Meeting report - shining light on septins.
Septins are enigmatic proteins; they bind GTP and assemble together like molecular Lego blocks to form intracellular structures of varied shapes such as filaments, rings and gauzes. To shine light on the biological mysteries of septin proteins, leading experts in the field came together for the European Molecular Biology Organization (EMBO) workshop held from 8-11 October 2017 in Berlin. Organized by Helge Ewers (Freie Universität, Berlin, Germany) and Serge Mostowy (Imperial College, London, UK), the workshop convened at the Harnack-Haus, a historic hub of scientific discourse run by the Max Planck Society.


Shock melting of a two-dimensional complex (dusty) plasma.
Shock waves with a linear front were experimentally studied in a monolayer hexagonal Yukawa lattice which was formed from charged monodisperse plastic microspheres and levitated in the sheath of a radio-frequency discharge. It was found that the shock can cause phase transitions from a crystalline to gaslike and liquidlike states. Melting occurred in two stages. First, the lattice was compressed in the direction of shock propagation and second, the particle velocities were randomized a few lattice lines downstream. The Mach number of the shock reached 2.7.


Shared decision-making for biologic treatment of autoimmune disease: influence on adherence, persistence, satisfaction, and health care costs.
Shared decision-making (SDM), a process whereby physicians and patients collaborate to select interventions, is not well understood for biologic treatment of autoimmune conditions. This was a cross-sectional survey of adults initiating treatment for Crohn's disease or ulcerative colitis (inflammatory bowel disease, IBD) or psoriatic arthritis or rheumatoid arthritis (RA/PA). Survey data were linked to administrative claims for 6 months before (baseline) and after (follow-up) therapy initiation. Measures included the Shared Decision Making Questionnaire, Patient Activation Measure (PAM), Morisky Medication Adherence Scale (MMAS), general health, and treatment satisfaction. Claims-based Quan-Charlson comorbidity scores, persistence, medication possession ratio (MPR), and health care costs were examined. Patients were compared by participation (SDM) and nonparticipation (non-SDM) in SDM. Among 453 respondents, 357 were eligible, and 306 patients (204 RA/PA and 102 IBD) were included in all analyses. Overall (n=357), SDM participants (n=120) were more often females (75.0% vs 62.5%, P=0.018), had lower health status (48.0 vs 55.4, P=0.005), and higher Quan-Charlson scores (1.0 vs 0.7, P=0.035) than non-SDM (n=237) participants. Lower MMAS scores (SDM 0.17 vs non-SDM 0.41; P<0.05) indicated greater likelihood of adherence; SDM participants also reported higher satisfaction with medication and had greater activation (PAM: SDM vs non-SDM: 66.9 vs 61.6; P<0.001). Mean MPR did not differ, but persistence was longer among SDM participants (111.2 days vs 102.2 days for non-SDM; P=0.029). Costs did not differ by SDM status overall, or among patients with RA/PA. The patients with IBD, however, experienced lower (P=0.003) total costs ($9,404 for SDM vs $25,071 for non-SDM) during follow-up. This study showed greater likelihood of adherence and satisfaction for patients who engaged in SDM and reduced health care costs among patients with IBD who engaged in SDM. This study provides a basis for defining SDM participation and detecting differences by SDM participation for biologic treatment selection for autoimmune conditions.


Persistence and degradation of coumaphos in model cattle dipping vats.
The stability of coumaphos [O,O-diethyl O-(3-chloro-4-methyl-2-oxo-2H-1-benzopyran-7 yl)phosphorothioatel was studied in model dipping vats under field conditions using 14C-labelled and unlabelled coumaphos, with or without additives. The stability of coumaphos in model vats increased significantly by maintaining a pH of 5 by addition of superphosphate. Copper sulphate amendment did not seem to have any additional effect on stability. Potasan was the major metabolite in addition to chlorferon and 4-methylumbelliferone. Coumaphos concentration was doubled in the sediment of vat treated with copper sulphate as compared to the control vat as a result of emulsion breakdown. Chlorferon was the only metabolite detected in the sediment of the former vat indicating inhibition of the anaerobic degradation.


Family history of myocardial infarction is a risk factor for venous thromboembolism among whites but not among blacks.
In addition to potentially sharing common pathogenesis and clinical manifestations, venous and arterial thromboses might have overlapping risk factors. To evaluate the family history of myocardial infarction (MI) as a risk factor for venous thromboembolism (VTE) among whites and blacks, we analyze data from the Genetic Attributes and Thrombosis Epidemiology (GATE) study. Results indicate that the association between VTE and a family history of MI is statistically significant only among whites (odds ratio [OR] = 1.3; 95% confidence interval [CI] = 1.03-1.8), particularly when they have diabetes mellitus (OR = 3.1; 95% CI = 1.2-8.0). Among blacks, the association between VTE and a family history of MI is not statistically significant (OR = 1.2; 95% CI = 0.89-1.5) either among those with diabetes or those without diabetes. We conclude that a family history of MI is a risk factor for VTE among certain populations stratified by race and comorbid conditions.


Korean ctenostome bryozoans-observations on living colonies, new records, five new species, and an updated checklist.
This paper describes 12 species of ctenostomes from the marine waters of the Korean Peninsula, including five new species-Alcyonidium bullitum n. sp., Alcyonidium pulposum n. sp., Alcyonidium busanensis n. sp., Immergentia cheongpodensis n. sp. and Penetrantia taeanata n. sp., the latter two constituting shell-borers that ramify within dead mollusk shells. Three previously described species are also newly added to the Korean fauna-Amathia acervata Lamouroux, 1824, Amathia medullaris Mawatari, 1972 and Walkeria prorepens Kubanin, 1992. The nomenclature of known species is updated and a revised checklist of all Korean Ctenostomata is included. New biological information is provided based on observation and microphotography of living and preserved colonies, SEM images of dry material and resin casts of ctenostome borings in mollusk shells.


Results of surgical treatment of neonatal necrotizing enterocolitis.
In 6 years, 74 neonates were treated for necrotizing enterocolitis, 40 (54 per cent) of whom underwent operation. The early postoperative mortality rate was 23 per cent and there was one late death. Of the 30 postoperative survivors, two have short bowel syndrome, two have chronic respiratory disease, two are partially sighted, and five (17 per cent) have severe developmental delay. Only 15 (50 per cent) survivors enjoy good health.


Face masks and speaking style affect audio-visual word recognition and memory of native and non-native speech.
Though necessary, protective mask wearing in response to the COVID-19 pandemic presents communication challenges. The present study examines how signal degradation and loss of visual information due to masks affects intelligibility and memory for native and non-native speech. We also test whether clear speech can alleviate perceptual difficulty for masked speech. One native and one non-native speaker of English recorded video clips in conversational speech without a mask and conversational and clear speech with a mask. Native English listeners watched video clips presented in quiet or mixed with competing speech. The results showed that word recognition and recall of speech produced with a mask can be as accurate as without a mask in optimal listening conditions. Masks affected non-native speech processing at easier noise levels than native speech. Clear speech with a mask significantly improved accuracy in all listening conditions. Speaking clearly, reducing noise, and using surgical masks as well as good signal amplification can help compensate for the loss of intelligibility due to background noise, lack of visual cues, physical distancing, or non-native speech. The findings have implications for communication in classrooms and hospitals where listeners interact with teachers and healthcare providers, oftentimes non-native speakers, through their protective barriers.


Large and Long-Range Dynamic Correlations in Supercooled Fluids Revealed via Four-Point Correlation Functions.
Dynamic heterogeneity in supercooled fluids is often monitored using four-point structure factors. We show that a four-point structure factor slightly different from the one commonly used reveals the existence of large and long-range dynamic correlations, which dominate the correlations identified with dynamic heterogeneity. Specifically, we compare two four-point structure factors that differ by the mobility field used in their definition. The mobility field used in our alternative four-point structure factor, which we denote as S4(F)(q, k; t), is the full microscopic self-intermediate scattering function, whereas the mobility field in the structure factor commonly used to examine dynamic heterogeneity, which we denote as S4(cos)(q, k; t), is is the real part of the microscopic self-intermediate scattering function. The susceptibility χ4(F)(t) and the correlation length ξ4(F)(t) associated with S4(F)(q, k; t) are both much larger than the susceptibility χ4(cos)(t) and the correlation length ξ4(cos)(t) associated with S4(cos)(q, k; t). In particular, we find that ξ4(F)(τα) grows proportionally with the relaxation time, ξ4(F)(τα) ∼ τα, while we previously established that ξ4(cos)(τα) has a much weaker dependence on the relaxation time. We argue that the growth of ξ4(F)(τα) is primarily due to the growth of a transient elastic response, while the growth of ξ4(cos)(τα) is due to spatially correlated domains of slow and fast particles, usually referred to as dynamic heterogeneity.


Vascular endothelial growth factor overexpression does not enhance adipose stromal cell-induced protection on muscle damage in critical limb ischemia.
Critical limb ischemia complicates peripheral artery disease leading to tissue damage and amputation. We hypothesized that modifying adipose stromal cells (ASCs) to overexpress human vascular endothelial growth factor 165 (VEGF) would limit ischemic muscle damage to a larger extent than nonmodified ASCs. Rabbits with critical hindlimb ischemia were injected with allogeneic abdominal fat-derived ASCs transfected with plasmid-VEGF165 (ASCs-VEGF; n=10). Additional rabbits received nontransfected ASCs (ASCs; n=10) or vehicle (placebo; n=10). One month later, ASCs-VEGF rabbits exhibited significantly higher density of angiographically visible collaterals and capillaries versus placebo (both P<0.05) but not versus ASCs (both P=NS). Arteriolar density, however, was increased in both ASCs and ASCs-VEGF groups (both P<0.05 versus placebo). ASCs-VEGF and ASCs showed comparable post-treatment improvements in Doppler-assessed peak systolic velocity, blood pressure ratio, and resistance index. Ischemic lesions were found in 40% of the muscle samples in the placebo group, 19% in the ASCs-VEGF group, and 17% in the ASCs groups (both P<0.05 versus placebo, Fisher test). In a rabbit model of critical limb ischemia, intramuscular injection of ASCs genetically modified to overexpress VEGF increase angiographically visible collaterals and capillary density. However, both modified and nonmodified ASCs increase arteriolar density to a similar extent and afford equal protection against ischemia-induced muscle lesions. These results indicate that modifying ASCs to overexpress VEGF does not enhance the protective effect of ASCs, and that arteriolar proliferation plays a pivotal role in limiting the irreversible tissue damage of critical limb ischemia.


[Polycystic disease of the salivary glands: report of an attack of the submaxillary glands].
Bilateral dysgenetic polycystic disease of parotid glands is an extremely rare pathologic condition in which salivary parenchyma is partially replaced by multiple epithelial-lined cysts arising from intercalated ducts. Review of the fourteen cases published in literature shows that it affects nearly exclusively women with an history of asymptomatic progressive enlargement of almost always both parotid glands. We report here the first case, to our knowledge, of a polycystic disease involving both submandibular salivary gland in a man.


[Leprosy reactions].
In leprosy, the causative bacteria, Mycobacterium leprae, will not threaten the lives of the hosts directly because they proliferate only slowly in the Schwann cells of the peripheral nerves. It is the "reactions" which give the patients irreversible morbidity through the inflammatory damages to the peripheral nerves. Physicians should be aware of the possibility of the state of the "reaction" when they examine leprosy patients. They also should be aware of the possibility of leprosy and the state of the "reaction" when they examine patients with cutaneous lesions and/or peripheral nerve disturbances, because it may be the first presenting symptom of the disease. In this review, recent advances on the issue about the reactions are discussed including pathogenesis, immunology, clinical features, pathology, treatment and prevention.


Tissue hemolysins as lysin-inhibitor complexes.
1. Lytic substances are enzymatically produced at 37 degrees C. from tissue slices or homogenates (mouse liver, kidney, etc.) and appear in the medium in which the tissue fragments are suspended. Their concentration increases with the time during which the tissue is kept at 37 degrees C. (preincubation), and is accompanied by pH changes, so that the lytic activity as finally measured is a function of both the time of preincubation and of the pH. The optimum pH for lysin production is above 7.0, but the lysins, once produced, hemolyze red cells more rapidly at low pH's than at high ones. The enzyme system which produces the lysins is inactivated by heating to 100 degrees C. for 5 minutes. Sodium iodoacetate and fluoride interfere with lysin production principally by reducing the concomitant pH shift; KCN accelerates the production of lytic material in mouse liver homogenates. 2. Comparison of the lytic activity of the supernatant fluid of a preincubated homogenate with the much greater lytic activity of the substances which can be extracted from the same supernatant fluid by alcohol and ether points to these extractable substances existing in the supernatant fluid as lysin-inhibitor complexes of relatively low lytic activity. These complexes are formed enzymatically during preincubation from non-lytic complexes in the tissue. The latter may be lipoproteins, and the highly lytic ether-extractable substances may be fatty acids or their soaps. 3. The diffusibility of the lysin-inhibitor complexes is small. 4. Lytic substances which are ether-insoluble can be extracted with alcohol from tissues as well as from serum. These "lysolecithin-like" substances exist in the supernatant fluids of homogenates as lysin-inhibitor complexes. 5. Lysis of mouse red cells by substances contained in mouse tissue (liver and kidney) is often accompanied by the formation of methemoglobin and choleglobin. Mouse red cells containing choleglobin are abnormally fragile both osmotically and mechanically, and it is possible that a process involving the production of choleglobin, accompanied or followed by globin denaturation, is one which contributes towards the hemolysis which occurs in systems containing tissue slices or homogenates.


Identifying Biopsychosocial Characteristics of Emergency Youth Shelter Residents With Psychiatric Diagnoses.
Background In the United States, homelessness is an issue that may affect a significant portion of the adolescent population. There is no consensus on the extent to which this population has been impacted by poor mental health and lack of resources. This study aimed to characterize trends among those who struggle with housing insecurity and mental illness to provide a clearer picture of mental health needs among this population. Methods Data from 641 adolescents who presented to a local adolescent homeless shelter between 2015 and 2021 were utilized to determine if there were significant associations between specific mental illness diagnoses and biopsychosocial characteristics. A chi-square test of independence was performed on demographic and psychosocial variables for categories with a frequency greater than five. For continuous variables, an unpaired t-test was utilized to assess significance (p<0.05). Results Among the study population, 61.3% (369) had at least one psychiatric diagnosis, which is higher than even the most conservative estimates of mental illness among the general public. Having one or more psychiatric diagnoses was significantly associated with suicide attempts, documented aggressive behavior, and tobacco use. Contrary to our initial hypothesis, there were no significant correlations between psychiatric diagnoses and demographic characteristics or drug use other than tobacco. Conclusions Our findings indicate that though the particular reasons for homelessness among adolescents may vary, the prevalence of mental illness among these young individuals was roughly uniformly distributed and vastly above normal levels. Future research must focus on developing interventions to mitigate the effects of mental illness among homeless adolescents, as they are at a vulnerable point in their formative years.


[Comparison of seed oil physicochemical characteristics among three cultivars of Jatropha curcas L].
Taking the cultivars Nanyou 1, 2, and 3 of barbadosnut (Jatropha curcas L. ) with different genotypes that can grow and seed normally at the inshore land in Hainan as test materials, the characters of their seeds and the physicochemical characteristics of their seed oils were analyzed and compared. No significant differences were observed in the seed length, width, thickness, and surface area among the cultivars, but Nanyou 2 had greater 1000 seed mass and lower unsound kernel percentage than Nanyou 1 and Nanyou 3, suggesting that the seed satiation of Nanyou 2 was good and the fecundity was excellent. The kernel oil content of Nanyou 3 was significantly higher than that of Nanyou 1 and Nanyou 2, and there was no significant difference between Nanyou 1 and Nanyou 2. The seed oil peroxide value, refractive index, and saponification value of the three cultivars had no significant differences, but the acid value for Nanyou 2 was much lower than that for Nanyou 1 and Nanyou 3. The seed oil iodine value of the three cultivars was all below 100, and was significantly lower for Nanyou 2 than for Nanyou 1 and Nanyou 3. The fatty acids in the three cultivars seed oils were mainly oleic acid, palmitic acid, linoleic acid, stearic acid, and margaric acid, and dominated by unsaturated fatty acids. The contents of saturated fatty acids in Nanyou 2 seed oil were relatively higher than those in Nanyou 1 and Nanyou 3 seed oils, indicating that comparing with Nanyou 1, cultivars Nanyou 2 and Nanyou 3 had relatively good potential for application.


Skin manifestations and biophysical changes following weight reduction induced by bariatric surgery: A 2-year prospective study.
Skin manifestations and biophysical changes are observed in patients with morbid obesity. However, reports of changes after significant weight loss, particularly through post-bariatric surgery (BaS), are limited. The aim of this 2-year prospective study was to evaluate the prevalence of skin signs and their changes in patients with morbid obesity who underwent BaS. Thirty-one patients were recruited for the study, with a mean age of 38.35 (SD 10.61) years and a male preponderance (male = 19 [61.29%], female = 12 [38.71%]). Patients were scheduled for multiple visits at months 0, 3, 6, 12, 18, and 24 post-BaS. Each subject had a thorough skin examination, biophysical measurements, and laboratory tests at each visit. Striae, acanthosis nigricans (AN), and plantar hyperkeratosis were the most common skin findings (n = 30 [96.77%], 29 [93.55%], 29 [93.55%], respectively). BaS provided improvements in many skin manifestations, namely striae, AN, acrochordons, plantar hyperkeratosis, hirsutism, lymphedema, pruritus, acne, finger pebbles, and chronic venous insufficiency with varied cumulative rates of improvements. However, acute telogen effluvium was observed in 17 (54.84%) patients. Regarding skin biophysical properties, transepidermal water loss, skin hydration, and pH did not change, while sebum production on the face significantly decreased at months 3 and 6, and elasticity decreased at months 6 and 24. In conclusion, weight reduction by BaS provided improvements in various skin signs, although telogen effluvium was a common sequelae.


Availability and readiness of the health facilities to provide HIV counseling and testing and prevention of mother-to-child transmission services in Burkina Faso: a trend analysis from 2012 to 2018.
Provider-Initiated HIV Testing and Counseling (PITC) and Prevention of Mother-To-Child Transmission (PMTCT) are key services for achieving the goal of complete elimination of HIV. However, there is limited evidence on the ability of health facilities to provide these services in Burkina Faso. Therefore, we aimed to assess the trends and disparities in the availability and readiness of health facilities to provide PITC and PMTCT services in Burkina Faso between 2012 and 2018. We performed a secondary analysis of facility-level data from the World Health Organization's Service Availability and Readiness Assessment (SARA) surveys conducted in 2012, 2014, 2016, and 2018 in Burkina Faso. The availability and readiness of health facilities were assessed using SARA's manual, and linear regressions were used to examine trends. Between 2012 and 2018, the mean proportion of health facilities providing PITC services increased, but not significantly, from 82.9% to 83.4% (p = 0.11), with the mean readiness index significantly decreasing from 71.5% to 65.4% (p < 0.001). This decrease concerned the staff and guidelines (73.8% to 50.5%; p < 0.001), equipment (79.0% to 77.4%; p < 0.001), and medicines and commodities (54.2% to 45.2%; p < 0.001) domains. Regarding the PMTCT services, the mean proportion of health facilities globally providing the service significantly decreased from 83.7% in 2012 to 67.7% (p = 0.030) in 2018, and the mean readiness significantly decreased from 53.2% in 2012 to 50.9% in 2018 (p = 0.004). This decreasing trend was related to the staff and training (80.3% to 57.6%; p < 0.001) and medicines and commodities (9.2% to 6.5%; p < 0.001) domains. The global significant negative trend of readiness was mainly observed at the primary level of healthcare (52.7% to 49.4%; p = 0.030). Four regions experienced a significant decrease in the readiness index of health facilities to provide PMTCT services: Cascades, Centre, Centre-Sud, and Sud-Ouest, while Haut-Bassins and Nord regions showed increasing trends. Availability and readiness of health facilities to provide PITC and PMTCT remain suboptimal in Burkina Faso. Actions to strengthen the skills of professionals and enhance the availability of medicines and commodities while focusing more on health regions with significant decreasing trends are urgently needed to improve the quality of services for HIV.


Effectiveness of and obstacles to antibiotic streamlining to amoxicillin monotherapy in bacteremic pneumococcal pneumonia.
Antibiotic streamlining is pivotal to reduce the emergence of resistant bacteria. However, whether streamlining is frequently performed and safe in difficult situations, such as bacteremic pneumococcal pneumonia (BPP), has still to be assessed. All adult patients admitted to Dijon Hospital (France) from 2005 to 2013 who had BPP without complications, and were alive on the third day were enrolled. Clinical, biological, radiological, microbiological and therapeutic data were recorded. A first analysis was conducted to assess factors associated with being on amoxicillin on the third day. A second analysis, adjusting for a propensity score, was performed to determine whether 30-day mortality was associated with streamlining to amoxicillin monotherapy. Of the 196 patients hospitalized for BPP, 161 were still alive on the third day and were included in the study. Treatment was streamlined to amoxicillin in 60 patients (37%). Factors associated with not streamlining were severe pneumonia (OR 3.11, 95%CI [1.23-7.87]) and a first-line antibiotic combination (OR 3.08, 95%CI [1.34-7.09]). By contrast, starting with amoxicillin monotherapy correlated inversely with the risk of subsequent treatment with antibiotics other than amoxicillin (OR 0.06, 95%CI [0.01-0.30]). The Cox model adjusted for the propensity-score analysis showed that streamlining to amoxicillin during BPP was not significantly associated with a higher risk of 30-day mortality (HR 0.38, 95%CI [0.08-1.87]). Streamlining to amoxicillin is insufficiently implemented during BPP. This strategy is safe and potentially associated with ecological and economic benefits; therefore, it should be further encouraged, particularly when antibiotic combinations are started for severe pneumonia.


Student nurses' experiences during clinical practice in the Limpopo Province.
A qualitative, exploratory, descriptive and contextual study was conducted to explore student nurses' experiences during clinical practice at a nursing college in the Limpopo Province. Purposive sampling was used and phenomenological interviews were held with eleven (11) student nurses who were in their final year of the four year basic nursing programme. The interviews were analysed by using Tesch's method of data analysis for qualitative research. The findings indicate that there are aspects which impact negatively on student nurses' clinical learning experiences, such as lack of teaching and learning support, lack of opportunities for learning, poor theory-practice integration, and poor interpersonal relationships between the students, college tutors and ward staff. Recommendations to enhance the clinical learning experiences of student nurses were outlined.


Protective effect of 3-hydroxybutyrate against endoplasmic reticulum stress-associated vascular endothelial cell damage induced by low glucose exposure.
The aim of this study was to elucidate the mechanism by which severe hypoglycemia accelerates vascular complications. Furthermore, we assessed the possible protective effect of ketone bodies against the endothelial cell damage caused by glucose deficiency. Human umbilical vein endothelial cells (HUVECs) were cultured at a glucose level of either 0.56 or 5.6 mmol/L with or without 3-hydroxybutyrate (3-HB) supplementation. Cell viability was assessed with a CCK-8 assay and a lactate dehydrogenase (LDH) release assay. The activity of caspases was measured using fluorogenic substrates. The expression of genes associated with endothelial cell function and endoplasmic reticulum (ER) stress was evaluated by real-time quantitative PCR. Protein levels of ER stress-related molecules were assessed by Western blotting. Culture of HUVECs in low-glucose medium for 24 or 48 h resulted in reduction of cell viability accompanied by activation of caspase-3/7 and caspase-8. The addition of a pan caspase inhibitor attenuated the cell death. After incubation in the low-glucose medium, we found reduced mRNA and protein levels of endothelial nitric oxide synthase. ER stress responses mediated by phosphorylation of protein kinase RNA-like ER kinase (PERK) and cleavage of activating transcription factor 6 (ATF6) were augmented, but X-box binding protein 1 (Xbp1) splicing was reduced. Most of these responses to glucose deficiency were significantly attenuated by supplementation with 3-HB. These observations showed that exposure to low glucose induces ER stress, caspase activation, endothelial cell dysfunction and cell death. The beneficial effects of 3-HB shown in this study suggest that hypoketonemic severe hypoglycemia induced by insulin injections or insulin secretagogue administration may be more harmful than hyperketonemic severe hypoglycemia.


A multicenter survey on distal pancreatectomy in Italy: results of minimally invasive technique and variability of perioperative pathways.
No data are available about distal pancreatectomy (DP) in Italy, regarding variability of care among centers, and outcome of minimally invasive distal pancreatectomy (MIDP) in a multicenter setting. Hence, a survey was conducted among 20 institutions experienced in pancreatic surgery by the Italian Association for Study of Pancreas. Centers were asked to fill in two questionnaires about (1) general approach and perioperative protocols for DP and MIDP; (2) detailed operative results of MIDP in the period 2010-2011. Results of questionnaire 1: a great variability in perioperative approach was observed: octreotide was used in 50 % of centers, enzyme supplementation in 35 %, postoperative gastric suction in 80 % and oral liquids on day 1 in 55 %. All hospitals used at least one drain and its removal ranged between days 3-5 (in absence of fistula). Differences in type and timing of post-splenectomy vaccinations were recorded. As regards MIDP, 17/20 centers performed laparoscopic or robotic DP. MIDP rate on overall DP varied among centers (range 0-82 %) and it doubled from 2007 (14 %) to 2011 (28 %). Results of questionnaire 2: in the period 2010-2011, 171 MIDP were performed (140 laparoscopic, 31 robotic). Overall conversion rate was 17 %, mean operative time was 230 min and blood loss 285 ml. Mortality was nil and morbidity was 62 %, with 4 % relaparotomies. Pancreatic fistula occurred in 49 % (grade A 35 %, B 14 %). Mean postoperative stay was 9.8 days with 10 % readmission rate. The results indicated a great variability in DP management among Italian centers. Most centers performed MIDP, but MIDP rate on overall DP largely varied among centers. As regards MIDP conversion rate and hospital stay were found to be worse than those in single-institution series.


Antigen restricted hybridization between antigen primed macrophage and thymic RNA.
Antigen primed macrophage and thymic RNAs hybridize in an antigen restricted fashion. Thymic RNA complementary to the antigen-stimulated macrophage RNA can be obtained from mice inoculated with antigen three days earlier. The ratio of thymic and macrophage RNAs for the optimal hybridization was 50 : 1. The optimal incubation time for hybridization was 32 hours at 60 degrees C. Hybrid RNAs were insensitive to various kinds of RNase. The 31p Fourier transform nuclear magnetic resonance spectra identified hybrid RNA molecules form single stranded RNS's.


Novel Cortical Thickness Pattern for Accurate Detection of Alzheimer's Disease.
Brain network occupies an important position in representing abnormalities in Alzheimer's disease (AD) and mild cognitive impairment (MCI). Currently, most studies only focused on morphological features of regions of interest without exploring the interregional alterations. In order to investigate the potential discriminative power of a morphological network in AD diagnosis and to provide supportive evidence on the feasibility of an individual structural network study, we propose a novel approach of extracting the correlative features from magnetic resonance imaging, which consists of a two-step approach for constructing an individual thickness network with low computational complexity. Firstly, multi-distance combination is utilized for accurate evaluation of between-region dissimilarity; and then the dissimilarity is transformed to connectivity via calculation of correlation function. An evaluation of the proposed approach has been conducted with 189 normal controls, 198 MCI subjects, and 163 AD patients using machine learning techniques. Results show that the observed correlative feature suggests significant promotion in classification performance compared with cortical thickness, with accuracy of 89.88% and area of 0.9588 under receiver operating characteristic curve. We further improved the performance by integrating both thickness and apolipoprotein E ɛ4 allele information with correlative features. New achieved accuracies are 92.11% and 79.37% in separating AD from normal controls and AD converters from non-converters, respectively. Differences between using diverse distance measurements and various correlation transformation functions are also discussed to explore an optimal way for network establishment.


Altered nucleotide sugar metabolism in Streptococcus thermophilus interferes with nitrogen metabolism.
Exopolysaccharide (EPS)-producing Streptococcus thermophilus strains have attracted interest recently, since the EPSs act as natural viscosifiers and texture enhancers of fermented foods. We have previously reported that the low level of EPS production by S. thermophilus LY03 could be improved by altering the activities of enzymes in the central carbon metabolism involved in the nucleotide sugar metabolism. In this study, we observed a reduced growth in milk for the strains with increased UDP-glucose pyrophosphorylase (GalU) activity together with either enhanced phosphoglucomutase activity, and/or enhanced activity of the Leloir enzymes. Rapid growth of these mutants in milk could be restored by the addition of four specific amino acids, i.e. Glu, His, Met, and Val. This amino acid requirement was confirmed in a defined medium. Furthermore, the 31P NMR spectra showed higher levels of the GalU reactants pyrophosphate (PPi) and UDP-glucose in the engineered strain, TMB 6013, compared to the parent strain, LY03. These products plus Glu and the GalU reactant UTP are known to be involved in the nitrogen regulatory system in many bacteria. Thus, these results suggest that the reaction catalyzed by GalU is connected to the nitrogen demand of these engineered strains.


An Alport syndrome boy with Van Wyk-Grumbach syndrome induced by prolonged untreated congenital hypothyroidism.
Alport syndrome (AS) is a rare genetic disorder that causes progressive nephritis and is more common among males. Studies have reported an association between thyroid antibodies and hypothyroidism in patients with AS, but the relevance of this relationship is under debate. Prolonged untreated hypothyroidism induces short stature, abnormal pubertal development, and various other symptoms. However, children with long-standing hypothyroidism rarely present with signs of precocious puberty, or Van Wyk-Grumbach syndrome (VWGS). We report the case of a boy, 8 years and 4 months old, who had VWGS caused by prolonged untreated congenital hypothyroidism and AS. The boy had repeated gross hematuria and proteinuria and was diagnosed with AS by renal biopsy and genetic testing. He had normal renal function but severe growth retardation and hypothyroidism. Obesity, bone age delay, hyperlipidemia, and abnormal increased testicle size were also present due to prolonged untreated hypothyroidism. His thyroid antibody titer elevation was unclear, although ultrasonography and thyroid scanning showed a decrease in thyroid volume. We diagnosed the patient with congenital hypothyroidism caused by thyroid dysgenesis. VWGS was diagnosed due to hypothyroidism, delayed bone age, and pseudoprecocious puberty. To the best of our knowledge, this is the first report of a prepubertal Korean boy with prolonged untreated congenital hypothyroidism complicated by VWGS in AS.


Auditory musical hallucinations associated with extended-release pramipexole in an elderly patient with Parkinson's disease.
Auditory musical hallucinations (AMHs) are rare complex auditory hallucinations in Parkinson's disease (PD) that have been limited previously. The characteristics of AMHs in PD remain uncertain. We describe a 72-year-old woman with PD who presented with AMHs. The AMHs occurred after immediate-release pramipexole was switched to extended-release pramipexole. The AMHs were a quiet piano or often songs on a loud radio or background music over other sounds. The music was unpleasant, but not objectionable, threatening, or ego-syntonic, and it did not interrupt her daily activities. AMHs in PD were non-threatening, and dopaminergic treatment may predispose patients to AMHs or be a unique possible cause of AMHs. The hallucinations can occur after immediate-release pramipexole was switched to extended-release pramipexole.


Small-Molecule Inhibitor Prevents Insulin Fibrillogenesis and Preserves Activity.
Amyloidosis is a well-known but poorly understood phenomenon caused by the aggregation of proteins, often leading to pathological conditions. For example, the aggregation of insulin poses significant challenges during the preparation of pharmaceutical insulin formulations commonly used to treat diabetic patients. Therefore, it is essential to develop inhibitors of insulin aggregation for potential biomedical applications and for important mechanistic insights into amyloidogenic pathways. Here, we have identified a small molecule M1, which causes a dose-dependent reduction in insulin fibril formation. Biophysical analyses and docking results suggest that M1 likely binds to partially unfolded insulin intermediates. Further, M1-treated insulin had lower cytotoxicity and remained functionally active in regulating cell proliferation in cultured Drosophila wing epithelium. Thus, M1 is of great interest as a novel agent for inhibiting insulin aggregation during biopharmaceutical manufacturing.


Metastatic potential of mouse skin carcinomas produced by different protocols of chemical carcinogenesis.
Squamous cell carcinomas (SCC) of the mouse skin were produced by three different protocols of chemical carcinogenesis, i.e., complete carcinogenesis with 7,12-dimethylbenz(a)anthracene (DMBA) two-stage carcinogenesis with DMBA as initiator, 12-O-tetradecanoyl-phorbol-13-acetate (TPA) as promoter and three stage carcinogenesis with DMBA, TPA and N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) as third-stage agent or progressor. Tumors were sequentially studied at weeks 38-52 of treatment. Although no significant differences in the rate of appearance of gamma-glutamyl transpeptidase (GGT) could be seen, a larger number of SCC produced by complete carcinogenesis protocols were GGT-negative. This coincided with the higher grade of malignancy of these tumors as evaluated by histopathology. In general terms high-grade tumors were seen more frequently in the complete carcinogenesis experiment than in the other two protocols. SCC produced by complete carcinogenesis also exhibited a markedly higher DNA index than the SCC from the other experimental groups. All three protocols were very effective in producing late metastasizing tumors, and no significant differences could be established in the incidence of spontaneous lung metastasis. This shows that, contrary to general knowledge, if adequately observed for more than 40 weeks, SCC of the murine skin is able to metastasize in the lung in approximately 30% of cases. Nevertheless, complete carcinogenesis-induced SCC were usually of higher histological grade, a proportion of these were GGT-negative and produced more multiple or diffuse metastases than the tumors induced by the multistage protocols.


Hyperopia is predominantly axial in nature.
Myopia has been found to be predominantly axial in nature, i.e. myopic eyes have longer than normal axial lengths, with corneal radius variations having only a small influence on the magnitude of the refractive error. In this study we assess whether a similar relationship exists for hyperopia. Biometric data were collected on 57 subjects with either emmetropic or hyperopic refractive errors ranging in magnitude from -0.37 D to +17.25 D. Our main analysis concentrated on subjects with less than +10 D of hyperopia (group 1, n = 53), as subjects with +10 D of hyperopia or more (group 2, n = 4) exhibited marked differences in their biometric characteristics. Analysis of group 1 data revealed a significant relationship (r2 = 0.611, p = 0.0001) between the degree of hyperopia and the measured axial lengths. A weak but statistically significant relationship (r2 = 0.128, p = 0.009) was also found between mean corneal radius measures and mean spherical refractive errors, with the mean corneal radius flattening with increasing hyperopia. In group 2, three of the four subjects exhibited much steeper corneal characteristics than predicted from the group 1 data. Our results suggest that hyperopia, like myopia, is predominantly axial in nature, although the corneal radius also plays a role in determining refractive error magnitude. These results have implications for refractive surgery and visual performance in hyperopic eyes.


Spoilage microflora of broiler carcasses washed with electrolyzed oxidizing or chlorinated water using an inside-outside bird washer.
The effect of acidic, electrolyzed oxidizing (EO) water and chlorinated water on the spoilage microflora of processed broiler carcasses was examined. Carcasses were sprayed for 5 s at 80 psi with tap, chlorinated, or EO water in an inside-outside bird washer. Treated carcasses were then stored at 4 degrees C for 0, 3, 7, or 14 d, and the microbial flora of the carcasses was sampled using the whole-carcass rinse procedure. Populations of psychrotrophic bacteria and yeasts in the carcass rinsates were enumerated. Results indicated that immediately after spraying the carcasses, significantly fewer psychrotrophic bacteria were recovered from carcasses sprayed with chlorinated or EO water than from carcasses sprayed with tap water. Furthermore, significantly fewer yeasts were recovered from carcasses sprayed with EO water than from carcasses sprayed with tap or chlorinated water. The population of psychrotrophic bacteria and yeasts increased on all carcasses during refrigerated storage. However, after 14 d of storage, significantly fewer psychrotrophic bacteria and yeasts were recovered from carcasses sprayed with EO water than from carcasses sprayed with tap or chlorinated water, and significantly fewer microorganisms were recovered from carcasses sprayed with chlorinated water than from carcasses sprayed with tap water. Pseudomonas spp. and Candida spp. were the primary microbial isolates recovered from the broiler carcasses. Findings from the present study indicate that EO water can effectively be used in inside-outside bird washers to decrease the population of spoilage bacteria and yeasts on processed broiler carcasses.


Fluid for thought: availability of drinks in primary and secondary schools in Cardiff, UK.
While adequate hydration is undoubtedly essential for health, well-being, performance and learning, it is important to recognize that drinks may also have significant detrimental effects on both general and dental health. Since, on weekdays, at least half of a child's recommended daily fluid intake must be imbibed at school, this study sought to examine the provision of drinks in that setting. A self-administered postal questionnaire study was sent to the head teachers of all 107 state primary and all 20 state secondary schools in Cardiff, South Wales, UK. A response rate of 81.9% was achieved. Children had access to fresh drinking water from at least one drinking fountain in 69 (66.4%) of the 104 schools who responded to the questionnaire. Milk was available in 69 (80.2%) of the 86 responding primary schools. Sixteen (88.9%) of the 18 responding secondary schools had drink vending machines selling a variety of drinks. While primary schools appear largely to restrict the availability of drinks to those conducive to the maintenance of good general and dental health, secondary schools appear to foster the use of vending machines. Guidelines should be developed on the use and content of vending machines in schools in order to both meet school objectives and promote healthy choices.


Genesis and genetics of retinoblastoma.
1. The increase in frequency of retinoblastoma reported during the last decades [FRANCOIS, 1968] is probably not due to an increase of the mutation rate but to a lowered mortality rate and also to improvement in methods of investigation and diagnosis. (2) In 95% of all affected families retinoblastoma occurs just once, the familial occurrence being +/- 5%. (3) The hereditary retinoblastomas (whether sporadic or familial) represent about 40% of all cases. (4) A new dominant mutation is judged to be responsible for 100% of the bilateral sporadic cases and for 10-20% of the unilateral sporadic cases. (5) Genetic susceptibility to cancer can be chromosomal, mendelian dominant, mendelian recessive, or polygenic. Retinoblastoma belongs to the first as well as to the second category. The fact that the chromosomal aberration of the D deletion syndrome antedates the appearance of the tumour suggests that chromosomal change may be the primary cause of the tumour formation. (6) Although several problems remain an unsolved question, attention must be given to KNUDSON'S working hypothesis [KNUDSON et al., 1973], according to which all childhood tumours fit at least a two-mutational aetiology, involving a two-step process. It remains an open question whether both events occur at different genetic loci or at alleles of the same locus. (7) The main difficulty in genetic counseling is the lack of means for identifying which sporadic retinoblastoma cases are due to new germinal mutations.


Imaging pulse wave propagation in human retinal vessels using full-field swept-source optical coherence tomography.
We demonstrate a new noninvasive method to assess biomechanical properties of the retinal vascular system. Phase-sensitive full-field swept-source optical coherence tomography (PhS-FF-SS-OCT) is used to investigate retinal vascular dynamics at unprecedented temporal resolution. The motion of retinal tissue that is induced by expansion of the vessels therein is measured with an accuracy of about 10 nm. The pulse shapes of arterial and venous pulsations, their temporal delays, as well as the frequency-dependent pulse propagation through the capillary bed, are determined. For the first time, imaging speed and motion sensitivity are sufficient for a direct measurement of pulse waves propagating with more than 600 mm/s in retinal vessels of a healthy young subject.


Indium-111 labeled leukocyte imaging following hepatic artery embolization.
The use of In-111 labeled leukocytes for abscess localization is becoming well established. The first report of In-111 imaging following hepatic embolization is presented. A 45-year-old man with adenocarcinoma of the colon and metastatic liver disease was treated for intractable pain using particulate embolization of the hepatic artery. In-111 leukocyte imaging was performed to rule out abscess formation. The distribution of the labeled leukocytes demonstrated hepatic uptake commensurate with Tc-99m sulfur colloid (SC) images. Areas of embolization did not accumulate tracer. Pathologic examination at autopsy correlated with the distribution of the labeled leukocytes. Thus, therapeutic embolization did not alter the normal distribution of this tracer in functional hepatic tissue.


Effects of vasopressin V1b receptor deficiency on adrenocorticotropin release from anterior pituitary cells in response to oxytocin stimulation.
Oxytocin (OT) is one of the secretagogues for stress-induced ACTH release. OT-induced ACTH release is reported to be mediated by the vasopressin V1b receptor in the rat pituitary gland, which contains both OT and V1b receptors. We examined OT-induced ACTH release using primary cultures of anterior pituitary cells from wild-type (V1bR+/+) and V1b receptor knockout (V1bR-/-) mice. OT stimulated similar levels of ACTH release from pituitary cells of V1bR+/+ and V1bR-/- mice. OT-induced ACTH release was significantly inhibited by the selective V1b receptor antagonist SSR149415 and the OT receptor antagonist CL-14-26 in V1bR+/+ mice. In addition, cotreatment with SSR149415 at 10(-6) m and CL-14-26 at 10(-6) m inhibited OT-induced ACTH release to the control level inV1bR+/+ mice. In V1bR-/- mice, OT-induced ACTH release was significantly inhibited by CL-14-26 at 10(-8) m and completely inhibited at 10(-7)m. These results indicate that OT induces the ACTH response via OT and V1b receptors inV1bR+/+ mice but via only OT receptors in V1bR-/- mice. The gene expression level of the OT receptor was significantly higher in the anterior pituitary gland of V1bR-/- mice than in that of V1bR+/+ mice, suggesting that the OT receptor is up-regulated to compensate for ACTH release under conditions of V1b receptor deficiency.


Low levels of monkeypox virus-neutralizing antibodies after MVA-BN vaccination in healthy individuals.
In July 2022, the ongoing monkeypox (MPX) outbreak was declared a public health emergency of international concern. Modified vaccinia Ankara-Bavarian Nordic (MVA-BN, also known as Imvamune, JYNNEOS or Imvanex) is a third-generation smallpox vaccine that is authorized and in use as a vaccine against MPX. To date, there are no data showing MPX virus (MPXV)-neutralizing antibodies in vaccinated individuals nor vaccine efficacy against MPX. Here we show that MPXV-neutralizing antibodies can be detected after MPXV infection and after historic smallpox vaccination. However, a two-shot MVA-BN immunization series in non-primed individuals yields relatively low levels of MPXV-neutralizing antibodies. Dose-sparing of an MVA-based influenza vaccine leads to lower MPXV-neutralizing antibody levels, whereas a third vaccination with the same MVA-based vaccine significantly boosts the antibody response. As the role of MPXV-neutralizing antibodies as a correlate of protection against disease and transmissibility is currently unclear, we conclude that cohort studies following vaccinated individuals are necessary to assess vaccine efficacy in at-risk populations.


[SHOULD EVERY WOMAN AFTER LABOR BE OFFERED PELVIC FLOOR PHYSIOTHERAPY?]
Pelvic floor damage can occur during pregnancy, during childbirth or post-partum, and may be expressed by symptoms such as urinary incontinence, fecal and gas incontinence, sexual dysfunction, pelvic organ prolapse and chronic pelvic pain. Some of the symptoms, which manifest later in a woman's life, will go unrecognized in the immediate postpartum period. Most women do not mention their general health, unless specifically asked. Physiotherapists, who are adept with the anatomy of the musculoskeletal system and the ability to diagnose unique differences, can aid in the diagnosis and treatment of postpartum pelvic floor problems. Monitoring, pelvic floor physiotherapy and exercise can be effective both in treatment and prevention of functional disorders of the pelvic floor. In this article, we will discuss pelvic floor problems and their appearance throughout pregnancy and childbirth, and the means of treatment from the physiotherapist's perspective. We raise the question as to whether all postpartum women could benefit from a training program of pelvic floor muscles. Pelvic floor physiotherapy is included in the health basket in Israel, but is not broadly utilized. Wider use could be adopted, particularly in light of the latest research evidence.


The effect of induction of CYP3A4 by St John's wort on ambrisentan plasma pharmacokinetics in volunteers of known CYP2C19 genotype.
To evaluate the impact of CYP2C19 polymorphisms on ambrisentan exposure and to assess its modification by St. John's wort (SJW), 20 healthy volunteers (10 CYP2C19 extensive, four poor and six ultrarapid metabolizers) received therapeutic doses of ambrisentan (5 mg qd po) for 20 days and concomitantly SJW (300 mg tid po) for the last 10 days. To quantify changes of CYP3A4 activity, midazolam (3 mg po) as a probe drug was used. Ambrisentan pharmacokinetics was assessed on days 1, 10 and 20, and midazolam pharmacokinetics before and on days 1, 10, 17 and 20. At steady state, ambrisentan exposure was similar in extensive and ultrarapid metabolizers but 43% larger in poor metabolizers (p < 0.01). In all volunteers, SJW reduced ambrisentan exposure and the relative change (17-26%) was similar in all genotype groups. The extent of this interaction did not correlate with the changes in CYP3A activity (midazolam clearance) (rs = 0.23, p = 0.34). Ambrisentan had no effect on midazolam pharmacokinetics. In conclusion, SJW significantly reduced exposure with ambrisentan irrespective of the CYP2C19 genotype. The extent of this interaction was small and thus likely without clinical relevance.


Heparin centenary - an ever-young life-saving drug.
On the centenary of the discovery of heparin, the International Journal of Cardiology agreed to publish a collection of mini reviews that summarize the historical development of this ever-young life-saving drug. The present articles deal not only with the historical milestones, but also with current and future perspectives regarding the development of heparin in terms of its structure, as well as on-going biochemical, biological and clinical research. Attention is focused on recent applications of heparin derivatives to non-anticoagulant or antithrombotic therapies, providing particular emphasis on their inhibitory activities, including their potential as anti-cancer agents. In the Chapter, entitled 'Recent innovations in the structural analysis of heparin', some recent technological advances are described for the problem of monitoring the purity and reproducibility of pharmaceutical heparin. These now permit sensitive detection of non-heparin impurities, as well as the detection of heparin from different animal sources, to be made in pharmaceutical heparin samples. In 'Past, present, and future perspectives of heparin in clinical settings and the role of impaired renal function', the author traces the history of heparin and the development of low molecular weight heparin, highlighting the large number of clinical trials in which it has been involved, and reviewing its efficacy among patients with impaired renal function. In the final chapter, 'Old and new applications of non-anticoagulant heparin', the authors survey some of the many non-anticoagulant activities of heparin and its derivatives, including glycol-split heparin, which has demonstrated promising activities in a wide-range of situations.


Air pollution by concrete dust from the Great Hanshin Earthquake.
Air pollution in the areas affected by the Great Hanshin Earthquake (Hyogo, Japan) of 17 Jan. 1995 was quite serious. We performed three investigations of dust. In the first investigation, we measured the total suspended particulate (TSP) concentration in the greatly damaged areas, located around the Sannomiya Station where a few hundred thousand people walked by during the daytime of 3 February. The maximum concentration at five points reached 150 microg/m3. In the second investigation, eight samples, which were classified into three groups (concrete, mortar, and soil dusts) as sources, were analyzed elementally by X-ray fluorescence. The elements found in concrete dust (Ca and S) were similar to those found in mortar dust. These differed from those found in soil dust (Ti, Fe, and Zr). The elements found in soil dust were important from the viewpoint of heavy metal contamination. In the third investigation, the alkalinity of concrete dust was observed by dissolution. This solution was equivalent to pH 11 to 12 and electrical conductivity 20 to 30 microS/m. We suspect that the alkaline component in the dust from debris in all the devastated areas was approximately comparable with the alkaline solution by which the acid rain falling over the Hanshin district of Osaka Megalopolis in one year could be neutralized into water of pH 7.0.


The FSSP database of structurally aligned protein fold families.
FSSP (families of structurally similar proteins) is a database of structural alignments of proteins in the Protein Data Bank (PDB). The database currently contains an extended structural family for each of 330 representative protein chains. Each data set contains structural alignments of one search structure with all other structurally significantly similar proteins in the representative set (remote homologs, < 30% sequence identity), as well as all structures in the Protein Data Bank with 70-30% sequence identity relative to the search structure (medium homologs). Very close homologs (above 70% sequence identity) are excluded as they rarely have marked structural differences. The alignments of remote homologs are the result of pairwise all-against-all structural comparisons in the set of 330 representative protein chains. All such comparisons are based purely on the 3D co-ordinates of the proteins and are derived by automatic (objective) structure comparison programs. The significance of structural similarity is estimated based on statistical criteria. The FSSP database is available electronically from the EMBL file server and by anonymous ftp (file transfer protocol).


Single-molecule evaluation of fluorescent protein photoactivation efficiency using an in vivo nanotemplate.
Photoswitchable fluorescent probes are central to localization-based super-resolution microscopy. Among these probes, fluorescent proteins are appealing because they are genetically encoded. Moreover, the ability to achieve a 1:1 labeling ratio between the fluorescent protein and the protein of interest makes these probes attractive for quantitative single-molecule counting. The percentage of fluorescent protein that is photoactivated into a fluorescently detectable form (i.e., the photoactivation efficiency) plays a crucial part in properly interpreting the quantitative information. It is important to characterize the photoactivation efficiency at the single-molecule level under the conditions used in super-resolution imaging. Here, we used the human glycine receptor expressed in Xenopus oocytes and stepwise photobleaching or single-molecule counting photoactivated localization microcopy (PALM) to determine the photoactivation efficiency of fluorescent proteins mEos2, mEos3.1, mEos3.2, Dendra2, mClavGR2, mMaple, PA-GFP and PA-mCherry. This analysis provides important information that must be considered when using these fluorescent proteins in quantitative super-resolution microscopy.


Changes in the Cross-Sectional Profile of Treated Anterior Cruciate Ligament Within 2 Years After Surgery.
The cross-sectional area (CSA) of the anterior cruciate ligament (ACL) and reconstructed graft has direct implications on its strength and knee function. Little is known regarding how the CSA changes along the ligament length and how those changes vary between treated and native ligaments over time. It was hypothesized that (1) the CSA of reconstructed ACLs and restored ACLs via bridge-enhanced ACL restoration (BEAR) is heterogeneous along the length. (2) Differences in CSA between treated and native ACLs decrease over time. (3) CSA of the surgically treated ACLs is correlated significantly with body size (ie, height, weight, body mass index) and knee size (ie, bicondylar and notch width). Cohort study; Level of evidence, 2. Magnetic resonance imaging scans of treated and contralateral knees of 98 patients (n = 33 ACL reconstruction, 65 BEAR) at 6, 12, and 24 months post-operation were used to measure the ligament CSA at 1% increments along the ACL length (tibial insertion, 0%; femoral insertion, 100%). Statistical parametric mapping was used to evaluate the differences in CSA between 6 and 24 months. Correlations between body and knee size and treated ligament CSA along its length were also assessed. Hamstring autografts had larger CSAs than native ACLs at all time points (P < .001), with region of difference decreasing from proximal 95% of length (6 months) to proximal 77% of length (24 months). Restored ACLs had larger CSAs than native ACLs at 6 and 12 months, with larger than native CSA only along a small midsubstance region at 24 months (P < .001). Graft CSA was correlated significantly with weight (6 and 12 months), bicondylar width (all time points), and notch width (24 months). Restored ACL CSA was significantly correlated with bicondylar width (6 months) and notch width (6 and 12 months). Surgically treated ACLs remodel continuously within the first 2 years after surgery, leading to ligaments/grafts with heterogeneous CSAs along the length, similar to the native ACL. While reconstructed ACLs remained significantly larger, the restored ACL had a CSA profile comparable with that of the contralateral native ACL. In addition to size and morphology differences, there were fundamental differences in factors contributing to CSA profile between the ACL reconstruction and BEAR procedures. NCT02664545 (ClinicalTrials.gov identifier).


Novel Approach for Simultaneous Recording of Renal Sympathetic Nerve Activity and Blood Pressure with Intravenous Infusion in Conscious, Unrestrained Mice.
Renal sympathetic nerves contribute significantly to both physiological and pathophysiological phenomena. Evaluating renal sympathetic nerve activity (RSNA) is of great interest in many areas of research such as chronic kidney disease, hypertension, heart failure, diabetes and obesity. Unequivocal assessment of the role of the sympathetic nervous system is thus imperative for proper interpretation of experimental results and understanding of disease processes. RSNA has been traditionally measured in anesthetized rodents, including mice. However, mice usually exhibit very low systemic blood pressure and hemodynamic instability for several hours during anesthesia and surgery. Meaningful interpretation of RSNA is confounded by this non-physiological state, given the intimate relationship between sympathetic nervous tone and cardiovascular status. To address this limitation of traditional approaches, we developed a new method for measuring RSNA in conscious, freely-moving mice. Mice were chronically instrumented with radio-telemeters for continuous monitoring of blood pressure as well as a jugular venous infusion catheter and custom-designed bipolar electrode for direct recording of RSNA. Following a 48-72 hour recovery period, survival rate was 100% and all mice behaved normally. At this time-point, RSNA was successfully recorded in 80% of mice, with viable signals acquired up to 4 and 5 days post-surgery in 70% and 50% of mice, respectively. Physiological blood pressures were recorded in all mice (116±2 mmHg; n=10). Recorded RSNA increased with eating and grooming, as well-established in the literature. Furthermore, RSNA was validated by ganglionic blockade and modulation of blood pressure with pharmacological agents. Herein, an effective and manageable method for clear recording of RSNA in conscious, freely-moving mice is described.


Recognizing and managing on toxicities in cancer immunotherapy.
Over the past 4 years, cancer immunotherapy has significantly prolonged survival time of patients with prostate cancer, melanoma, lung cancer, and liver cancer, but its side effects are also impressive. Different types of the immune therapeutic agents have different on-target or off-target toxicity due to high affinity or weak specificity, respectively. Treatment toxicity spectrums vary greatly even in patients with the same type of cancer. Common toxicities are fevers, chills, diarrhea colitis, maculopapular rash, hepatitis, and hormone gland disorder; therefore, routine monitoring of thyroid function, liver function, renal function, and complete blood count are absolutely necessary once treatment begins. Some side effects are reversible, and can be processed through the standard medicines. However, serious toxicities are lethal, which should be frequently followed-up, identified at an early stage and immediately symptomatic treated by high-dose immunosuppressors. In this case, thereafter, the same agent should not be challenged again.


Acute renal failure in essential mixed cryoglobulinemia: precipitation and reversal by plasma exchange.
A patient with essential mixed cryoglobulinemia was treated for deteriorating renal function with plasma exchange alone. This therapy was immediately followed by acute oliguric renal failure due to precipitation of the cryoglobulin within glomerular capillary loops, probably as a result of infusion of cold plasma. The composition of the intracapillary deposits reflected not only the variety of cryoglobulin but the rapidity of deposition. Renal function returned when plasma exchange was re-introduced, this time coupled with immunosuppressive chemotherapy. After resolution of the intraluminal caogula a membranoproliferative picture remained. Replacement fluids for plasma exchange in cryoglobulinemia should be warmed before infusion.


Biophysical and biochemical heterogeneity of purified hepatitis B antigen.
Hepatitis B antigen of the D (a+, d+, y-) subtype was purified from plasma of apparently healthy persons and from hepatitis patients. The original samples contained 20- and 42-nm particles and tubular forms (20-nm diameter). Ultracentrifugation during the purification procedure yielded pellets which were then treated at pH 2.4. Both the large, 42-nm Dane particles and the tubular forms were lost during the acid treatment of the pelleted particles, yielding a preparation containing a mixture of particles approximately 20 and 25 nm in diameter. This difference in size was substantiated in that two distinct molecular weights were calculated from high-speed equilibrium data, 3.6 x 10(6) and 4.5 x 10(6). Further heterogeneity was observed in that hepatitis B antigenic activity was present in purified particles with an isoelectric pH of 4.0 and also in those with a pH of 4.4. No significant differences were observed in the gross amino acid composition of purified antigen obtained from plasma of three different persons. (125)I-labeled, purified antigen was found to contain six distinct polypeptides with molecular weights ranging from 10,000 to 39,000.


The 12-hour control of intraocular pressure on carteolol 2% twice daily.
In a 15-week double-blind, placebo-controlled, crossover study both carteolol 2% and timolol 0.5% produced a significant reduction in intraocular pressure. Twelve hours after administration the reduction in pressure on carteolol was significantly less than that obtained on timolol.


Lysosomal Regulation of mTORC1 by Amino Acids in Mammalian Cells.
The mechanistic target of rapamycin complex 1 (mTORC1) is a master regulator of cell growth in eukaryotic cells. The active mTORC1 promotes cellular anabolic processes including protein, pyrimidine, and lipid biosynthesis, and inhibits catabolic processes such as autophagy. Consistent with its growth-promoting functions, hyper-activation of mTORC1 signaling is one of the important pathomechanisms underlying major human health problems including diabetes, neurodegenerative disorders, and cancer. The mTORC1 receives multiple upstream signals such as an abundance of amino acids and growth factors, thus it regulates a wide range of downstream events relevant to cell growth and proliferation control. The regulation of mTORC1 by amino acids is a fast-evolving field with its detailed mechanisms currently being revealed as the precise picture emerges. In this review, we summarize recent progress with respect to biochemical and biological findings in the regulation of mTORC1 signaling on the lysosomal membrane by amino acids.


Characterization of three Staphylococcus aureus isolates from a 17-year-old female who died of tampon-related toxic shock syndrome.
We report the identification and characterization of three Staphylococcus aureus isolates recovered from throat and vaginal cultures, as well as from an axillary abscess, of a 17-year-old female who died of tampon-related toxic shock syndrome. The three S. aureus isolates were unrelated as determined by pulsed-field gel electrophoresis. The vaginal isolate was mecA, Panton-Valentine leukocidin, and staphylococcal enterotoxin B and C negative, toxic shock syndrome toxin 1 positive, and staphylococcal cassette chromosome mec element (SCCmec) untypeable, which was consistent with the clinical and autopsy findings that death was due to tampon-related toxic shock syndrome.


Global chemokine expression in systemic sclerosis (SSc): CCL19 expression correlates with vascular inflammation in SSc skin.
To characterise global chemokine expression in systemic sclerosis (SSc) skin in order to better understand the relationship between chemokine expression and vascular inflammation in this disease. We investigated chemokine mRNA expression in the skin through quantitative PCR analysis comparing patients with diffuse cutaneous (dcSSc) or limited cutaneous (lcSSc) disease with healthy controls. We tested correlations between the most regulated chemokines and vascular inflammation and macrophage recruitment. CCL19 expression was examined in human primary immune cells treated with innate immune activators. The chemokines, CCL18, CCL19 and CXCL13, were upregulated in dcSSc skin, and CCL18 in lcSSc skin. Expression of CCL19 in dcSSc skin correlated with markers of vascular inflammation and macrophage recruitment. Immunofluorescence data showed CCL19 colocalisation with CD163 macrophages in dcSSc skin. In vitro studies on human primary cells demonstrated that CCL19 expression was induced after toll-like receptor activation of peripheral blood mononuclear cells and separated populations of CD14 monocytes. CCL18, CCL19 and CXCL13-chemoattractants for macrophage and T cell recruitment-were three of six chemokines with the highest expression in dcSSc skin. Increased CCL19 expression in the skin suggests a role for CCL19 in the recruitment of immune cells to the peripheral tissue. Induction of CCL19 in macrophages but not structural cells indicates a role for skin-resident or recruited immune cells in perivascular inflammation. This study demonstrates that CCL19 is a sensitive marker for the perivascular inflammation and immune cell recruitment seen in dcSSc skin disease.


MCI-9042: high affinity for serotonergic receptors as assessed by radioligand binding assay.
This study assessed the specificity of displacement potencies of MCI-9042 [+/-)-1-[o-[2-(m-methoxyphenyl)ethyl]phenoxy]-3-(dimethylamino)- 2-propyl hydrogen succinate hydrochloride), a new antiplatelet agent, in [3H]4-bromo-2,5-dimethoxyphenylisopropylamine [( 3H]DOB) and [3H]ketanserin binding to 5HT2-serotonergic receptors in the rat brain using the radioligand binding method. The assessment of the displacement potencies of MCI-9042 and its derivative, M-1 [(+/-)-1-[2-[2-(3-methoxyphenyl)ethyl]phenoxy]-3-(dimethylamino)- 2-propanol)hydrochloride), on alpha 1-, alpha 2-, beta-adrenergic, 5HT1-, 5HT2- and muscarinic receptors were also studied. MCI-9042 and M-1 showed strong displacement effects to 5HT2-receptors using [3H]DOB binding and [3H]ketanserin binding sites, and weak displacement effects for [3H]serotonin, [3H]prazosin, [3H]p-aminoclonidine, [3H]CGP12177 and [3H]QNB binding to 5HT1-, alpha 1-, alpha 2-, beta-adrenergic receptors and muscarinic receptors. Thus, the present study suggests that MCI-9042 is specific for 5HT2-receptor.


Adipose Mesenchymal Stem Cell-Derived Exosomes Promote the Regeneration of Corneal Endothelium Through Ameliorating Senescence.
Human corneal endothelial cells (hCECs) have been considered unable to regenerate in vivo, resulting in corneal decompensation after significant loss of hCECs. adipose-derived mesenchymal stem cell (ASC)-derived exosomes can regenerate tissues and organs. In this study, we investigated whether ASC-derived exosomes could protect and regenerate CECs. We performed cell viability and cell-cycle analyses to evaluate the effect of ASC-derived exosomes on the regeneration capacity of cultured hCECs. Transforming growth factor-β (TGF-β) and hydrogen peroxide (H2O2) were used to induce biological stress in CECs. The effect of ASC-derived exosomes on CECs was investigated in vivo. ASC-derived exosomes were introduced into rat CECs using electroporation, and rat corneas were injured using cryoinjury. Next-generation sequencing analysis was performed to compare the differentially expressed microRNAs (miRNAs) between ASC-derived and hCEC-derived exosomes. ASC-derived exosomes induced CEC proliferation and suppressed TGF-β- or H2O2-induced oxidative stress and senescence. ASC-derived exosomes protect hCECs against TGF-β- or H2O2-induced endothelial-mesenchymal transition and mitophagy. In an in vivo study, ASC-derived exosomes promoted wound healing of rat CECs and protected the corneal endothelium against cryoinjury-induced corneal endothelium damage. Next-generation sequencing analysis revealed differentially expressed miRNAs for ASC-derived and hCEC-derived exosomes. They are involved in lysine degradation, adherens junction, the TGF-β signaling pathway, the p53 signaling pathway, the Hippo signaling pathway, the forkhead box O (FoxO) signaling pathway, regulation of actin cytoskeleton, and RNA degradation based on Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. ASC-derived exosomes promoted wound healing and regeneration of endothelial cells by inducing a shift in the cell cycle and suppressing senescence and autophagy.


Successful pregnancy after myomectomy using preoperative adjuvant uterine artery embolization.
A 30-year-old-woman presented with uterine leiomyoma and primary sterility. Abdominal myomectomy was performed; however, one cervical leiomyoma was not resected because of a risk of excessive blood loss. Two years after the procedure, a secondary myomectomy using preoperative adjuvant uterine artery embolization (UAE) was performed because of pronounced menorrhagia and her hope for bearing children. The patient's dysmenorrhea disappeared postoperatively and she conceived spontaneously 3 years after the secondary myomectomy. This case suggests that myomectomy using preadjuvant UAE may be an another approach for the treatment of leiomyoma in patients who wish bear children in the future.


[Effect of pilsicainide hydrochloride on atrial fibrillation threshold].
To inventigate the electrophysiologic effects of pilsicainide hydrochloride on atrial fibrillation, we compared atrial fibrillation threshold (AFT), right atrial effective refractory period (RAERP), and inter-atrial conduction time (Inter-ACT) before and after the administration of pilsicainide in 12 patients with lone paroxysmal atrial fibrillation. The following electrophysiologic study was performed before and after the administration of the drug as the paced cycle length of 500msec. First, RAERP was measured. Secondly, Inter-ACT from the stimulating artifact to the initial deflection in the elecrocardiograms of the coronary sinus was measured. Thirdly, high-frequency (50Hz) stimulation was given at right atrial appendage continuously for one second just after the eighth basic paced beat. The stimulation current was increased by 1mA in a stepwise fashion from 2mA until atrial fibrillation ensued. AFT was defined as the lowest intensity of the current that induced atrial fibrillation or flutter of more than 30 seconds. Pilsicanide significantly increased AFT and Inter-ACT, but did not change RAERP. In conclusion, it is suggested that pilsicainide might decrease atrial vulnerability mainly by its effect on inter-atrial conduction delay and by the resulting increase in AFT.


[Target gene sequence capture and next generation sequencing technology to diagnose four children with Alagille syndrome].
To make genetic diagnosis of Alagille syndrome (ALGS) patients using target gene sequence capture and next generation sequencing technology. Target gene sequence capture and next generation sequencing were used to detect ALGS gene of 4 patients. They were hospitalized at the Affiliated Hospital, Capital Institute of Pediatrics between January 2014 and December 2015, referred to clinical diagnosis of ALGS typical and atypical respectively in 2 cases. Blood samples were collected from patients and their parents and genomic DNA was extracted from lymphocytes. Target gene sequence capture and next generation sequencing was detected. Sanger sequencing was used to confirm the results of the patients and their parents. Cholestasis, heart defects, inverted triangular face and butterfly vertebrae were presented as main clinical features in 4 male patients. The first hospital visiting ages ranged from 3 months and 14 days to 3 years and 1 month. The age of onset ranged from 3 days to 42 days (median 23 days). According to the clinical diagnostic criteria of ALGS, patient 1 and patient 2 were considered as typical ALGS. The other 2 patients were considered as atypical ALGS. Four Jagged 1(JAG1) pathogenic mutations were detected. Three different missense mutations were detected in patient 1 to patient 3 with ALGS(c.839C>T(p.W280X), c. 703G>A(p.R235X), c. 1720C>T(p.V574M)). The JAG1 mutation of patient 3 was first reported. Patient 4 had one novel insertion mutation (c.1779_1780insA(p.Ile594AsnfsTer23)). Parental analysis verified that the JAG1 missense mutation of 3 patients were de novo. The results of sanger sequencing was consistent with the results of the next generation sequencing. Target gene sequence capture combined with next generation sequencing can detect two pathogenic genes in ALGS and test genes of other related diseases in infantile cholestatic diseases simultaneously and presents a high throughput, high efficiency and low cost. It may provide molecular diagnosis and treatment for clinicians with good clinical application prospects.


Percutaneous coronary intervention versus coronary artery bypass grafting as primary revascularization in patients with acute coronary syndrome.
New European Society of Cardiology/American College of Cardiology guidelines classify patients with acute coronary syndrome and increased cardiac troponins as non-ST-segment elevation myocardial infarction (NSTEMI) who would have been classified as unstable angina pectoris (UAP) using the older World Health Organization (WHO) definition. The optimal revascularization strategy in these patients is poorly defined. This prospective cohort study included 1,024 consecutive patients with acute coronary syndrome classified as UAP, NSTEMI according to the WHO definition (WHO NSTEMI), and NSTEMI additionally identified by the novel European Society of Cardiology/American College of Cardiology definition (additional NSTEMI). All patients underwent coronary angiography within 24 hours and were treated with immediate percutaneous coronary intervention (PCI) or early coronary artery bypass grafting (CABG). The primary end point was all-cause mortality during follow-up of 36 months. Patients with additional NSTEMI showed excessive cumulative 3-year mortality if undergoing CABG (hazard ratio 5.9, 95% confidence interval 2.7 to 13.1, p <0.001). In patients with UAP or WHO NSTEMI, mortality was similar in the CABG and PCI groups. In conclusion, in the absence of randomized trials specifically including patients with additional NSTEMI, the excessive mortality observed with CABG in this cohort study suggested that PCI may be the preferable revascularization strategy in this subgroup.


Evaluating the use of commercial drug-specific calibrators for determining PT and APTT reagent sensitivity to dabigatran and rivaroxaban.
Suitable laboratory methodologies for quantifying the non-vitamin K oral anticoagulants (NOAC) include liquid chromatography-tandem mass spectrometry (LC-MS/MS) or drug-calibrated assays such as the dilute thrombin time for dabigatran or anti-Xa measurements for rivaroxaban. In situations when these tests are unavailable, it has been suggested that using commercial drug calibrators on APTT and PT assays would theoretically provide reagent sensitivity to these drugs. The purpose of this study was to determine whether commercial drug calibrators deliver similar reagent sensitivity information as samples from patients receiving dabigatran or rivaroxaban as part of their routine care. Two laboratory sites tested commercial calibrator material for dabigatran and rivaroxaban (Hyphen Biomedical) using PT and APTT reagents and data was compared to samples collected from patients taking NOACs that were quantified by LC-MS/MS. Correlation statistics and calculating the amount of drug required to double the clotting time of normal plasma were performed. All drug calibrator material correlated more strongly (R²> 0.95) for any reagent/drug combination than patient samples (R² ranged from 0.29-0.86). Dabigatran calibrator results and patient data were equivalent for SynthASil and PTT-A APTT reagents. The dabigatran and rivaroxaban calibrator material over-estimated drug sensitivity for all PT reagents when compared to sensitivity data calculated based on drug levels obtained by LC-MS/MS from patient samples. In conclusion, drug-specific calibrators overestimated reagent sensitivity which may underestimate in vivo drug concentration in a given patient. Further studies are required to assess whether this method of determining relative sensitivity of NOAC on routine coagulation assays should be recommended.


Screening for cancer. Is it useful?
Screening is a diagnostic activity applied to whole populations, or defined sections of populations, with the aim to detect the signs of disease before there are symptoms. The rationale of screening is the assumption that disease detected early is likely to be curable. When setting up a screening programme, the epidemiology and the natural history of the condition to be screened for, the sensitivity and specificity of the screening method, its acceptability, its dangers and its costs need to be taken into account. Examining in the light of these criteria, the screening programmes advocated for the six commonest cancers one comes to the following conclusions: skin cancers can be detected by regular inspection early; for colorectal cancer, the best screening method seems to be to offer one colonoscopy at the age of 55 and so identify those at risk; there is no useful screening method for bronchial carcinoma; serum prostate specific antigen estimation augmented by rectal ultrasound examination of the prostate will detect most prostatic cancers early but it is difficult to know what the therapeutic consequences are; carcinoma of the uterine cervix has been eliminated in middle class western society possibly with the help of cytological screening and; the early detection of cancer of the female breast by means of mammography has proven to benefit women between the ages 50 and 65. In conclusion, some present cancer screening programmes are of definite but limited value.


The origins of new pandemic viruses: the acquisition of new host ranges by canine parvovirus and influenza A viruses.
Transfer of viruses between hosts to create a new self-sustaining epidemic is rare; however, those new viruses can cause severe outbreaks. Examples of such viruses include three pandemic human influenza A viruses and canine parvovirus in dogs. In each case one virus made the original transfer and spread worldwide, and then further adaptation resulted in the emergence of variants worldwide. For the influenza viruses several changes were required for growth and spread between humans, and the emergence of human H2N2 and H3N2 strains in 1957 and 1968 involved the acquisition of three or two new genomic segments, respectively. Adaptation to humans involved several viral genes including the hemagglutinin, the neuraminidase, and the replication proteins. The canine adaptation of the parvoviruses involved capsid protein changes altering the recognition of the host transferrin receptors, allowing canine transferrin receptor binding and its use as a receptor for cell infection.


A novel form of the RelA nuclear factor kappaB subunit is induced by and forms a complex with the proto-oncogene c-Myc.
Members of both Myc and nuclear factor kappaB (NF-kappaB) families of transcription factors are found overexpressed or inappropriately activated in many forms of human cancer. Furthermore, NF-kappaB can induce c-Myc gene expression, suggesting that the activities of these factors are functionally linked. We have discovered that both c-Myc and v-Myc can induce a previously undescribed, truncated form of the RelA(p65) NF-kappaB subunit, RelA(p37). RelA(p37) encodes the N-terminal DNA binding and dimerization domain of RelA(p65) and would be expected to function as a trans-dominant negative inhibitor of NF-kappaB. Surprisingly, we found that RelA(p37) no longer binds to kappaB elements. This result is explained, however, by the observation that RelA(p37), but not RelA(p65), forms a high-molecular-mass complex with c-Myc. These results demonstrate a previously unknown functional and physical interaction between RelA and c-Myc with many significant implications for our understanding of the role that both proteins play in the molecular events underlying tumourigenesis.


NMDA receptor antagonist treatment at the time of nerve injury prevents injury-induced changes in spinal NR1 and NR2B subunit expression and increases the sensitivity of residual pain behaviours to subsequently administered NMDA receptor antagonists.
Spinal NMDA receptors (NMDA R) are important in neuropathic sensitisation and acute administration of antagonists can provide temporary attenuation of sensitisation. If establishment of the chronic pain state could be prevented by brief administration of such agents at or around the time of nerve injury (pre-emptive analgesia) it might be possible to avoid many of the unacceptable side effects associated with repeated administration of these or other antagonists. Several reports describe aspects of effective pre-emptive analgesia from NMDA R antagonists in animal models of neuropathic pain. The first aim of the present study was to make a direct comparison of changes in mechanical allodynia, cold allodynia and thermal hyperalgesia following nerve injury, demonstrating their increasing degree of susceptibility to pre-emptive NMDA R antagonist treatment. Secondly, we used immunoblotting and immunohistochemistry to investigate the effects of nerve injury on NMDA receptor subunit expression, revealing increased expression of NR2B, but not NR2A and reduced NR1 in the superficial dorsal horn. These changes were attenuated following NMDA receptor antagonist pre-treatment. Thirdly, we investigated the pharmacological properties of residual mechanical allodynia and cold allodynia that remained after pre-emptive treatment and revealed a greater sensitivity to NMDA R antagonists. These findings indicate that in addition to a marked suppression of thermal hyperalgesia and cold allodynia, pre-emptive treatment with NMDA R antagonist causes a lasting change in spinal NMDA R complexes such that remaining mechanical allodynia should be more effectively targeted by NMDA R antagonists.


Comparison of oxidative stress/DNA damage in semen and blood of fertile and infertile men.
Abnormal spermatozoa frequently display typical features of oxidative stress, i.e. excessive level of reactive oxygen species (ROS) and depleted antioxidant capacity. Moreover, it has been found that a high level of oxidatively damaged DNA is associated with abnormal spermatozoa and male infertility. Therefore, the aim of our study was the comparison of oxidative stress/DNA damage in semen and blood of fertile and infertile men. The broad range of parameters which describe oxidative stress and oxidatively damaged DNA and repair were analyzed in the blood plasma and seminal plasma of groups of fertile and infertile subjects. These parameters include: (i) 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) and 8-oxo-7,8-dihydroguanine (8-oxoGua) levels in urine; (ii) 8-oxodG level in DNA isolated from leukocytes and spermatozoa; (iii) antioxidant vitamins (A, C and E) and uric acid. Urinary excretion of 8-oxodG and 8-oxoGua and the level of oxidatively damaged DNA in leukocytes as well as the level of antioxidant vitamins were analyzed using HPLC and HPLC/GC/MS methods. The results of our study demonstrate that 8-oxodG level significantly correlated with every parameter which describe sperm quality: sperm count, motility and morphology. Moreover, the data indicate a higher level of 8-oxodG in sperm DNA compared with DNA of surrogate tissue (leukocytes) in infertile men as well as in healthy control group. For the whole study population the median values of 8-oxodG/10(6) dG were respectively 7.85 and 5.87 (p=0.000000002). Since 8-oxodG level in sperm DNA is inversely correlated with urinary excretion rate of 8-oxoGua, which is the product of OGG1 activity, we hypothesize that integrity of spermatozoa DNA may be highly dependent on OGG1 activity. No relationship between the whole body oxidative stress and that of sperm plasma was found, which suggests that the redox status of semen may be rather independent on this characteristic for other tissues.


The anesthetic efficacy of diphenhydramine and the combination diphenhydramine/lidocaine for the inferior alveolar nerve block.
The purpose of this prospective, randomized study was to compare the degree of pulpal anesthesia obtained in vital, asymptomatic teeth using 1.8 mL of 1% diphenhydramine with 1:100,000 epinephrine, 1.8 mL of 2% lidocaine with 1:100,000 epinephrine, and 3.6 mL of 2% lidocaine with 1:100,000 epinephrine combined with 1% diphenhydramine with 1:100,000 epinephrine in inferior alveolar nerve blocks. We also studied injection and postinjection pain. An electric pulp tester was used to test for anesthesia, in 4-minute cycles for 60 minutes, of the second molars through the central incisor. Anesthesia was considered successful when two consecutive 80 readings were obtained within 15 minutes and the 80 reading was continuously sustained through the 60th minute. Using the lidocaine solution, successful pulpal anesthesia ranged from 12% to 84%. For the combined lidocaine/diphenhydramine solution, successful pulpal anesthesia ranged from 4% to 60%. The diphenhydramine solution was irritating on injection and postinjection and resulted in a very low level of anesthetic success. We concluded that a 1% diphenhydramine solution should be used cautiously for the inferior alveolar nerve block. The combination lidocaine/diphenhydramine solution was irritating postinjection and was not as effective as a lidocaine solution for pulpal anesthesia.


Purification and immunological characterization of the major internal protein of the RD-114 virus.
The major internal protein of the RD-114 virus that appeared in a human tumor cell line, RD, after passage through fetal cats was purified by isoelectric focusing and used to prepare antibody in guinea pigs. The protein has a molecular weight of 33,500 in sodium dodecyl sulfate-polyacrylamide gels, and an isoelectric point of 9.1. This contrasts with a molecular weight of 25,000 and an isoelectric point of 8.3 obtained previously for the major protein of cat C-type viruses. The RD-114 protein carries determinants common to all mammalian C-type viruses but not species-specific determinants of cat, mouse, rat, hamster, chicken, and Russell's viper C-type viruses. In addition, two other proteins found in cat virus preparations were not detected in the RD-114 virus. Antiserum to the RD-114 viral protein proved highly specific by complement fixation and gel diffusion assays, and, in addition to the above mentioned mammalian viruses, the antiserum did not react with concentrates of the Woolly monkey or Gibbon ape C-type viruses. These data support the conclusion that the RD-114 virus is a human virus activated by passage through fetal cats. Until additional isolates are made from human cells, the identity of the RD-114 virus as a human C-type tumor virus cannot be fully established; however, this would be highly probable if the species-specific group-specific antigen of RD-114 could be demonstrated in human tumor and/or embryonic tissues.


Gastrin-releasing peptide stimulates glycoconjugate release from feline trachea.
The effect of gastrin-releasing peptide (GRP) on respiratory glycoconjugate (RGC) secretion was investigated in a feline tracheal organ culture model. RGC secretion was stimulated by GRP in a dose-dependent fashion at concentrations from 10(-8) to 10(-5) M (range 15-38% increase above control) with a peak effect within 0.5-1 h of incubation. GRP-(14-27), the receptor binding portion of GRP, and the related molecule, bombesin, also stimulated RGC secretion by approximately 20% above control. Acetyl-GRP-(20-27) stimulated RGC release by 10%, whereas GRP-(1-16) was inactive. Autoradiographic studies with 125I-GRP revealed that specific binding was restricted to the submucosal glands and the surface epithelium. A specific radioimmunoassay showed the content of GRP in feline trachea after extraction with ethanol-acetic acid to be 156 +/- 91 fmol/g wet wt. Indirect immunohistochemistry indicated that ganglion cells located just outside the cartilage contained GRP-immunoreactive materials. GRP is a novel mucus secretagogue that may participate in regulating airway mucosal gland secretion.


Life after colectomy.
Animal and epidemiologic studies have raised hopes that effective chemoprevention of colorectal cancer in very high-risk patients may be possible in the future, but at present, the only established effective way to prevent colorectal cancer in very high-risk patients is colectomy. Advances in surgical techniques, improvements in stomal appliances, and medical advances in dealing with complications of colorectal surgery (such as impotence) have contributed to substantial improvements in the quality of life of patients who have had prophylactic colectomies.


Partial splenic ablation in preparation for renal transplantation in children.
Patients with end-stage renal disease who develop hypersplenism, patients with mild neutropenia, and those patients whose WBC fails to increase in response to cortisol administration will develop significant neutropenia following transplantation with routine doses of azathioprine. This "intolerance" of azathioprine mandates a reduction in the dose of azathioprine often resulting in allograft rejection. Splenectomy will prevent azathioprine-induced neutropenia, but the hazards of splenectomy in these immunosuppressed patients have led to attempts to salvage at least part of the spleen. Partial splenic ablation by embolization has been utilized in adults prior to transplantation to prevent azathioprine-induced neutropenia while preserving the spleen's protective mechanisms against infection. Eight children in our series of transplant candidates required a reduction of splenic function to prevent azathioprine induced neutropenia. One child had a functioning renal allograft but had recurrent neutropenia limiting the azathioprine dose. Partial splenic embolization was attempted in four children and was initially successful in two. Both patients later developed recurrent neutropenia and needed partial splenectomy. The two patients in whom partial splenic embolization was unsuccessful and five further patients in whom embolization was not attempted also underwent partial splenectomy. Approximately 75% to 80% of the spleen was resected. Six children have since undergone renal transplantation and one child had a transplant with chronic rejection at the time of partial splenectomy. Routine doses of azathioprine have been used in these children with no episodes of neutropenia or sepsis observed. We recommend partial splenectomy in those children requiring renal transplantation who are at risk for development of azathioprine induced neutropenia.


Consideration of grip and push forces for the assessment of vibration exposure.
There is much influence of the coupling forces between hand and grip of the vibrating tool on the measuring results as well as on the vibration effects on the hand-arm system. In a research project the effects of grip and push forces on acute responses of the hand-arm system under vibration conditions have been studied. Using these results of the biodynamic response, vibration perception threshold and subjective vibration sensation a bonus/malus system for a correction of the measured frequency-weighted r.m.s. acceleration was drafted, to assess the hand-arm vibration at the workplace: Since there is no difference between the acute effects of grip force and push force, so the forces have to be added and defined as coupling force Fcf. On the basis of this coupling force Fcf a correcting factor cF may be calculated. The factor amounts to 0.6 at Fcf = 20 N, 1.0 at Fcf = 120 N and 1.1 at Fcf = 200 N. To correct the measured weighted r.m.s. acceleration ahwz (Fcf) depending on coupling forces the r.m.s. acceleration has to be multiplied with the correcting factor cF. The drafted procedure enables to assess vibration exposure depending on coupling forces in a standardized way.


[Characteristics of the heat-stable enterotoxin of NAG vibrios].
Cell-free culture filtrate boiled for 15 minutes has been found to retain its biological activity in various experimental models used for the determination of the toxicogenicity of cholera vibrio filtrates. During gel filtration of the concentrated filrate o. NAG vibrio, strain NO. 9852, through Sephadex G-75 toxic activity could be detected in the free volume of the column, which was indicative of the fact that the molecular weight of the thermostable enterotoxin was about 70,000 daltons and greater. The methods of gel diffusion and aggregated hemagglutination have been used to show that the thermostable enterotoxin of NAG vibrio No. 9852 is immunologically unrelated to cholerogen. Some data obtained in experimental models suggest that the thermostable enterotoxin probably differs from cholera enterotoxin in the mechanism of its action.


The existence of protein kinase C in cone photoreceptors in the rat retina.
We examined the localization of protein kinase C (PKC) in the rat retina, using a monoclonal antibody against PKC. The PKC immunoreactivities were localized in bipolar cells as reported previously, and also in outer and inner segments (OS and IS) of photoreceptor cells. As the PKC immunoreactive OS and IS of photoreceptors were very few in number, we hypothesized that these were of cone photoreceptors. We then examined whether the PKC immunoreactive OS and IS were bound by peanut agglutinin which was shown to bind specifically to cone OS and IS. Almost all of the PKC immunoreactive OS and IS showed PNA binding.