Issues and barriers to development of clinically useful tumor markers: a development pathway proposal.
There are few tumor markers that are clinically useful in predicting therapeutic responses or patient outcomes despite nearly 20 years of advances in molecular biology. We discuss a variety of issues and barriers that have affected movement of clinical tests from research into clinical practice. Studies of new markers frequently lack clear hypotheses and are generally underpowered to reach statistically valid conclusions. Relevant clinical endpoints may not be possible to evaluate, often leading to suboptimal study designs. Major stumbling blocks exist because studies are rarely comparable. This makes it difficult to determine why results vary from study to study. It also prevents pooling of small datasets for analysis. We propose a tumor marker development pathway that we think will be more efficient and effective. The pathway depends on developing statistically valid study designs, focusing on assay refinement and standardization early in the process, including assay details in publications, and providing data in a format that allows comparison with other studies. The process described should be applicable to development of new technologies that include analysis and interpretation of large, complex datasets. The proposed marker development pathway will require thoughtful refinement and expansion, but it should begin a productive dialog.


Evidence for an association of the big five personality factors with recurrence of depressive symptoms in later life.
Although it is well known that recurrence of late-life depression is very common, little is known about the characteristics of older people who are vulnerable for recurrence. In order to identify characteristics of those who are at risk, the present study aimed to investigate the strength of the associations of the big five personality factors with recurrence in later life. Secondly, we studied whether there are gender and age differences in the strength of these associations. Using data from the longitudinal aging study Amsterdam (LASA) a subsample with clinically relevant depressive symptoms at one or more of the first three LASA-cycles, but who had recovered at the fourth cycle, was approached to participate in a fifth cycle to determine recurrence (n = 92). Respondents completed self-report questionnaires on personality (NEO-FFI) and depression (CES-D). By means of logistic regression analyses the associations between the Big Five and recurrence of depression at fifth cycle was investigated. 58 (63%) had a recurrence of depressive symptoms. A high level of neuroticism was significantly associated with recurrence. No gender differences or age-related differences in strength of the associations of personality with recurrence were found. In later life, neuroticism still is associated with the recurrence of depression. Efforts to prevent recurrence of late-life depression should focus on those with high levels of neuroticism and future research should aim at further unravelling the association between depression and personality in later life.


Carnosic acid: a potent chemopreventive agent against oral carcinogenesis.
The present study was aimed to investigate the chemopreventive potential of carnosic acid in 7,12-dimethylbenz(a)anthracene (DMBA)-induced hamster buccal pouch carcinogenesis. The chemopreventive potential was assessed by analyzing the tumor incidence, tumor volume and burden as well as by measuring the status of lipid peroxidation, non-enzymatic and enzymatic antioxidants and phase I and phase II detoxification enzymes. Oral squamous cell carcinoma was developed in the buccal pouch of golden Syrian hamsters by painting with 0.5% DMBA in liquid paraffin three times a week for 14 weeks. In the present study, 100% tumor formation was observed in hamsters treated with DMBA alone. Also, the status of lipid peroxidation, antioxidants and phase I and phase II detoxification enzymes were significantly altered during DMBA-induced oral carcinogenesis. Oral administration of carnosic acid at a dose of 10mg/kg body weight/day to DMBA-treated animals completely prevented the tumor formation in the hamsters' buccal pouches. Also, carnosic acid exerted potent anti-lipid peroxidative function and stimulated the detoxification cascade during DMBA-induced hamster buccal pouch carcinogenesis. The results of the present study suggest that the chemopreventive potential of carnosic acid is probably due to its anti-lipid peroxidative potential and modulating effect on carcinogen detoxification enzymes during DMBA-induced oral carcinogenesis.


The pigmentary system of developing axolotls. II. An analysis of the melanoid phenotype.
The melanoid mutant in the Mexican axolotl (Ambystoma mexicanum) is analysed with respect to the differentiation of pigment cells. Pigment cells were observed with the transmission electron microscope in order to determine any unusual structural characteristics and to determine what happens to each of the cell types as development proceeds. Chemical analysis of pteridine pigments was also carried out, and changes in pteridine biosynthesis were found to correlate well with changes in xanthophore morphology and number. In melanoid axolotls, as development proceeds, melanophore numbers increase, xanthophores decrease, and iridophores fail to differentiate at all. This is considered to result from: (a) conversion of xanthophores (that are present in young larvae) to melanophores; (b) the gradual programming of the majority of chromatoblasts to become, exclusively, melanophores, and (c) the failure of some chromatoblasts (possibly iridoblasts) to differentiate altogether. The ultrastructural and chemical evidence presented in this study is compared to similar data for wild-type axolotls, and a mechanism regarding how the melanoid gene might act is suggested.


Cavotricuspid isthmus mapping to assess bidirectional block during common atrial flutter radiofrequency ablation.
We sought to compare published methods to an alternative approach ascertaining cavotricuspid isthmus (CTI) block during atrial flutter ablation. In 39 consecutive patients who underwent an atrial flutter ablation procedure, a 24-pole mapping catheter was positioned so that 2 adjacent dipoles were bracketing the targeted CTI line of block (LOB), with proximal dipoles lateral to the LOB and distal dipoles in the coronary sinus. Two pacing sites were lateral (positions A and B) and 2 were septal (positions C and D) to the LOB, with locations A and D closest to the LOB. A resulting CTI block was accepted when 3 criteria were fulfilled: (1) complete reversal of the right atrial depolarization on the 24-pole catheter when pacing in the coronary sinus, (2) conduction delays from A to D greater than from B to D, and (3) conduction delays from D to A greater than from C to A. A successful CTI block was obtained in all patients. Before CTI block was obtained, a progressive CTI conduction delay was observed in 11 patients (28.2%). During the procedure, the 3 criteria defined above were either all present or all absent. This study establishes that reversal of the atrial depolarization sequence up to the LOB is a definitive and mandatory criteria of successful atrial flutter ablation.


Crack Length Effect on the Fracture Behavior of Single-Crystals and Bi-Crystals of Aluminum.
Molecular dynamics simulations of cracked nanocrystals of aluminum were performed in order to investigate the crack length and grain boundary effects. Atomistic models of single-crystals and bi-crystals were built considering 11 different crack lengths. Novel approaches based on fracture mechanics concepts were proposed to predict the crack length effect on single-crystals and bi-crystals. The results showed that the effect of the grain boundary on the fracture resistance was beneficial increasing the fracture toughness almost four times for bi-crystals.


Impact of High Diversity of Achromobacter Populations within Cystic Fibrosis Sputum Samples on Antimicrobial Susceptibility Testing.
Chronic colonization by opportunistic environmental bacteria is frequent in the airways of cystic fibrosis (CF) patients. Studies of Pseudomonas aeruginosa evolution during persistence have highlighted the emergence of pathoadaptive genotypes and phenotypes, leading to complex and diversified inpatient colonizing populations also observed at the intraspecimen level. Such diversity, including heterogeneity in resistance profiles, has been considered an adaptive strategy devoted to host persistence. Longitudinal genomic diversity has been shown for the emergent opportunistic pathogen Achromobacter, but phenotypic and genomic diversity has not yet been studied within a simple CF sputum sample. Here, we studied the genomic diversity and antimicrobial resistance heterogeneity of 132 Achromobacter species strains (8 to 27 strains of identical or distinct colonial morphotypes per specimen) recovered from the sputum samples of 9 chronically colonized CF patients. We highlighted the high within-sample and within-morphotype diversity of antimicrobial resistance (disk diffusion) and genomic (pulsed-field gel electrophoresis) profiles. No sputum sample included strains with identical pulsotypes or antibiotic susceptibility patterns. Differences in clinical categorization were observed for the 9 patients and concerned 3 to 11 antibiotics, including antibiotics recommended for use against Achromobacter Within-sample antimicrobial resistance heterogeneity, not predictable from colonial morphology, suggested that it may represent a selective advantage against antibiotics in an Achromobacter persisting population and potentially compromise the antibiotic management of CF airway infections.


Ligature-Induced Experimental Peri-Implantitis-A Systematic Review.
This systematic review sought to analyze different experimental peri-implantitis models, their potential to induce marginal bone resorption (MBR) and the necessity of bacteria for bone loss to occur in these models. An electronic search in PubMed/Medline, Web of Science, and ScienceDirect was undertaken. A total of 133 studies were analyzed. Most studies induced peri-implantitis with ligatures that had formed a biofilm, sometimes in combination with inoculation of specific bacteria but never in a sterile environment. Most vertical MBR resulted from new ligatures periodically packed above old ones, followed by periodically exchanged ligatures and ligatures that were not exchanged. Cotton ligatures produced the most MBR, followed by steel, "dental floss" (not further specified in the studies) and silk. The amount of MBR varied significantly between different animal types and implant surfaces. None of the analyzed ligature studies aimed to validate that bacteria are necessary for the inducement of MBR. It cannot be excluded that bone loss can be achieved by other factors of the model, such as an immunological reaction to the ligature itself or trauma from repeated ligature insertions. Because all the included trials allowed plaque accumulation on the ligatures, bone resorbing capacity due to other factors could not be excluded or evaluated here.


Association of ambulatory blood pressure and heart rate with advanced white matter lesions in ischemic stroke patients.
White matter lesions (WMLs) are a common finding in stroke patients, and the most important risk factors are old age and hypertension. Although many studies have described the association between WMLs and ambulatory blood pressure monitoring (ABPM) parameters in healthy subjects and hypertensive patients, little is known about the association in hypertensive ischemic stroke patients. From July 2009 to June 2012, 169 consecutive hypertensive noncardioembolic ischemic stroke patients were recruited within 1 week of suffering a stroke, and ABPM was applied 1 or 2 weeks after stroke onset. The subjects were classified into 2 groups according to the presence of advanced WMLs, and their ABPM parameters were compared. Finally, multivariable logistic regression analyses were performed to investigate the independent relationships between WMLs and ABPM parameters. Seventy (41%) patients had advanced WMLs. In univariable analysis, higher 24-hour, awake, and asleep systolic blood pressure (SBP)/diastolic blood pressure levels and 24-hour pulse pressure were associated with advanced WMLs. However, circadian blood pressure parameters such as 24-hour BP variability, morning surge, and nocturnal dipping pattern were not associated with advanced WMLs. After adjustments, old age (odds ratio (OR) = 1.063; 95% confidence interval (CI) = 1.024-1.104; P = 0.002), high 24-hour SBP levels (OR = 1.055; 95% CI = 1.028-1.082; P < 0.001), and high 24-hour heart rate (OR = 1.041; 95% CI = 1.006-1.078; P = 0.023) were independently associated with advanced WMLs. In addition to old age and elevated 24-hour SBP, increased heart rate is associated with advanced WMLs in ischemic stroke patients. Heart rate deserves more attention in predicting advanced WMLs in those patients.


Arterial stiffening following eccentric exercise-induced muscle damage.
Acute inflammatory responses are linked to a transient increase in risk of a cardiovascular event, and this risk may be mediated by a concomitant reduction in vascular function. Humans experience an acute inflammatory response as a consequence of infection, injury, or muscle damage. We measured macrovascular function before and after eccentric exercise to determine whether muscle damage from unaccustomed exercise has an unfavorable effect on the large elastic arteries. A total of 27 healthy sedentary or recreationally active men (age 18-38 years) participated in either bilateral leg press eccentric exercise or unilateral elbow flexor eccentric exercise. Postexercise muscle damage was confirmed by significant reductions in isometric strength and increases in muscle soreness (P < 0.05). Carotid-femoral pulse-wave velocity was significantly elevated 48 h after leg exercise (808 ± 31 vs. 785 ± 30 cm/s; P < 0.05) and arm exercise (790 ± 28 vs. 755 ± 24 cm/s; P < 0.05). There were no changes in mean arterial pressure. C-reactive protein was elevated after leg exercise but not after arm exercise. The increase in carotid-femoral pulse wave velocity 48 h after arm exercise was associated with muscle strength (r = -0.47; P < 0.05) and creatine kinase concentrations (r = 0.70; P < 0.01). We concluded that eccentric exercise in both small and large muscle mass translates to transient, unfavorable changes in central macrovascular function and that the increase in central arterial stiffness after small muscle eccentric exercise is associated with indicators of muscle damage.


Co-Mutation of FAT3 and LRP1B in Lung Adenocarcinoma Defines a Unique Subset Correlated With the Efficacy of Immunotherapy.
Immunotherapy based on immune checkpoint inhibitors (ICIs) have demonstrated remarkable survival benefits and gained regulatory approval in non-small cell lung cancer (NSCLC) patients without an actionable driver mutation, but currently there is no well-established standard for how to screen the most suitable population for ICIs treatment. Here, we conducted a comprehensive analysis of the somatic mutation landscape of lung adenocarcinoma (LUAD) samples. After the stepwise screening of high-frequency mutated genes, two genes with prominent significance, FAT3 and LRP1B, were finally screened out. Through further analysis, we discovered that the co-mutation of FAT3 and LRP1B was associated with an earlier age of onset and occurred more frequently in Black/African American. Furthermore, co-mutation defines a unique subgroup of lung adenocarcinoma that can increase tumor mutational burden (TMB), boost cytotoxicity and tumor immunogenicity, and facilitate lymphocyte infiltration. The results of gene set enrichment analysis (GSEA) indicated that co-mutation can influence tumorigenesis through a variety of mechanisms. More strikingly, the subset of LUAD with co-mutation of FAT3 and LRP1B exhibited significantly prolonged immunotherapy progression free survival (PFS). In summary, co-mutation of FAT3 and LRP1B is a promising useful biomarker for predicting the efficacy of immunotherapy, which can improve the clinical efficiency of practicing precision medicine in lung adenocarcinoma patients.


Knee kinematic coupling in males and females: open and closed-chain tasks.
The purpose of this study was to compare the magnitude of knee kinematic coupling between genders and among open- and closed-chain tasks. A secondary purpose was to compare the consistency of knee kinematic coupling between genders and among open- and closed-chain tasks. Vector-coding methods were used to quantify coupling in the sagittal and transverse planes of the knee between full extension and 20 degrees of flexion as 10 males and 10 females walked, ascended and descended stairs, and performed a passive pendulum leg drop. An ANOVA showed no main effect of gender. There was a main effect of task, where coupling during the stance phase of walking was significantly greater than each of the other tasks. Intraclass correlation values suggested that males were slightly more consistent than females. A general lack of divergence between genders may be related to the tasks analyzed in this study. It is possible that more strenuous tasks may elicit larger differences.


Activation of Type 4 Metabotropic Glutamate Receptor Attenuates Oxidative Stress-Induced Death of Neural Stem Cells with Inhibition of JNK and p38 MAPK Signaling.
Promoting both endogenous and exogenous neural stem cells' (NSCs) survival in the hostile host environments is essential to cell replacement therapy for central nervous system (CNS) disorders. Type 4 metabotropic glutamate receptor (mGluR4), one of the members of mGluRs, has been shown to protect neurons from acute and chronic excitotoxic insults in various brain damages. The present study investigated the preventive effects of mGluR4 on NSC injury induced by oxidative stress. Under challenge with H2O2, loss of cell viability was observed in cultured rat NSCs, and treatment with selective mGluR4 agonist VU0155041 conferred protective effects against the loss of cellular viability in a concentration-dependent manner, as shown by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Pretreatment of VU0155041 (30 μM) also inhibited the excessive NSC death induced by H2O2, and group III mGluRs antagonist (RS)-a-methylserine-O-phosphate (MSOP) or gene-targeted knockdown abolished the protective action of mGluR4, indicated by propidium iodide-Hoechst and terminal deoxynucleotidyl transferase-mediated UTP nick end labeling (TUNEL) staining. Western blot assay demonstrated that mGluR4 activation reversed the decreased procaspase-8/9/3and the destructed Bcl-2/Bax expressing balance, and likewise, MSOP and mGluR4 knockdown abrogated the action of mGluR4 activity. Furthermore, inhibition of JNK and p38 mitogen-activated protein kinases (MAPKs) were observed after mGluR4 activation, and as paralleling control, JNK-specific inhibitor SP600125 and p38-specific inhibitor SB203580 significantly rescued the H2O2-mediated NSC apoptosis and cleavage of procaspase-3. We suggest that activation of mGluR4 prevents oxidative stress-induced NSC death and apoptotic-associated protein activities with involvement of inhibiting the JNK and p38 pathways in cell culture. Our findings may help to develop strategies for enhancing the resided and transplanted NSC survival after oxidative stress insult of CNS.


Campus Reset: Dynamic Planning and Response to SARS-CoV-2 Infections at the US Air Force Academy.
Predominantly asymptomatic infections, such as those for SARS-CoV-2, require robust surveillance testing to identify people who are unknowingly spreading the virus. The US Air Force Academy returned to in-person classes for more than 4000 cadets aged 18-26 years during the fall 2020 semester to meet graduation and leadership training requirements. To enable this sustained cadet footprint, the institution developed a dynamic SARS-CoV-2 response plan using near-real-time data to inform decisions and trigger policies. A surveillance testing program based on mathematical modeling and a policy-driven campus reset option provided a scaled approach to react to SARS-CoV-2 conditions. This program adequately controlled the spread of the virus for the first 2 months of the academic semester but failed to predict or initially mitigate a significant outbreak in the second half of the semester. Although this approach did not completely eliminate SARS-CoV-2 infections in the population, it served as an early warning system to alert public health authorities to potential issues, which allowed timely responses while containment was still possible.


Detection of vaccine-derived poliovirus circulation by environmental surveillance in the absence of clinical cases.
On August 25, 2019, the Noguchi Memorial Institute for Medical Research notified the confirmation of a circulating-vaccine-derived poliovirus type-2 (cVDPV2) from the Agbogbloshie environmental surveillance (AES) site, in the Greater Accra Region. A field investigation of the outbreak was conducted to describe the results of epidemiological and laboratory investigations, and control efforts. We conducted a descriptive investigation, records review, and active-case-search. Caregivers were interviewed on the vaccination status of their children; knowledge, attitude, and practices on polio prevention; water, sanitation and hygiene practices, and health-seeking behaviors. Stool from healthy children <5 y and sewage samples were taken for laboratory diagnosis. cVDPV2 genetically similar to the cVDPV2 diagnosed recently in the Northern Region of Ghana and Nigeria was identified. 2019 half-year coverage of OPV and IPV was 22%. Fully immunized children were 49% (29/59). Most health workers (70%) had a fair knowledge of polio and acute flaccid paralysis (AFP). Forty-six percent of care-givers admitted to using the large drain linked to the site where the cVDPV2 was isolated as their place of convenience and disposing of the fecal matter of their children. No AFP case was identified. Stool samples from 40 healthy children yielded non-polio enteroviruses while 75% (3/4) of the additional sewage samples yielded cVDPV2. cVDPV2 was isolated from the AES site. No AFP or poliovirus was identified from healthy children. There is a need to improve health workers' knowledge on AFP and to address the dire sanitation conditions in the Agbogbloshie market and its environs.


Enzyme immunoassay of neocarzinostatin using beta-galactosidase as label.
A novel convenient procedure was introduced for enzyme labelling of neocarzinostatin (NCS) with beta-D-galactosidase using a new hetero-bifunctional reagent N-(gamma-maleimidobutyryloxy) succinimide (GMBS). With the enzyme labelled NCS and a rabbit anti-NCS serum which was elicited in a rabbit immunized with NCS in complete Freund's adjuvant, a highly sensitive enzyme immunoassay which can quantify a femto mol order NCS was developed. Accuracy and precision of the assay as well as a comparison with the immunodiffussion method were studied with satisfactory results. This enzyme immunoassay of NCS was applicable to monitoring serum NCS levels of patients in clinical treatment of the antitumor agent.


Implant Failure Rates and Cost Analysis of Contoured Locking Versus Conventional Plate Fixation of Distal Fibula Fractures.
The authors analyzed 330 consecutive Weber B distal fibula fractures that occurred during a 3-year period and were treated with either a contoured locking plate or a conventional one-third tubular plate to compare the cost and failure rates of the 2 constructs. The primary outcomes were failure of the distal fibular implant and loss of reduction. Secondary outcomes were surgical wound infection requiring surgical debridement and/or removal of the fibular implant, and removal of the fibular plate for persistent implant-related symptoms. No failure of the fibular plates or distal fibular fixation occurred in either group. A total of 5 patients required surgical revision of syndesmotic fixation within 4 weeks of the index surgery. Of these patients, 1 was in the contoured locking plate group and 4 were in the one-third tubular plate group (P=.610). The rate of deep infection requiring surgical debridement and/or implant removal was 6.2% in the contoured locking plate group and 1.4% in the one-third tubular plate group (P=.017). The rate of lateral implant removal for either infection or symptomatic implant was 9.3% in the contoured locking plate group and 2.3% in the one-third tubular plate group (P=.005). A typical contoured locking plate construct costs $800 more than a comparable one-third tubular plate construct. Based on a calculated estimate of 60,000 locking plates used annually in the United States, this difference translates to a potential avoided annual cost of $50 million nationally. This study demonstrates that it is possible to treat Weber B distal fibula fractures with one-third tubular plates at a substantially lower cost than that of contoured locking plates without increasing complications. [Orthopedics. 2017; 40(6):e1024-e1029.].


Dehumanization and burden of care among caregivers of terminally ill patients.
This article considers the relationship between dehumanization, ontological representation of death, trust in physicians, and burden of care on the part of caregivers of terminally ill patients. One hundred informal caregivers (relatives and friends) of patients hospitalized in four hospice facilities in northern Italy were involved. Of these, 77% were primary caregivers (those who mostly helped the patient). All of the participants were given a questionnaire comprising the Caregiver Burden Inventory (CBI) to determine caregivers' burden in their roles, the questionario post mortem (QPM) (post mortem questionnaire) for the effectiveness of and their trust in the medical nursing team of palliative care services, the Testoni death representation scale (TDRS) to detect their ontological representations of death and the humanity attribution test (HAT) to investigate their attributions of humanity to terminally ill patients. Per the literature, the present results demonstrated higher burden levels for female caregivers and primary caregivers. In informal caregiving, the dehumanization of patients does not have any advantage in reducing the burden of care. Further studies are required to compare formal and informal caregivers concerning the effect of dehumanization.


Selective sulfonylation and diazotization of indoles.
A metal-free synthesis of bifunctionalized indole derivatives was developed through a novel TBHP/TBAI-mediated oxidative coupling of C2,C3-unsubstituted indoles with arylsulfonyl hydrazide. Under the same conditions C3-methyl substituted indoles underwent a diazotization process, affording 2-sulfonyldiazenyl-1H-indoles. The former reaction simultaneously established C-S and C-N bonds through selective sulfonylation and diazotization of the indole framework, enabling a mild and practical access to polyfunctionalized indoles with good to excellent yields.


Glomangiosarcoma Arising from a Prior Biopsy Site.
Glomangiosarcoma represents a rare malignant variant of the benign glomus tumor that typically presents as a tender, slowly growing nodule with a predilection for the lower extremities. Unlike their benign counterparts, glomangiosarcomas may display aggressive characteristics such as large size, local invasion, and a tendency to recur after excision. Although wide local excision remains the treatment of choice, rare cases of systemic metastasis have been previously reported. We present a case of glomangiosarcoma arising at a prior biopsy site after excision of an unknown soft tissue lesion.


Porocarcinoma arising in pigmented hidroacanthoma simplex.
Hidroacanthoma simplex (HAS) is a rare benign tumor that is also known as intraepidermal poroma. While there have been a few reports of HAS with malignant transformation (porocarcinoma), we report an unusual case of porocarcinoma, arising in a pigmented HAS, the latter also showing secondary amyloid deposits. An 80-year-old Japanese man presented with a cutaneous tumor on his left buttock, which had first been noticed in his childhood. The tumor consisted of flat pigmented plaque and a depigmented papule with erosion. Histologic analysis revealed many pigmented and well-defined nests within the epidermis of the flat pigmented portion. The nests were composed of cuboidal to oval and occasionally elongated, bland, basaloid cells with numerous melanin granules. In addition, there were infrequently ductal structures and small clusters of sebocytes, and abundant amyloid deposits in the upper dermis. These findings were consistent with pigmented HAS with amyloid deposition. In the depigmented portion, markedly atypical cells with occasional ductal structures and intracytoplasmic lumina extended throughout the entire thickness of the epidermis, with minimal invasion of the dermis. We considered this portion of the tumor to be a porocarcinoma. Since the two portions of the tumor were continuous, we made a final diagnosis of porocarcinoma arising in pre-existing pigmented HAS with amyloid deposition.


Is consanguinity prevalence decreasing in Saudis?: A study in two generations.
Saudi population is unique in that there is a strong preference for cousin marriages in the general population. We studied the prevalence of consanguinity in educated Saudi females and compared the results with the results obtained in their parents, to access if a generation difference in which extensive educational activities have prevailed to inform the people of the influence of cousin marriages on health, has made any difference in prevalence of consanguineous marriages. A total of 600 Saudi women (421 university students and 179 women attending outpatients' clinics) were interviewed about their own and their parents' consanguinity. The total consanguinity (first and second cousins) was 29.7% in the parents. Consanguinity was significantly higher among the daughters than the parents, where 37.9% of the 293 married women had consanguineous marriages. The prevalence of consanguinity was studied in different age groups, though no significant pattern was observed. A strong correlation was found between consanguinity of parents and their daughters; consanguinity was highest (52.3%) in the daughters of parents who were themselves consanguineous. The results did not reveal any decrease in the prevalence of consanguinity over a generation. This shows that the tradition of marrying within the family is a preferred practice, despite the awareness that certain genetic disorders occur at a higher frequency in cousin marriages. There is a need at the primary health care level to inform the public of the consequences of this common practice.


Spatial reversal learning in preclinical scrapie-inoculated mice.
Acquisition and reversal of a two-choice spatial discrimination were tested in scrapie-inoculated mice. Both acquisition and reversal were normal in mice tested 138 and 103 days prior to the onset of clinical symptoms. At 65 days before onset of clinical symptoms, scrapie-inoculated mice required more trails to criterion in reversal learning, but this effect was not significant in a second experiment (68 days preclinical) and was transient: no effect was seen 33 days before symptoms. However, the course of reversal learning was abnormal in all three late preclinical groups (68, 65 and 33 days before symptoms). Reversal learning in these three groups was characterized by a rapid extinction of the original discrimination, followed by a period, absent in controls, during which performance showed no further improvement. This effect corresponds in time of onset to the appearance of characteristic neuropathological features.


[Pharmacological approach to cerebral cortical development].
In the developing mammalian cerebral cortex, newly generated neurons migrate toward the pial surface to form a mammalian-specific six-layered cerebral cortex. Genetic studies of human neurological diseases have suggested the involvement of several molecules in cortical neuronal migration. In vivo electroporation is another powerful tool for understanding the molecular mechanisms of neuronal migration. By using these techniques, however, it is difficult to understand molecular basis of time-dependent changes of neuronal morphologies. Here, we introduce a pharmacological approach to cerebral cortical development. Major advantages of the pharmacological approach include the transient suppression of molecules of interest and analyzing time-dependent changes of neuronal morphologies. It also allows us to search molecules regulating neuronal migration with comparative ease. We propose the complementarity between the pharmacological approach and genetics or in vivo electroporation experiments.


Renal complications of polyarteritis nodosa: CT findings.
Three patients with multiple renal complications of polyarteritis nodosa are described. The arteriographic findings in these patients were multiple arterial aneurysms and tissue infarctions, with the development of spontaneous intrarenal, subcapsular, and perirenal hemorrhagic complications. Computed tomography showed the hemorrhages and multiple renal infarcts; acute and late forms could be differentiated on the basis of contrast medium enhancement and scar formation.


Proteo-dendrimers designed for complementary recognition of cytochrome c: dendrimer architecture toward nanoscale protein complexation.
"Proteo-dendrimers" in which polyanionic hepta(glutamic acids), fluorescent zinc porphyrinate cores, hydrophilic polyether surfaces, and nonpeptide hydrophobic dendrons are combined, were developed as a new series of synthetic receptors for protein recognition. They have polyanionic "patch" structures on their surfaces and undergo complementary electrostatic interactions with a positively charged cytochrome c patch, as observed in biological protein-protein recognition systems. Stability constants of the resulting supramolecular complexes were determined in phosphate buffer (pH 7) by monitoring the fluorescence quenching of the zinc porphyrinates. These proteo-dendrimer receptors exhibited higher affinities with cytochrome c proteins in aqueous solutions than with biological cytochrome b5. Furthermore, they effectively blocked complexation of biological cytochrome b5 with cytochrome c, indicating that the proteo-dendrimers and cytochrome b5 similarly occupy the polycationic patch of cytochrome c.


Fetal demise by umbilical cord around abdomen and stricture.
Umbilical cord abnormalities are accepted as conditions associated with intrauterine fetal demise (IUFD), and umbilical cord stricture is most frequently encountered. In addition, although cord entanglement with multiple loops rarely increases the perinatal mortality, it is associated with a significant increase in variable kind of morbidity such as growth restriction. We describe a 27-year-old woman, with a missed abortion history at about 10 weeks' gestation in her first pregnancy, who presented to our outpatient department at 34 4/7 weeks of gestation due to decreased fetal activity during the preceding week. No fetal heart activity and blood flow had been detected by ultrasonography and pulsed-wave Doppler. A demised fetus with umbilical cord stricture and three loops around abdomen was delivered and was weighted 1,830 g that was below the tenth percentile for the gestational age. Either umbilical cord stricture or entanglement around the body can affect the development of the fetus and even be lethal. The former might play a more important role in this case. Their etiology and the sequence of the events are still undetermined, and additional evaluation such as autopsy and further research may be needed. In addition, counsel and frequent fetal surveillance should be done in patients with previous IUFD attributed to cord stricture during next pregnancy because of undetermined risk of recurrence.


Identification of Surface Antigens That Define Human Pluripotent Stem Cell-Derived PRRX1+Limb-Bud-like Mesenchymal Cells.
Stem cell-based therapies and experimental methods rely on efficient induction of human pluripotent stem cells (hPSCs). During limb development, the lateral plate mesoderm (LPM) produces limb-bud mesenchymal (LBM) cells that differentiate into osteochondroprogenitor cells and form cartilage tissues in the appendicular skeleton. Previously, we generated PRRX1-tdTomato reporter hPSCs to establish the protocol for inducing the hPSC-derived PRRX1+ LBM-like cells. However, surface antigens that assess the induction efficiency of hPSC-derived PRRX1+ LBM-like cells from LPM have not been identified. Here, we used PRRX1-tdTomato reporter hPSCs and found that high pluripotent cell density suppressed the expression of PRRX1 mRNA and tdTomato after LBM-like induction. RNA sequencing and flow cytometry suggested that PRRX1-tdTomato+ LBM-like cells are defined as CD44high CD140Bhigh CD49f-. Importantly, other hPSC lines, including four human induced pluripotent stem cell lines (414C2, 1383D2, HPS1042, HPS1043) and two human embryonic stem cell lines (SEES4, SEES7), showed the same results. Thus, an appropriate cell density of hPSCs before differentiation is a prerequisite for inducing the CD44high CD140Bhigh CD49f- PRRX1+ LBM-like cells.


A Novel Antimicrobial Peptide Scyreprocin From Mud Crab Scylla paramamosain Showing Potent Antifungal and Anti-biofilm Activity.
Natural antimicrobial peptides (AMPs) are potential antibiotic alternatives. Marine crustaceans are thought to generate more powerful and various AMPs to protect themselves from infections caused by pathogenic microorganisms in their complex aquatic habitat, thus becoming one of the most promising sources of AMPs or other bioactive substances. In the study, a novel protein was identified as an interacting partner of male-specific AMP SCY2 in Scylla paramamosain and named scyreprocin. The recombinant product of scyreprocin (rScyreprocin) was successfully expressed in Escherichia coli. rScyreprocin exerted potent, broad-spectrum antifungal, antibacterial, and anti-biofilm activity (minimum inhibitory concentrations from 0.5 to 32 μM) through differential modes of action, including disruption of cell membrane integrity and induction of cell apoptosis, and has rapid bactericidal (in 0.5-2 h) and fungicidal (in 8-10 h) kinetics. In addition to its fungicidal activity against planktonic fungi, rScyreprocin also prevented the adhesion of fungal cells, inhibited biofilm formation, and eradicated the mature biofilms. Moreover, rScyreprocin showed a profound inhibitory effect on spore germination of Aspergillus spp. (minimum inhibitory concentrations from 4 to 8 μM). This peptide was not cytotoxic to murine and mammalian cells and could increase the survival rate of Oryzias melastigma under the challenge of Vibrio harveyi. Taken together, the novel AMP scyreprocin would be a promising alternative to antibiotics used in aquaculture and medicine.


Regulation of asparaginase, glutamine synthetase, and glutamate dehydrogenase in response to medium nitrogen concentrations in a euryhaline chlamydomonas species.
The ammonium assimilatory enzymes glutamine synthetase (EC 220.127.116.11) and glutamate dehydrogenase (EC 188.8.131.52) were investigated for a possible role in the regulation of asparaginase (EC 184.108.40.206) in a Chlamydomonas species isolated from a marine environment. Cells grown under nitrogen limitation (0.1 millimolar NH(4) (+), NO(3) (-), or l-asparagine) possessed 6 times the asparaginase activity and approximately one-half the protein of cells grown at high nitrogen levels (1.5 to 2.5 millimolar). Biosynthetic glutamine synthetase activity was 1.5 to 1.8 times greater in nitrogen-limited cells than cells grown at high levels of the three nitrogen sources.Conversely, glutamate dehydrogenase (both NADH- and NADPH-dependent activities) was greatest in cells grown at high levels of asparagine or ammonium, while nitrate-grown cells possessed little activity at all concentrations employed. For all three nitrogen sources, glutamate dehydrogenase activity was correlated to the residual ammonium concentration of the media after growth (r = 0.88 and 0.94 for NADH- and NADPH-dependent activities, respectively).These results suggest that glutamate dehydrogenase is regulated in response to ambient ammonium levels via a mechanism distinct from asparaginase or glutamine synthetase. Glutamine synthetase and asparaginase, apparently repressed by high levels of all three nitrogen sources, are perhaps regulated by a common mechanism responding to intracellular nitrogen depletion, as evidenced by low cellular protein content.


Improvement of cardiac contractile response to beta-adrenergic stimulation in normal and two-kidney, one-clip hypertensive rats treated with nitrendipine.
To evaluate the effect of nitrendipine on cardiac hypertrophy and inotropic response to isoproterenol in two-kidney, one-clip (2K,1C) renovascular hypertension, male Wistar rats (n = 56) were divided into a clipped group (K) (n = 28) and a sham group (S) (n = 28). Twenty-one days after surgery, the rats were placed in metabolic cages where they received either a normal diet (S and K rats) or a similar one containing N (18 mg/day) (SN and KN rats) during 3 weeks. Arterial pressure and body weight were measured twice a week. At the end of the experimental period, the hearts were excised, the ventricles were weighed, and cardiac proteins were measured by the Lowry method in seven hearts of each group. In the remaining hearts, the left ventricular papillary muscle was excised and mounted in a bath where the developed tension (dt) and the maximal rate of rise of developed tension (+T) were recorded in basal conditions, and after cumulative doses of isoproterenol (10(-11) to 10(-4) M). The arterial pressure, ventricular weight, and cardiac proteins were similar in S, SN, and KN groups and were significantly higher in the K group (p less than 0.05). No difference in the basal values of +T and dt were observed among the four groups. The increment in +T after isoproterenol was higher in treated (KN and SN) than in nontreated (K and S) groups (p less than 0.05). In conclusion, nitrendipine enhances the contractile response to isoproterenol and reverses the cardiac hypertrophy in the 2K, 1C model.


Enhanced oxidative stress as a potential mechanism underlying the programming of hypertension in utero.
Maternal undernutrition during critical periods of organ development is known to impair fetal growth and predispose to the development of adulthood diseases, such as hypertension, coronary heart disease and type II diabetes that are linked to low birth weight and are characterized by endothelial dysfunction. Increased oxidative stress, in rats submitted to intrauterine undernutrition, provides a potential explanation for the endothelial dysfunction development. The aim of this study was to determine the oxidative stress and its consequence on mesenteric arteriolar responses to vasoactive agents in offspring from diet-restricted dams. For this, female pregnant Wistar rats were fed either normal or 50% of normal intake diets, during the whole gestational period. In male offspring, arterial blood pressure was determined by the tail cuff method in anesthetized rats, mesenteric arteriolar reactivity and superoxide anion generation were studied using intravital microscopy and superoxide dismutase activity was determined in mesentery by spectrophotometric assay. Intrauterine undernutrition induced hypertension, decreased vasodilation to acetylcholine and bradykinin but did not alter the responses to sodium nitroprusside. Topical application of superoxide dismutase and superoxide dismutase mimetic manganese (III) tetrakis (1-methyl-4-pyridyl) porphyrin significantly improved the altered arteriolar responses to acetylcholine and bradykinin. A decreased superoxide dismutase activity and an increased superoxide anion concentration were observed in the offspring of diet-restricted dams. This study shows for the first time that intrauterine undernutrition enhances oxidative stress in vivo and relates this to the impaired endothelium-dependent vasodilation.


Synthetic photoplethysmography (PPG) of the radial artery through parallelized Monte Carlo and its correlation to body mass index (BMI).
Cardiovascular disease is one of the leading causes of death in the United States and obesity significantly increases the risk of cardiovascular disease. The measurement of blood pressure (BP) is critical in monitoring and managing cardiovascular disease hence new wearable devices are being developed to make BP more accessible to physicians and patients. Several wearables utilize photoplethysmography from the wrist vasculature to derive BP assessment although many of these devices are still at the experimental stage. With the ultimate goal of supporting instrument development, we have developed a model of the photoplethysmographic waveform derived from the radial artery at the volar surface of the wrist. To do so we have utilized the relation between vessel biomechanics through Finite Element Method and Monte Carlo light transport model. The model shows similar features to that seen in PPG waveform captured using an off the shelf device. We observe the influence of body mass index on the PPG signal. A degradation the PPG signal of up to 40% in AC to DC signal ratio was thus observed.


Steady-state pharmacokinetics of corticosteroid delivery from glucuronide prodrugs in normal and colitic rats.
Ulcerative colitis and Crohn's colitis are chronic intestinal diseases usually treated with various nonsteroidal antiinflammatory agents to maintain remission. Corticosteroids, while useful in acute treatment of these diseases, present side-effects generally too serious to allow maintenance therapy. Colon-specific drug delivery may permit use of corticosteroids for maintenance therapy if doses can be reduced while maintaining efficacy. In this study, two prodrugs (dexamethasone-beta-D-glucuronide (DXglrd) and budesonide-beta-D-glucuronide (BUDglrd)) were administered by intragastric (i.g.) infusion to conventional and colitic rats. In addition, dexamethasone (DX) and budesonide (BUD) were administered either i.g. or subcutaneously (s.c.) to healthy and colitic rats. Colon-specific delivery was assessed using the drug delivery index (DDI). In conventinal rats, DDIs for DXglrd ranged from about five to as high as 11 in the luminal contents relative to DX administered sc or i.g. DDI values were also elevated in the mucosa of both healthy and colitic rats following i.g. administration of DXglrd. BUD was delivered somewhat less effectively from BUDglrd to the rat large intestine than was DX from DXglrd. The data are consistent with efficacy studies and support the conclusion that local delivery of corticosteroids to the large intestine is due, at least in part, to higher levels of drug delivery into the mucosal tissues.


[Current status of chondrosarcoma in a reference center].
Chondrosarcoma is the second most frequent primary malignant bone tumor, is characterized by the formation of a chondroid matrix, is more frequent in advanced ages and its prognosis is directly related to the histological subtype and tumor grade. The objective of this article is to present the demographic and treatment profile in a reference center. A retrospective, descriptive, observational, analytical cohort-type study was conducted in a reference center between January 1, 2005 and December 31, 2019, the demographic variables were analyzed and contrasted with the outcome variables that were overall survival and disease-free period. 85 patients were included, the mean age of presentation was 43.3 years, the clinical stage at the time of presentation was mostly IIa and IIb covering 54.1%. 35 patients had metastases with clinical stage IV. The main site of metastasis was the lung in 32 patients (37.6%). Thirty-six patients (42.3%) had specific mortality. 35 patients (41.2%) died as a result of metastatic disease to lung or other organs, one patient died from direct extension of the tumor in the cervical spine. The important prognostic factors found in our series that influence global survival are: the presence of metastases and axial localization.


β-Cyclodextrin-Stabilized Biosynthesis Nanozyme for Dual Enzyme Mimicking and Fenton Reaction with a High Potential Anticancer Agent.
The myth of inactivity of inorganic materials in a biological system breaks down by the discovery of nanozymes. From this time, the nanozyme has attracted huge attention for its high durability, cost-effective production, and easy storage over the natural enzyme. Moreover, the multienzyme-mimicking activity of nanozymes can regulate the level of reactive oxygen species (ROS) in an intercellular system. ROS can be generated by peroxidase (POD), oxidase (OD), and Fenton-like catalytic reaction by a nanozyme which kills the cancer cells by oxidative stress; therefore, it is important in CDT (chemo dynamic therapy). Our current study designed to investigate the enzyme mimicking behavior and anticancer ability of cerium-based nanomaterials because the cerium-based materials offer a high redox ability while maintaining nontoxicity and high stability. Our group synthesized CeZrO4 nanoparticles by a green method using β-cyclodextrin as a stabilizer and neem leaf extract as a reducing agent, exhibiting POD- and OD-like dual enzyme activities. The best enzyme catalytic activity is shown in pH = 4, indicating the high ROS generation in an acidic medium (tumor microenvironment) which is also supported by the Fenton-like behavior of CeZrO4 nanoparticles. Inspired by the high ROS generation in vitro method, we investigated the disruption of human kidney cells by this nanoparticle, successfully verified by the MTT assay. The harmful effect of ROS in a normal cell is also investigated by the in vitro MTT assay. The results suggested that the appreciable anticancer activity with minimal side effects by this synthesized nanomaterial.


Purification and characterization of human biliverdin reductase.
Conversion of biliverdin to bilirubin is catalyzed by the cytosolic enzyme biliverdin reductase. We have purified and characterized the human liver reductase and find it to differ extensively from the previously described rat enzyme (H. Fakhrai and M. D. Maines, 1992, J. Biol. Chem. 267, 4023-4029) in its primary structure/composition, yet share kinetic properties. The human enzyme is substantially larger than the rat enzyme (approximately 41,000-42,000 versus 33,000-34,000), is dual cofactor and dual pH dependent, and requires free-SH groups. At pH 6.0-7.0 the NADH was the more effective cofactor, whereas at pH 8.5-8.75 NADPH was the preferred cofactor. The activity was inhibited by-SH reagents, 5'-dithiobis(2-nitrobenzoic acid) and p-chloromercuribenzoic acid, and protected from these reagents by cofactors and substrate. On two-dimensional electrophoresis, the purified protein resolved into four distinct isoelectric zones (pI 6.03, 5.83, 5.68, and 5.55) and two molecular weight forms (approximately 40,700 and approximately 39,600). Variants with similar pI values were detected in the purified human kidney reductase, although their relative tissue abundance varied. The tryptic map, amino acid composition, and sequence of NH2 terminus and four tryptic peptides of human reductase were compared with those of the rat. The HPLC profile and amino acid composition of the human and the rat enzymes differed vastly, and two tryptic peptides were present in the human that could not be detected in the predicted amino acid sequence of the rat enzyme. At the same time, the first 21 amino acids of the NH2 terminus of rat and human, except for the substitution of glutamic acid in human for lysine (amino acid 4) in the rat, were found identical and two peptides with 78-87% similarity to the rat reductase were found in the human reductase. Of the seven cysteine residues present in the human, four or five were titratable with 5'-dithiobis(2-nitrobenzoic acid).


C1222C Deletion in Exon 8 of ABL1 Is Involved in Carcinogenesis and Cell Cycle Control of Colorectal Cancer Through IRS1/PI3K/Akt Pathway.
Colorectal cancer (CRC) is one of the most commonly diagnosed cancers worldwide. ABL1 (c-Abl) is a non-receptor tyrosine kinase, whose role, and molecular mechanism in CRC remain largely unclear. The aim of this study was to elucidate the role of ABL1 to obtain information on colon cancer gene mutation. We analyzed the tissue samples obtained from patients with CRC, CRC cell lines, and the immunodeficient mice. The proliferation, cell cycle, and apoptosis of CRC cells were examined. IPA software was used to analyze the molecules involved in CRC after ABL1 RNA interference. We found ABL1 was highly expressed in CRC tissues and cells. This high expression was associated with the TNM stage of CRC patients. In exon 8 of the ABL1 gene, we identified a novel mutation of C1222C deletion, which was related to the CRC stage. Depletion of ABL1 resulted in the inhibition of proliferation and escalation of apoptosis in two CRC cell lines, SW480, and HCT-116. Our in vivo study also demonstrated that depletion of ABL1 reduced CRC tumor progression. The results of the ingenuity pathway analysis indicated that the expression of 732 genes was upregulated and that of 691 genes was downregulated in mice transplanted with ABL1-downregulated CRC cells, among which we confirmed that depletion of ABL1 inhibited TGF-β1 via IRS1/PI3K/AKT pathway in CRC progression. These findings demonstrated that ABL1 plays an important role and that it can be a potential molecular target for CRC therapy.


Preparation of Li-Excess and Li-Deficient Antiperovskite Structured Li2+xOH1-xBr and Their Effects on Total Ionic Conductivity.
This paper describes about the effect of Li-H exchange amount on total lithium-ion (Li+) conductivity of Li2+xOH1-xBr (x = -0.5 to +0.4). These samples are systematically prepared at room temperature by a dry ball-milling process using LiOH, LiOH·H2O, Li2O, and LiBr as starting materials. Synchrotron X-ray diffraction analysis reveals that single-phase Li2+xOH1-xBr samples are formed within x = -0.5 to +0.35. For improving total Li+ conductivity (σt), a larger x value increases both the Li carrier density and lattice constant as positive factors, while that decreases both the crystallite size and OH rotational unit possibly assisting Li+ conduction as negative factors. This trade-off provides an optimized σt of 3.6 × 10-6 S cm-1 at the Li-excess Li2.2OH0.8Br composition, which is ca. 3 times higher than pristine Li2OHBr (1.1 × 10-6 S cm-1). The hydrogen incorporation into the lattice is confirmed by neutron diffraction analysis, and the refined composition is almost consistent with the prepared composition.


From Amsterdam to Bamako: a qualitative case study on diffusion entrepreneurs' contribution to performance-based financing propagation in Mali.
For the past 15 years, several donors have promoted performance-based financing (PBF) in Africa for improving health services provision. European and African experts known as 'diffusion entrepreneurs' (DEs) assist with PBF pilot testing. In Mali, after participating in a first pilot PBF in 2012-13, the Ministry of Health and Public Hygiene included PBF in its national strategic plan. It piloted this strategy again in 2016-17. We investigated the interactions between foreign experts and domestic actors towards PBF diffusion in Mali from 2009 to 2018. Drawing on the framework on DEs (Gautier et al., 2018), we examine the characteristics of DEs acting at the global, continental and (sub)national levels; and their contribution to policy framing, emulation, experimentation and learning, across locations of PBF implementation. Using an interpretive approach, this longitudinal qualitative case study analyses data from observations (N = 5), interviews (N = 33) and policy documentation (N = 19). DEs framed PBF as the logical continuation of decentralization, contracting policies and existing policies. Policy emulation started with foreign DEs inspiring domestic actors' interest, and succeeded thanks to longstanding relationships and work together. Learning was initiated by European DEs through training sessions and study tours outside Mali, and by African DEs transferring their passion and tacit knowledge to PBF implementers. However, the short-time frame and numerous implementation gaps of the PBF pilot project led to incomplete policy learning. Despite the many pitfalls of the region-wide pilot project, policy actors in Mali decided to pursue this policy in Mali. Future research should further investigate the making of successful African DEs by foreign DEs advocating for a given policy.


Biological control to diminish dangers of urolithiasis.
High concentrations of phosphate, oxalate, and calcium ions in urine may cause formation of mineral deposits, i.e., urolithiasis. This is prevented in healthy individuals by substances present in trace quantities. However, there is no recognizable difference between normals and stone formers in urinary substance content. The enzymes glutamic oxaloacetic transaminase and glutamic pyruvic transaminase produce glutamic acid which retards calcium oxalate crystallization. The combined transaminase activity in 70 stone former urine samples was 12.2 +/- 4.1 IU and 31.9 +/- 10.7 IU for 47 normal controls. Incubation of stone former urine with glutamic oxaloacetic transaminase improved overall inhibitory potential, raised glutamic acid levels, and decreased aspartic acid concentrations. Correlation was established between the success of therapeutic treatment and the improvement of enzyme activity. The relative content of glutamic acid is low stone former urines and high in active inhibitory fractions of urinary materials. It is suggested that part of the mechanisms of prevention of stone formation is subjected to biological control.


Long-term efficacy and safety of polyalkylimide gel for the treatment of HIV-associated lipoatrophy.
The long-term safety and efficacy of products used in the correction of HIV-associated facial lipoatrophy (FLA) are largely unknown. The purpose of this study was to describe the long-term efficacy and safety of polyalkylimide gel (PAIG) in the treatment of HIV-associated FLA. In this open-label, randomized, single-center study, 31 HIV-positive individuals (median age 48 years (interquartile ranges (IQR) 45, 55, 97% male) with FLA were randomized to immediate (week 0 and six) or delayed (week 12 and 18) PAIG injections. Week 96 endpoints included change in FLA severity scores (FLSS) (five-point scale), proportion of patients with adverse events, and changes in quality of life, depression and anxiety using validated surveys. Results at week 96 were available for 28 patients. Adverse events, including swelling, redness, bruising and pain, were mild, and resolved after a median of three days following the injection. At week 96, median changes in physician and patient FLSS scores were -2 (IQR -3, -1; p<0.001 vs. baseline) and -2 (IQR -2, -1; p<0.001 vs. baseline), respectively. Physician and patient FLSS scores were not significantly different between the groups at week 96. Significant improvements in patient's anxiety (p<0.001), depression (p<0.001) and mental health (p=0.01) were observed from baseline to week 96. In conclusion, treatment with PAIG was associated with sustained improvements in both the physical and psychological components of FLA through 96 weeks of follow-up.


Phasic cutaneous input facilitates locomotor recovery after incomplete spinal injury in the chick.
1. Walking and swimming of hatchling chicks was videotaped before hemisection of the left thoracic cord and thereafter at regular intervals, for up to 2 wk. With the use of kinematic techniques, movements of the left knee and ankle were quantified to assess recovery of the ipsilateral leg during walking and swimming trials. To study the effects of exteroceptive (cutaneous) feedback in the absence of limb loading, one group of animals was also provided with cutaneous stimulation during swimming in the form of neutrally buoyant tubes that only contacted the foot during the retraction (extension) phase of the swim cycle. 2. One day after hemisection, for both swimming and walking, the left knee failed to extend normally, and the ankle joint remained hyperextended. During walking, all chicks adopted an asymmetric gait, whereas during swimming the left leg remained retracted and motionless. 3. Over the next 2 wk, knee extension and ankle flexion during walking recovered to normal preoperative values, but neither measure returned to preoperative values during swimming trials. However, when chicks were provided with phasic cutaneous stimulation during swimming trials, they showed improvements in leg motion as soon as 5 days after hemisection. Temporary removal of the cutaneous stimulation during swimming (5 days after hemisection) resulted in reduced limb action. However, 14 days after hemisection, the improvement in limb motion was retained even when the cutaneous stimulation was not provided. 4. Improvement in leg motion after swim training with phasic cutaneous stimulation took the form of increased extension of the limb during retraction. Possible neurophysiological mechanisms for this behavior include reflex reinforcement of limb extensor activity in response to cutaneous stimulation of the foot. Repeated exposure to phasic stimulation during swimming trials results in a permanent alteration in limb action. Thus increased cutaneous afferent inputs, even in the absence of limb loading, can facilitate locomotor recovery after spinal cord injury.


Inflammasome Activation in Myeloid Malignancies-Friend or Foe?
Myeloid malignancies including myelodysplastic syndromes, myeloproliferative neoplasms and acute myeloid leukemia are heterogeneous disorders originating from mutated hematopoietic stem and progenitor cells (HSPCs). Genetically, they are very heterogeneous and characterized by uncontrolled proliferation and/or blockage of differentiation of abnormal HSPCs. Recent studies suggest the involvement of inflammasome activation in disease initiation and clonal progression. Inflammasomes are cytosolic innate immune sensors that, upon activation, induce caspase-1 mediated processing of interleukin (IL) -1-cytokine members IL-1β and IL-18, as well as initiation of gasdermin D-dependent pyroptosis. Inflammasome activation leads to a pro-inflammatory microenvironment in the bone marrow, which drives proliferation and may induce clonal selection of mutated HSPCs. However, there are also contradictory data showing that inflammasome activation actually counteracts leukemogenesis. Overall, the beneficial or detrimental effect of inflammasome activation seems to be highly dependent on mutational, environmental, and immunological contexts and an improved understanding is fundamental to advance specific therapeutic targeting strategies. This review summarizes current knowledge about this dichotomous effect of inflammasome activation in myeloid malignancies and provides further perspectives on therapeutic targeting.


The effects of instruction and hand dominance on grip-to-load force coordination in manipulation tasks.
The force applied upon a vertically oriented hand-held object could be decomposed into two orthogonal and highly coordinated components: the grip force (GF; the component perpendicular to the hand-object contact area that provides friction) and the load force (LF; the parallel component that can move the object or support the body). The aim of this study was to investigate the underexplored effects of task instruction and hand dominance on GF-LF coordination. Sixteen right-handed subjects performed bimanual manipulation against a horizontally oriented instrumented device under different sets of instructions. The tasks involved exertion of ramp-and-hold or oscillation patterns of LF performed symmetrically with two hands, while the instructions regarding individual actions were either similar (pull with both hands) or dissimilar (pull with one hand and hold with another). The results revealed that the instruction "to pull" leads to higher indices of GF-LF coordination than the instruction "to hold", as evidenced by a lower GF-LF ratio, higher GF-LF coupling, and higher GF modulation. The only effect of hand dominance was a moderate time lag of GF relative to LF changes observed in the non-dominant hand. We conclude that the instructions could play an important role in GF-LF coordination and, therefore, they should be taken into account when exploring or routinely testing hand function. Additionally, the results suggest that the neural control of GF of the non-dominant hand could involve some feedback mechanisms.


Inhibitors of collagenase IV and cell adhesion reduce the invasive activity of malignant tumour cells.
The invasive activities of some malignant cells appear to be activated by contact with laminin. This protein occurs solely in basement membranes and the binding of malignant cells to the surface of this extracellular matrix initiates the invasion process. Passage of the cells across basement membrane requires degradative activity and laminin induces the production of collagenase IV which lyses the collagen IV network. The motility of the cells is enhanced by chemo-attractants and by matrix molecules. Peptides that inhibit the binding of tumour cells to laminin, inhibitors of collagenase IV, and inhibitors of specific pathways of arachidonic acid metabolism prevent invasion as well as the metastasis of malignant cells and could be employed to stop the spread of cancer.


DNA detective: a review of molecular approaches to wildlife forensics.
Illegal trade of wildlife is growing internationally and is worth more than USD$20 billion per year. DNA technologies are well suited to detect and provide evidence for cases of illicit wildlife trade yet many of the methods have not been verified for forensic applications and the diverse range of methods employed can be confusing for forensic practitioners. In this review, we describe the various genetic techniques used to provide evidence for wildlife cases and thereby exhibit the diversity of forensic questions that can be addressed using currently available genetic technologies. We emphasise that the genetic technologies to provide evidence for wildlife cases are already available, but that the research underpinning their use in forensics is lacking. Finally we advocate and encourage greater collaboration of forensic scientists with conservation geneticists to develop research programs for phylogenetic, phylogeography and population genetics studies to jointly benefit conservation and management of traded species and to provide a scientific basis for the development of forensic methods for the regulation and policing of wildlife trade.


Placebo in Surgical Research: A Case-Based Ethical Analysis and Practical Consequences.
Placebo is a form of simulated medical treatment intended to deceive the patient/subject who believes that he/she received an active therapy. In clinical medicine, the use of placebo is allowed in particular circumstances to assure a patient that he is taken care of and that he/she receives an active drug, even if this is not the case. In clinical research placebo is widely used, as it allows a baseline comparison for the active intervention. If the use of placebo is highly regulated in pharmacological trials, surgery studies have a series of particularities that make its use extremely problematic and regarded less favorably. The purpose of this paper is to present three famous cases of placebo use in surgical trials and to perform an ethical analysis of their acceptability using the Declaration of Helsinki as a main regulatory source.


The Role of Cannabinoids in Bone Metabolism: A New Perspective for Bone Disorders.
Novel interest has arisen in recent years regarding bone, which is a very complex and dynamic tissue deputed to several functions ranging from mechanical and protective support to hematopoiesis and calcium homeostasis maintenance. In order to address these tasks, a very refined, continuous remodeling process needs to occur involving the coordinated action of different types of bone cells: osteoblasts (OBs), which have the capacity to produce newly formed bone, and osteoclasts (OCs), which can remove old bone. Bone remodeling is a highly regulated process that requires many hormones and messenger molecules, both at the systemic and the local level. The whole picture is still not fully understood, and the role of novel actors, such as the components of the endocannabinoids system (ECS), including endogenous cannabinoid ligands (ECs), cannabinoid receptors (CBRs), and the enzymes responsible for endogenous ligand synthesis and breakdown, is extremely intriguing. This article reviews the connection between the ECS and skeletal health, supporting the potential use of cannabinoid receptor ligands for the treatment of bone diseases associated with accelerated osteoclastic bone resorption, including osteoporosis and bone metastasis.


[Sensitivity of HIV infection screening assays in 2001].
Early detection of human immunodeficiency virus (HIV) infection is critical for clinical diagnosis and treatment of patients, as well as for ensuring the safety of blood products. Recently, fourth-generation HIV screening assays have been developed with the objective to offer an increased sensitivity by combining detection of anti-HIV antibodies (Ab) with detection of the p24 viral antigen (Ag). Eight different HIV assays commercially available in France (five fourth-generation HIV screening assays and three third-generation HIV Ab-only assays) were compared in a broad number of seroconversion panels (n = 27). This extensive analysis highlights: 1) the importance of p24 Ag detection for early diagnosis; 2) the improved sensitivity of fourth-generation assays over Ab-only tests. In conclusion, these results emphasize the detection limitations of the different assays and suggest improvements for future HIV screening assays.


Specification of ectoderm restricts the size of the animal plate and patterns neurogenesis in sea urchin embryos.
The animal plate of the sea urchin embryo becomes the apical organ, a sensory structure of the larva. In the absence of vegetal signaling, an expanded and unpatterned apical organ forms. To investigate the signaling that restricts the size of the animal plate and patterns neurogenesis, we have expressed molecules that regulate specification of ectoderm in embryos and chimeras. Enhancing oral ectoderm suppresses serotonergic neuron differentiation, whereas enhancing aboral or ciliary band ectoderm increases differentiation of serotonergic neurons. In embryos in which vegetal signaling is blocked, Nodal expression does not reduce the size of the thickened animal plate; however, almost no neurons form. Expression of BMP in the absence of vegetal signaling also does not restrict the size of the animal plate, but abundant serotonergic neurons form. In chimeras in which vegetal signaling is blocked in the entire embryo, and one half of the embryo expresses Nodal, serotonergic neuron formation is suppressed in both halves. In similar chimeras in which vegetal signaling is blocked and one half of the embryo expresses Goosecoid (Gsc), serotonergic neurons form only in the half of the embryo not expressing Gsc. We propose that neurogenesis is specified by a maternal program that is restricted to the animal pole by signaling that is dependent on nuclearization of beta-catenin and specifies ciliary band ectoderm. Subsequently, neurogenesis in the animal plate is patterned by suppression of serotonergic neuron formation by Nodal. Like other metazoans, echinoderms appear to have a phase of neural development during which the specification of ectoderm restricts and patterns neurogenesis.


Proteinuria in amyloidosis correlates with epithelial detachment and distortion of amyloid fibrils.
To obtain a better understanding of the mechanism of proteinuria in amyloidosis we investigated the ultrastructural distribution and pattern of arrangement of glomerular amyloid fibrils in 15 patients. Clinical data were correlated with the ultrastructural features and with the morphometric indices obtained from composite electron micrographs of whole glomeruli. The epithelial side of amyloid deposits showed spicules (discrete pointed bundles of fibrils) and apparent fraying which we termed tufts (ill-defined and diffuse expansion of fibrils). The extent of mesangial amyloid deposition and total GBM deposition did not correlate with proteinuria. Current data suggest the importance of partial detachment of epithelial cells in pathogenesis of proteinuria, which in turn correlates with distortion of amyloid fibrils, spicules and tufts.


Three-dimensional digital mouse atlas using high-resolution MRI.
We present an archetypal digital atlas of the mouse embryo based on microscopic magnetic resonance imaging. The atlas is composed of three modules: (1) images of fixed embryos 6 to 15.5 days postconception (dpc) [Theiler Stages (TS) 8 to 24]; (2) an annotated atlas of the anterior portion of a 13.5 dpc (TS 22) mouse with anatomical structures delineated and linked to explanatory files; and (3) three-dimensional renderings of the entire 13.5 dpc embryo and specific organ systems. The explanatory files include brief descriptions of the structure at each volume element in the image and links to 3D reconstructions, allowing visualization of the shape of the isolated structures. These files can also contain or be linked to other types of information and data including detailed anatomical and physiological information about structures with pointers to online references, relationships between structures, temporal characteristics (cell lineage patterns, size, and shape changes), and gene expression patterns (both spatial and temporal). As an example, we have "painted" in the expression pattern of Dlx5/Dlx6 genes. This digital atlas provides a means to put specific data within the context of normal specimen anatomy, to analyze the information in 3D, and to examine relationships between different types of information.


Integrating REDCap patient-reported outcomes with the HealtheIntent population health platform: proof of concept.
Patient-reported outcome measures (PROMs) are critical to drive patient-centered care and to understanding patients' perspectives on their health status, quality of life, and the overall effectiveness of the care they receive. PROMs are increasingly being used in clinical and research settings, but the mechanisms to aggregate data from different systems can be cumbersome. As part of an FDA Real-World Evidence demonstration project, we enriched routine care clinical data from our Cerner electronic health record (EHR) with PROMs collected using REDCap. We used SSIS, sFTP, and the REDCap Application Programming Interface to aggregate both data sources into the Cerner HealtheIntent Population Health Platform. We successfully built dashboards, reports, and datasets containing both REDCap and EHR data collected prospectively. This technically straightforward approach using commonly available clinical and research tools can be readily adopted and adapted by others to better integrate PROMs with clinical data sources.


Coronary intervention in patients>or=75 years old with ST-elevation myocardial infarction: in-hospital and 6-month clinical outcomes.
ST-segment elevation myocardial infarction (STEMI) in elderly patients presents specific clinical characteristics. The study on percutaneous coronary intervention (PCI) in elderly patients (>or=75 years) with STEMI, however, has less been performed. In the present study, 522 consecutive STEMI patients undergoing PCI within 12 hours from symptom onset were investigated, and clinical characteristics and in-hospital and 6-month outcomes of 66 elderly patients (>or=75 years, group A) were compared to those of 456 younger patients (<75 years, group B). Compared to younger patients, elderly ones had more females (42.4% vs. 17.8%, P<0.005), a history of cerebral vascular events (7.6% vs. 0.9%, P<0.05), higher serum creatinine level ((96.48+/-31.65) mmol/L vs. (84.87+/-19.81) mmol/L, P<0.005) and fewer smokers (28.8% vs. 45.4%, P<0.05). The elderly ones had worse Killip class (Killip I class: 69.7% vs. 85.7%, P<0.05), less drug-eluting stent implantation and lower rates of TIMI flow 3 following PCI (33.3% vs. 47.1%, and 84.8% vs. 94.7%, P<0.05 respectively). Additionally, both in-hospital mortality and myocardial infarction rate were found to be higher in elderly patients (16.7% vs. 1.5%, and 7.6% vs. 2.6%, P<0.05 respectively), which were also observed until 6-month follow-up (9.1% vs. 0, and 6.1% vs. 0, P<0.05 respectively). In multivariable Cox regression analysis, serum creatinine level, history of hypertension, left anterior descending coronary artery as infarct-related artery and Killip class were independent predictors of 6-month overall death in elderly patients. The clinical characteristics of elderly patients with STEMI after PCI are different from those of younger patients. Although PCI in this population is with a low rate of PCI failure, it is still associated with a worse outcome.


A study of the structure and dynamics of complexes between polymyxin B and phosphatidylglycerol in monolayers by fluorescence.
Interactions between the antibiotic polymyxin B and monolayers of dipalmitoylglycerophosphoglycerol have been reinvestigated through a study of the structure and dynamics of the complexes by means of an interface fluorimeter of our fabrication. A fluorescence technique has been developed where the use of linearly polarized incident beams gives the simultaneous determination of the orientation and the lateral diffusion rate of a fluorescent probe inserted in the film. The present investigation was carried out with 12-(9-anthroyloxy)-stearic acid, a fluorescent compound which forms non-fluorescent photodimers upon illumination. Orientation of the probe was studied by computing the ratio of the two dimerization constants KD and the ratio of the fluorescence intensities obtained with crossed linearly polarized incident lights. The lateral diffusion rate of the probe was obtained by measuring fluorescence recovery after photobleaching (photodimerization) of the probe. Control experiments, carried out with dimyristoylglycerophosphocholine, a lipid which does not interact with polymyxin B, show that the antibiotic does not significantly modify the behaviour of the probe. Both in terms of orientation and dynamics, with respect to dipalmitoylglycerophosphoglycerol, when the antibiotic is present in the subphase (1 microM, saturating conditions), data indicate that the lipid remains in a liquid-expanded state. This is true even at a high surface pressure (pi approximately equal to 37 mN X m-1), above the apparent 'transition' which can be observed at 30-35 mN X m-1 on its compression isotherm. Computation of the contribution of polymyxin B to the film expansion to the conclusion that this 'transition' would be a structural transition between two models of interaction: one, below the 'transition', where the polypeptide ring penetrates between the film-forming lipid molecules and another one, above the 'transition', were the antibiotic is adsorbed at the lipid-water interface with only its hydrocarbon chain penetrating the film.


New approaches to antiplatelet therapy.
The importance of platelets in the pathophysiology of cardiovascular and cerebrovascular disorders has been demonstrated by numerous clinical and pathological studies. Conventional antiplatelet agents are effective in both the primary and secondary prevention of vascular disorders, but suffer for the combined shortcomings of lack of selectivity and relative potency. Many new antiplatelet agents have been developed to overcome these differences, including serotonin receptor antagonists, prostanoid derivatives and antagonists, fibrinogen receptor antagonists, and selective thrombin inhibitors. In this review we consider the mechanisms by which these novel antiplatelet agents impair platelet function and their potential clinical utility.


Relations of proliferative activities of gastric carcinoma cells to lymphatic involvement, venous invasion and prognosis.
This study was to evaluate bivariate bromodeoxyuridine (BrdUrd)/DNA flow cytometric analysis in detection of gastric carcinoma and to study the relations of cellular BrdUrd labeling indices (LI), G2/M-phase fraction (G2/MPF) and DNA ploidy pattern to lymphatic involvement, venous invasion and prognosis. Fresh tumor samples from 60 patients with gastric carcinoma were analyzed by bivariate BrdUrd/DNA flow cytometry. The results were correlated with lymphatic vessel invasion, lymphatic node metastasis, the number of metastatic lymphatic nodes, and venous invasion. Propidium iodide (PI) was used as a fluorescent probe for total cellular DNA, and a monoclonal antibody against BrdUrd was used as a probe for BrdUrd incorporated into DNA. Fluorescent-labeled goat anti-mouse antibody was used as a second antibody. S-phase fractions were measured by in vitro BrdUrd labeling, and DNA ploidy and G2/MPF were also measured. Comparison of survival was performed with the log-rank test, the Chi-square test for qualitative data, and Student's t test for quantu data. BrdUrd LI and G2/MPF values were significantly higher in tumors with lymphatic vessel invasion than in those without invasion respectively (P < 0.01); the patients who had tumors with lymphatic vessel invasion showed a significantly poor prognosis (P < 0.01). Both BrdUrd LI and G2/MPF values were significantly higher in tumors with lymphatic node metastasis than in those without metastasis (P < 0.01). A statistical significant difference was noted in the 5-year survival rates between the patients with lymph node metastasis and those without metastasis. Compared with diploid carcinoma, the incidence of lymph node metastasis was significantly higher in aneuploid carcinoma (P < 0.05), and the patients with aneuploid carcinoma showed a significantly poor prognosis (P < 0.05). BrdUrd LI was significantly higher in patients with more than 5 metastatic lymph nodes than those with 1 - 4 metastatic lymph nodes (P < 0.05) and those without metastasis (P < 0.01). G2/MPF values in those patients either with more than 5 metastatic lymph nodes or 1 - 4 metastatic lymph nodes were higher than those without metastasis (P < 0.01 and P < 0.05). A statistical significance was seen in the 5-year survival rates among the patients with no metastatic lymph node, 1 - 4 metastatic nodes and more than 5 metastatic nodes (P < 0.01). G2/MPF values were significantly higher in patients with venous invasion than in those without invasion (P < 0.01). Positive correlations exist between cellular BrdUrd LI, G2/MPF with lymphatic involvement and prognosis, and DNA aneuploid with lymphatic involvement and prognosis. The same was true between G2/MPF value and venous invasion in gastric carcinoma.


Immunopathogenesis of Behçet's disease.
A working hypothesis is postulated for the immunopathogenesis of Behçet's disease (BD). Microbial infection induced stress upregulates HSP65 and MICA gene product which stimulate gamma delta and alpha beta T cell receptor positive cells, to generate effector and suppressor T cells (fig. 1). These activated T cells, antibodies and neutrophils induce cytokines to modulate the immune response. The end result of these complex cellular and cytokine immune interactions in HLA-B51 or genetically related subjects is to induce pathological changes consistent with BD.


Male infertility.
With infertile couples, one third of cases can be due to a male factor. It is important to have a comprehensive yet efficient approach to identifying potential causes for appropriate counseling and treatment. We combined the experience of a urologist and a gynecologist, both fertility subspecialists, and reviewed current literature on the investigation of male infertility. A history and physical examination supplemented by relevant investigations will help unravel any significant diseases that can be associated with male subfertility or any conditions that may be transmitted to future offspring. Semen analysis is a common, convenient measure of assessing the male. It should precede any invasive tests of the female. While reference values are important in standardization, the current trend in using sperm morphology alone in predicting male fertility remains problematic. Overreliance on this reference can lead to misdiagnosis and unnecessary invasive treatment with intracytoplasmic sperm injection (ICSI). In those who have no identifiable or correctable causes, ICSI provides new hope for couples with male infertility who, in the past, could only choose among therapeutic donor insemination, adoption or voluntary childlessness. With increasing application of these assisted reproductive technologies, it is important to rule out genetic causes, such as cystic fibrosis, and to provide appropriate genetic counseling before embarking on these invasive and costly procedures.


Evidence for energy conservation during pubertal growth. A 10-year longitudinal study (EarlyBird 71).
Diabetes is closely linked to obesity, and obesity rates climb during adolescence for reasons that are not clear. Energy efficiency is important to obesity, and we describe a temporary but substantial fall in absolute energy expenditure, compatible with improved energy efficiency, during the rapid growth phase of puberty. In a longitudinal cohort study lasting 10 years, we measured voluntary energy expenditure as physical activity (PA) by accelerometry, involuntary energy expenditure as resting energy expenditure (REE) by oxygen consumption, body mass index (BMI) and body composition by dual energy X-ray absorptiometry annually on 10 occasions from 7 to 16 years in the 347 children of the EarlyBird study. We used mixed effects modelling to analyse the trends in REE and their relationship to BMI, lean mass (LM), fat mass (FM), age, PA and pubertal stage. Relative REE and total PA fell during puberty, as previously described, but the longitudinal data and narrow age-range of the cohort (s.d.±4m) revealed for the first time a substantial fall in absolute REE during the period of maximum growth. The fall became clearer still when adjusted for FM and LM. The fall could not be explained by fasting insulin, adiponectin, leptin, luteinising hormone or follicle stimulating hormone. There appears to be a temporary but substantial reduction in energy expenditure during puberty, which is unrelated to changes in body composition. If it means higher energy efficiency, the fall in REE could be advantageous in an evolutionary context to delivering the extra energy needed for pubertal growth, but unfavourable to weight gain in a contemporary environment.


Continuous traumatic stress as a mental and physical health challenge: Case studies from South Africa.
This article discusses the condition of continuous traumatic stress as common on the African continent and in other international settings characterised by very high levels of ongoing violence and threat of community, political or war-related origin. Through consideration of three case studies from South Africa, contexts of continuous traumatic stress are described, and the mental health and physical health effects of living in such contexts are elaborated. Cautions are raised about attempting to transpose existing posttraumatic stress models onto individuals exposed to situations of continuous traumatic stress, and guidelines for optimal interventions with such cases are proposed.


Effect of exercise timing on postprandial lipemia in hypertriglyceridemic men.
We investigated the effect of exercise timing on attenuation of postprandial hyper-triglyceridemia (PHTG) in individuals with hypertriglyceridemia (HTG). Subjects were 10 males (TG = 290.1 +/- 28.5 mg/dl). Each subject performed a control trial (Ctr), 12-hr premeal exercise trial (12-hr Pre), and 24-hr premeal exercise trial (24-hr Pre). In each trial, subjects had a fat-rich meal. In the exercise trials they jogged on a treadmill at 60% of their VO2max for 1 hr at a designated time. Blood samples were taken at 0 (immediately before the fat meal), and at 2, 4, 6, 8, and 24 hrs after the meal. The results indicated that plasma TG concentrations in 12-hr Pre were lower than in Ctr and 24-hr Pre (p < 0.03). The area score under the TG concentration curve (TG AUC score) in 12-hr Pre was 37% and 33% lower than in 24-hr Pre and Ctr (p < 0.02), respectively. Insulin concentrations in 12-hr Pre were lower than Ctr and 24-hr Pre (p < 0.001). The plasma nonesterified fatty acid (NEFA) concentration was higher in 12-hr Pre than in both 24-hr Pre and Ctr (p < 0.003). There were no trial differences in both HDLtot-Ch and HDL2-Ch. These results suggest that exercising 12 hrs prior to a fat-meal intake significantly reduces PHTG response whereas exercising 24 hrs prior to the meal does not attenuate PHTG in hypertriglyceridemic men. The effect of an acute exercise bout on PHTG lowering may be short-lived and diminished by 24 hrs.


Uterine surveillance of asymptomatic postmenopausal women taking tamoxifen.
The aim of this study was to assess the ability of transvaginal sonography (TVS) and office hysteroscopy with sharp curettage to characterize the morphological changes in the uteri of asymptomatic postmenopausal women taking long-term tamoxifen for breast cancer. The overall acceptability of a single-visit screening clinic for these women was also evaluated. Fifty-eight women were recruited from patients undergoing regular follow-up at the Leicester Royal Infirmary for breast cancer. A single-visit clinic was acceptable to 94.8% of these women. Transvaginal sonography detected endometrial thickness of greater than 5 mm in 84.5% of cases, but there was no relationship between total tamoxifen exposure and endometrial thickness. Transvaginal sonography also detected uterine lesions such as fibroids and endometrial cysts in 34.5% of cases. Hysteroscopy detected the latter uterine lesions in 53.4% of cases, with three cases (5.2%) of endometrial polyps also being identified in these women. Sharp curettage sampling of the endometrium produced specimens sufficient for diagnosis in 84.5% of cases; 70.7% of specimens were reported as showing types of 'quiescent' endometrium with 13.8% of specimens showing 'active' endometrium. In the latter group, there was a case of complex hyperplasia detected and also a case with granulomatous endometritis. For each histopathological diagnosis identified, there was a wide range of endometrial thickness recorded by TVS. A single-visit screening clinic involving TVS and hysteroscopy with sharp curettage was acceptable to asymptomatic women taking tamoxifen. However, hysteroscopy was more effective than TVS in detecting endometrial lesions such as polyps, fibroids and cystic areas. Although TVS detected endometrial thickness greater than 5 mm in the majority of cases, there were no malignancies detected and, for each histopathological classification, there was a wide range of endometrial thickness associated. Thus, the isolated use of TVS is insufficient for screening the endometria of these women.


Sexual concerns and sexual counseling in heart failure.
Patients with heart failure (HF) face significant challenges in maintaining quality of life (QOL), particularly for sexual intimacy. Although recommended for all cardiac patients, it has been suggested that few HF patients receive sexual counseling. This study explored sexual counseling needs, sexual concerns, and sexual activity using a descriptive survey with HF patients (n = 45), recruited from a HF clinic or cardiology office. Most (77%) had not discussed sexual concerns with a health care professional (HCP). Sexual concerns that were rated as occurring 'occasionally/frequently' included partner overprotectiveness (63%), partner fear of sex (36%), lack of sexual interest (42%), erectile problems (74%), orgasmic difficulties (51%). Frequency of sexual intercourse before HF to present was striking, with 53% reporting no sexual activity in the last 2 months compared with 11% before diagnosis of HF. HCPs must provide sexual counseling to HF patients and partners to enhance QOL and to assist in any adaptations to sexual activity.


Angiotensin II receptor subtypes in rat brain and peripheral tissues.
Angiotensin II (Ang II) receptor binding was localized in rat adrenal gland, kidney, and brain by in vitro autoradiography using the antagonist analogue 125I-[Sar1, Ile8]Ang II and differentiated into type I (AT-1) and type II (AT-2) subtypes using unlabelled non-peptide antagonists specific for Ang II subtypes. AT-1 binding was determined as that remaining in the presence of an excess of the AT-2 antagonist, PD 123177 (10 microM), and AT-2 binding as that remaining in the presence of an excess of the AT-1 antagonist, DUP753 (10 microM). The reducing agent dithiothreitol decreased the binding to AT-1 receptors and enhanced the binding to AT-2 receptors. The rat adrenal gland contains both AT-1 and AT-2 receptors in the ratio of approximately 3:2 in the cortex and 1:9 in the medulla. By contrast, in the kidney only AT-1 receptors were evident in glomeruli, proximal tubule, and inner stripe of the outer medulla. In the brain, the pattern of Ang II receptor subtypes varies greatly from region to region. Many brain structures known to be involved in blood pressure regulation and fluid and electrolyte balance, such as circumventricular organs (including vascular organ of the lamina terminalis, subfornical organ, median eminence, and area postrema), median preoptic nucleus, hypothalamic paraventricular nucleus, and regions in the medulla oblongata involved in autonomic control (nucleus of the solitary tract, dorsal motor nucleus of the vagus, and intermediate reticular nucleus), contain exclusively AT-1 receptors. By contrast, locus coeruleus, lateral septal nuclei, superior colliculus, subthalamic nucleus, many nuclei of the thalamus, and nuclei of the inferior olive contain predominantly AT-2 receptors. The detailed binding characteristics of each subtype were determined by competition studies with a series of antagonists. The pharmacological specificity obtained in kidney, adrenal cortex and adrenal medulla, superior colliculus, and nucleus of the solitary tract produces specificity patterns which confirm the assignments of AT-1 and AT-2 receptors described above. The present study reveals important pharmacological heterogeneity of Ang II receptors in key target organs. The subtype-specific receptor mapping described here is relevant to the understanding of the role of angiotensin peptides in peripheral organs and in the central nervous system and is relevant to the actions of non-peptide Ang II receptor antagonists.


At short telomeres Tel1 directs early replication and phosphorylates Rif1.
The replication time of Saccharomyces cerevisiae telomeres responds to TG1-3 repeat length, with telomeres of normal length replicating late during S phase and short telomeres replicating early. Here we show that Tel1 kinase, which is recruited to short telomeres, specifies their early replication, because we find a tel1Δ mutant has short telomeres that nonetheless replicate late. Consistent with a role for Tel1 in driving early telomere replication, initiation at a replication origin close to an induced short telomere was reduced in tel1Δ cells, in an S phase blocked by hydroxyurea. The telomeric chromatin component Rif1 mediates late replication of normal telomeres and is a potential substrate of Tel1 phosphorylation, so we tested whether Tel1 directs early replication of short telomeres by inactivating Rif1. A strain lacking both Rif1 and Tel1 behaves like a rif1Δ mutant by replicating its telomeres early, implying that Tel1 can counteract the delaying effect of Rif1 to control telomere replication time. Proteomic analyses reveals that in yku70Δ cells that have short telomeres, Rif1 is phosphorylated at Tel1 consensus sequences (S/TQ sites), with phosphorylation of Serine-1308 being completely dependent on Tel1. Replication timing analysis of a strain mutated at these phosphorylation sites, however, suggested that Tel1-mediated phosphorylation of Rif1 is not the sole mechanism of replication timing control at telomeres. Overall, our results reveal two new functions of Tel1 at shortened telomeres: phosphorylation of Rif1, and specification of early replication by counteracting the Rif1-mediated delay in initiation at nearby replication origins.


Phyllanthus emblica L. fruit extract induces chromosomal instability and suppresses necrosis in human colon cancer cells.
Phyllanthus emblica L. (PE) is an edible fruit indigenous to Southeast Asia. It has been considered as a potent functional food due to its numerous pharmacological applications, such as anti-oxidant, antimicrobial, anti-diabetic and protection for multiple organs. The aim of this study was to evaluate the effects of a water extract of PE fruit on genomic damage and cell death in the human colon adenocarcinoma cell line COLO320 using the cytokinesis-block micronucleus cytome assay. Cells were exposed to RPMI-1640 medium containing 0, 20, 40, 80, or 160 μg/mL PE for 24, 48, 72, or 96 hours. The results showed that PE induced a significant decrease in necrosis (p < 0.001) and nuclear division index (NDI) (p < 0.001) in a dose- and time-dependent manner, and there was a highly significant correlation between the reduction of necrosis and NDI (r = 0.820, p < 0.001). Dose- and time-dependent increases (p < 0.001) in the frequency of chromosomal instability (CIN) were observed after PE exposure, and the frequency of CIN was negatively correlated with NDI (r = - 0.640, p < 0.001). PE also significantly increased apoptosis (p < 0.001), and there was a significant correlation of apoptosis with CIN (r = 0.566, p < 0.001). In conclusion, PE suppresses necrosis and delays mitotic progression, which results in massive CIN followed by apoptosis in COLO320 cells.