycin. Mutant strains were isolated, which were highly sensitive to streptomycin group aminoglycosides, had no measurable phosphotransferase activity and were unable to form detectable amounts of hydroxystreptomycin. This suggests either a correlation between phosphotransferase activity, streptomycin resistance and hydroxystreptomycin formation or defects in more than one gene in the strS mutant strains tested.


Bictegravir/Emtricitabine/Tenofovir Alafenamide Treatment: Efficacy and Tolerability in Clinical Practice.
Analysis of bictegravir/emtricitabine/tenofovir alafenamide (BFTAF) efficacy and safety in virologically suppressed people living with HIV (PLWH) in clinical practice. The retrospective cohort study, which included adult treatment-experienced and virologically suppressed PLWH, switched to BFTAF from June 2019 to June 2021. Efficacy and safety were evaluated as virological failure (VF=2 consecutive HIV-RNA>50 copies/mL or a single HIV-RNA>400 copies/mL) and treatment failure (TF=VF or discontinuation for any reason) until data freezing (August 2022). Of the 1040 PLWH included, 67.8% switched from elvitegravir/cobicistat/FTAF. VF occurred in 4.2% (n=44), with incidence rate of 1.63 per 1000 person-months of follow-up (PMFU) and probability at 24-30 months of 3.8%-4.0%, respectively. Out of the 44 VF, in 75% virological re-suppression was achieved while maintaining BFTAF. Discontinuation occurred in 15% after a median time of 13.5 months of follow-up, with an incidence rate of 5.67 per 1000 PMFU, and a probability at 24-30 months of 11.9%-15.3%, respectively. Main discontinuation reasons were simplification (51.3%) and toxicity (21.8%, involving CNS in half of cases). TF occurred in 18.6% with an incidence rate of 7.01 per 1000 PMFU after a median time of 13.6 observation months; probability at 24-30 months was 14.8%-18.4%, respectively. BFTAF has proven effective and well tolerated in clinical practice.


Modulation of calcium-activated potassium channels from rat brain by protein kinase A and phosphatase 2A.
By incorporating plasma membrane vesicles into planar lipid bilayers, we previously characterized a family of four types of Ca(2+)-activated K+ channels from rat brain (Reinhart et al., 1989). Two of these are "large-conductance" or "maxi"-K+ channels, which differ in their gating kinetics and toxin sensitivity and are henceforth referred to as "type 1" and "type 2" channels. Here we show that the gating of these two channel types can be modulated by phosphorylation and dephosphorylation. The effects of cAMP-dependent protein kinase catalytic subunit (PK-A) on type 1 maxi-K+ channels are complex in that, while half of these channels are upregulated by the kinase, about one out of seven channels is downregulated. Thus, there may be several distinct channels within the type 1 category. Type 2 maxi-K+ channels are consistently downregulated by PK-A. The effects of PK-A on both channel types are reversed by the catalytic subunit of protein phosphatase 2A (PP-2A), but not by protein phosphatase 1 (PP-1). Furthermore, some of the type 1 maxi-K+ channels can be modulated by PP-2A, even without any prior PK-A treatment, indicating they are in a phosphorylated state when they are incorporated into the bilayer. The results demonstrate that (1) type 1 and type 2 maxi-K+ channels are substrates for PK-A; (2) phosphorylation can shift the open probability of channels in either direction, by a mechanism involving multiple phosphorylation sites; (3) phosphorylation alters the Ca2+/voltage sensitivity of these channels; and (4) dephosphorylation of type 1 and type 2 channels is catalyzed by specific phosphatases.


Corticosterone concentrations in blood and excretion in faeces after ACTH administration in male Sprague-Dawley rats.
The aim of the present study was to analyse the corticosterone response to exogenous ACTH in the circulation of catheterised male rats and to investigate the sensitivity of faecal corticosterone output as a measure of preceding elevated levels in the circulation. A total of 21 adult male Sprague-Dawley rats permanently catheterised (v. jugularis externa for intravenous administration of ACTH and a. carotis communis for blood sampling), were used. Administration of both 10 and 100 microg/kg ACTH resulted in a rapid and pronounced corticosterone increase three minutes after injection (226 and 220 ng/ml, respectively), but the duration of the response was different. In the 10 microg/kg group, corticosterone levels were significantly elevated for 3-90 min after injection, while in the 100 microg/kg group, the levels remained elevated for 240 min after injection. In faeces, a significant increase during eight hours after ACTH injection was found in the group treated with 100 microg/kg, but not in the group treated with 10 microg/kg. In conclusion, quantification of faecal excretion of corticosteroids is a useful non-invasive measure of prior substantial stress (e.g. surgery), but not sufficiently sensitive to reveal minor stress or acute stress of short duration.


Valproic acid-associated vanishing bile duct syndrome.
Hepatotoxicity as a result of valproic acid therapy is well documented. Elevation in aminotransferase activities is rarely associated with symptoms. It sometimes manifests as acute liver failure. Here, we report a 8-year-old girl who was referred for unresolving jaundice and itching for 3 months. Past history revealed afebrile convulsion 5 months previously and beginning of valproic acid treatment. Valproic acid was discontinued after the development of jaundice. Physical examination revealed ichterus, xanthomas on extensor surfaces of extremities, and hepatomegaly without any sign of chronic liver disease. Total and direct bilirubin levels were 20.2 and 12.9 mg/dL, respectively. Enzyme activities indicating cholestasis were increased together with blood cholesterol. Tests for infectious and autoimmune, metabolic, and genetic disorders were not informative. Liver biopsy revealed portal inflammation, severe bile duct loss, and cholestasis. The patient was considered to have valproic acid-associated vanishing bile duct syndrome, which has not been reported previously.


Audit of referral of diabetic patients.
To determine the referral, the feedback rates and to find out the prevalence of diabetic retinopathy among diabetics who were referred from a primary health care center, to Abha General Hospital in Aseer Region. This study was conducted at Wasat Abha Primary Health Care Center in Aseer region at the end of 1996 and 1997. All the diabetic files were reviewed and evaluated for referral to the eye clinic, their feedback and the prevalence of diabetic retinopathy among diabetics for whom fundoscopy was carried out. The related data was entered and analyzed by using the statistical package for social sciences program. A total of 203 diabetic files were reviewed. Forty percent were referred during 1996 while 68.5% were referred during 1997. The rates of feedback were 72% in 1996 and 71.2% in 1997. Twenty percent were referred twice during these 2 years. The prevalence of diabetic retinopathy was 11.3%. The referral rate and the feedback from the eye clinic was lower than the national target. A poor referral system was responsible for such a low rate. Either reorganization of the referral system or calling the ophthalmologists to attend Primary Health Care Centers monthly will increase the rate of eye examination among diabetics and will help in early detection of diabetic retinopathy and in prevention of blindness.


Experiences of family caregivers the first six months after patient diagnosis of necrotising soft tissue infection: A thematic analysis.
Necrotising soft tissue infection, or necrotising fasciitis, is a rapidly progressing disease requiring immediate diagnosis and treatment consisting of antimicrobial therapy, hyperbaric oxygen, debridement surgery and treatment in the intensive care unit. The harrowing illness trajectory affects the family caregivers potentially producing long-term psychological issues. We aimed to explore the experiences and coping strategies of family caregivers during the first six months after patient diagnosis of necrotising soft tissue infection. Our study had a prospective, explorative, qualitative design using semi-structured interviews and thematic analysis to understand necrotising soft tissue infection as an intrinsic and instrumental case. Family caregivers (n = 25) were recruited at three university hospitals in Denmark and Sweden. We identified three chronological themes describing issues of importance to the family caregivers. In the intensive care unit: Coping with illness and intensive care; In the ward: Coping with injury and post-intensive care and At home: Coping with recovery and new home life. Challenges facing family caregivers of necrotising soft tissue infections survivors are still under-recognised. Healthcare professionals need to ensure that families and stakeholders throughout the patient trajectory have access to and co-create timely information and care plans to bridge the knowledge gap across care environments and to relieve family responsibility.


Meningococcal disease in Italy: public concern, media coverage and policy change.
Between 2015 and 2017 six deaths due to meningitis in the Lombardy Region, Northern Italy, caught the attention of media and increased concern among the population, with a consequent increase in demand for vaccination. Considering the evidence about the impact of media coverage of health issues on public behaviour, this paper investigates the trend of media coverage and internet searches regarding meningitis in the Lombardy Region. Content analysis of online articles published from January 2015 to May 2017 and analysis of Google Trends were carried out. A codebook was created in order to assess the content of each article analysed, based on six areas: article characteristics, information about meningococcal disease and vaccination, Local Health Authority activities, accuracy of information and tone of the message. Both public interest and media attention peaked in December 2016 and January 2017, when the Lombardy Regional Authority changed its policy by offering co-payment to adults with a saving of 50%. The frequency of meningitis coverage decreased after the announcement of policy change. For example, articles containing new information on meningitis or meningococcal vaccine (76 to 48%, p = 0.01) and preventive recommendations (31% down to 10%, p = 0.006) decreased significantly. An alarmist tone appeared in 21% of pre-policy articles that decreased to 5% post-policy (p = 0.03). The findings suggest a role for the media in fostering public pressure towards health services and policy-makers. A collaboration between Public Health institutions and the media would be beneficial in order to improve communication with the public.


Shedding light on selenium biomineralization: proteins associated with bionanominerals.
Selenium-reducing microorganisms produce elemental selenium nanoparticles with particular physicochemical properties due to an associated organic fraction. This study identified high-affinity proteins associated with such bionanominerals and with nonbiogenic elemental selenium. Proteins with an anticipated functional role in selenium reduction, such as a metalloid reductase, were found to be associated with nanoparticles formed by one selenium respirer, Sulfurospirillum barnesii.


Oncolytic Immunotherapy: Dying the Right Way is a Key to Eliciting Potent Antitumor Immunity.
Oncolytic viruses (OVs) are novel immunotherapeutic agents whose anticancer effects come from both oncolysis and elicited antitumor immunity. OVs induce mostly immunogenic cancer cell death (ICD), including immunogenic apoptosis, necrosis/necroptosis, pyroptosis, and autophagic cell death, leading to exposure of calreticulin and heat-shock proteins to the cell surface, and/or released ATP, high-mobility group box 1, uric acid, and other damage-associated molecular patterns as well as pathogen-associated molecular patterns as danger signals, along with tumor-associated antigens, to activate dendritic cells and elicit adaptive antitumor immunity. Dying the right way may greatly potentiate adaptive antitumor immunity. The mode of cancer cell death may be modulated by individual OVs and cancer cells as they often encode and express genes that inhibit/promote apoptosis, necroptosis, or autophagic cell death. We can genetically engineer OVs with death-pathway-modulating genes and thus skew the infected cancer cells toward certain death pathways for the enhanced immunogenicity. Strategies combining with some standard therapeutic regimens may also change the immunological consequence of cancer cell death. In this review, we discuss recent advances in our understanding of danger signals, modes of cancer cell death induced by OVs, the induced danger signals and functions in eliciting subsequent antitumor immunity. We also discuss potential combination strategies to target cells into specific modes of ICD and enhance cancer immunogenicity, including blockade of immune checkpoints, in order to break immune tolerance, improve antitumor immunity, and thus the overall therapeutic efficacy.


Development of indicators for quality assurance in public health medicine.
To develop structure, process, and outcome indicators within a quality rating index for audit of public health medicine. Development of an audit matrix and indicator of quality through a series of group discussions with public health physicians, from which self administered weighted questionnaires were constructed by a modified Delphi technique. Five Scottish health boards. Public health physicians in the five health boards. Indicators of quality and a quality rating index for seven selected service categories for each of seven agreed roles of public health medicine: assessment of health and health care needs in information services, input into managerial decision making in health promotion, fostering multisectoral collaboration in environmental health services, health service research and evaluation for child services, lead responsibility for the development and/or running of screening services, and public health medicine training and staff development in communicable disease. Indicators in the form of questionnaires were developed for each topic. Three types of indicator emerged: "global," "restricted," and "specific." A quality rating index for each topic was developed on the basis of the questionnaire scores. Piloting of indicators showed that they are potentially generalisable; evaluation of the system is under way across all health boards in Scotland. Measurable indicators of quality for public health medicine can be developed.


Extracorporeal repair of the renal artery for renovascular hypertension: a case report.
A case of successful correction of renovascular hypertension by means of extracorporeal repair of fibroplasia of the main trunk and an aneurysm of a secondary division branch of the left renal artery is presented. The ureter was not transected and a segment of the internal ilac artery with two branches was used for the substitution of the diseased renal artery segments. The kidney was preserved by intermittent flushing with iced Ringer's lactate solution. A probable post-operative arterial thrombosis of the repaired secondary branch was successfully treated with intravenous streptokinase. One year after the operation the patient is normotensive.


Extent of possible rehabilitation of service personnel with ischaemic heart disease.
Sixty unselected survivors from ischaemic heart disease, all male service personnel, were treated, within 1 to 6 months of recovery from the acute episode, by promotion of physical activity, correction of coronary risk factors, and routine use of nicoumalone and prenylamine lactate. The results showed that this approach was conducive to more rewarding rehabilitation than was obtained in 89 patients who, in the past, were on conventional treatment based on advice regarding weight, diet, and physical and mental activity. Thus within 1 to 30 months of treatment by this approach, out of 60 patients, 12 were fit for medical category A active service duties in operational areas in any part of the world in any terrain, including altitudes between 10,000 and 18,000 feet, 30 were fit for medical category B service duties in non-operational communication zones in any part of the world including altitudes below 10,000 feet, and 17 were fit for medical category C sedentary duties in non-operational areas in India only. One patient, who initially recovered from congestive heart failure, died while in a state of temporary unfitness for service. No patient was released from service on account of ischaemic heart disease. Against this, by the conventional approach, out of 89 patients, 81 were fit for medical category C sedentary duties in non-operational areas in India only, 4 were released from service, and 4 died.


Microstimulation of auditory nerve for estimating cochlear place of single fibers in a deaf ear.
Multielectrode cochlear prostheses seek to approximate the cochlea's normal frequency-place mapping through spatial segregation of stimulus currents. Various electrode configurations have been employed to achieve such segregation. Direct measurements of stimulation regions among single auditory nerve (AN) fibers has been possible only when normal hearing is preserved, such that each fiber's cochlear place can be inferred from its tuning curve. This precludes measurements in deafened ears, or ears compromised by implantation of the electrodes. Data presented here demonstrate that the cochlear place of an AN fiber can be estimated without acoustic sensitivity, using electrical microstimulation through a recording pipette in the AN bundle. The procedure exploits cochleotopic projection to isofrequency laminae within the contralateral inferior colliculus (IC). Microstimulation excites a small group of fibers neighboring the recorded fiber, generating centrally propagated volleys along a narrow frequency-specific pathway. Evoked potential recordings at varying depths are made to identify the ICC lamina where the response to AN microstimulation is greatest. Preliminary data are also presented for an alternative method of identifying the lamina using a frequency domain measure of binaural interactions within the IC.


"Keep It Simple"-Co-Creation of a Tailored Newborn Resuscitation Course for Midwifery Students.
To develop a tailored newborn resuscitation course for midwifery students. A qualitative study using an explorative, abductive approach was applied. Co-creation through workshops was facilitated to develop a tailored newborn resuscitation course for midwifery students. Four workshops with midwifery students and midwives were conducted from May to October 2020. Twenty participants attended one workshop of their choice. Five out of Norway's six midwifery education programmes participated, and included midwives from across Norway. All workshops were held digitally via the Zoom platform. A seven-step framework analysis method was applied to analyse the workshop data. We identified four themes: 1) practice guidance, 2) technical skills 3) non-technical skills 4) innovative methods. Findings emphasize the importance of practice guidance, technical skills, non-technical skills and innovative methods to facilitate the learning process. However, these skills cannot be acquired without the context to facilitate them, and thus a supportive culture is essential to sustain newborn resuscitation expertise as a midwife/midwifery student. We found that midwives expressed the same need to learn, train and prepare themselves for newborn resuscitation as midwifery students. The importance of facilitating the learning of newborn resuscitation with low-dose, high-frequency training in a supportive culture thus matters to both midwifery students and expert midwives.


[Diagnostic algorithm for von Willebrand Disease (vWD) in a Mexican population].
The diagnosis of von Willebrand disease (vWD) is complex and requires several screening and confirmation tests, such as the analysis of vWF multimers, which is considered the gold standard for vWD subtyping; however, it only discriminates 2A subtype while the 2B, 2M, and 2N subtypes require additional tests and even genetic testing for final confirmation. It is important to consider the patients' hemotype for the vWD diagnosis, particularly in Mexico where hemotype "O" predominates and may entail a 20-25% decreased level of plasma vWF and increased bleeding tendency.


Aspalathin ameliorates doxorubicin-induced oxidative stress in H9c2 cardiomyoblasts.
Aspalathin (ASP) is a C-dihydrochalcone abundantly found in Aspalathus linearis. While we have provide evidence that ASP can protect H9c2 cardiomyoblasts against doxorubicin (Dox)-induced apoptosis through regulation of autophagy, the complete mechanism involved in the cardioprotective effect of this dihydrochalcone remains to be explored. Here we provide evidence that ASP reverses Dox-induced apoptosis through the amelioration of oxidative stress in H9c2 cardiomyoblasts. Cultured cells were treated with 0.2 μM Dox or co-treated with either 20 μM dexrazoxane (Dexra) or 0.2 μM ASP daily for five days, to a final dose of 1 μM Dox, 100 μM Dexra and 1 μM ASP, respectively. Superoxide dismutase, catalase, glutathione, malondialdehyde and dichloro-dihydro-fluorescein diacetate fluorescence were used as end-point measurements for oxidative stress, while JC-1 and TUNEL labeling were performed to assess mitochondria depolarization and apoptosis. Co-treatment with ASP attenuated Dox-induced cardiotoxicity by improving endogenous antioxidant levels and mitochondrial membrane potential, while inhibiting reactive oxygen species production and cellular apoptosis. These findings suggested that ASP can prevent Dox-induced oxidative stress and apoptosis and needs further assessment to confirm its therapeutic potential to prevent Dox-induced cardiotoxicity.


Solute-solvent cavity and bridge functions. I. Varying size of the solute.
In this work we present the results of the extensive molecular simulations of solute-solvent cavity and bridge functions. The mixtures of Lennard-Jones solvent with Lennard-Jones solute at infinite dilution are considered for different solute-solvent size ratios-up to 4:1. The Percus-Yevick and hypernetted chain closures deviate substantially from simulation results in the investigated temperature and density ranges. We also find that the behavior of the indirect and cavity correlation functions is non-monotonous within the hard-core region, but the latter can be successfully approximated by mean-field theory if the solute-solvent interaction energy is divided into repulsive and attractive contribution, according to Weeks-Chandler-Andersen theory. Furthermore, in spite of the non-monotonous behavior of logarithm of the cavity function and the indirect correlation function, their difference, i.e., the bridge function remains constant within the hard-core region. Such behavior of the bridge and indirect correlation functions at small distances and for small values of indirect correlation function is well known from the Duh-Haymet plots, where the non-unique relationship results in loops of the bridge function vs. indirect correlation function graphs. We show that the same pathological behavior appears also when distance is small and indirect correlation function is large. We further show that the unique functional behavior of the bridge function can be established when bridge is represented as a function of the renormalized, repulsive indirect correlation function.


In vivo interaction between mutated tryptophan repressors of Escherichia coli.
By expressing a mutant trpR gene in an Escherichia coli strain that is trpR and has beta-galactosidase activity fused to the trp promoter/operator, thus putting the beta-galactosidase activity under the control of the Trp repressor, we can determine quantitatively the relative repression activity of such mutant(s). We used this technique to analyse the biological consequences of substituting certain amino acid residues in only one of the two corepressor binding pockets. By combining two compatible plasmids in this strain, one expressing the mutant T44M and the other expressing only one substitution at a time at position 85, we analysed the repression activity of the resulting interactions in vivo. This approach allowed us to engineer active dimer repressors made of two inactive or partially active monomers. Amino acid substitutions at position 85 with a positive or with an indole ring (W) appeared to complement T44M, which amino acids with a negative charge did not. Only L substitution at position 85 appeared to restore activity among the hydrophobic amino acids tested. Similar to the wild-type repressor activity, the successful mutant-mutant interactions were L-tryptophan dependent. In vivo regulation by three known L-tryptophan analogues demonstrated the same trend of regulation among the wild-type repressors and the active mutant-mutant combinations.


Endothelial cell phenotypes in the rheumatoid synovium: activated, angiogenic, apoptotic and leaky.
Endothelial cells are active participants in chronic inflammatory diseases. These cells undergo phenotypic changes that can be characterised as activated, angiogenic, apoptotic and leaky. In the present review, these phenotypes are described in the context of human rheumatoid arthritis as the disease example. Endothelial cells become activated in rheumatoid arthritis pathophysiology, expressing adhesion molecules and presenting chemokines, leading to leukocyte migration from the blood into the tissue. Endothelial cell permeability increases, leading to oedema formation and swelling of the joints. These cells proliferate as part of the angiogenic response and there is also a net increase in the turnover of endothelial cells since the number of apoptotic endothelial cells increases. The endothelium expresses various cytokines, cytokine receptors and proteases that are involved in angiogenesis, proliferation and tissue degradation. Associated with these mechanisms is a change in the spectrum of genes expressed, some of which are relatively endothelial specific and others are widely expressed by other cells in the synovium. Better knowledge of molecular and functional changes occurring in endothelial cells during chronic inflammation may lead to the development of endothelium-targeted therapies for rheumatoid arthritis and other chronic inflammatory diseases.


Validating a comprehensive plan scoring system for healthy community design in League City, Texas.
Comprehensive plans provide an overall vision for a city's land use and development and influence community health conditions. As part of a community-engaged research project, a Healthy Living and Active Design Scorecard was applied to the comprehensive plan of League City, Texas, to identify health-promoting design and planning strategies. Although the plan scored similarly to published examples, several areas of improvement aligned with priorities from a community health assessment were identified, including improving public transportation and access to exercise and recreation. Future revisions of the plans and neighbourhood scaled designs targeting these issues could improve the health of the community.


β-blockers in the treatment of dilated cardiomyopathy: which is the best?
Among the multiple pharmacologic and non-pharmacologic therapies, beta-blockers seem to be the mainstay of heart failure management. Although the first large studies failed to demonstrate a significant improvement in mortality, the results from subsequent studies revealed significant risk reductions in mortality with metoprolol, carvedilol and bisoprolol. Decision of the appropriate therapeutic agent depends on the existence of some specific conditions complicating dilated cardiomyopathy. This article reviews the impact of beta-blocker therapy in patients with dilated cardiomyopathy of ischemic or non-ischemic etiology.


The lifetime and past-year prevalence of dual diagnosis in people with schizophrenia across Europe: findings from the European Schizophrenia Cohort (EuroSC).
Relatively little is known about rates of comorbid drug and alcohol problems in people with schizophrenia outside the USA. Most studies have recruited from single countries. Newly available data provided an unmatched opportunity to investigate the prevalence of comorbid dependence on alcohol and other psychoactive substances in people with schizophrenia in France, Germany and the UK at the same time. The European Schizophrenia Cohort study data set used semi-structured clinical interviews to establish DSM-IV diagnoses. 1,208 patients were interviewed in nine centres. The lifetime rate for comorbid dependence on any substance was highest in the UK (35 %), but considerably lower in Germany (21 %) and in France (19 %), and generally more than double the past-year rates. Dependence on alcohol and on other psychoactive substances showed similar variations (comorbid alcohol dependence: UK 26 %; Germany 18 %; France 14 %; comorbid drug dependence: UK 18 %; Germany 8 %; France 7 %). Differences within countries persisted after controlling for individual characteristics. The relative odds of dependence were higher than in the general population, but varied between countries and centres. Dependence disorders are a common problem in people with schizophrenia in Western Europe, although effective service configurations have yet to be developed. Overall, these European rates are less than those reported from the USA. Research comparing people with current comorbidity with those who are no longer dependent is needed.


Characterisation of the unsteady transport of labelled species in permeable capillaries: role of convective dispersion.
A mathematical model of transcapillary exchange has been developed which considers in detail the contribution of Taylor dispersion (i.e., non-uniform velocity and inequalities in radial concentration) to the uptake and wash-out of multiple tracers from single capillaries. A numerical solution to the two-dimensional unsteady-state species continuity equation for a single capillary is obtained (by the use of an array processor) which simulates the convective dispersion of labelled indicators in permeable capillaries. Particular attention is directed toward an analysis of the roles of capillary permeability, transport regime as characterised by the Peclet number, velocity profile and the length of the exchange region on the development of time-concentration profiles and on the importance of convective dispersion. For the physiologic range of parameters consistent with pulmonary capillary transport of multiple indicators (e.g., labelled urea, albumin, etc.), we find that the magnitude of convective dispersion is insufficient to markedly affect the shapes of simulated tracer concentration profiles. The implication in these circumstances is that the use of one-dimensional models, which do not account for Taylor dispersion, does not lead to significant errors in parameter estimates derived from data of multiple indicator dilution.


Exposure to the three structurally different PCB congeners (PCB 118, 153, and 126) results in decreased protein expression and altered steroidogenesis in the human adrenocortical carcinoma cell line H295R.
Polychlorinated biphenyls (PCB), synthetic, persistent organic pollutants (POP), are detected ubiquitously, in water, soil, air, and sediments, as well as in animals and humans. PCB are associated with range of adverse health effects, such as interference with the immune system and nervous system, reproductive abnormalities, fetotoxicity, carcinogenicity, and endocrine disruption. Our objective was to determine the effects of three structurally different PCB congeners, PCB118, PCB 126, and PCB 153, each at two concentrations, on the steroidogenic capacity and proteome of human adrenocortical carcinoma cell line cultures (H295R) . After 48 h of exposure, cell viability was monitored and estradiol, testosterone, cortisol and progesterone secretion measured to quantify steroidogenic capacity of the cells. Two-dimensional (2D) gel-based proteomics was used to screen for proteome alterations in H295R cells in response to the PCB. Exposure to PCB 118 increased estradiol and cortisol secretion, while exposure to PCB 153 elevated estradiol secretion. PCB 126 was the most potent congener, increasing estradiol, cortisol, and progesterone secretion in exposed H295R cells. Seventy-three of the 711 spots analyzed showed a significant difference in normalized spot volumes between controls (vehicle only) and at least one exposure group. Fourteen of these protein spots were identified by liquid chromatography with mass spectroscopy (LC-MS/MS). Exposure to three PCB congeners with different chemical structure perturbed steroidogenesis and protein expression in the H295R in vitro model. This study represents an initial analysis of the effects on proteins and hormones in the H295R cell model, and additional studies are required in order to obtain a more complete understanding of the pathways disturbed by PCB congeners in H295R cells. Overall, alterations in protein regulation and steroid hormone synthesis suggest that exposure to PCB disturbs several cellular processes, including protein synthesis, stress response, and apoptosis.


A Whole-Brain Cell-Type-Specific Sparse Neuron Labeling Method and Its Application in a Shank3 Autistic Mouse Model.
Single neurons, as the basic unit of the brain, consist of a cell body and processes, including dendrites and axons. Even neurons of the same type show various subtle process characteristics to fit into the diverse neural circuits. Different cell types of neurons form complicated circuits in the brain. Therefore, detailed neuronal morphology is required to understand normal neuronal function and pathological mechanisms, such as those that occur in autism. Here, we developed a strategy to sparsely label the same type of neurons throughout the whole brain and tested its application in an autistic animal model-Shank3 knockout (KO) mice. To achieve this, we designed an adeno-associated virus (AAV) that expresses Cre recombinase-dependent regular and membrane-targeted enhanced green fluorescent protein (EGFP) under a human synapsin 1 promoter and verified it in several Cre transgenic mice. We could sparsely label the projection neurons in multiple brain areas by retro-ocular injection of the virus into CaMKIIα-Cre mice. Then, we analyzed the morphology of the projection neurons in Shank3 KO mice with this method. We found differential dendritic complexity and dendritic spine changes in projection neurons in Shank3 KO mice crossed with CaMKIIα-Cre mice compared with littermate control mice in the striatum, cortex, and hippocampus. By combining this method with various Cre mouse lines crossed with mouse models of disease, we can screen the morphological traits of distinct types of neurons throughout the whole brain that will help us to understand the exact role of the specific cell types of neurons not only in autism spectrum disorder (ASD) mouse models but also in other psychiatric disorder mouse models.


The motor-evoked potential threshold evaluated by tractography and electrical stimulation.
To validate the corticospinal tract (CST) illustrated by diffusion tensor imaging, the authors used tractography-integrated neuronavigation and direct fiber stimulation with monopolar electric currents. Forty patients with brain lesions adjacent to the CST were studied. During the operation, the motor responses (motor evoked potential [MEP]) elicited at the hand by the cortical stimulation to the hand motor area were continuously monitored, maintaining the consistent stimulus intensity (mean 15.1 +/- 2.21 mA). During lesion resection, direct fiber stimulation was applied to elicit MEP (referred to as fiber MEP) to identify the CST functionally. The threshold intensity for the fiber MEP was determined by searching for the best stimulus point and changing the stimulus intensity. The minimum distance between the resection border and illustrated CST was measured on postoperative isotropic images. Direct fiber stimulation demonstrated that tractography accurately reflected anatomical CST functioning. There were strong correlations between stimulus intensity for the fiber MEP and the distance between the CST and the stimulus points. The results indicate that the minimum stimulus intensity of 20, 15, 10, and 5 mA had stimulus points approximately 16, 13.2, 9.6, and 4.8 mm from the CST, respectively. The convergent calculation formulated 1.8 mA as the electrical threshold of the CST for the fiber MEP, which was much smaller than that of the hand motor area. The investigators found that diffusion tensor imaging-based tractography is a reliable way to map the white matter connections in the entire brain in clinical and basic neuroscience applications. By combining these techniques, investigating the cortical-subcortical connections in the human CNS could contribute to elucidating the neural networks of the human brain and shed light on higher brain functions.


The use of the case-crossover design in a continuous common source food-borne outbreak.
When applicable, case-crossover studies may be quicker and cheaper to complete than case-control studies. Because time is a major issue in outbreak investigations, we evaluated the interest of this design during a continuous common source food-borne outbreak of salmonellosis for which the vehicle (hamburgers) was also implicated by environmental and laboratory investigations. Seventeen of the 35 cases identified in the outbreak were included in the study according to the availability of menu records with detailed information on food consumed at each meal. Food exposures during a 3-day risk period before onset of illness were compared to those of a control time-interval of the same duration that preceded the risk period by two days. Seventy-seven per cent of the cases (13/17) had consumed hamburgers in the three days preceding onset of illness compared with 29% (5/17) during the control period (P = 0.04, odds ratio = 5, 95% CI: 1.1-46.9). In this investigation the case-crossover design proved to be a useful and efficient alternative to the case-control approach. However, further evaluation of this design in outbreak investigation is needed.


Impaired Pain Processing at a Brainstem Level Is Involved in Maladaptive Neuroplasticity in Patients with Chronic Complex Regional Pain Syndrome.
Neuroinflammatory mechanisms and maladaptive neuroplasticity underlie the progression of complex regional pain syndrome (CRPS), which is prototypical of central neuropathic pain conditions. While cortical maladaptive alterations are well described, little is known about the contribution of the brainstem to the pathophysiology. This study investigates the role of pain-modulatory brainstem pathways in CRPS using the nociceptive blink reflex (nBR), which not only provides a direct read-out of brainstem excitability and habituation to painful stimuli but may also be suitable for use as a diagnostic biomarker for CRPS. Thirteen patients with CRPS and thirteen healthy controls (HCs) participated in this prospective case-control study investigating the polysynaptic trigemino-cervical (R2) nBR response. The R2 area and its habituation were assessed following repeated supraorbital electrical stimulation. Between-group comparisons included evaluations of diagnostic characteristics as a potential biomarker for the disease. Patients with CRPS showed a substantial decrease in habituation on the stimulated (Cohen's d: 1.3; p = 0.012) and the non-stimulated side (Cohen's d: 1.1; p = 0.04). This is the first study to reveal altered nBR habituation as a pathophysiological mechanism and potential diagnostic biomarker in CRPS. We confirmed previous findings of altered nBR excitability, but the diagnostic accuracy was inferior. Future studies should investigate the nBR as a marker of progression to central mechanisms in CRPS and as a biomarker to predict treatment response or prognosis.


[Cross resistance to pyrethroids in Aedes aegypti from Cuba induced by the selection with organophosphate malathion].
A strain from Aedes aegypti (L) collected in the municipality of Santiago de Cuba with low levels of resistance to malathion (1.79x) was subjected to selection pressure aimed at evaluating the evolution of the resistance to this organophosphate and its usefulness for control. After 5 generations of selection (SAN-F5) with malathion, it was not possible to increase the resistance and it was obtained a value of resistance factor (FR50) of 2.22x. None or little cross resistance to the following organophosphate insecticides was observed: fenthion, temephos and fenitrotion; however, it was found an elevated cross resistance to deltamethrin pyrethroid, with a value of 287.5x in the third generation of selection. It was also observed cross resistance to other pyrethroids (lambda-cyhalothrin, cypermethrin and ciflutrine). The mechansim of elevated esterases did not generate resistance to pyrethroids, which was corroborated through inhibition gel studies and through the calculation of the frequency in microtitering plaques. Nevertheless, there was an increase in the frequency of the glutathione-s-transferase mechanism from 0.049 in Santiago de Cuba to 0.42 in SAN-F5, which may be associated with the resistance to pyrethroids. The cross resistance to pyrethroids, mainly to deltamethrin, resulting from the selection with malathion may limit the use of these insecticides in the control unless a good strategy for their use is laid down.


Ku70 can translocate to the nucleus independent of Ku80 translocation and DNA-PK autophosphorylation.
Ku plays an important role in multiple nuclear processes, e.g., DNA repair, chromosome maintenance, and transcriptional regulation. Although some evidence suggests that the nuclear translocation of Ku plays a key role in regulating the function of Ku, the mechanism is poorly understood. Using the site-directed mutagenesis technique, we demonstrate here that Ku70 can translocate to the nucleus without heterodimerization with Ku80. The nuclear accumulation of Ku70 mutants of the nuclear localization signal, which retained their binding ability with Ku80, was diminished. On the other hand, Ku70 mutants which lacked the ability to bind with Ku80 could translocate to the nuclei. Human Ku70, when transfected, accumulated within the nuclei of hamster xrs-6 cells which had undetectable DNA-PK activity and Ku80. Ku70 and Ku80 mutants of DNA-PK phosphorylation sites showed normal heterodimerization and nuclear translocation. These findings also support the idea that Ku70 can translocate to the nucleus independent of DNA-PK autophosphorylation.


The impact of the Mississippi Improved Child Health Project on prenatal care and low birthweight.
A quasi-experimental, nonequivalent control group design was used to evaluate the Improved Child Health Projects in northwest Mississippi (ICHP1 and ICHP2). Control counties were selected for each project that on average were similar to ICHP counties on racial composition, median family income in 1970 and 1980, and number of births in 1978-79. The study population comprised all resident births in the ICHP and control counties during a pre-ICHP period (1975-78) and the ICHP period (1979-81). The percentage of women with adequate prenatal care rose between the two periods for all counties; the rise was greater for the study than for the control counties for ICHP1; the reverse was found for ICHP2. For both projects, the low birthweight rate remained constant in the pre-ICHP and ICHP periods for the study and control counties. Adjustment for changes in the childbearing characteristics between the two periods did not alter these results. Community involvement in its development and coordination may explain ICHP1's impressive rise in the use of prenatal care.


[Safety and efficacy of specific sublingual immunotherapy in patients with asthma and allergy to Dermatophagoides pteronyssinus].
First reports on sublingual immunotherapy were published in 1980. To compare safety and effectiveness of sublingual immunotherapy, as compared with placebo, in asthmatic patients. In a blinded randomized controlled trial asthmatic patients with positive skin prick tests to Dermatophagoides pteronyssinus, and with serum IgE at least 200 UI were included. According to GINA, asthma severity was mild persistent and moderate. All patients improved their baseline FEV1 at least by 14% after inhaled albuterol. Spirometry was performed again after three and six months after initiating treatment. Patients were randomized to receive for six months either sublingual immunotherapy with Der p 1 standardized allergens (IPI-ASAC, México) at a total dose of 10,469 UBE or identically looking and tasting placebo. Both groups received conventional pharmacological therapy. Sixty four patients enter the study; four were excluded because of systemic oral steroid therapy. Sixty patients underwent randomization. Both groups (30 patients in each one) were similar in their baseline characteristics. After six months, patients that received sublingual immunotherapy had less exacerbations than those in the control group (61 vs 123, T 2.61, p<0.001, IC 1.8-7.2), better FEV1 as compared with baseline values (25% vs 9%, Z=0.66, p=0.03), and less need of albuterol (50% of initial dose, vs 21% (Z=1.4, p=0.03, IC 1.39-1.49). Sublingual immunotherapy improves patient symptoms and pulmonary functional tests, makes exacerbations less frequent, and reduces albuterol needs. It may improve asthma related quality of life.


The fusion of mental imagery and sensation in the temporal association cortex.
It is well understood that the brain integrates information that is provided to our different senses to generate a coherent multisensory percept of the world around us (Stein and Stanford, 2008), but how does the brain handle concurrent sensory information from our mind and the external world? Recent behavioral experiments have found that mental imagery--the internal representation of sensory stimuli in one's mind--can also lead to integrated multisensory perception (Berger and Ehrsson, 2013); however, the neural mechanisms of this process have not yet been explored. Here, using functional magnetic resonance imaging and an adapted version of a well known multisensory illusion (i.e., the ventriloquist illusion; Howard and Templeton, 1966), we investigated the neural basis of mental imagery-induced multisensory perception in humans. We found that simultaneous visual mental imagery and auditory stimulation led to an illusory translocation of auditory stimuli and was associated with increased activity in the left superior temporal sulcus (L. STS), a key site for the integration of real audiovisual stimuli (Beauchamp et al., 2004a, 2010; Driver and Noesselt, 2008; Ghazanfar et al., 2008; Dahl et al., 2009). This imagery-induced ventriloquist illusion was also associated with increased effective connectivity between the L. STS and the auditory cortex. These findings suggest an important role of the temporal association cortex in integrating imagined visual stimuli with real auditory stimuli, and further suggest that connectivity between the STS and auditory cortex plays a modulatory role in spatially localizing auditory stimuli in the presence of imagined visual stimuli.


Vertical transmission of arboviruses in mosquitoes: a historical perspective.
Arthropod-borne viruses (arboviruses) are mainly transmitted horizontally among vertebrate hosts by blood-feeding invertebrate vectors, but can also be transmitted vertically in the vector from an infected female to its offspring. Vertical transmission (VT) is considered a possible mechanism for the persistence of arboviruses during periods unfavorable for horizontal transmission, but the extent and epidemiological significance of this phenomenon have remained controversial. To help resolve this question, we reviewed over a century of published literature on VT to analyze historical trends of scientific investigations on experimental and natural occurrence of VT in mosquitoes. Our synthesis highlights the influence of major events of public health significance in arbovirology on the number of VT publications. Epidemiological landmarks such as emergence events have significantly stimulated VT research. Our analysis also reveals the association between the evolution of virological assays and the probability of VT detection. Increased sensitivity and higher-throughput of modern laboratory assays resulted in enhanced VT detection. In general, VT contribution to arbovirus persistence is likely modest because vertically infected mosquitoes are rarely observed in nature. Taken together, however, our results call for caution when interpreting VT studies because their conclusions are context- and method-dependent.


Novel chelate-setting calcium-phosphate cements fabricated with wet-synthesized hydroxyapatite powder.
The hydroxyapatite (HAp) powder preparation process was optimized to fabricate inositol phosphate-HAp (IP6-HAp) cement with enhanced mechanical properties. Starting HAp powders were synthesized via a wet chemical process. The effect of the powder preparation process on the morphology, crystallinity, median particle size, and specific surface area (SSA) of the cement powders was examined, together with the mechanical properties of the resulting cement specimens. The smallest crystallite and median particle sizes, and the highest SSA were obtained from ball-milling of as-synthesized HAp powder under wet conditions and then freeze-drying. IP6-HAp cement fabricated with this powder had a maximum compressive strength of 23.1 ± 2.1 MPa. In vivo histological studies using rabbit models revealed that the IP6-HAp cements were directly in contact with newly formed and host bones. Thus, the present chelate-setting HAp cement is promising for application as a novel paste-like artificial bone.


Adrenomedullin(22-52) combats inflammation and prevents systemic bone loss in murine collagen-induced arthritis.
Adrenomedullin(22-52) is a truncated peptide derived from adrenomedullin, a growth factor with antiapoptotic and immunoregulatory properties. It can act as an agonist or an antagonist depending on cell type. Its in vivo effects are unknown, but adrenomedullin(22-52) could possess immunomodulatory properties. This study was undertaken to evaluate the effect of adrenomedullin(22-52) in a mouse model of arthritis. DBA/1 mice with collagen-induced arthritis (CIA) were treated with 1.2 μg/gm adrenomedullin(22-52) , adrenomedullin, or saline at arthritis onset. Bone mineral density was measured at the beginning of the experiment and when mice were killed. Mouse joints were processed for histologic analysis and protein studies, and spleens were examined for Treg cell expression. Cytokine expression was studied in mouse joint tissue and serum. In mice with CIA, adrenomedullin and adrenomedullin(22-52) reduced clinical and histologic arthritis scores and shifted the pattern of articular and systemic cytokine expression from Th1 to Th2, as compared to untreated mice with CIA (controls). Tumor necrosis factor α, interleukin-6 (IL-6), and IL-17A levels were significantly decreased in the joints of mice with CIA treated with adrenomedullin or adrenomedullin(22-52) as compared to controls, whereas IL-4 and IL-10 levels were increased. Adrenomedullin(22-52) was more effective than adrenomedullin in modulating cytokine content and enhanced Treg cell function without changing Treg cell expression compared to controls. Adrenomedullin receptor binding and transcriptional adrenomedullin receptor expression were markedly increased in joints from controls, whereas adrenomedullin receptor binding was considerably decreased in treated animals. Mice with CIA treated with adrenomedullin or adrenomedullin(22-52) had considerably fewer apoptotic chondrocytes and diminished cartilage degradation. Adrenomedullin(22-52) completely prevented systemic bone loss by preserving osteoblastic activity, but without changes in osteoclastic activity. Our findings indicate that adrenomedullin(22-52) , which has no vasoactive or tumor-inducing effects, is a potent antiinflammatory and bone-protective agent in this arthritis model.


Intervention Effect of Group Counseling Based on Positive Psychology on Psychological Crisis of College Student.
To analyze the intervention effect of group counseling based on positive psychology on psychological crisis of college student. SCL-90 mental health screening was performed on second-year students in a college by cluster stratification. Among the detected students, 210 were included into the group after brief interview and randomly assigned to the experimental group and the control group. The control group was given conventional intervention measures, and the experimental group was given group counseling according to the interview results. One week before the start, on the day of the end of the intervention and 3 months after the end of the intervention, the mental health level of all subjects was evaluated by symptom self-rating scale, general well-being scale, and adolescent mental resilience scale. The scores of two groups were different at different time points during the intervention. With the extension of time, the score of the self-rating symptom scale in the experimental group decreased significantly, while the total score of the self-rating symptom scale in the control group increased, with statistical significance (P < 0.05). Before the intervention, there was no significant difference in general well-being between the two groups (P > 0.05). At 3 months after the end of intervention, the total score of the general well-being scale in the experimental group increased, while that in the control group decreased, with statistical significance (P < 0.05). Three months after the end of the intervention, the total score of adolescent mental resilience scale in the experimental group increased. Group counseling from the perspective of positive psychology can effectively improve the mental health status of medical students with psychological crisis and improve their mental resilience.


Use of purified Clostridium difficile spores to facilitate evaluation of health care disinfection regimens.
Clostridium difficile is a major cause of antibiotic-associated diarrheal disease in many parts of the world. In recent years, distinct genetic variants of C. difficile that cause severe disease and persist within health care settings have emerged. Highly resistant and infectious C. difficile spores are proposed to be the main vectors of environmental persistence and host transmission, so methods to accurately monitor spores and their inactivation are urgently needed. Here we describe simple quantitative methods, based on purified C. difficile spores and a murine transmission model, for evaluating health care disinfection regimens. We demonstrate that disinfectants that contain strong oxidizing active ingredients, such as hydrogen peroxide, are very effective in inactivating pure spores and blocking spore-mediated transmission. Complete inactivation of 10⁶ pure C. difficile spores on indicator strips, a six-log reduction, and a standard measure of stringent disinfection regimens require at least 5 min of exposure to hydrogen peroxide vapor (HPV; 400 ppm). In contrast, a 1-min treatment with HPV was required to disinfect an environment that was heavily contaminated with C. difficile spores (17 to 29 spores/cm²) and block host transmission. Thus, pure C. difficile spores facilitate practical methods for evaluating the efficacy of C. difficile spore disinfection regimens and bringing scientific acumen to C. difficile infection control.


Hexokinase II: the integration of energy metabolism and control of apoptosis.
Hexokinase II is often highly expressed in poorly differentiated and rapidly growing tumors that exhibit a high rate of aerobic glycolysis. Hexokinase II binds to the mitochondrial membrane through its interaction with the outer membrane voltage-dependent anion channel (VDAC), preferentially at contact sites between the outer and inner mitochondrial membrane. This location is thought to be important for the integration of glycolysis with mitochondrial energy metabolism. VDAC is a critical component of the mitochondrial phase of apoptosis and its interaction with Bcl-2 family proteins controls the rate of release of mitochondrial intermembrane space proteins that activate the execution phase of apoptosis. The proteins involved in the contact sites also constitute the mitochondrial permeability transition, one of the mechanisms by which mitochondrial protein release can be mediated. Hexokinase II binding to VDAC suppresses the release of intermembrane space proteins and inhibits apoptosis, thereby contributing to the survival advantage of tumor cells. This interaction places hexokinase II in a position to integrate glycolytic metabolism of the tumor cell with the control of apoptosis at the mitochondrial level. Mitochondrial binding of hexokinase II may constitute an attractive target for therapeutic intervention to suppress tumor growth.


Expression and secretion of cathelicidin LL-37 in human epithelial cells after infection by Mycobacterium bovis Bacillus Calmette-Guérin.
The antimicrobial cathelicidin LL-37 is considered to play an important role in the innate immune response to tuberculosis infection. However, little is known about the induction and secretion of this antimicrobial peptide in A549 epithelial cells after infection with Mycobacterium bovis bacillus Calmette-Guérin (BCG), the world's most widely used tuberculosis vaccine. In this study, we investigated the effect of M. bovis BCG on LL-37 mRNA levels in A549 cells by real-time PCR and on protein levels by Western blotting. Treatment of cells with M. bovis BCG upregulates LL-37 mRNA expression in a dose- and time-dependent manner. The quantitative analysis of LL-37 gene expression correlated with our Western blotting results. Moreover, our results demonstrated that treatment of cells with the transcriptional inhibitor actinomycin D effectively inhibited in a concentration-dependent manner the ability of M. bovis BCG to induce LL-37 mRNA expression. Finally, inhibition of the MEK1/2 and p38 mitogen-activated protein kinase (MAPK) signaling pathways reduced M. bovis BCG-mediated LL-37 mRNA expression, a reduction that correlated with the observed high level of downregulation of LL-37 protein induction. Thus, these results indicate that the MEK1/2 and p38 MAPK signaling pathways play a critical role in the regulation of inducible LL-37 gene expression in A549 cells infected with M. bovis BCG.


[The effect of resuscitation on the results obtained in identification of enterobacteriaceae by the API system (author's transl)].
During studies on the reliability of the API system, the question arose whether the small proportion of adequate results in cultures stored at 4 degrees C could be attributable to this method of storage. Therefore, the reactions of those cultures which had caused most problems were studied following periods of storage ofone day, six weeks and nine weeks at 4 degrees C, and in the freeze-dried state. Storage in particular conditions was found to have a definite effect on the reliability of the API system so that a period of resuscitation of forty-eight hours in a bouillon at 37 degrees C is required to restore the cultures to normal.


An fMRI study of the interface between affective and cognitive neural circuitry in pediatric bipolar disorder.
The pathophysiology of pediatric bipolar disorder (PBD) impacts both affective and cognitive brain systems. Understanding disturbances in the neural circuits subserving these abilities is critical for characterizing developmental aberrations associated with the disorder and developing improved treatments. Our objective is to use functional neuroimaging with pediatric bipolar disorder patients employing a task that probes the functional integrity of attentional control and affect processing. Ten euthymic unmedicated pediatric bipolar patients and healthy controls matched for age, sex, race, socioeconomic status, and IQ were scanned using functional magnetic resonance imaging. In a pediatric color word matching paradigm, subjects were asked to match the color of a word with one of two colored circles below. Words had a positive, negative or neutral emotional valence, and were presented in 30-s blocks. In the negative affect condition, relative to the neutral condition, patients with bipolar disorder demonstrated greater activation of bilateral pregenual anterior cingulate cortex and left amygdala, and less activation in right rostral ventrolateral prefrontal cortex (PFC) and dorsolateral PFC at the junction of the middle frontal and inferior frontal gyri. In the positive affect condition, there was no reduced activation of PFC or increased amygdala activation. The pattern of reduced activation of ventrolateral PFC and greater amygdala activation in bipolar children in response to negative stimuli suggests both disinhibition of emotional reactivity in the limbic system and reduced function in PFC systems that regulate those responses. Higher cortical cognitive areas such as the dorsolateral PFC may also be adversely affected by exaggerated emotional responsivity to negative emotions. This pattern of functional alteration in affective and cognitive circuitry may contribute to the reduced capacity for affect regulation and behavioral self-control in pediatric bipolar disorder.


Understanding Retroperitoneal Anatomy for Lateral Approach Spine Surgery.
Lateral approach spine surgery provides effective interbody stabilization, and correction and indirect neural decompression with minimal-incision and less invasive surgery compared with conventional open anterior lumbar fusion. It may also avoid the trauma to paraspinal muscles or facet joints found with transforaminal lumbar interbody fusion and posterior lumbar interbody fusion. However, because lateral approach surgery is fundamentally retroperitoneal approach surgery, it carries potential risk to intra- and retroperitoneal structures, as seen in a conventional open anterior approach. There is an innovative lateral approach technique that reveals different anatomical views; however, it requires reconsideration of the traditional surgical anatomy in more detail than a traditional open anterior approach. The retroperitoneum is the compartmentalized space bounded anteriorly by the posterior parietal peritoneum and posteriorly by the transversalis fascia. The retroperitoneum is divided into three compartments by fascial planes: anterior and posterior pararenal spaces and the perirenal space. Lateral approach surgery requires mobilization of the peritoneum and its content and accurate exposure to the posterior pararenal space. The posterior pararenal space is confined anteriorly by the posterior renal fascia, anteromedially by the lateroconal fascia, and posteriorly by the transversalis fascia. The posterior renal fascia, the lateroconal fascia or the peritoneum should be detached from the transversalis fascia and the psoas fascia to allow exposure to the posterior pararenal space. The posterior pararenal space, however, does not allow a clear view and identification of these fasciae as this relationship is variable and the medial extent of the posterior pararenal space varies among patients. Correct anatomical recognition of the retroperitoneum is essential to success in lateral approach surgery. Spine surgeons must be aware that the retroperitoneal membrane and fascia is multilayered and more complex than is commonly understood. Preoperative abdominal images would facilitate more efficient surgical considerations of retroperitoneal membrane and fascia in lateral approach surgery.


Conserved arginines on the rim of Hfq catalyze base pair formation and exchange.
The Sm-like protein Hfq is required for gene regulation by small RNAs (sRNAs) in bacteria and facilitates base pairing between sRNAs and their mRNA targets. The proximal and distal faces of the Hfq hexamer specifically bind sRNA and mRNA targets, but they do not explain how Hfq accelerates the formation and exchange of RNA base pairs. Here, we show that conserved arginines on the outer rim of the hexamer that are known to interact with sRNA bodies are required for Hfq's chaperone activity. Mutations in the arginine patch lower the ability of Hfq to act in sRNA regulation of rpoS translation and eliminate annealing of natural sRNAs or unstructured oligonucleotides, without preventing binding to either the proximal or distal face. Stopped-flow FRET and fluorescence anisotropy show that complementary RNAs transiently form a ternary complex with Hfq, but the RNAs are not released as a double helix in the absence of rim arginines. RNAs bound to either face of Hfq quench the fluorescence of a tryptophan adjacent to the arginine patch, demonstrating that the rim can simultaneously engage two RNA strands. We propose that the arginine patch overcomes entropic and electrostatic barriers to helix nucleation and constitutes the active site for Hfq's chaperone function.


[Ophthalmo-rhinosurgical decompression operation in malignant endocrine exophthalmus].
The thyroid metabolism has to be treated first. If this is not followed by regression of the exophthalmos, treatment with prednisone over a period of several weeks is indicated. If this does not result in permanent improvement, surgical decompression of the orbit is indicated, especially if the exposition of the cornea or compression of the optic nerve represent an imminent danger. In such cases surgical decompression is preferable to multiple series of prednisone or radiation treatment (Wright 1981, Trokel 1982). Both the Krönlein-Berke operation for resection of the lateral orbital wall as well as neurosurgical resection of the orbital roof have proved to be excellent for removal of correspondingly located orbital tumors; effective decompression of the orbit in malignant endocrine exophthalmos, however, cannot be achieved this way, as the temporal muscle and the brain, respectively, prevent protrusion of the swollen orbital tissues through these apertures. Resection of the orbital floor is somewhat more effective. The mucosa of the maxillary sinus can easily be moved by the protruding orbital tissues, resulting in a reduction in the size of the maxillary sinus. This decompression, however, is effective only in the anterior two-thirds of the orbit, while the optic nerve must be relieved from the pressure of the swollen muscles in particular in the posterior one-third of the orbit. The only effective way is to remove the medical orbital wall up to the tip of the orbit. Surgical techniques for this purpose have already been recommended, but appeared unsatisfactory from a rhinological point of view.(ABSTRACT TRUNCATED AT 250 WORDS)


Report of Workshop on Euthanasia for Zebrafish-A Matter of Welfare and Science.
The increasing importance of zebrafish as a biomedical model organism is reflected by the steadily growing number of publications and laboratories working with this species. Regulatory recommendations for euthanasia as issued in Directive 2010/63/EU are, however, based on experience with fish species used for food production and do not take the small size and specific physiology of zebrafish into account. Consequently, the currently recommended methods of euthanasia in the Directive 2010/63/EU are either not applicable or may interfere with research goals. An international workshop was held in Karlsruhe, Germany, March 9, 2017, to discuss and propose alternative methods for euthanasia of zebrafish. The aim was to identify methods that adequately address the physiology of zebrafish and its use as a biomedical research model, follow the principles of the 3Rs (Replacement, Reduction, and Refinement) in animal experimentation and consider animal welfare during anesthesia and euthanasia. The results of the workshop are summarized here in the form of a white paper.


Targeting of nuclear factor kappaB Pathways by dehydroxymethylepoxyquinomicin, a novel inhibitor of breast carcinomas: antitumor and antiangiogenic potential in vivo.
We previously designed and synthesized the new nuclear factor kappaB (NF-kappaB) inhibitor dehydroxymethylepoxyquinomicin (DHMEQ) derived from the structure of the antibiotic epoxyquinomicin C. We looked into the effect of DHMEQ on cellular phenotypes and tumor growth in mice injected with human breast carcinoma cell line MDA-MB-231 or MCF-7. In estrogen-independent breast adenocarcinoma cell line MDA-MB-231, NF-kappaB is constitutively activated. The addition of DHMEQ (10 microg/mL) completely inhibited the activated NF-kappaB for at least 8 hours. On the other hand, NF-kappaB is not activated in estrogen-dependent MCF-7 cells. In this cell line, DHMEQ completely inhibited the tumor necrosis factor-alpha-induced activation of NF-kappaB. DHMEQ did not inhibit the degradation of IkappaB but inhibited the nuclear translocation of NF-kappaB by both p65/p50 and RelB/p52 pathways. MDA-MB-231 cells secrete interleukin (IL)-6 and IL-8 without stimulation, and DHMEQ decreased the secretion levels of both cytokines. When MDA-MB-231 or MCF-7 cells were stimulated by tumor necrosis factor-alpha, the inhibitory effects of DHMEQ were still maintained. I.p. administration of DHMEQ (thrice a week) significantly inhibited the tumor growth of MDA-MB-231 (12 mg/kg) or MCF-7 (4 mg/kg) in severe combined immunodeficiency mice. No toxicity was observed during the experiment, including the loss of body weight. An immunohistological study on resected MCF-7 tumors showed that DHMEQ inhibited angiogenesis and promoted apoptosis. Furthermore, in Adriamycin-resistant MCF-7 cells highly expressing multidrug resistance gene-1, DHMEQ also exhibited the above capability, including down-regulation of IL-8. Thus, DHMEQ might be a potent drug for the treatment of various breast carcinomas by inhibiting the NF-kappaB activity.


Mutation analysis of the carbohydrate sulfotransferase gene in Vietnamese with macular corneal dystrophy.
Mutations in a new carbohydrate sulfotransferase gene (CHST6) encoding corneal N-acetylglucosamine-6-sulfotransferase (C-GlcNac-6-ST) have been identified as the cause of macular corneal dystrophy (MCD) in various ethnicities. This study was conducted to examine the CHST6 gene in Vietnamese with MCD. Nineteen unrelated families, including 35 patients and 38 unaffected relatives were examined clinically. Blood samples were collected. Fifty normal Vietnamese individuals served as control subjects. Genomic DNA was extracted from leukocytes. Analysis of the CHST6 gene was performed with polymerase chain reaction and direct sequencing. Corneal buttons were studied histopathologically. A slit lamp examination revealed clinical features of MCD with gray-white opacities and stromal haze between. On histopathology, corneal sections showed positive staining with colloidal iron. Sequencing of the CHST6 gene revealed six homozygous and three compound heterozygous mutations. The homozygous mutations, including L59P, V66L, R211Q, W232X, Y268C, and 1067-1068ins(GGCCGTG) were detected, respectively, in two, one, eight, one, one, and two families. Compound heterozygous mutations R211Q/Q82X, S51L/Y268C, and Y268C/1067-1068ins(GGCCGTG) were identified, each in one family. A single heterozygous change at codon 76 (GTG-->ATG) was detected in family L, resulting in a valine-to-methionine substitution (V76M). None of these mutations was detected in the control group. Mutations identified in the CHST6 gene cosegregated with the disease phenotype in all but one family studied and thus caused MCD. Among these, the R211Q detected in 9 of 19 families may be the most common mutation in Vietnamese. These data also indicate that significant allelic heterogeneity exists for MCD.


A Canadian programme for early intervention in non-affective psychotic disorders.
To provide a brief overview of the development of clinical services and research for early intervention in psychotic disorders in Canada; to describe components of a comprehensive clinical/research programme for nonaffective psychotic disorders; and to present a summary of results of clinical and social outcomes achieved. This is a descriptive paper providing some details of how clinical services are being developed in Canada and concentrating on one particular early intervention programme, Prevention and Early intervention Programme for Psychoses (PEPP) London, Ontario, which is using a historical control design to evaluate the impact of an assertive approach to community case detection. Components of a phase-specific treatment programme and early case detection are described followed by results based on clinical and psychosocial data collected according to a defined protocol. One year outcome for patients treated in PEPP shows use of low dose, pre-dominantly novel antipsychotics and high (81.5%) retention and remission (75%) rates. Highly significant improvements were also reported for self-rated quality of life and cognition. Duration of untreated psychosis (DUP) and premorbid adjustment were associated with improvement in positive and negative symptoms, respectively. Systemic changes to improve access to the service resulted in substantial increases in number of cases treated and a> 50% decline in DUP. Phase-specific treatment approach and case identification strategies to reduce delay in treatment are likely to substantially improve outcome in nonaffective psychotic disorders compared with what has been reported with traditional approaches.


Potent morphiceptin analogs: structure activity relationships and morphine-like activities.
Morphiceptin (Tyr-Pro-Phe-Pro-NH2), a nonenkephalin peptide, is an opioid agonist highly selective for mu opiate receptors. Chemical modification of Tyr-Pro-Phe-Pro-NH2 was carried out by substituting structurally related amino acids at residues 2, 3 and 4. The morphiceptin analogs synthesized were then examined for receptor binding activities using 125I-labeled FK 33,824 (Tyr-D-Ala-Gly-NMePhe-Met(O)-ol) as the mu-ligand and 125I-labeled D-Ala2,D-Leu5-enkephalin as the delta-ligand, and for inhibitory activities on electrically evoked smooth muscle contraction of mouse vas deferens and isolated myenteric plexus-longitudinal muscle strips of guinea-pig ileum. All of these analogs showed virtually no activity at delta opiate receptor binding sites. Methylation of the nitrogen atom of phenylalanine and the substitution at the C-terminal of L-proline by D-proline produced potent mu-agonists, the prototype analog being Tyr-Pro-NMePhe-D-Pro-NH2 (PL017). The IC50 values of morphiceptin and its analogs for mu receptor binding were correlated to the ED50 values in the guinea-pig ileum assay, suggesting that the ileum effects were mediated by mu receptor interactions. A similar correlation between mu receptor binding activity and the ED50 values in the mouse vas deferens assay suggested that morphiceptin and its analogs also acted on mu receptors in this tissue. This idea is supported by the observation that naloxone has a high pA2 value of 8.71 against PL017 in mouse vas deferens. In in vivo studies, PL017 administered centrally into the rostral portion of the 4th ventricle produced long-lasting, naloxone-reversible analgesia in rats.(ABSTRACT TRUNCATED AT 250 WORDS)