Impact of human papillomavirus (HPV) 16 and 18 vaccination on prevalent infections and rates of cervical lesions after excisional treatment.
Human papillomavirus vaccines prevent human papillomavirus infection and cervical precancers. The impact of vaccinating women with a current infection or after treatment for an human papillomavirus-associated lesion is not fully understood. To determine whether human papillomavirus-16/18 vaccination influences the outcome of infections present at vaccination and the rate of infection and disease after treatment of lesions. We included 1711 women (18-25 years) with carcinogenic human papillomavirus infection and 311 women of similar age who underwent treatment for cervical precancer and who participated in a community-based trial of the AS04-adjuvanted human papillomavirus-16/18 virus-like particle vaccine. Participants were randomized (human papillomavirus or hepatitis A vaccine) and offered 3 vaccinations over 6 months. Follow-up included annual visits (more frequently if clinically indicated), referral to colposcopy of high-grade and persistent low-grade lesions, treatment by loop electrosurgical excisional procedure when clinically indicated, and cytologic and virologic follow-up after treatment. Among women with human papillomavirus infection at the time of vaccination, we considered type-specific viral clearance, and development of cytologic (squamous intraepithelial lesions) and histologic (cervical intraepithelial neoplasia) lesions. Among treated women, we considered single-time and persistent human papillomavirus infection, squamous intraepithelial lesions, and cervical intraepithelial neoplasia 2 or greater. Outcomes associated with infections absent before treatment also were evaluated. Infection-level analyses were performed and vaccine efficacy estimated. Median follow-up was 56.7 months (women with human papillomavirus infection) and 27.3 months (treated women). There was no evidence of vaccine efficacy to increase clearance of human papillomavirus infections or decrease incidence of cytologic/histologic abnormalities associated with human papillomavirus types present at enrollment. Vaccine efficacy for human papillomavirus 16/18 clearance and against human papillomavirus 16/18 progression from infection to cervical intraepithelial neoplasia 2 or greater were -5.4% (95% confidence interval -19,10) and 0.3% (95% confidence interval -69,41), respectively. Among treated women, 34.1% had oncogenic infection and 1.6% had cervical intraepithelial neoplasia 2 or greater detected after treatment, respectively, and of these 69.8% and 20.0% were the result of new infections. We observed no significant effect of vaccination on rates of infection/lesions after treatment. Vaccine efficacy estimates for human papillomavirus 16/18 associated persistent infection and cervical intraepithelial neoplasia 2 or greater after treatment were 34.7% (95% confidence interval -131, 82) and -211% (95% confidence interval -2901, 68), respectively. We observed evidence for a partial and nonsignificant protective effect of vaccination against new infections absent before treatment. For incident human papillomavirus 16/18, human papillomavirus 31/33/45, and oncogenic human papillomavirus infections post-treatment, vaccine efficacy estimates were 57.9% (95% confidence interval -43, 88), 72.9% (95% confidence interval 29, 90), and 36.7% (95% confidence interval 1.5, 59), respectively. We find no evidence for a vaccine effect on the fate of detectable human papillomavirus infections. We show that vaccination does not protect against infections/lesions after treatment. Evaluation of vaccine protection against new infections after treatment and resultant lesions warrants further consideration in future studies.


Uptake and toxicity of copper oxide nanoparticles in cultured primary brain astrocytes.
To test for consequences of an exposure of brain cells to copper oxide nanoparticles (CuO-NPs), we synthesised and characterised dimercaptosuccinate-coated CuO-NPs. These particles had a diameter of around 5 nm as determined by transmission electron microscopy, while their average hydrodynamic diameter in aqueous dispersion was 136 ± 4 nm. Dispersion in cell-culture medium containing 10% fetal calf serum increased the hydrodynamic diameter to 178 ± 12 nm and shifted the zeta potential of the particles from -49 ± 7 mV (in water) to -10 ± 3 mV. Exposure of cultured primary brain astrocytes to CuO-NPs increased the cellular copper levels and compromised the cell viability in a time-, concentration- and temperature-dependent manner. Application of CuO-NPs in concentrations above 100 µM copper (6.4 µg/ml) severely compromised the viability of the cells, as demonstrated by a lowered 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide reduction capacity, a lowered cellular lactate dehydrogenase activity and an increased membrane permeability for the fluorescent dye propidium iodide. Copper internalisation as well as cell toxicity of astrocytes exposed to CuO-NPs were similar to that observed for cells that had been incubated with copper salts. The CuO-NP-induced toxicity was accompanied by an increase in the generation of reactive oxygen species (ROS) in the cells. Both, ROS formation and cell toxicity in CuO-NP-treated astrocytes, were lowered in the presence of the cell-permeable copper chelator tetrathiomolybdate. These data demonstrate that CuO-NPs are taken up by cultured astrocytes and suggest that excess of internalised CuO-NPs cause cell toxicity by accelerating the formation of ROS.


Phylogenetic analysis of rabbit haemorrhagic disease and European brown hare syndrome viruses by comparison of sequences from the capsid protein gene.
A 398 bp fragment of the capsid protein (VP60) gene of 39 clinical samples of rabbit haemorrhagic disease virus (RHDV) and 17 of European brown hare syndrome virus (EBHSV), collected between 1981 and 1995 from 17 countries, was amplified by PCR and directly sequenced. The alignment of the nucleotide sequences and the subsequently constructed phylogenetic tree clearly separated RHDV from EBHSV as phylogenetic entities. The nucleotide homology rates between the RHDV and EBHSV groups ranged between 52.6% and 60.0%. The homology rates within the groups were much higher, 89.4% to 100% for the RHDV samples, and 89.4% to 100% for the EBHSV specimens. No intermediate viruses were found. Despite the high homology, three main branches could be identified in the phylogenetic tree of the RHDV samples, corresponding to the epizootiological data, while the EBHSV dendrogram did not show such well defined branches. The present results support the classification of RHDV and EBHSV as two distinct members of the Caliciviridae family. Nevertheless, a comparison with previously determined sequences of other caliciviruses shows that RHDV and EBHSV are more closely related to each other than to any other calicivirus.


Change in Patient MELD-Na and Albumin Level From the Time of Celiac Disease Diagnosis to Six Months Later After Gluten-Free Diet.
Background & aims Celiac disease (CD) is a multisystem disorder triggered by dietary gluten in genetically predisposed individuals that may affect any organ system, including the liver. We evaluated a change in patient model for end-stage liver disease (MELD)-Na and albumin level from the time of celiac disease diagnosis to six months later, after implementing a gluten-free diet. Methods A retrospective study was conducted from January 1, 2006, to June 30, 2018. CD was diagnosed based on celiac antibodies and/or histopathological data. MELD-Na and albumin were calculated at the start of the gluten-free diet and six months later. Additional variables like gender, ethnicity, serum IgA level, serum IgG level, human leukocyte antigen (HLA) type, and markers of end-stage liver disease were collected. Descriptive statistics, including means, were reported with the standard deviation for the continuous variables along with frequencies and percentages for all categorical variables. Results A total of 18 patients (55.6% male) were identified as having both cirrhosis and CD. The mean age at the time of celiac diagnosis was 53.6, and 94.4% were Caucasian. CD was diagnosed using celiac antibodies (100%) and histopathological data (44.4%). Most common celiac antibodies include anti-tissue transglutaminase antibodies (77.8%). End-stage liver disease markers like abdominal ascites (55.6%), variceal bleed (50.0%), acute or chronic kidney injury (16.7%), hepatocellular carcinoma (HCC) (11.1%), hepatic encephalopathy (HE) (50.0%), spontaneous bacterial peritonitis (SBP) (5.6%), and liver transplant (0.0%) were seen. The mean baseline MELD-Na score was 11.8, and albumin was 3.5 at the time of celiac diagnosis and mean MELD-Na was 11.8, and albumin was 3.5 six months after a gluten-free diet. Conclusion It is difficult to conclude any exact relationship between change in MELD-Na score after gluten-free diet, but an improving trend is noted in patients with higher MELD-Na score such as 17 or higher. There is no change or worsening of MELD-Na score in patients with lower MELD-Na score. There was no change in mean MELD-Na and albumin level after gluten-free diet.


Pain and electrophysiological parameters are improved by combined 830-1064 high-intensity LASER in symptomatic carpal tunnel syndrome versus Transcutaneous Electrical Nerve Stimulation. A randomized controlled study.
The aim of the study was to compare LASER versus transcutaneous electrical nerve stimulation (TENS) in reducing pain and paraesthesia; and in improving motor and sensory median nerve conduction parameters in mild to moderate carpal tunnel syndrome (CTS). Randomised blinded pilot study. Patients and staff administered treatments and outcome measures were blinded. Outpatient; Research and Care Rehabilitation Institute. Twenty CTS symptomatic patients. Fifteen sessions of: 1) 100 Hz TENS (30 minutes; rectangular waves; 80 ms width, intensity below muscle contraction); 2) combined 830-1064 nm LASER (radiating dose: 250 J cm-2 delivered to the skin overlying the course of the median nerve at the wrist for 100 s at 25 W (18 W [1064 nm] + 7 W [830 nm]) via a fiber-optic probe with a spot size of ~1 cm2). Outcome measures. Visual analogue scale (VAS) for pain and paresthesia; median nerve distal motor latency and sensory nerve conduction velocity. LASER improved both positive and negative sensory symptoms. TENS induced clinical improvement but this was not statistically significant and was limited to pain reduction. LASER but not TENS favourably modified the neurophysiological parameters. High-intensity combined LASER wavelengths of 830 nm and 1064 nm, which produce a better transparency with less scattering and a high energy transfer, are better than TENS in improving both pain and paraesthesia as well as neurophysiological parameters in CTS.


Exercise program affects body composition but not weight in postmenopausal women.
The aim of this study was to investigate the effect of a 12-month moderate-to-vigorous exercise program combining aerobic and muscle strength training on body composition among sedentary, postmenopausal women. A randomized controlled trial was conducted. A total of 189 sedentary postmenopausal women (age 50-69 y, body mass index 22-40 kg/m2) were randomly assigned to an exercise (n = 96) or a control group (n = 93). Study parameters measured at baseline, 4 months, and 12 months were as follows: body weight and body height (body mass index), waist and hip circumference (body fat distribution), and dual-energy x-ray absorptiometry (total body fat and lean mass). Differences in changes in study parameters between exercise and control group were examined with generalized estimating equations analysis. The exercise program did not result in significant effects on weight, body mass index, and hip circumference. The exercise group experienced a statistically significant greater loss in total body fat, both absolute (-0.33 kg) (borderline) as in a percentage (-0.43%) compared with the control group. In addition, lean mass increased significantly (+0.31 kg), whereas waist circumference (-0.57 cm) decreased significantly compared with the control group. We conclude that a 12-month exercise program combining aerobic and muscle strength training did not affect weight but positively influenced body composition of postmenopausal women. Affecting body fat distribution and waist circumference may have important health implications because it is an independent risk factor in obese but also in nonobese people. Therefore, this study gives further credence to efforts of public health and general practitioners aiming to increase physical activity levels of postmenopausal women.


[Infiltrating lobular carcinoma of the breast: clinical stage, radiological and histological features].
The characteristics of the infiltrating lobular carcinoma of the breast are not clear, this is caused by special histological and biological features. To identify the clinical stage and correlate it with the radiological and histological features of the disease to make an opportune diagnosis. From January 1st, 2003 to July 31st, 2004 we identified 68 cases of infiltrating lobular carcinoma at the Luis Castelazo Ayala Hospital. The cases were classified by clinical, radiological and histological features. The results were expressed in media and standard deviation. The difference between the mixed and pure lobular carcinoma types were compared by chi-square test. From 68 patients, 31 had mixed lobular carcinoma, 20 pure, 12 were eliminated because the patient's records were not found and five because the definitive histological report was not lobular carcinoma. The median age for both groups was 50 years old. The most affected region was the upper one (> 80%). The hormonal receptor was more frequently expressed in the pure subtype (p < 0.04), this latter was associated to a delayed diagnosis (p < 0.03). Mammography was considered useful for diagnosis. The histological behavior of lobular carcinoma is related to non clear characteristics and to delayed diagnosis; however, with a careful evaluation these features can be recognized and make possible an opportune mammography.


Using Machine Learning to Identify Metabolomic Signatures of Pediatric Chronic Kidney Disease Etiology.
Untargeted plasma metabolomic profiling combined with machine learning (ML) may lead to discovery of metabolic profiles that inform our understanding of pediatric CKD causes. We sought to identify metabolomic signatures in pediatric CKD based on diagnosis: FSGS, obstructive uropathy (OU), aplasia/dysplasia/hypoplasia (A/D/H), and reflux nephropathy (RN). Untargeted metabolomic quantification (GC-MS/LC-MS, Metabolon) was performed on plasma from 702 Chronic Kidney Disease in Children study participants (n: FSGS=63, OU=122, A/D/H=109, and RN=86). Lasso regression was used for feature selection, adjusting for clinical covariates. Four methods were then applied to stratify significance: logistic regression, support vector machine, random forest, and extreme gradient boosting. ML training was performed on 80% total cohort subsets and validated on 20% holdout subsets. Important features were selected based on being significant in at least two of the four modeling approaches. We additionally performed pathway enrichment analysis to identify metabolic subpathways associated with CKD cause. ML models were evaluated on holdout subsets with receiver-operator and precision-recall area-under-the-curve, F1 score, and Matthews correlation coefficient. ML models outperformed no-skill prediction. Metabolomic profiles were identified based on cause. FSGS was associated with the sphingomyelin-ceramide axis. FSGS was also associated with individual plasmalogen metabolites and the subpathway. OU was associated with gut microbiome-derived histidine metabolites. ML models identified metabolomic signatures based on CKD cause. Using ML techniques in conjunction with traditional biostatistics, we demonstrated that sphingomyelin-ceramide and plasmalogen dysmetabolism are associated with FSGS and that gut microbiome-derived histidine metabolites are associated with OU.


Pneumothorax-Time for New Guidelines?
Pneumothorax is a common pathology, but optimal management strategies are not yet defined. There are significant differences in international guidelines and therefore variation in clinical practice.There is increasing interest in pneumothorax research, particularly primary spontaneous pneumothorax (PSP), with evidence of lung abnormalities in this group without clinically apparent lung disease and recently completed clinical trials aiming to optimize management. The most robust evidence base is that of the equivalence of needle aspiration and chest tube insertion for initial management of PSP; although, patients with secondary spontaneous pneumothorax may also benefit. A convincing case for surgical intervention or thoracoscopy and talc poudrage to prevent recurrence at first episode in PSP has yet to be made. Clinicians should be vigilant for PSP being the first manifestation of a systemic disease, and should have a low threshold for onward referral. Time to change guidelines? First, chest tube drainage and hospitalization without recurrence prevention should no longer be standard treatment, as this has no advantage over the less invasive manual aspiration, which moreover can be performed on an outpatient basis in an important number of patients. The results of recent trials in conservative and ambulatory management are eagerly awaited. Second, shared decision-making should become more important with the well-informed patient, who may want to avoid a 1 in 3 recurrence rate and therefore will have the possibility to choose treatment including recurrence prevention even after the first episode of PSP. Third, surgical research should urgently make clear if the current practice of resection of emphysema-like changes is routinely necessary, alongside pleurodesis. Future studies should utilize risk stratification by clinical and radiological parameters (e.g., high-resolution computed tomography scanning and digital air leak monitoring) to predict short- and long-term outcomes, and hence personalize management.


[A comparative analysis of immunoenzyme methods of determining bradykinin].
Various modifications of enzyme immunoassay (EIA) for measuring kallikrein-kinin system components (bradykinin, kininogen, kallikrein) are considered. Bradykinin assays are analyzed in detail: double antibodies and competitive solid-phase EIA techniques with the use of various enzymic labels. Special attention is paid to the solid-phase variant in polymeric microplates, the simplest technique that holds good promise for clinical application. The authors have used this method for bradykinin measurements and tried to improve its sensitivity.


Transforming Lindblad Equations into Systems of Real-Valued Linear Equations: Performance Optimization and Parallelization of an Algorithm.
With their constantly increasing peak performance and memory capacity, modern supercomputers offer new perspectives on numerical studies of open many-body quantum systems. These systems are often modeled by using Markovian quantum master equations describing the evolution of the system density operators. In this paper, we address master equations of the Lindblad form, which are a popular theoretical tools in quantum optics, cavity quantum electrodynamics, and optomechanics. By using the generalized Gell-Mann matrices as a basis, any Lindblad equation can be transformed into a system of ordinary differential equations with real coefficients. Recently, we presented an implementation of the transformation with the computational complexity, scaling as O(N5logN) for dense Lindbaldians and O(N3logN) for sparse ones. However, infeasible memory costs remains a serious obstacle on the way to large models. Here, we present a parallel cluster-based implementation of the algorithm and demonstrate that it allows us to integrate a sparse Lindbladian model of the dimension N=2000 and a dense random Lindbladian model of the dimension N=200 by using 25 nodes with 64 GB RAM per node.


[A determination of iodides in salts: a validation of methods].
The studies reported were aimed at finding a simple analytical method enabling quantitative determination of iodide in table iodised salt and in therapeutic iodide-bromide salts. The analytical procedure proposed is a modification of spectrophotometric method recommended in the Polish Standards. The method based on the reaction of iodide oxidation by sodium nitrite was validated by determining its precision, accuracy and linearity. Statistical analysis has shown that the coefficient of variation varies between 2.73 and 4.82%, recovery is from 99.17 to 101.83% and falls within the confidence interval for the mean recovery at the assumed level of significance. The method can be used for controlling the technology of table salt iodisation.


Financial stress and mental health among higher education students in the UK up to 2018: rapid review of evidence.
In the United Kingdom and many other countries, debt accrued during higher education has increased substantially in recent decades. The prevalence of common mental health problems has also increased alongside these changes. However, it is as yet unclear whether there is an association between financial stress and mental health among higher education students. We conducted a rapid review of the peer-reviewed scientific literature. Eligible studies were English-language publications testing the association between any indicator of financial stress and mental health among higher education students in the UK. Papers were located through a systematic search of PsychINFO, PubMed and Embase up to November 2018. The search strategy yielded 1272 studies-9 met the inclusion criteria. A further two were identified through hand-searching. The median sample size was 408. Only three of seven studies found an association between higher debt and worse mental health. There was a consistent cross-sectional relationship between worse mental health and both experience of financial difficulties (seven of seven studies) and debt worry/financial concern (four of five studies), though longitudinal evidence was mixed and limited to six studies. Among higher education students in the UK, there is little evidence that the amount of debt is associated with mental health. However, more subjective measures of increased financial stress were more consistently associated with worse mental health outcomes. Nevertheless, the identified evidence was judged to be weak; further research is required to examine whether links between financial stress and mental health outcomes are robust and causal in nature.


Probiotics for the Prevention of Antibiotic-Associated Diarrhea in Outpatients-A Systematic Review and Meta-Analysis.
A common adverse effect of antibiotic use is diarrhea. Probiotics are living microorganisms, which, upon oral ingestion, may prevent antibiotic-associated diarrhea (AAD) by the normalization of an unbalanced gastrointestinal flora. The objective of this systematic review was to assess the benefits and harms of probiotics used for the prevention of AAD in an outpatient setting. A search of the PubMed database was conducted and yielded a total of 17 RCTs with 3631 participants to be included in the review. A meta-analysis was conducted for the primary outcome: the incidence of AAD. The pooled results found that AAD was present in 8.0% of the probiotic group compared to 17.7% in the control group (RR 0.49, 95% CI 0.36 to 0.66; I² = 58%), and the species-specific results were similar regarding the probiotic strains L. rhamnosus GG and S. boulardii. However, the overall quality of the included studies was moderate. A meta-analysis of the ten trials reporting adverse events demonstrated no statistically significant differences in the incidence of adverse events between the intervention and control group (RD 0.00, 95% CI -0.02 to 0.02, 2.363 participants). The results suggests that probiotic use may be beneficial in the prevention of AAD among outpatients. Furthermore, the use of probiotics appears safe.


Inversion of the two segments of the herpes simplex virus genome in intertypic recombinants.
We have analysed by restriction site mapping the structures of the termini and L-S joint in several HSV-1/HSV-2 intertypic recombinants, including Bx1(28-1), the virion DNA of which has a marked overabundance of one orientation of the L segment, and subclones of Bx1(28-1). All recombinants with both orientations of L present in equal amounts contain TRL and IRL regions derived at least in part from the same parent (HSV-1 or HSV-2) as a result of previously undetected crossovers in these regions. Recombinants with a predominance of one orientation of L have TRL and IRL regions derived from different parents. Homology between a sequences alone at the L terminus and L-S joint is sufficient for normal inversion of L. Analysis of another recombinant, RE4, which fails to invert normally in both L and S, suggests that normal inversion of S is dependent upon the presence of TRS and IRS regions derived at least in part from the same parent. We conclude that segment inversion specifically depends upon the a sequence, that the process of DNA replication and maturation does not necessarily produce molecules with identical a sequences, and that direct ligation of termini may occur during DNA replication.


Oviposition of diamondback moth in the presence and absence of a novel host plant.
The diamondback moth (DBM, Plutella xylostella L. (Lepidoptera: Plutellidae)) consumes a wide variety of brassicaceous host plants and is a common pest of crucifer crops worldwide. A highly unusual infestation of a sugar pea crop was recorded in Kenya in 1999, which persisted for two consecutive years. A strain (DBM-P) from this population was established in the laboratory and is the only one of several strains tested that can complete larval development on sugar peas. The oviposition acceptance and preference of the DBM-P strain was assessed in the presence of cabbage plants, sugar pea plants or both, in comparison to another strain (DBM-Cj) that was collected from cabbage and is unable to grow on pea plants. As expected, DBM-Cj females preferred to oviposit on cabbage plants. Surprisingly, DBM-P females also laid most eggs on cabbage and very few on peas. However, they laid significantly more eggs on the cabbage plant when pea plants were present. Our findings suggest that DBM-P manifested the initial stages of an evolutionary host range expansion, which is incomplete due to lack of oviposition fidelity on pea plants.


The relationship between hegemonic norms of masculinity and men's conceptualization of sexually coercive acts by women in South Africa.
While sexual abuse against women and girls in South Africa has generated much deserved attention, the awareness of men's experiences of sexual coercion is limited, and often restricted to a homosexual context. This article illuminates men's experiences of pressurized sex in a heterosexual context, which were revealed in a broader men's sexual health study. Fifty sexual history narrative interviews were conducted with men purposively sampled from three age categories: (18-24, 25-54, and 55+), a wide range of cultural and racial backgrounds, and in urban and rural sites across five provinces in South Africa. Narrative interviews began with accounts of early knowledge of sex and sexual experimentation and explored the range of sexual relationships and experiences through adulthood. The narratives privileged the diversity of men's conceptualizations of and the impact of reportedly sexually coercive experiences by women. Many men described feeling unready for their first sexual experiences but pressured to do so by their peers and female partners, who were often older. There were also some instances of sexual coercion by women against men, some of which would constitute a criminal offense in South Africa. Due to the pressure for men to always be responsive to women's sexual desires, these experiences were often not framed as sexual coercion. Nevertheless, for many of these men, such experiences were uncomfortable and unrewarding. Men's negative responses to such experiences appeared to be linked to the fact that they did not fit social stereotypes of masculine sexuality as being initiative and dominant. Such coercive experiences could influence men's sexual risk-taking, including their use of sexual coercion against women. Research on sexual abuse should not be limited to male against male sexual abuse, but needs to explore the meanings and experiences associated with reported coercion against men by women to more comprehensively prevent and respond to sexual violence.


An open-label, randomized trial indicates that everolimus with tacrolimus or cyclosporine is comparable to standard immunosuppression in de novo kidney transplant patients.
This is a randomized trial (ATHENA study) in de novo kidney transplant patients to compare everolimus versus mycophenolic acid (MPA) with similar tacrolimus exposure in both groups, or everolimus with concomitant tacrolimus or cyclosporine (CsA), in an unselected population. In this 12-month, multicenter, open-label study, de novo kidney transplant recipients were randomized to everolimus with tacrolimus (EVR/TAC), everolimus with CsA (EVR/CsA) or MPA with tacrolimus (MPA/TAC), with similar tacrolimus exposure in both groups. Non-inferiority of the primary end point (estimated glomerular filtration rate [eGFR] at month 12), assessed in the per-protocol population of 338 patients, was not shown for EVR/TAC or EVR/CsA versus MPA/TAC. In 123 patients with TAC levels within the protocol-specified range, eGFR outcomes were comparable between groups. The mean increase in eGFR during months 1 to 12 post-transplant, analyzed post hoc, was similar with EVR/TAC or EVR/CsA versus MPA/TAC. The incidence of treatment failure (biopsy proven acute rejection, graft loss or death) was not significant for EVR/TAC but significant for EVR/CsA versus MPA/TAC. Most biopsy-proven acute rejection events in this study were graded mild (BANFF IA). There were no differences in proteinuria between groups. Cytomegalovirus and BK virus infection were significantly more frequent with MPA/TAC. Thus, everolimus with TAC or CsA showed comparable efficacy to MPA/TAC in de novo kidney transplant patients. Non-inferiority of renal function, when pre-specified, was not shown, but the mean increase in eGFR from month 1 to 12 was comparable to MPA/TAC.


Stiffness of the inferior oblique neurofibrovascular bundle.
To assess the mechanical ability of the inferior oblique neurofibrovascular bundle (NFVB) to act as an ancillary origin for the inferior oblique muscle after anterior transposition. Stress-strain relations and Young's modulus of elasticity, a measure of tissue stiffness, were determined for the NFVB in vitro, in situ, and in vivo in dynamic and static conditions. For comparison, similar studies were performed in vitro on the superior oblique tendon (SOT). Young's moduli for NFVB in situ (6.3 MPa [megapascals]) and in vivo (11.8 MPa) were approximately 2 and 4 times greater (P < 0.05), respectively, than those of isolated NFVB in vitro at 5% to 10% dynamic strain (3 MPa). In dynamic conditions, Young's moduli in vitro for the NFVB and the SOT were similar. The NFVB is a biomaterial that has stiffness properties similar to the SOT. Within the range of forces typical of normal eye movements (79 to 393 mN), the NFVB alone can tolerate forces of 98 mN at 0% to 10% strain and 393 mN at 15% to 20% strain, based on dynamic in vitro analysis. The greater measured stiffness in situ and in vivo suggest that the NFVB in the intact orbit potentially has a resting strain of 15% to 20%, and additional tissues in parallel with the NFVB also contribute to total stiffness. These data support the hypothesis that the NFVB, acting alone or in concert with adjacent orbital tissues, may form an ancillary origin for the inferior oblique muscle after anterior transposition.


Challenges for Targeting SARS-CoV-2 Proteases as a Therapeutic Strategy for COVID-19.
Two proteases produced by the SARS-CoV-2 virus, the main protease and papain-like protease, are essential for viral replication and have become the focus of drug development programs for treatment of COVID-19. We screened a highly focused library of compounds containing covalent warheads designed to target cysteine proteases to identify new lead scaffolds for both Mpro and PLpro proteases. These efforts identified a small number of hits for the Mpro protease and no viable hits for the PLpro protease. Of the Mpro hits identified as inhibitors of the purified recombinant protease, only two compounds inhibited viral infectivity in cellular infection assays. However, we observed a substantial drop in antiviral potency upon expression of TMPRSS2, a transmembrane serine protease that acts in an alternative viral entry pathway to the lysosomal cathepsins. This loss of potency is explained by the fact that our lead Mpro inhibitors are also potent inhibitors of host cell cysteine cathepsins. To determine if this is a general property of Mpro inhibitors, we evaluated several recently reported compounds and found that they are also effective inhibitors of purified human cathepsins L and B and showed similar loss in activity in cells expressing TMPRSS2. Our results highlight the challenges of targeting Mpro and PLpro proteases and demonstrate the need to carefully assess selectivity of SARS-CoV-2 protease inhibitors to prevent clinical advancement of compounds that function through inhibition of a redundant viral entry pathway.


Platelet function testing in pediatric patients.
Platelets play a key role in primary hemostasis and are also intricately linked to secondary hemostasis. Investigation of platelet function in children, especially in neonates, is seriously challenged by the volumes required to perform the majority of platelet function tests and due to the lack of standardization of these tests for use in children. Areas covered: The present review summarizes developmental hemostasis with a focus on the differences in platelet adhesion, activation and aggregation, between preterm neonates, full-term neonates, during childhood and in adults. Some of the most widely used platelet function tests are presented, including novel tests requiring only a small blood volume. Expert commentary: Currently available platelet function tests are limited as regards to investigation of neonates due to difficulties in obtaining adequate blood volume, poor standardization, lack of reference intervals for neonates and children, and an incomplete understanding of the functional phenotype of neonatal platelets, especially preterm neonatal platelets.


The Impact of Machine Learning on 2D/3D Registration for Image-Guided Interventions: A Systematic Review and Perspective.
Image-based navigation is widely considered the next frontier of minimally invasive surgery. It is believed that image-based navigation will increase the access to reproducible, safe, and high-precision surgery as it may then be performed at acceptable costs and effort. This is because image-based techniques avoid the need of specialized equipment and seamlessly integrate with contemporary workflows. Furthermore, it is expected that image-based navigation techniques will play a major role in enabling mixed reality environments, as well as autonomous and robot-assisted workflows. A critical component of image guidance is 2D/3D registration, a technique to estimate the spatial relationships between 3D structures, e.g., preoperative volumetric imagery or models of surgical instruments, and 2D images thereof, such as intraoperative X-ray fluoroscopy or endoscopy. While image-based 2D/3D registration is a mature technique, its transition from the bench to the bedside has been restrained by well-known challenges, including brittleness with respect to optimization objective, hyperparameter selection, and initialization, difficulties in dealing with inconsistencies or multiple objects, and limited single-view performance. One reason these challenges persist today is that analytical solutions are likely inadequate considering the complexity, variability, and high-dimensionality of generic 2D/3D registration problems. The recent advent of machine learning-based approaches to imaging problems that, rather than specifying the desired functional mapping, approximate it using highly expressive parametric models holds promise for solving some of the notorious challenges in 2D/3D registration. In this manuscript, we review the impact of machine learning on 2D/3D registration to systematically summarize the recent advances made by introduction of this novel technology. Grounded in these insights, we then offer our perspective on the most pressing needs, significant open problems, and possible next steps.


Photobleaching of melanosomes from retinal pigment epithelium: I. Effects on protein oxidation.
Melanin in the long-lived melanosomes of the retinal pigment epithelium (RPE) may undergo photobleaching with aging, which appears to diminish the antioxidant function of melanin and could make photobleached melanosomes less efficient in protecting biomolecules from oxidative modification. Here we analyzed whether photobleaching of melanosomes affects their ability to modify the oxidation state of nearby protein. As conventional methods developed to study soluble antioxidants are not well suited for analysis of granules such as melanosomes, we developed a new analytic method to focus on particle surfaces that involves experimentally coating granules with the cytoskeletal protein beta-actin to serve as a reporter for local protein oxidation. Isolated porcine RPE melanosomes were photobleached with visible light to simulate aging, then photobleached melanosomes, untreated melanosomes and control particles (black latex beads) were actin coated and illuminated in a photosensitized cell free system. Protein was re-stripped from particles and analyzed for carbonylation by Western blotting. Quantitative densitometry showed no reproducible differences for protein associated with untreated melanosomes when compared with control particles. Melanin has both anti- and pro-oxidant functions when light irradiated, but neither of these functions predominated in the protein oxidation assay when untreated melanosomes were used. However, protein extracted from photobleached melanosomes showed markedly increased carbonylation, both of associated actin and of endogenous melanosomal protein(s), and the effect increased with extent of granule photobleaching. Photobleaching of RPE melanosomes therefore changes the oxidation state of protein endogenous to the organelle and reduces the ability of the granule to modify the oxidation of exogenous protein near the particle surface. The results support the growing body of evidence that photobleaching of RPE melanosomes, which is believed to occur with aging, changes the physicochemical properties of the organelle and reduces the likelihood that the granules perform an antioxidant function.


[The morphological changes in the eye tissues of rabbits in transconjunctival-scleral cyclocoagulation by the radiation of a krypton laser operating continuously].
The paper describes a morphologic picture of changes in eye tissues of rabbits in transconjunctival scleral action on the ciliary body by means of krypton laser (wavelength--647.1 nm) working in a continuous regime. The applications were located 1.0-1.5 mm from the limbus with energy--0.8--3.0 Wt, exposition time--2.0 sec., spot diameter--1000 mcm. The degree of necrosis of ciliary epithelium and atrophy of ciliary processes are directly dependent on the dose of action. Thus, the greatest part of laser radiation with 647.1 nm wavelength is absorbed at the level of the ciliary body. In perspective, this can lead to development of new methods for treatment of some eye diseases, such as glaucoma, tumors of the uveal tract, etc.


Identification of an osmo-dependent and an osmo-independent choline transporter in Acinetobacter baylyi: implications in osmostress protection and metabolic adaptation.
Members of the genus Acinetobacter are well known for their metabolic versatility that allows them to adapt to different ecological niches. In previous studies, we have demonstrated that Acinetobacter baylyi ADP1 can cope with high salinities by uptake and accumulation of the well-known compatible solute glycine betaine. Here, we demonstrate that addition of choline restores growth at high salinities. We further show that choline was actively taken up by the cells and converted to glycine betaine. Uptake of choline was induced by high salinity and the presence of choline in the growth medium. At high salinities, glycine betaine was accumulated in the cells whereas in the absence of osmotic stress it was exported. Inspection of the genome sequence followed by mutant studies led to the identification of two genes encoding secondary transporters (BetT1 and BetT2) of the betaine-choline-carnitine transporter (BCCT) family. The BetT1 transporter lacks an extended C-terminal domain usually found in osmoregulated choline BCCTs. BetT1 was found to facilitate osmolarity-independent choline transport most likely by a uniport mechanism. We propose that BetT1 does not primarily function in osmoadaptation but might play a role in metabolic adaptation to choline-rich environments.


Swelling kinetics of microgels embedded in a polyacrylamide hydrogel matrix.
Composite hydrogels--macroscopic hydrogels with embedded microgel particles--are expected to respond to external stimuli quickly because microgels swell much faster than bulky gels. In this work, the kinetics of the pH-induced swelling of a composite hydrogel are studied using turbidity measurements. The embedded microgel is a pH- and thermosensitive poly(N-isopropylacrylamide-co-acrylic acid) microgel and the hydrogel matrix is polyacrylamide. A rapid pH-induced swelling of the embedded microgel particles is observed, confirming that composite hydrogels respond faster than ordinary hydrogels. However, compared with the free microgels, the swelling of the embedded microgel is much slower. Diffusion of OH(-) into the composite hydrogel film is identified as the main reason for the slow swelling of the embedded microgel particles, as the time of the pH-induced swelling of this film is comparable to that of OH(-) diffusion into the film. The composition of the hydrogel matrix does not significantly change the characteristic swelling time of the composite hydrogel film. However, the swelling pattern of the film changes with composition of the hydrogel matrix.


Gastruloid-derived primordial germ cell-like cells develop dynamically within integrated tissues.
Primordial germ cells (PGCs) are the early embryonic precursors of gametes - sperm and egg cells. PGC-like cells (PGCLCs) can currently be derived in vitro from pluripotent cells exposed to signalling cocktails and aggregated into large embryonic bodies, but these do not recapitulate the native embryonic environment during PGC formation. Here, we show that mouse gastruloids, a three-dimensional in vitro model of gastrulation, contain a population of gastruloid-derived PGCLCs (Gld-PGCLCs) that resemble early PGCs in vivo. Importantly, the conserved organisation of mouse gastruloids leads to coordinated spatial and temporal localisation of Gld-PGCLCs relative to surrounding somatic cells, even in the absence of specific exogenous PGC-specific signalling or extra-embryonic tissues. In gastruloids, self-organised interactions between cells and tissues, including the endodermal epithelium, enables the specification and subsequent maturation of a pool of Gld-PGCLCs. As such, mouse gastruloids represent a new source of PGCLCs in vitro and, owing to their inherent co-development, serve as a novel model to study the dynamics of PGC development within integrated tissue environments.


Dietary aspects related to health and obesity in Williams syndrome, Down syndrome, and Prader-Willi syndrome.
Dietary aspects that might contribute to development of obesity and secondary conditions are not well documented in genetic subgroups associated with intellectual disability. To describe the intake frequencies of selected foods in participants with Prader-Willi syndrome (PWS), Down syndrome (DS), and Williams syndrome (WS), and investigate the association with body mass index (BMI). To explore food-related autonomy and intake frequencies among persons with DS in different living arrangements. Self-reported intake frequencies and measurement of plasma carotenoids and erythrocyte content of omega-3 fatty acids (FAs) were investigated in persons aged 16-42 years, with WS (n=21), DS (n=40), and PWS (n=20). A larger proportion of participants with PWS showed high-frequency intake of fruits (p=0.012) and vegetables (p=0.004), and had higher plasma carotenoids (p<0.001) compared to participants with DS and WS. Furthermore, a larger proportion of participants with WS were low-frequency consumers of fish (p=0.005), less likely to use omega-3 FA supplements (p=0.023), and had reduced erythrocyte concentrations of long-chain omega-3 FAs (p<0.001), compared to participants with PWS and DS. In DS, BMI was negatively associated with plasma carotenoids. Increased proportions of participants living in communities showed high-frequency intake of precooked meals (p=0.030), and a tendency toward high-frequency consumption of soft drinks (p=0.079), when compared to peers living with relatives. Participants in community residences were also more likely to participate frequently in food-related decisions and preparations. Persons with WS had a less-favorable dietary pattern when compared to persons with PWS. A larger proportion of persons living in communities frequently consumed precooked meals and showed a tendency of high-frequency soft drink consumption. Otherwise, their intake frequencies of the investigated foods were similar to those living with relatives, but they participated more frequently in decisions and preparations of foods.


Glucose tolerance and antioxidant activity of spent brewer's yeast hydrolysate with a high content of Cyclo-His-Pro (CHP).
To elevate the Cyclo-His-Pro (CHP) content in yeast, the yeast hydrolysate that was obtained from enzymatic hydrolysis was subjected to various treatments. Flavourzyme-treated hydrolysate showed the highest CHP content (674.0 μg/g) among the various proteases treatments. Ultrafiltration was selected as the best method for concentrating CHP in yeast hydrolysate, based on the yields and CHP contents. In addition, we evaluated the radical scavenge and glucose tolerance of yeast hydrolysate with a high content of CHP. Yeast hydrolysate showed intense scavenging abilities of both 1, 1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) radicals. The IC(50) values of yeast hydrolysate on DPPH and ABTS radicals were 1.9 and 0.9 mg/mL, respectively. There were significant differences in glucose level between the diabetes-control and yeast hydrolysate group at 30, 60, 90, and 120 min after injection in a type 1 diabetes model (P < 0.01). Also, there were significant differences in blood glucose levels between the 2 groups at 30, 60, and 100 min after injection in the type 2 diabetes group (P < 0.05). Therefore, it is possible to use the yeast hydrolysate with high levels of CHP as an antioxidative and/or antidiabetic material for the preparation of functional foods. This study tried to develop a material containing a high content of CHP using yeast for possible applications of this cyclic dipeptide in the therapy of metabolic disorders. The yeast hydrolysate prepared with Flavourzyme showed a high level of CHP. The hydrolysate with a high content of CHP showed high levels of radical scavenging activities and oral glucose tolerance activity. Therefore, it is possible to use the yeast hydrolysate with high levels of CHP as an antioxidative and/or antidiabetic material for the preparation of functional foods.


Plant hormones increase efficiency of reprogramming mouse somatic cells to induced pluripotent stem cells and reduce tumorigenicity.
Reprogramming of somatic cells into induced pluripotent stem (iPS) cells by defined pluripotency and self-renewal factors has taken stem cell technology to the forefront of regenerative medicine. However, a number of challenges remain in the field including efficient protocols and the threat of cancer. Reprogramming of plant somatic cells to plant embryonic stem cells using a combination of two plant hormones was discovered in 1957 and has been a routine university laboratory practical for over 30 years. The plant hormones responsible for cell reprogramming to pluripotency, indole-3-acetic acid (IAA) and isopentenyl adenosine (IPA), are present in human cells, leading to the exciting possibility that plant hormones might reprogram mammalian cells without genetic factors. We found that plant hormones on their own could not reprogram mammalian cells but increase the efficiency of the early formation of iPS cells combined with three defined genetic factors during the first 3 weeks of reprogramming by accelerating the cell cycle and regulating pluripotency genes. Moreover, the cytokinin IPA, a known human anticancer agent, reduced the threat of cancer of iPS cell in vitro by regulating key cancer and stem cell-related genes, most notably c-Myc and Igf-1. In conclusion, the plant hormones, auxin and cytokinin, are new small chemicals useful for enhancing early reprogramming efficiency of mammalian cells and reducing the threat of cancer from iPS cells. These findings suggest a novel role for plant hormones in the biology of mammalian cell plasticity.


Do ovarian steroid hormones control the resumption of embryonic growth following the period of diapause in roe deer (Capreolus capreolus)?
Embryonic diapause in the European roe deer includes a period of five months from August to December in which embryonic development is extremely decelerated. Following exit from diapause, the embryo rapidly elongates and subsequently implants. In diapausing carnivores and marsupials, resumption of embryonic growth is regulated by ovarian steroid hormones. In the roe deer, the role of steroid hormones is not known to date. In the present study, progesterone (P4), estradiol-17β (E2) and total estrogens (Etot) were determined in blood plasma and endometrium of roe deer shot in the course of regular huntings between September and December. Steroid hormone concentrations were correlated to the corresponding size of the embryo derived from ex vivo uterine flushing and to the date of sampling. The mean plasma concentrations of P4 (5.4 ± 0.2 ng/ml, mean ± SE, N = 87), E2 (24.3 ± 2.6 pg/ml, N = 86) and Etot (21.7 ± 2.6 pg/ml, N = 78) remained constant over the sampling period and were not correlated to embryonic size. Likewise, endometrial concentrations of P4 (66.1 ± 6.5 ng/g), E2 (284.0 ± 24.43 pg/g) and, Etot (440.9 ± 24.43 pg/g) showed no changes over time [corrected]. Therefore, it was concluded that ovarian steroid hormones do not play a determining role in resumption of embryonic growth following the period of diapause in the roe deer.


Evaluation of orthodontic mini-implant anchorage in premolar extraction therapy in adolescents.
The purpose of this study was to determine the success rate, positional stability, and patient evaluation of orthodontic mini-implants (OMIs). Thirteen patients (8 girls, 5 boys; average age, 14 years 10 months) were treated with 82 OMIs measuring 1.6 mm in diameter and 6 mm in length placed in the buccal alveoli (1 unloaded OMI and 1 loaded OMI per quadrant). The right or left side of each arch was randomly selected for immediate loading with up to 250 g of direct force; the contralateral side was loaded 3 to 5 weeks later. Serial impressions, clinical observations, and orthodontic maintenance were performed until adequate space closure was achieved. The overall OMI success rate was 70.73%. As calculated with a mixed-model analysis, there was no statistically significant difference between the success rates of immediately loaded OMIs (80.0%) and delayed loaded OMIs (80.95%). The combined success rate for loaded OMIs (80.49%) was significantly higher than that of unloaded OMIs (60.98%). Patients' motivation for OMI treatment was primarily the desire to avoid headgear. Using a 100-mm visual analog scale, the patients indicated average scores of 54.77 for the amount of pain during OMI placement and 27.10 for the amount of pain during OMI removal. OMIs are a predictable, effective, and well-tolerated anchorage source for adolescents. Neither the timing of force application nor the force itself precipitated failure of the OMIs. Orthodontic forces can be applied immediately to OMIs. Various anatomic and behavioral conditions unique to adolescents and a clinical learning curve can affect the success rate of OMIs.


Marginally compact phase and ordered ground states in a model polymer with side spheres.
We present the results of a quantitative study of the phase behavior of a model polymer chain with side spheres using two independent computer simulation techniques. We find that the mere addition of side spheres results in key modifications of standard polymer behavior. One obtains a marginally compact phase at low temperatures; the structures in this phase are reduced in dimensionality and are ordered, they include strands assembled into sheets and a variety of helices, and at least one of the transitions on lowering the temperature to access these ordered states is found to be first order. Our model serves to partially bridge conventional polymer phases with biomolecular phases.


Modeling the environmental fate of perfluorooctanoic acid and perfluorooctanoate: An investigation of the role of individual species partitioning.
A multimedia multi-species environmental fate model was developed for the conjugate pair perfluorooctanoic acid (PFOA):perfluorooctanoate (PFO). The model allows assessment of the relative contribution of each individual species, in equilibrium with each other, to the overall environmental movement of the pair. The Lake Ontario (Canada/USA) watershed system was selected for this investigation and is simulated in a single-region, seven-compartment model, including a water surface microlayer, and aqueous aerosol generation and redeposition. Results indicate that in the equilibrated presence of both PFOA and PFO, the environmental fate of the pair can be accomplished by consideration of the physical properties of the neutral acid, which govern the intermedia distribution of the pair, coupled with processes of media advection, such as air or water flow. The role of the anion, while the most populous species in the aqueous phase, appears merely to be as a source of the neutral acid for subsequent partitioning. Thus, when only the bulk aqueous phase anion concentrations are of interest a multimedia fate model is not required because these concentrations are largely predictable from the magnitude of emissions to and the advection of the phase. With neutral species partitioning, all local field measurement concentrations of the conjugate pair, PFO(A), are explained by the model to within approximately an order of magnitude, with the exception of lake sediment solids. Model results indicate that bulk aqueous phase PFO acts as a net source for PFOA to the atmosphere, where it may be subject to long-range transport (LRT). Initial calculations suggest an atmospheric LRT potential for PFO(A) of thousands of kilometers, rendering it comparable to hexachlorobenzene.


Similar hibernation physiology in bats across broad geographic ranges.
Species with broad geographic ranges may experience varied environmental conditions throughout their range leading to local adaptation. Variation among populations reflects potential adaptability or plasticity, with implications for populations impacted by disease, climate change, and other anthropogenic influences. However, behavior may counteract divergent selection among populations. We studied intraspecific variation in hibernation physiology of Myotis lucifugus (little brown myotis) and Corynorhinus townsendii (Townsend's big-eared bat), two species of bats with large geographic ranges. We studied M. lucifugus at three hibernacula which spanned a latitudinal gradient of 1500 km, and C. townsendii from 6 hibernacula spread across 1200 km latitude and 1200 km longitude. We found no difference in torpid metabolic rate among populations of either species, nor was there a difference in the effect of ambient temperature among sites. Evaporative water loss was similar among populations of both species, with the exception of one C. townsendii pairwise site difference and one M. lucifugus site that differed from the others. We suggest the general lack of geographic variation is a consequence of behavioral microhabitat selection. As volant animals, bats can travel relatively long distances in search of preferred microclimates for hibernation. Despite dramatic macroclimate differences among populations, hibernating bats are able to find preferred microclimate conditions within their range, resulting in similar selection pressures among populations spread across wide geographic ranges.


Dietary Patterns are Associated with Leukocyte LINE-1 Methylation in Women: A Cross-Sectional Study in Southern Italy.
Bioactive food compounds have different effects on global DNA methylation, an epigenetic mechanism associated with chromosomal stability and genome function. Since the diet is characterized by a mixture of foods, we aimed to identify dietary patterns in women, and to evaluate their association with long interspersed nuclear elements (LINE-1) methylation, a surrogate marker of global DNA methylation. We conducted an observational cross-sectional study of 349 women from Southern Italy, with no history of severe diseases. Dietary patterns were derived by food frequency questionnaire and principal component analysis. LINE-1 methylation of leukocyte DNA was assessed by pyrosequencing. We observed that intake of wholemeal bread, cereals, fish, fruit, raw and cooked vegetables, legumes, soup, potatoes, fries, rice, and pizza positively correlated with LINE-1 methylation levels. By contrast, vegetable oil negatively correlated with LINE-1 methylation levels. Next, we demonstrated that adherence to a prudent dietary pattern-characterized by high intake of potatoes, cooked and raw vegetables, legumes, soup and fish-was positively associated with LINE-1 methylation. In particular, women in the 3rd tertile exhibited higher LINE-1 methylation level than those in the 1st tertile (median = 66.7 %5mC; IQR = 4.67 %5mC vs. median = 63.1 %5mC; IQR = 12.3 %5mC; p < 0.001). Linear regression confirmed that women in the 3rd tertile had higher LINE-1 methylation than those in the 1st tertile (β = 0.022; SE = 0.003; p < 0.001), after adjusting for age, educational level, employment status, smoking status, use of folic acid supplement, total energy intake and body mass index. By contrast, no differences in LINE-1 methylation across tertiles of adherence to the Western dietary pattern were evident. Interestingly, women who exclusively adhered to the prudent dietary pattern had a higher average LINE-1 methylation level than those who exclusively or preferably adhered to the Western dietary pattern (β = 0.030; SE = 0.004; p < 0.001; β = 0.023; SE = 0.004; p < 0.001; respectively), or those with no preference for a specific dietary pattern (β = 0.013; SE = 0.004; p = 0.002). Our study suggested a remarkable link between diet and DNA methylation; however, further mechanistic studies should be encouraged to understand the causal relationship between dietary intake and DNA methylation.


Development of an upper extremity FES system for individuals with C4 tetraplegia.
The application of functional electrical stimulation (FES) to provide upper extremity function for individuals with C4 tetraplegia is under investigation. In this study, a FES system was designed that allowed one individual with complete C4 tetraplegia to coordinate stimulated lateral or palmar prehension with stimulated arm movements using contralateral shoulder position. The system consisted of percutaneous intramuscular electrodes implanted to muscles for hand grasp and release, supination, elbow flexion, and extension and arm adduction. Due to peripheral denervation, transposition and subsequent stimulation of the paralyzed latissimus dorsi muscle provided elbow flexion and transfer and stimulation of the paralyzed extensor carpi ulnaris muscle provided forearm supination. A suspended sling provided shoulder joint stability. The subject controlled stimulation proportionally using contralateral shoulder motion sensed by a position transducer. Control of stimulated hand grasp and release were coupled with stimulated arm motions so that hand-to-mouth activities could be accomplished with one motion of the contralateral shoulder. With this system, the subject was able to grasp a milk carton placed in the hand and lift and lower it from his mouth to drink from a short straw. He could also scoop a semisolid substance with an adapted spoon and lift and lower it from the work surface to his mouth to eat. The subject required assistance to place the milk carton or spoon in his hand and position the plate for scooping. Further investigation is needed to generate the necessary arm movements to make completion of these tasks possible without assistance and to expand the range of activities possible with the FES system.


The glycobiome of the rumen bacterium Butyrivibrio proteoclasticus B316(T) highlights adaptation to a polysaccharide-rich environment.
Determining the role of rumen microbes and their enzymes in plant polysaccharide breakdown is fundamental to understanding digestion and maximising productivity in ruminant animals. Butyrivibrio proteoclasticus B316(T) is a gram-positive, butyrate-forming rumen bacterium with a key role in plant polysaccharide degradation. The 4.4 Mb genome consists of 4 replicons; a chromosome, a chromid and two megaplasmids. The chromid is the smallest reported for all bacteria, and the first identified from the phylum Firmicutes. B316 devotes a large proportion of its genome to the breakdown and reassembly of complex polysaccharides and has a highly developed glycobiome when compared to other sequenced bacteria. The secretion of a range of polysaccharide-degrading enzymes which initiate the breakdown of pectin, starch and xylan, a subtilisin family protease active against plant proteins, and diverse intracellular enzymes to break down oligosaccharides constitute the degradative capability of this organism. A prominent feature of the genome is the presence of multiple gene clusters predicted to be involved in polysaccharide biosynthesis. Metabolic reconstruction reveals the absence of an identifiable gene for enolase, a conserved enzyme of the glycolytic pathway. To our knowledge this is the first report of an organism lacking an enolase. Our analysis of the B316 genome shows how one organism can contribute to the multi-organism complex that rapidly breaks down plant material in the rumen. It can be concluded that B316, and similar organisms with broad polysaccharide-degrading capability, are well suited to being early colonizers and degraders of plant polysaccharides in the rumen environment.


Nurse-led self-management group programme for patients with congestive heart failure: randomized controlled trial.
This paper reports on of the effects of the Chronic Disease Self-Management Programme on psychosocial attributes, self-care behaviour and quality of life among congestive heart failure patients who experienced slight to marked limitation of physical activity. Most self-management programmes for congestive heart failure patients emphasize the medical aspects of this chronic condition, without incorporating psychosocial aspects of self-management. The programme has been used with various patient groups, but its effectiveness with congestive heart failure patients when led by pairs of cardiac nurse specialists and peer leaders is unknown. A randomized controlled trial with 12 months of follow-up from start of the programme was conducted with 317 patients. Control group patients (n = 131) received usual care, consisting of regular outpatient checkups. Intervention group patients (n = 186) received usual care and participated in the six-week self-management programme. The programme teaches patients medical, social and emotional self-management skills. Twenty-one classes were conducted in six hospitals in the Netherlands, and data were collected between August 2004 and January 2007. Directly after the programme, statistically significant effects were found for cognitive symptom management (P < 0.001), self-care behaviour (P = 0.008) and cardiac-specific quality of life (P = 0.005). No effects were found at 6- and 12-month follow-up. Further research is necessary to study how long-term effectiveness of the programme with patients with congestive heart failure can be achieved, and how successful adaptations of the programme can be integrated into standard care.


Repeated dose toxicity and relative potency of 1,2,3,4,6,7-hexachloronaphthalene (PCN 66) 1,2,3,5,6,7-hexachloronaphthalene (PCN 67) compared to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) for induction of CYP1A1, CYP1A2 and thymic atrophy in female Harlan Sprague-Dawley rats.
In this study we assessed the relative toxicity and potency of the chlorinated naphthalenes 1,2,3,4,6,7-hexachloronaphthalene (PCN 66) and 1,2,3,5,6,7-hexachloronaphthalene (PCN 67) relative to that of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Chemicals were administered in corn oil:acetone (99:1) by gavage to female Harlan Sprague-Dawley rats at dosages of 0 (vehicle), 500, 1500, 5000, 50,000 and 500,000 ng/kg (PCN 66 and PCN 67) and 1, 3, 10, 100, and 300 ng/kg (TCDD) for 2 weeks. Histopathologic changes were observed in the thymus, liver and lung of TCDD treated animals and in the liver and thymus of PCN treated animals. Significant increases in CYP1A1 and CYP1A2 associated enzyme activity were observed in all animals exposed to TCDD, PCN 66 and PCN 67. Dose response modeling of CYP1A1, CYP1A2 and thymic atrophy gave ranges of estimated relative potencies, as compared to TCDD, of 0.0015-0.0072, for PCN 66 and 0.00029-0.00067 for PCN 67. Given that PCN 66 and PCN 67 exposure resulted in biochemical and histopathologic changes similar to that seen with TCDD, this suggests that they should be included in the WHO toxic equivalency factor (TEF) scheme, although the estimated relative potencies indicate that these hexachlorinated naphthalenes should not contribute greatly to the overall human body burden of dioxin-like activity.


Nonenhancing peritumoral hyperintense lesion on diffusion-weighted imaging in glioblastoma: a novel diagnostic and specific prognostic indicator.
OBJECTIVE Glioblastoma differentials include intracranial tumors, like malignant lymphomas and metastatic brain tumors with indiscernible radiological characteristics. The purpose of this study was to identify a distinct radiological feature for the preoperative differentiation of glioblastoma from its differentials, which include malignant lymphomas and metastatic brain tumors. METHODS Preoperative MR images, including diffusion-weighted imaging (DWI) studies (b = 1000 and 4000 sec/mm2), obtained in patients with newly diagnosed malignant tumor, were analyzed retrospectively after receiving approval from the institutional review board. Sixty-four patients with histologically confirmed glioblastoma, 32 patients with malignant lymphoma, and 46 patients with brain metastases were included. The presence of a nonenhancing peritumoral DWI high lesion (NePDHL, i.e., hyperintense lesion in a nonenhancing peritumoral area on DWI) was confirmed in both DWI sequences. Gray matter lesions were excluded. Lesions were termed "definite" if present within 3 cm of the hyperintense tumor border with a signal intensity ratio ≥ 30% when compared with the contralateral normal white matter in both sequences. Discriminant analysis between the histological diagnosis and the presence of Definite-NePDHL was performed, as well as Kaplan-Meier survival analysis incorporating the existence of Definite-NePDHL. RESULTS In 25% of glioblastoma patients, Definite-NePDHL was present, while it was conspicuously absent in patients with malignant lymphoma and metastatic brain tumors. The specificity and positive predictive value were 100%. In the glioblastoma subset, a higher preoperative Karnofsky Performance Scale score (p = 0.0028), high recursive partitioning analysis class (p = 0.0006), and total surgical removal (p = 0.0012) were associated with better median overall survival. Patients with Definite-NePDHL had significantly early local (p = 0.0467) and distant/dissemination recurrence (p < 0.0001) and poor prognosis (p = 0.0007). CONCLUSIONS The presence of Definite-NePDHL is very specific for glioblastoma and indicates poor prognosis. Definite-NePDHL is a significant indicator of early local and distant/dissemination recurrence in patients with glioblastoma. Studying peritumoral DWI and high-b-value DWI is useful for tumor differentiation.


[Effect of cyclic changes in culture conditions on the growth kinetics and physiological characteristics of yeasts].
Candida utilis, Saccharomyces cerevisiae and Candida scottii were used to study the effect of cyclic changes in the pH and pO2 within a range of 1 to 60 min on their growth kinetics and physiological properties. These changes were shown to increase the specific growth rate from 0.33 to 0.5-0.6 h-1 without decreasing the economic coefficient and the quantity of budding cells during 2-3 generations of the exponentially growing batch culture of C. utilis. Optimal conditions of cyclic changes in the pO2 (minutes) were found to increase the specific growth rate of C. scottii and S. cerevisiae. The authors discuss a hypothesis for the formation of intermediate products in the substrate oxidation in the course of pulse aeration by the yeasts during the aerobic stage and the utilization of the products at the anaerobic stage of cyclic regimes. The paper describes a mathematical model for the yeast growth under the nonsteady-state conditions of pH and pO2, which accounts for the formation and utilization of possible intermediate biosynthetic products within the studied time intervals.


[Behavioral pharmacology of a new antidepressant, lopramine].
The behavioral effects of lopramine [N-methyl-N-(4-chlorobenzoyl-methyl)-3-(10, 11-dihydro-5H-dibenz (b,f) azepin-5-yl) propylamine hydrochloride] were investigated in mice and rats and compared with those of amitriptyline and imipramine. Lopramine inhibited reserpine hypothermia and haloperidol catalepsy in mice and tetrabenazine ptosis in rats. In addition the drug potentiated the effects of methamphetamine, and DOPA- or apomorphine-induced stereotypy in mice, whereas it suppressed muricide of the rat induced by either olfactory bulbectomy or delta9-tetrahydrocannabinol, similar to the responses seen with imipramine and amitriptyline. On the other hand,lopramine increased spontaneous motor activity and markedly potentiated methamphetamine hyperactivity. In contrast to imipramine and amitriptyline, lopramine failed to counteract both the lethal effect of physostigmine and oxotremorine tremor in mice, indicating that the drug had no central anticholinergic effect. Lopramine, even at such a large dose as 5,000 mg/kg p.o., caused neitherimpairment of coordinated motor activity nor muscle relaxation. It is concluded that lopramine is a new type of tricyclic antidepressant with extremely low toxicity and without central anticholinergic action.


The Role of Alternative Lengthening of Telomeres Mechanism in Cancer: Translational and Therapeutic Implications.
Telomere maintenance mechanisms (i.e., telomerase activity (TA) and the alternative lengthening of telomere (ALT) mechanism) contribute to tumorigenesis by providing unlimited proliferative capacity to cancer cells. Although the role of either telomere maintenance mechanisms seems to be equivalent in providing a limitless proliferative ability to tumor cells, the contribution of TA and ALT to the clinical outcome of patients may differ prominently. In addition, several strategies have been developed to interfere with TA in cancer, including Imetelstat that has been the first telomerase inhibitor tested in clinical trials. Conversely, the limited information available on the molecular underpinnings of ALT has hindered thus far the development of genuine ALT-targeting agents. Moreover, whether anti-telomerase therapies may be hampered or not by possible adaptive responses is still debatable. Nonetheless, it is plausible hypothesizing that treatment with telomerase inhibitors may exert selective pressure for the emergence of cancer cells that become resistant to treatment by activating the ALT mechanism. This notion, together with the evidence that both telomere maintenance mechanisms may coexist within the same tumor and may distinctly impinge on patients' outcomes, suggests that ALT may exert an unexpected role in tumor biology that still needs to be fully elucidated.


Growth, carcass traits, and fatty acid profiles of adipose tissues from steers fed whole cottonseed.
To investigate the impact of dietary whole cottonseed (WCS) level on fatty acid composition, growth, and carcass traits, 45 Hereford steers were assigned to diets containing 0, 15, or 30% dietary WCS. The 15 and 30% WCS contributed an estimated 3.3 and 6.6% additional lipid, respectively, to the diets. After being fed for 54 d, all animals were weighed and slaughtered, and carcass measurements were obtained. There were no differences (P greater than .05) among dietary treatment groups in live weight or ADG for the 54-d feeding period. Control steers had larger (P less than .05) longissimus muscle areas than steers fed 30% WCS, which accounted for the advantage in yield grade (P less than .05) exhibited by the control group. Feeding of 30% WCS resulted in minor increases in linoleic and total polyunsaturated fatty acid content of perinephric fat expressed in both normalized (area percentage) and gravimetric (g/100 g of fresh tissue) formats. There were no significant differences in the monounsaturated or saturated fatty acid content of adipose tissues from animals fed the different diets. Subcutaneous adipose tissue samples were higher (P less than .01) in total unsaturates but had lower (P less than .05) proportions of C18:0 and C18:1 than perinephric samples. Feeding WCS at the levels reported herein only had minor effects on fatty acid composition of beef adipose tissues.


Conformational equilibria in butane-1,4-diol: a benchmark of a prototypical system with strong intramolecular H-bonds.
Explicitly correlated CCSD(T) valence basis limit relative energies were obtained for the 65 conformers of butane-1,4-diol, a prototypical system with a strong internal hydrogen bond. The performance of a variety of ab initio and DFT methods (with and without empirical dispersion corrections) was assessed in detail. Consideration of all pairwise conformer energies provides a performance gauge for both H-bonds and van der Waals interactions, aside from internal strain of angles and bonds. In the post-HF realm, it was found that SCS(MI)CCSD-F12/cc-pVDZ-F12 can be a cost-effective alternative to CCSD(T)/CBS, almost without any loss in accuracy. In the DFT arena, the double-hybrid DSD-PBEP86-D3BJ surpasses the accuracy of all other methods (except for SCS(MI)CCSD, but at a small fraction of its cost). Several hybrid functionals provide an acceptable accuracy with the def2-QZVP basis set, especially BMK, M06, LC-ωPBE-D3, and TPSS0-D3. With the more modest 6-311+G(d,p) basis set, the H-bonds are far from basis set completeness and, due to error compensation, the inclusion of a dispersion correction is generally counterproductive. Some functionals that represent "Pauling points" at this level are LC-ωPBE, TPSS0, B1B95, BMK, TPSSh, PBE0, TPSS, and ωB97X.


Role of marrow-derived monocytes and mesangial cells in removal of immune complexes from renal glomeruli.
Phagocytosis of intravenously administered immune complexes by cells in the mesangium was investigated. The model used was that of exchange marrow transplantation between Chediak-Higashi (CH) mice and syngeneic partners after X-irradiation. This model was chosen since marrow-derived macrophages could be differentiated from resident mesangial cells by the presence of the characteristic giant lysosomes in phagocytic cells of the CH mice. Injected immune complexes were cleared normally and localized in the glomerular mesangium in CH or C57BL/6J mice receiving either C57BL/6J or CH marrow. C57BL/6J mice with CH marrow injected with immune complexes prepared with reduced and alkylated antibodies accumulated many cells within the mesangium that contained both giant lysosomes and electron dense deposits. Deposits were not found in cells with subplasmalemmal microfilaments and perpheral dense bodies. Conversely, the cells in the mesangium of CH mice with C57BL/6J marrow that contained electron dense deposits were devoid of giant lysosomes. Based on these observations, we concluded that (a) marrow-derived monocytes contribute to mesangial hypercellularity after deposition of immune complexes and (b) phagocytosis of immune complexes localized in the glomerular mesangium was by marrow-derived monocytes rather than by mesangial cells.


Low-dose sodium nitrite vasodilates hypoxic human pulmonary vasculature by a means that is not dependent on a simultaneous elevation in plasma nitrite.
Inorganic nitrite has recently been recognized to possess vascular activity that is enhanced in hypoxia. This has been demonstrated in humans in the forearm vascular bed. In animal models nitrite reduces pulmonary vascular resistance, but its effects upon the pulmonary circulation of humans have not yet been demonstrated. This paradigm is of particular interest mechanistically since the pulmonary vasculature is known to behave differently to the systemic. To investigate, 18 healthy volunteers were studied in a hypoxic chamber (inspired oxygen, 12%) or while breathing room air. Each received an infusion of sodium nitrite (1 micromol/min) or 0.9% saline. Three protocols were performed: nitrite/hypoxia (n = 12), saline/hypoxia (n = 6), and nitrite/normoxia (n = 6). Venous blood was sampled for plasma nitrite, forearm blood flow was measured by strain-gauge plethysmography, and pulmonary arterial pressure was measured by transthoracic echocardiography. Plasma nitrite doubled and clearance kinetics were similar whether nitrite was infused in hypoxia or normoxia. During hypoxia, nitrite increased forearm blood flow (+36%, P < 0.001) and reduced three separate indirect indexes of pulmonary arterial pressure by 16%, 12%, and 17% (P < 0.01). Pulmonary, but not systemic, arterial effects persisted 1 h after stopping the infusion, at a time when plasma nitrite had returned to baseline. No effects were observed during normoxia. Therefore, in hypoxic but not normoxic subjects, sodium nitrite causes arterial and pulmonary vasodilatation. In addition, hypoxia-induced pulmonary vasoconstriction was attenuated for a prolonged period and not dependent on a simultaneous elevation of plasma nitrite. This finding is consistent with the direct extravascular metabolism of nitrite to nitric oxide to effect hypoxia-associated bioactivity.


[The effect of latent carbohydrate metabolic disorders on the serum lipid spectrum in patients with ischemic heart disease and hypertension].
Overall eighty four patients suffering from ischemic heart disease (IHD) and hypertensive disease (HD) were evaluated for levels of glycated hemoglobin (gly-Hb) in blood and effects of glycating on blood serum lipid spectrum. Elevated gly-Hb levels were recordable in 51.1% of the patients. Such patients showed significant augmentation of content of total lipids, triglycerides, total cholesterin, Burstein's test, rise in the coefficient of atherogenicity as compared to those subjects having normal gly-Hb level. Positive correlation was established between gly-Hb, levels of total lipids, triglycerides. Given in this paper are the following recommendations for IHD and HD patients presenting with increased gly-Hb level: restricted consumption of easily assimilable carbohydrates, diet N 9, no diabetogenic drugs.


Cholesterol is required for infection by Semliki Forest virus.
Semliki Forest virus (SFV) and many other enveloped animal viruses enter cells by a membrane fusion reaction triggered by the low pH within the endocytic pathway. In vitro, SFV fusion requires cholesterol in the target membrane, but the role of cholesterol in vivo is unknown. In this paper, the infection pathway of SFV was studied in mammalian and inset cells substantially depleted of sterol. Cholesterol-depleted cells were unaltered in their ability to bind, internalize, and acidify virus, but were blocked in SFV fusion and subsequent virus replication. Depleted cells could be infected by the cholesterol-independent vesicular stomatitis virus, which also enters cells via endocytosis and low pH-mediated fusion. The block in SFV infection was specifically reversed by cholesterol but not by cholestenone, which lacks the critical 3 beta-hydroxyl group. Cholesterol thus is central in the infection pathway of SFV, and may act in vivo to modulate infection by SFV and other pathogens.


Advanced manufacturing of nanoparticle formulations of drugs and biologics using microfluidics.
Numerous innovative nanoparticle formulations of drugs and biologics, named nano-formulations, have been developed in the last two decades. However, methods for their scaled-up production are still lagging, as the amount needed for large animal tests and clinical trials is typically orders of magnitude larger. This manufacturing challenge poses a critical barrier to successfully translating various nano-formulations. This review focuses on how microfluidics technology has become a powerful tool to overcome this challenge by synthesizing various nano-formulations with improved particle properties and product purity in large quantities. This microfluidic-based manufacturing is enabled by microfluidic mixing, which is capable of the precise and continuous control of the synthesis of nano-formulations. We further discuss the specific applications of hydrodynamic flow focusing, a staggered herringbone micromixer, a T-junction mixer, a micro-droplet generator, and a glass capillary on various types of nano-formulations of polymeric, lipid, inorganic, and nanocrystals. Various separation and purification microfluidic methods to enhance the product purity are reviewed, including acoustofluidics, hydrodynamics, and dielectrophoresis. We further discuss the challenges of microfluidics being used by broader research and industrial communities. We also provide future outlooks of its enormous potential as a decentralized approach for manufacturing nano-formulations.


Proteus mirabilis phages 5006M, 5006M HFT k and 5006M HFT ak: physical comparison of genome characteristics.
The genomes of proteus mirabilis phages 5006M, kanamycin resistance transducing variant 5006M HFT k and kanamycin-ampicillin resistance transducing variant 5006M HFT ak have been compared. Homo- and heteroduplex and partial denaturation mapping analyses were performed. The results confirm a sequential headful packaging mechanism, facilitate mapping of the ampicillin resistance marker, demonstrate a hairpin loop structure in both variants, reveal a common insertion site for 8 X 10(6) mol. wt. non-phage DNA in both variants and implicate a role for the non-inducible cryptic host strain prophage 5006M in the generatin cycle of the variant phages.


Evaluation and Management of Endometriosis in the Adolescent.
Endometriosis is a chronic condition, with debilitating symptoms affecting all ages. Dysmenorrhea and pelvic pain often begin in adolescence, affecting school, daily activities, and relationships. Despite the profound burden of endometriosis, many adolescents experience suboptimal management and significant delay in diagnosis. The symptomatology and laparoscopic findings of endometriosis in adolescents are often different than in adults, and the medical and surgical treatments for adolescents may differ from those for adults as well. This Narrative Review summarizes the diagnosis, evaluation, and management of endometriosis in adolescents. Given the unique challenges and complexities associated with diagnosing endometriosis in this age group, it is crucial to maintain a heightened level of suspicion and to remain vigilant for signs and symptoms. By maintaining this lower threshold for consideration, we can ensure timely and accurate diagnosis, enabling early intervention and improved management in our adolescent patients.


A two-stage approach for measuring vascular water exchange and arterial transit time by diffusion-weighted perfusion MRI.
Changes in the exchange rate of water across the blood-brain barrier, denoted k(w), may indicate blood-brain barrier dysfunction before the leakage of large-molecule contrast agents is observable. A previously proposed approach for measuring k(w) is to use diffusion-weighted arterial spin labeling to measure the vascular and tissue fractions of labeled water, because the vascular-to-tissue ratio is related to k(w). However, the accuracy of diffusion-weighted arterial spin labeling is affected by arterial blood contributions and the arterial transit time (τ(a)). To address these issues, a two-stage method is proposed that uses combinations of diffusion-weighted gradient strengths and post-labeling delays to measure both τ(a) and k(w). The feasibility of this method was assessed by acquiring diffusion-weighted arterial spin labeling data from seven healthy volunteers. Repeat measurements and Monte Carlo simulations were conducted to determine the precision and accuracy of the k(w) estimates. Average grey and white matter k(w) values were 110 ± 18 and 126 ± 18 min(-1), respectively, which compare favorably to blood-brain barrier permeability measurements obtained with positron emission tomography. The intrasubject coefficient of variation was 26% ± 23% in grey matter and 21% ± 17% in white matter, indicating that reproducible k(w) measurements can be obtained.


Differences in antibody-dependent cellular cytotoxicity and activated killing of tumor cells by macrophage cell lines.
A series of murine macrophage cell lines was assayed for killing of B and T lymphocytic and myeloid tumor targets by radiolabel release at effector:target ratios of 20:1. One group of lines was inactive in all assays. Another group of lines showed moderate spontaneous cytotoxicity to lymphoid tumors that was greatly enhanced by inclusion of antibody, lipopolysaccharide, or phorbol myristate acetate in the 22-hr assays. Addition of lymphokine to the assays induced only moderate killing. Pretreatment of cell lines with lipopolysaccharide, phorbol myristate acetate, lymphokine, or nonlymphocyte sources of macrophage colony-stimulating factors did not activate "angry" or nonspecific killing. However, such pretreatment greatly stimulated antibody-dependent cellular cytotoxicity. Cytotoxicity was not due to crowding or poor culture conditions: macrophage line supernatants were not toxic; rapidly growing lymphoid lines used in place of macrophages did not kill; and high macrophage concentrations (10(6)/ml) had reduced cytotoxic activity. These experiments show that the same cell type can mediate both activated killing and antibody-dependent cellular cytotoxicity of tumor targets. The active macrophage lines appear to be qualitatively similar to each other and to peritoneal exudate populations in tumoricidal activity. We showed previously that these macrophage lines could also phagocytose and lyse antibody-coated red blood cells. The pretreatment experiments suggest that the antibody-dependent cellular cytotoxicity state can be regulated independently of macrophage nonspecific tumoricidal capacity. These culture line models of macrophages offer several advantages in the analysis of cytotoxicity.


Sequence analysis of the alpha 1 Na+,K(+)-ATPase gene in the Dahl salt-sensitive rat.
In the inbred Dahl salt-sensitive rat (SS/Jr strain), it has been proposed that a T for A transversion in the DNA sequence encoding amino acid 276 in the alpha 1 subunit isoform of Na+,K(+)-ATPase may impair ion transport and contribute to the pathogenesis of hypertension. This hypothesis is of major scientific interest because it represents the first attempt to explain the pathogenesis of salt-sensitive hypertension on the basis of a specifically defined mutation at the DNA level. We devised a polymerase chain reaction technique to screen the genomic DNA of multiple SS/Jr rats for the T for A transversion reported in the complementary DNA (cDNA) encoding the alpha 1 subunit of Na+,K(+)-ATPase. When eight Dahl SS/Jr rats from Harlan Sprague Dawley Inc. were tested with the polymerase chain reaction technique, we found no evidence of this mutation in the Na+,K(+)-ATPase gene. Direct sequence analysis of the gene in three SS/Jr rats also did not show the T for A transversion. These results 1) strongly suggest that commercially available Dahl SS/Jr rats do not carry a T for A transversion in the genomic DNA sequence encoding amino acid 276 in the alpha 1 subunit isoform of Na+,K(+)-ATPase and 2) raise the possibility that the previous finding of a mutation in the cDNA of the SS/Jr rat may have been due to a reverse transcriptase error during cDNA synthesis.


Construction of new ligand binding sites in proteins of known structure. I. Computer-aided modeling of sites with pre-defined geometry.
We have devised a molecular model building computer program (DEZYMER) which builds new ligand binding sites into a protein of known three-dimensional structure. It alters only the sequence and the side-chain structure of the protein, leaving the protein backbone fold intact by definition. The program searches for a constellation of backbone positions arranged such that if appropriate side-chains were placed there, they would bind the ligand according to a pre-defined geometry of interaction specified by the experimentalist. These binding sites are introduced by the program by taking into account simple rules such as steric hindrance, atomic close-packing and hydrogen bond patterns, which are known to maintain the integrity of a protein structure to a first approximation. A test case is presented in this paper where the copper binding site found in blue-copper proteins such as plastocyanin, azurin and cupredoxin is introduced into Escherichia coli thioredoxin. The model building of one of the solutions found by the program is presented in some detail. The experimental construction and properties of this new protein are described in an accompanying paper. It is hoped that this program provides a general method for the design of ligand binding sites and enzyme active sites, which can then be tested experimentally.


Chemotherapy near the end of life: a retrospective single-centre analysis of patients' charts.
Chemotherapy near the end of life is an issue frequently discussed nowadays. The concern is that chemotherapy could cause more harm than good in a palliative situation; this is even truer as the patient nears death. The objective of our study is to evaluate the aggressiveness of patient care near the end of life by determining how many cancer patients receive chemotherapy during their final weeks. In a retrospective analysis of patient charts, we investigated whether cancer patients had been treated with chemotherapy during the last four or two weeks of life. If they had, we looked at whether treatment was ongoing or newly initiated. Out of the 119 cancer patients who died in our hospital over two years, 14 (11.7%) received chemotherapy during the last four weeks of life, nine of whom (7.6%) in the last two weeks of life. Treatment had been ongoing in six (5%) and newly initiated for eight (6.7%) within four weeks of death. Corresponding figures for the last two weeks of life were seven patients (5.9%) who continued previously prescribed treatment and two (1.7%) who were started on chemotherapy. Patients given chemotherapy during the last four weeks of life were significantly younger than those who were not (p = 0.003). Cancer patient care in our hospital is not considered overly aggressive as only 7.6% of these patients receive chemotherapy within the last two weeks of life. To determine how aggressive care near the end of life really is, however, we suggest evaluating newly started chemotherapy alongside ongoing treatment. As the line between the effects (beneficience) and side effects (nonmaleficience) of chemotherapy is often very narrow, doctors and patients have to work together to find the best way of treading this fine line.


Pertussis in California: A Tale of 2 Epidemics.
California experienced pertussis epidemics in 2010 and 2014, with more than 9000 and 11,000 reported cases, respectively. Using an ecologic study design, we compared age-specific pertussis incidence for persons ≤20 years of age in California in 2010 and 2014 to describe any changes in disease burden that may be attributable to changes in pertussis vaccination policy that occurred during interepidemic years. We evaluated California pertussis surveillance data for reported pertussis cases 20 years of age and younger with disease onset from January 1, 2010 to December 31, 2010 or from January 1, 2014 to December 31, 2014. Age-specific pertussis rates were calculated for each of the epidemic years. Characteristics of each age group were compared across epidemic years and age-specific rate ratios (RRs) and associated 95% confidence intervals (CIs) were calculated. In both years, the highest disease rate was reported in infants <6 months of age; however, the incidence of pertussis declined significantly among infants <6 months of age in 2014 compared with 2010 (RR 0.58, 95% CI: 0.53-0.63). Incidence remained stable among infants ≥6 months of age, children and adolescents <12 years of age (RR 1.1, 95% CI: 0.5-1.3). Adolescents 12-18 years of age had significantly higher rates of pertussis in 2014 compared with 2010, with the highest rates observed among 14- to 16-year-olds (RR 6.5, 95% CI: 5.4-7.8). A 42% reduction (53%-63%) in pertussis incidence among infants <6 months of age in 2014 after the routine prenatal Tdap vaccination recommendation suggested that an increase in prenatal Tdap coverage has been having some impact on infant pertussis. The excess number of cases observed in 2014 compared with 2010 occurred almost exclusively among the 12- to 17-year-old age group. Among older children, the peak age of pertussis is shifting, consistent with aging of the acellular pertussis vaccine cohort.


Coagulation Parameters in Human Immunodeficiency Virus Infected Patients: A Systematic Review and Meta-Analysis.
Coagulation abnormalities are common complications of human immunodeficiency virus (HIV) infection. Highly active antiretroviral treatment (HAART) decreased the mortality of HIV but increased coagulopathies. HIV-related thrombocytopenia, prolonged prothrombin time (PT), activated partial thromboplastin time (APTT), and high D-dimer level commonly manifested in patients with HIV. Thus, this study is aimed to compare coagulation parameters of HAART-treated and HAART-naïve HIV-infected patients with HIV-seronegative controls. A systematic literature search was conducted using the databases PubMed/MEDLINE, Embase, Web of Science, and Google Scholar of studies published until July 2021. The primary outcome of interest was determining the pooled mean difference of coagulation parameters between HIV-infected patients and seronegative controls. The Joana Briggs Institute (JBI) critical appraisal tool was used for quality appraisal. Statistical analyses were performed using Stata11.0 software. The statistical results were expressed as the effect measured by standardized mean difference (SMD) with their related 95% confidence interval (CI). A total of 7,498 participants (1,144 HAART-naïve patients and 2,270 HAART-treated HIV-infected patients and 3,584 HIV-seronegative controls) from 18 studies were included. HIV-infected patients (both on HAART and HAART-naive) exhibited significantly higher levels of PT than HIV-seronegative controls (SMD = 0.66; 95% CI: 0.53-0.80 and SMD = 1.13; 95% CI: 0.60-2.0, respectively). The value of APTT was significantly higher in patients with HIV on HAART than in seronegative controls. However, the values of PLT count, APTT, and fibrinogen level were significantly higher in seronegative controls. Besides, the level of fibrinogen was significantly higher in HAART-treated than treatment-naïve patients (SMD = 0.32; 95%CI: 0.08, 0.57). Moreover, the level of APTT and PT had no statistical difference between HAART and HAART-naïve HIV-infected patients. This study identified that HIV-infected patients are more likely to develop coagulation abnormalities than HIV-seronegative controls. Therefore, coagulation parameters should be assessed regularly to prevent and monitor coagulation disorders in HIV-infected patients.


Rhinoplasty: the african american patient.
Over the past three decades, an increasing number of African American patients have undergone rhinoplasty, and many continue to present to surgeons for rhinoplasty evaluation. The reality is that rhinoplasty is no longer an uncommon procedure in the African American population. Most patients desire nasal refinement while preserving their cultural identity. The African American nose has many unique features that have to be appreciated and understood to provide the desired outcome. In this paper, we present an overview of the unique anatomic features of the African American nose, rhinoplasty techniques tailored to this patient population, and complications encountered postoperatively.