Contrasting effects of anti-Ly49A due to MHC class I cis binding on NK cell-mediated allogeneic bone marrow cell resistance.
NK subsets have activating and inhibitory receptors that bind MHC-I. Ly49A is a mouse inhibitory receptor that binds with high affinity to H2(d) in both a cis- and trans-manner. Ly49A cis-associations limit trans-interactions with H2(d)-expressing targets as well as mAb binding. We demonstrate that cis-interactions affect mAb effector functions. In vivo administration of anti-Ly49A depleted NK cells in H2(b) but not H2(d) mice. Despite lack of depletion, in vivo treatment with anti-Ly49A reduced NK killing capabilities and inhibited activation, partially due to its agonistic effect. These data explain the previously described in vivo effects on bone marrow allograft rejection observed with anti-Ly49A treatment in H2(d)-haplotype mice. However, prior treatment of mice with poly(I:C) or mouse CMV infection resulted in increased Ly49A expression and Ly49A(+) NK cell depletion in H2(d) mice. These data indicate that, although Ly49 mAbs can exert similar in vivo effects in mice with different MHC haplotypes, these effects are mediated via different mechanisms of action correlating with Ly49A expression levels and can be altered within the same strain contingent on stimuli. This illustrates the marked diversity of mAb effector functions due to the regulation of the level of expression of target Ags and responses by stimulatory incidents such as infection.


The characterisation of interstitial lung disease multidisciplinary team meetings: a global study.
Multidisciplinary team (MDT) diagnosis of interstitial lung disease (ILD) has been proposed as a gold standard, but there are no formal recommendations for MDT process or composition and limited knowledge regarding prevalence in routine practice. We performed a systematic evaluation of ILD diagnostic practice across a range of healthcare settings around the world. Electronic questionnaires were distributed across all global regions via society and collaborators networks. Responses from 457 unique centres across 64 countries were included in the analysis. Of the 350 (76.6%) centres holding formal meetings, the majority held face-to-face MDT meetings (80%), for a minimum of 30 min (93%), and discussed diagnosis (96.9%) and patient management (94.9%) at the meetings. Compared with non-academic and academic non-ILD centres, ILD academic centres reported a higher ILD caseload, held more formal MDT meetings, and were more likely to include histopathology and rheumatology specialists in their diagnostic team. Of the centres holding MDT meetings, 5.5% routinely discussed all new cases at such meetings. An MDT approach to ILD diagnosis is consistently interpreted and widely implemented across a range of routine care settings around the world. This observation will inform future ILD diagnostic agreement studies and diagnostic pathway recommendations.


Increased responsiveness to exogenous insulin administration in the hyperglycemic obese person with type II diabetes.
This study was undertaken to investigate whether obese persons with type II diabetes mellitus showed less of a response to the hypoglycemic effect of insulin treatment than did lean type II diabetic patients. Forty-three patients (lean, n = 17; obese, n = 22; obese on hypercaloric diets, n = 4) were treated with insulin and placed on a diet calculated to maintain ideal body weight. A rapid decrease in the mean serum glucose concentration was seen in both the lean and obese groups following the initiation of insulin therapy. The glucose concentration of the lean patients plateaued after the second day of insulin treatment, whereas the obese patients' levels continued to fall after the third and fourth days. A difference was noted in the caloric intake of the two groups. Lean patients ingested 28 kCal/kg/day; obese patients ingested 17 kCal/kg/day. An additional group of four obese diabetic patients who ingested 28 kCal/kg/day was studied to determine if the hypocaloric intake of the first group of obese patients had altered their responsiveness to insulin. Despite the higher caloric intake, the mean serum glucose concentration of this group of patients fell more rapidly than did that of the lean patients. Caloric intake during the initiation of insulin therapy does not seem to influence the increase in insulin responsiveness of obese insulin-requiring type II diabetic patients.


Oral and Topical Administration of ROQUETTE Schizochytrium sp. Alleviate Skin Inflammation and Improve Wound Healing in Mice.
The human body is constantly exposed to the risk of traumatic lesions. ROQUETTE Schizochytrium sp. (SCs) is a marine microalgae containing large amounts of health-valuable nutrients, more particularly polyunsaturated fatty acids such as docosahexaenoic acid. SCs was investigated by oral administration (125, 250 and 500 mg/kg) and cutaneous application (2.5, 5.0 and 10.0%) to evaluate its impact in two dermatological disorder models in mice: skin inflammation and wound healing. For skin inflammation, it was administered during 14 days starting one week before the induction of chronic skin inflammation by repeated cutaneous application of 12-O-tetradecanoylphorbol 13-acetate (TPA). For wound healing the microalgae was administered after incisional wound healing of the skin until complete wound healing. Results indicated that oral and topical administrations of the two higher doses of SCs had significant effects on macroscopic score of skin inflammation. It had also efficient effect on healing process and duration of wound healing with a dose-response by oral administration and a maximal effect observed from the lowest to the highest dose by topical application. These findings suggest that administration of SCs by both oral and topical routes appeared to have beneficial effects on skin lesions.


Association of Patients' Perception of Quality of Healthcare Received and Colorectal Cancer Screening Uptake: An Analysis of 2 National Surveys in the USA.
It is not known whether patients' ratings of the quality of healthcare services they receive truly correlate with the quality of care from their providers. Understanding this association can potentiate improvement in healthcare delivery. We evaluated the association between patients' ratings of the quality of healthcare services received and uptake of colorectal cancer (CRC) screening. We used 2 iterations of the Health Information National Trends Survey (HINTS) of adults in the USA. HINTS 2007 (4,007 respondents; weighted population = 75,397,128) evaluated whether respondents were up to date with CRC screening while HINTS 4 cycle 3 (1,562 respondents; weighted population = 76,628,000) evaluated whether participants had ever received CRC screening in the past. All included respondents from both surveys were at least 50 years of age, had no history of CRC, and had rated the quality of healthcare services that they had received at their healthcare provider's office in the previous 12 months. HINTS 2007 data showed that respondents who rated their healthcare as good or fair/poor were significantly less likely to be up to date with CRC screening compared to those who rated their healthcare as excellent. We found comparable results from analysis of HINTS 4 cycle 3 data with poorer uptake of CRC screening as the healthcare quality ratings of respondents reduced. Our study suggests that patients who reported receiving lower quality of healthcare services were less likely to have undergone and be compliant with CRC screening recommendations. It is important to pay close attention to patient feedback surveys in order to improve healthcare delivery.


Prognosis assessment in stroke patients at discharge from hospital.
accurate prognostic assessments of need of help, health and dwelling situation in stroke patients are important for patient management, rehabilitation, discharge planning, and for providing reliable information to patients and their relatives. to analyse factors affecting the accuracy of discharge prognosis assessments. prospective study of stroke patients discharged from hospital. two cities in central Sweden. three hundred and ninety stroke patients, 65 years or older, living in their own homes and having no dementia diagnosis prior to hospital admission. at discharge, physicians, nurses, occupational therapists and physiotherapists in the Departments of Internal Medicine and Geriatrics were asked to make an individual prognosis assessment regarding patients' need for help, health and dwelling situation at 3 and 12 months after admission to hospital. the prognosis assessments were on average accurate in 68.4% (3 months) and 61.5% (12 months), far better than chance (33.3%). There were no significant differences between staff categories. The accuracy was influenced by a number of patient linked factors, such as activity degree, household situation before admission, and Mini Mental State Examination level and need of help measured 1 week after discharge, and ranged from 22 to 89%, depending on factor combinations. prognosis assessments based on clinical judgement were on average quite accurate but the accuracy varied markedly with patient linked factors. Feedback of outcome might be one way to further improve the accuracy of prognosis assessment.


Ultraviolet B 308-nm excimer laser treatment of psoriasis: a new phototherapeutic approach.
Excimer laser-derived 308-nm ultraviolet (UV) B therapy is a new alternative for treating psoriasis by phototherapy. Some studies have been made showing the effectiveness of intralesional phototherapy technology in treating psoriasis. However, there has been no information available so far with regard to the cumulative dosage on a larger group of patients and on therapy optimized treatment strategies. One hundred and twenty psoriatic patients were treated according to standard protocol to define the effectiveness. Our aim was to develop new parameters and determine whether effectiveness could be improved and whether treatment exposure, the cumulative UVB dose and adverse effects could be minimized. Initially, the excimer laser's effectiveness in treating psoriasis was evaluated in an open prospective study according to standard protocol. This included 120 adult patients (67 female/53 male) with chronic plaque psoriasis and < 20% involved body surface. The initial dose was based on three multiples of a predetermined minimal erythema dose (MED). Patients were treated twice a week for the first 3 weeks, then once a week until clearance was achieved. The main parameters were the initial starting dose, psoriasis area and severity index (PASI), the number of treatments needed, the time in treatment and the cumulative dose needed to clear psoriatic plaques. Thereafter, 43 patients were treated as a second comparable group. Therapy began with a starter dose, defined as MED-I. MED-I was defined as a UVB 308-nm dose which provoked a visible increase in erythema after 24 h. In addition, the epidermal thickness of the plaques was measured on an individual basis by 20-MHz ultrasound and correlated to the MED-I. Of the patients who met the standard protocol, 65.7% were at least 90% clear after a maximum of 10 treatments; an even greater number (85.3%) showed a > or = 90% improvement in PASI after 13 sessions, while 14.7% of patients had only a < or = 50% PASI improvement. The cumulative UVB dose was 11.25 +/- 4.21 J cm-2 and the average treatment time was 7.2 weeks. Patients treated individually with the MED-I starter dose showed nearly identical rates of clearance (83.7%), but were clear in 7.07 +/- 2.15 sessions with a cumulative dose of 6.25 +/- 4.02 J cm-2. The majority of our patients benefited greatly from laser-derived 308-nm UVB therapy, which cleared skin lesions faster than conventional phototherapy. As this therapy targets only the involved skin, the thickness of the plaques and individual MED-I should determine the initial dose, thus increasing the effectiveness of the therapy. We propose that light therapy sparing uninvolved skin will become predominant in the future.


Fully endoscopic endonasal approach for the treatment of intrasellar arachnoid cysts.
To report our experience of endoscopic endonasal obliteration of symptomatic sellar arachnoid cyst (SAC). From 2002 to 2019, we retrospectively reported the data of 17 cases of SAC operated in Timone University Hospital, Marseille, France. Preoperative clinical findings were collected including main symptoms, visual function and endocrinological assessment. Surgical procedure was homogeneous and consisted in endonasal fully endoscopic surgical obliteration of the cyst cavity with fat graft. Post-operative outcomes, complication and follow-up was reported. Visual disorders and/or headaches were the main symptoms. Our technique provided improvement for 83.3% of the patients suffering from visual disturbance and for 87.5% of those suffering from headaches. We reported 2 cases of cerebrospinal fluid (CSF) leakage (11.8%), but no meningitis. One case of definitive diabetes insipidus occurred and one case of postoperative syndrome of inappropriate antidiuretic hormone secretion was temporary. There was one case of recurrence reported. The mean follow-up was 39 months. Patients with symptomatic SAC can be treated successfully by endoscopic endonasal obliteration of the cyst. This simple technique offers true benefits for the patients without craniotomy, but the complication rate remains high especially with the risk of CSF leakage. Special attention to skull base opening and closing could reduce this risk.


Ten-year progress of coronary artery lesions prior to Behçet disease diagnosis: A case report and care-compliant article.
Behçet disease is a multisystemic chronic inflammatory disorder characterized by recurrent oral aphthous ulcers, genital ulcers, uveitis, and skin lesions. However, particularly part of patients would present cardiovascular involvements and vascular lesions could be the presenting sign of Behçet disease preceding classical symptoms. We presented a middle aged male patient, in whom abdominal aorta pseudoaneurysm was as the first leading sign to reveal Behçet disease, and with his coronary artery lesions progress through a 10-year period before Behçet disease was diagnosed. Coronary artery involvement of Behçet disease warrants more attention and investigation; repeated in-stent stenosis, aggressive progress, and elevated inflammation markers should be regard with more care earlier in clinical practice.


Manifestations of Mucormycosis and Its Complications in COVID-19 Patients: A Case Series Study.
Mucormycosis is an opportunistic fungal disease that affects immunocompromised patients. With the advent of SARS-CoV-2, this opportunistic disease has increased. A case series of 47 patients with COVID-19 associated mucormycosis have been analyzed. Demographic information, signs, symptoms, laboratory investigations, imaging studies, and their association with ICU admission and 30-day mortality were assessed. Total number of 47 consecutive rhino-orbital cerebral mucormycosis (ROCM) cases were analyzed. Periorbital swelling was the most common sign among patients. Majority of cases had diabetes. All patients received liposomal Amphotericin B. Debridement was performed for all cases. SARS-CoV-2 increases the susceptibility to mucormycosis infection in various ways. Uncontrolled level of HbA1c in all patients, even non-diabetic individuals, indicates hyperglycemia over the past three months. Diabetes, orbital exenteration, ptosis, periorbital swelling, DKA, LOC, brain involvement, and mechanical ventilation all correlated with a higher rate of ICU admission and 30-day mortality. In addition, a higher white blood cell count is related to the higher probability of ICU admission. While considering all of the inflammatory laboratory data and HbA1c could help predict 30-day mortality.


Massive perivillous fibrinoid causing recurrent placental failure.
To establish the incidence, recurrence rate and consequences of massive perivillous fibrinoid. Retrospective analysis of the histology of all placentas with a diagnosis of massive perivillous fibrinoid between 1991 and 1998, together with the maternal case records. The histopathology department of the Rotunda Hospital, Dublin, Ireland. A relatively homogeneous group of pregnant women in the northern part of Dublin City, which is the catchment area for the Rotunda Hospital, delivered between 1991 and 1998. Retrospective review of archival placental pathology and maternal charts. The incidence of massive perivillous fibrinoid, perinatal outcome and recurrence rate. The incidence of massive perivillous fibrinoid was 0.028%, with a recurrence rate of approximately 18%. All the infants suffered intrauterine growth restriction; there was a 31% fetal loss rate and a 33% preterm delivery rate. Massive perivillous fibrinoid is associated with intrauterine death, intrauterine growth restriction and preterm delivery. It has a significant recurrence rate and both the clinical findings of intrauterine growth restriction and the postmortem findings imply a syndrome of chronic placental insufficiency.


An unrecognized cause of paralysis in ED: thyrotoxic normokalemic periodic paralysis.
Hypokalemic paralysis associated with hyperthyroidism (TPP) is a well-known acute electrolyte and muscle function disorder. Lesser known is normokalemic periodic paralysis associated with hyperthyroidism. We describe two cases of young men with acute muscular paralysis and bilateral impairment of sensation over the lower legs who had normal plasma potassium concentrations. They were initially misdiagnosed as having Guillain-Barré syndrome or hysterical paralysis. However, thyroid function tests showed elevated serum T(3) and T(4) and markedly depressed thyroid-stimulating hormone findings consistent with hyperthyroidism. Control of the hyperthyroidism completely abolished their periodic paralysis. Thyrotoxic normokalemic periodic paralysis (TNPP) should be kept in mind as a cause of acute muscle weakness to avoid missing a treatable and curable condition.


Head-to-Head Comparison of Two SARS-CoV-2 Serology Assays.
While molecular techniques remain the gold standard for diagnosis of acute SARS-CoV-2 infection, serological tests have the unique potential to ascertain how much of the population has been exposed to the COVID-19 pathogen. There have been limited published studies to date documenting the performance of SARS-CoV-2 antibody assays. We compared the DiaSorin Liaison SARS-CoV-2 S1/S2 IgG and Roche Diagnostics Elecsys Anti-SARS-CoV-2 assays using 228 samples spanning patients with positive PCR for SARS-CoV-2, patients with compatible symptoms but negative PCR, pre-COVID specimens, and potential cross-reactives. Both assays detected antibodies in 18/19 samples collected at least one week after a positive PCR result. Neither method consistently detected antibodies in specimens collected within one week of a positive PCR result (sensitivity < 50%), but antibodies were detected by only Roche in four samples in this time frame. Using 139 pre-COVID and 35 PCR-negative samples, the Roche and DiaSorin assays demonstrated specificities of 100.0% and 98.9%, respectively. Neither assay demonstrated cross-reactivity from other coronaviruses (229E, HKU1, NL63, OC43), respiratory pathogens (adenovirus, metapneumovirus, rhinovirus/enterovirus), or antibodies to other viruses (HIV, EBV, CMV, HBV, HCV, HAV). Overall, the qualitative interpretations afforded by the Roche and DiaSorin assays agreed for 97% of samples evaluated. Minor discrepancies in sensitivity and specificity were observed between methods, with the differences in specificity more clinically significant for our low-prevalence population. For the DiaSorin assay, all disagreements with the Roche assay occurred in samples with quantitative signals near the cut-off determining positivity.


Long-term results (≤18 years) of the edge-to-edge mitral valve repair without annuloplasty in degenerative mitral regurgitation: implications for the percutaneous approach.
To assess the long-term results of the edge-to-edge mitral repair performed without annuloplasty in degenerative mitral regurgitation (MR). From 1993 to 2002, 61 patients with degenerative MR were treated with an isolated edge-to-edge suture without any annuloplasty. Annuloplasty was omitted in 36 patients because of heavy annular calcification and in 25 for limited annular dilatation. A double-orifice repair was performed in 53 patients and a commissural edge-to-edge in 8. Hospital mortality was 1.6%. Follow-up was 100% complete (mean length, 9.2±4.21 years; median, 9.7; longest, 18.1). Survival at 12 years was 51.3±7.75%. At the last echocardiographic examination, MR ≥3+ was demonstrated in 33 patients (55%). At 12 years, freedom from reoperation was 57.8±7.21% and freedom from recurrence of MR ≥3+ was 43±7.6%. Residual MR >1+ at hospital discharge was identified as a risk factor for recurrence of MR ≥3+ (hazard ratio, 3.8; 95% confidence interval, 1.7-8.2; P=0.001). In patients with residual MR ≤1+ immediately after surgery, freedom from MR ≥3+ at 5 and 10 years was 80±6% and 64±7.58%, respectively. In degenerative MR, the overall long-term results of the surgical edge-to-edge technique without annuloplasty are not satisfactory. Early optimal competence (residual MR ≤1+) was associated with higher freedom from recurrent severe regurgitation.


Processing of the precursor of protamine P2 in mouse. Peptide mapping and N-terminal sequence analysis of intermediates.
Protamine P2, the major basic chromosomal protein of mouse spermatozoa, is synthesized as a precursor almost twice as long as the mature protein, its extra length arising from an N-terminal extension of 44 amino acid residues. This precursor is integrated into chromatin of spermatids, and the extension is processed during chromatin condensation in the haploid cells. We have studied processing in the mouse and have identified two intermediates generated by proteolytic cleavage of the precursor. H.p.l.c. separated protamine P2 from four other spermatid proteins, including the precursor and three proteins known to possess physiological characteristics expected of processing intermediates. Peptide mapping indicated that all of these proteins were structurally similar. Two major proteins were further purified by PAGE, transferred to poly(vinylidene difluoride) membranes and submitted to automated N-terminal sequence analysis. Both sequences were found within the deduced sequence of the precursor extension. The N-terminus of the larger intermediate, PP2C, was Gly-12, whereas the N-terminus of the smaller, PP2D, was His-21. Both processing sites involved a peptide bond in which the carbonyl function was contributed by an acidic amino acid.


Xpert MTB/RIF diagnosis of childhood tuberculosis from sputum and stool samples in a high TB-HIV-prevalent setting.
The Xpert MTB/RIF assay is a major advance for diagnosis of tuberculosis (TB) in high-burden countries but is limited in children by their difficulty to produce sputum. We investigated TB in sputum and stool from children with the aim of improving paediatric TB diagnosis. A prospective cohort of children with presumptive TB, provided two sputum or induced sputum at enrolment in a regional referral hospital in Uganda. Stool was collected from those started on TB treatment. All specimen were tested for Xpert MTB/RIF, mycobacteria growth indicator tube (MGIT), Lowenstein Jensen cultures and microscopy (except stool). We compared TB detection between age categories and assessed the performance of Xpert MTB/RIF in sputum and stool. Of the 392 children enrolled, 357 (91.1%) produced at least one sputum sample. Sputum culture yield was 13/357 (3.6%): 3/109 (2.6%), 3/89 (3.2%), 3/101 (2.6%) and 4/44 (8.2%) among children of < 2, 2-5, ≥ 5-10 and > 10 years, respectively (p = 0.599). Xpert MTB/RIF yield was 14/350 (4.0%): 3/104 (2.9%), 4/92 (4.3%), 3/88 (2.9%) and 4/50 (.0%), respectively (p = 0.283). Sensitivity and specificity of Xpert MTB/RIF in sputum against sputum culture were 90.9% (95% CI 58.7-99.8) and 99.1% (99.1-99.8). In stool, it was 55.6% (21.2-86.3) and 98.2% (98.2-100) against Xpert MTB/RIF and culture in sputum. Only a minority of children had microbiologically confirmed TB with a higher proportion in children above 10 years. Although sensitivity of Xpert MTB/RIF in stool was low, with good optimization, it might be a good alternative to sputum in children.


Probing the interaction of the multidrug-resistance phenotype with the polypeptide ionophore gramicidin D via functional channel formation.
It has been proposed that the multidrug resistance (MDR) transporter, P-glycoprotein (P-170), may be physiologically involved in the transport of polypeptides. As a step towards understanding the interaction of P-170 with polypeptides, we isolated various gramicidin-D-resistant mammalian cell lines. Gramicidin D is a hydrophobic pentadecapeptide ionophore that forms proton and alkali metal cation-permeable channels in lipid bilayers. Gramicidin-D-resistant cells displayed a prominent MDR gene amplification, P-170 overexpression, reduced drug accumulation, and consequent resistance to MDR-type cytotoxic agents. Modulators of the MDR phenotype, including verapamil, reserpine and quinidine, rendered these cells sensitive to gramicidin D. Using these cell lines, we established an assay that probes for the intra-membranal interaction between P-170 and gramicidin D. Gramicidin-D channel formation was followed by cellular accumulation of 86Rb+. Ionophore-resistant cells, and other MDR cells, did not show an appreciable increase in 86Rb+ influx rates, in the presence of increasing gramicidin-D concentrations. In contrast, parental cells displayed a dose-dependent increase in the 86Rb+ influx rates. Interestingly, in the absence of serum, gramicidin-D-resistant cells resumed the wild-type, ionophore-dose-dependent increase in 86Rb+ influx rates. MDR modulators caused a resumption of channel formation in ionophore-resistant cells. We conclude that acquisition of the MDR phenotype is an efficient means of cellular protection against gramicidin D. Hence, a new approach is offered in which P-170 interaction with gramicidin D is quantitatively followed by a rapid assessment of the biological activity (i.e. channel formation) of the substrate itself. Possible mechanisms of P-170 interaction with free ionophore monomers, and membrane-associated gramicidin D are discussed.


Hemodialysis removes uremic toxins that alter the biological actions of endothelial cells.
Chronic kidney disease is linked to systemic inflammation and to an increased risk of ischemic heart disease and atherosclerosis. Endothelial dysfunction associates with hypertension and vascular disease in the presence of chronic kidney disease but the mechanisms that regulate the activation of the endothelium at the early stages of the disease, before systemic inflammation is established remain obscure. In the present study we investigated the effect of serum derived from patients with chronic kidney disease either before or after hemodialysis on the activation of human endothelial cells in vitro, as an attempt to define the overall effect of uremic toxins at the early stages of endothelial dysfunction. Our results argue that uremic toxins alter the biological actions of endothelial cells and the remodelling of the extracellular matrix before signs of systemic inflammatory responses are observed. This study further elucidates the early events of endothelial dysfunction during toxic uremia conditions allowing more complete understanding of the molecular events as well as their sequence during progressive renal failure.


Present, future, and novel bioclimates of the San Francisco, California region.
Bioclimates are syntheses of climatic variables into biologically relevant categories that facilitate comparative studies of biotic responses to climate conditions. Isobioclimates, unique combinations of bioclimatic indices (continentality, ombrotype, and thermotype), were constructed for northern California coastal ranges based on the Rivas-Martinez worldwide bioclimatic classification system for the end of the 20(th) century climatology (1971-2000) and end of the 21(st) century climatology (2070-2099) using two models, Geophysical Fluid Dynamics Laboratory (GFDL) model and the Parallel Climate Model (PCM), under the medium-high A2 emission scenario. The digitally mapped results were used to 1) assess the relative redistribution of isobioclimates and their magnitude of change, 2) quantify the loss of isobioclimates into the future, 3) identify and locate novel isobioclimates projected to appear, and 4) explore compositional change in vegetation types among analog isobioclimate patches. This study used downscaled climate variables to map the isobioclimates at a fine spatial resolution -270 m grid cells. Common to both models of future climate was a large change in thermotype. Changes in ombrotype differed among the two models. The end of 20(th) century climatology has 83 isobioclimates covering the 63,000 km(2) study area. In both future projections 51 of those isobioclimates disappear over 40,000 km(2). The ordination of vegetation-bioclimate relationships shows very strong correlation of Rivas-Martinez indices with vegetation distribution and composition. Comparisons of vegetation composition among analog patches suggest that vegetation change will be a local rearrangement of species already in place rather than one requiring long distance dispersal. The digitally mapped results facilitate comparison with other Mediterranean regions. Major remaining challenges include predicting vegetation composition of novel isobioclimates and developing metrics to compare differences in climate space.


Transcranial ultrasound imaging with speed of sound-based phase correction: a numerical study.
This paper presents a numerical study for ultrasound transcranial imaging. To correct for the phase aberration from the skull, two critical steps are needed prior to brain imaging. In the first step, the skull shape and speed of sound are acquired by either CT scans or ultrasound scans. In the ultrasound scan approach, phased array and double focusing technique are utilized, which are able to estimate the thickness of the skull with a maximum error of around 10% and the average speed of sound in the skull is underestimated by less than 2%. In the second step, the fast marching method is used to compute the phase delay based on the known skull shape and sound speed from the first step, and the computation can be completed in seconds for 2D problems. The computed phase delays are then used in combination with the conventional delay-and-sum algorithm for generating B-mode images. Images of wire phantoms with CT or ultrasound scan-based phase correction are shown to have much less artifact than the ones without correction. Errors of deducing speed of sound from CT scans are also discussed regarding its effect on the transcranial ultrasound images. Assuming the speed of sound grows linearly with the density, this study shows that, the CT-based phase correction approach can provide clear images of wire phantoms even if the speed of sound is overestimated by 400 m s(-1), or the linear coefficient is overestimated by 40%. While in this study, ultrasound scan-based phase correction performs almost equally well with the CT-based approach, potential problems are identified and discussed.


Determinants recognized by human cytotoxic T cells on a natural hybrid class I HLA molecule.
The major histocompatibility complex class I HLA molecules are the primary determinants recognized by allogeneic cytotoxic T lymphocytes (CTL), and serve as restricting elements for CTL recognition of viral, chemical, or minor histocompatibility antigens. HLA-Aw69 is a naturally occurring hybrid class I molecule that we have used to investigate the regions of class I antigens involved in human CTL recognition. HLA-Aw69 appears to have resulted from an exon shuffle between two closely related class I genes: the alpha 1 domain of HLA-Aw69 is identical to that of HLA-Aw68, while the alpha 2 and alpha 3 domains are identical to HLA-A2. The determinants recognized by human allogeneic CTL clones specific for HLA-A2, -Aw68, and/or -Aw69 fall into three patterns: (a) CTL determinants are located on both the alpha 1 and alpha 2 domains; (b) interaction of the alpha 1 and alpha 2 domains results in new combinatorial determinants; (c) interaction of the alpha 1 and alpha 2 domains in the hybrid molecule results in the loss of CTL determinants that are present on both parental molecules. Thus, using human CTL clones, target cells, and HLA molecules, we show that the interaction of the alpha 1 and alpha 2 domains alters CTL determinants in ways not directly predictable from primary structure.


Influence of caregivers' experiences on service use among children with attention-deficit hyperactivity disorder.
This study examined how prior experiences of caregivers of children with attention-deficit hyperactivity disorder (ADHD) leading up to treatment related to later service use. The investigators interviewed caregivers of 48 children with ADHD recruited from outpatient clinic settings and recorded the children's medication use and clinic attendance six and 12 months later. Interview transcripts were analyzed to identify characteristic experiences, or themes, felt by the caregivers before seeking treatment. The investigators also looked for patterns in the way themes were endorsed. Caregivers' experiences with the children's ADHD were characterized by six main themes-caregiver strain, attribution of meaning, perception of responsibility, problem-solving approach, beliefs about ADHD treatment, and response to societal influences. Based on distinct patterns in which they endorsed the six themes, caregivers were classified as motivated by observation (high on theme of perception of responsibility and low on theme of caregiver strain; 27%), motivated by experience (positive attribution of meaning and beliefs about ADHD treatment and low on strain; 19%), motivated by strain (high on strain, uncertain attribution of meaning, and contradictory societal influences; 23%), and struggling with meaning (high on strain, nonmedical attribution of meaning; 31%). At 12 months, children of caregivers who were motivated by experience were the most likely and those struggling with meaning were the least likely to attend clinic appointments (89% and 53%, respectively, p=.017). The groups did not vary in medication use. Early identification of the factors influencing caregivers' use of services for children with ADHD may have implications for treatment retention.


HEMATOPOIETIC PROGENITORS CELLS IN PERIPHERAL BLOOD OF BALB/C MICE UNDER IONIZING RADIATION ACTION IN SUBLETHAL DOSE.
determination of the content of hematopoietic progenitor cells circulating in peripheral blood of Balb/Cmice, under ionizing radiation action in sublethal dose, at different periods after the irradiation, using cell culturein diffusion chambers in vivo. Peripheral blood smears of Balb/C mice were prepared and studied, its cellular composition was determined, as well as by cultivation of peripheral blood cells in diffusion chambers in vivo their colony-forming efficien-cy was determined on the 0th, 5th, and 30th day after external irradiation in sublethal dose 5.85 Gy. The content of myelocytes and metamyelocytes among blood nucleated cells of the irradiated animals wasincreased, compared to control, during the whole investigated period. In particular, on the 30th day after irradiationthe content of myelocytes in peripheral blood was 3.3 ± 0.7 % compared to (0.8 ± 0.4) % in control, and the content of metamyelocytes - (3.4 ± 0.7) % compared to (0.9 ± 0.3) % in control. A significant increase in the amountof circulating progenitor cells in the peripheral blood was observed in the early stages after irradiation (12.5 ± 1.6colony-forming units per 100,000 explanted cells, compared to 5.1 ± 0.8 in control). However, on the 5th day theircontent was slightly reduced compared to control (1.3 ± 0.9), and only to the 30th day a normalization of the amountof progenitor cells occurred in the peripheral blood (6.8 ± 0.7 colony-forming units per 100,000 explanted cells). The analysis of the obtained results revealed an increased level of immature forms of cells in theperipheral blood of irradiated animals, compared to control, in the early stages after irradiation, includinghematopoietic progenitor cells, which are able to colony forming in cell culture. Therefore, the action of ionizingradiation in sublethal dose had a critical effect on the proliferation of hematopoietic cells in bone marrow and provoked their increased migration into the bloodstream. Determination of the content of hematopoietic cells' immature forms in peripheral blood allowed assessing the degree of hematopoietic damage due to the action of ionizing radiation.


Development of Stem Cell-derived Antigen-specific Regulatory T Cells Against Autoimmunity.
Autoimmune diseases arise due to the loss of immunological self-tolerance. Regulatory T cells (Tregs) are important mediators of immunologic self-tolerance. Tregs represent about 5 - 10% of the mature CD4+ T cell subpopulation in mice and humans, with about 1 - 2% of those Tregs circulating in the peripheral blood. Induced pluripotent stem cells (iPSCs) can be differentiated into functional Tregs, which have a potential to be used for cell-based therapies of autoimmune diseases. Here, we present a method to develop antigen (Ag)-specific Tregs from iPSCs (i.e., iPSC-Tregs). The method is based on incorporating the transcription factor FoxP3 and an Ag-specific T cell receptor (TCR) into iPSCs and then differentiating on OP9 stromal cells expressing Notch ligands delta-like (DL) 1 and DL4. Following in vitro differentiation, the iPSC-Tregs express CD4, CD8, CD3, CD25, FoxP3, and Ag-specific TCR and are able to respond to Ag stimulation. This method has been successfully applied to cell-based therapy of autoimmune arthritis in a murine model. Adoptive transfer of these Ag-specific iPSC-Tregs into Ag-induced arthritis (AIA)-bearing mice has the ability to reduce joint inflammation and swelling and to prevent bone loss.


Workplace violence from the perspective of hospital ward managers in Sweden: A qualitative study.
The aims of the study are to explore workplace violence perpetrated by patients or visitors from the perspective of hospital ward managers and to describe how ward managers perceive their leadership role and manage related incidents. Few studies focus on workplace violence from the perspective of ward managers even though they are the closest managers to the operational staff. Fifteen semistructured interviews were analysed using qualitative content analysis. Four categories emerged: the face of workplace violence, a two-fold assignment, strive towards readiness to act, and managing incidents. While the most common acts of workplace violence are considered less serious and related to patients' medical conditions or dissatisfied visitors, hospital organizations focus on serious but rarely occurring incidents. Consequently, ward managers have limited opportunities to ensure a safe work environment on an everyday basis. To support ward managers' occupational safety and health management, workplace violence prevention and management should be acknowledged as an important responsibility for senior management in hospitals. It is important to identify incidents that most likely will occur at the wards and to create strategies related to those incidents. Strategies could include risk assessments, prevention, evaluation, education and reflection combined with, for example, scenario training.


Subcutaneous nodules caused by Pseudomonas aeruginosa without sepsis.
Pseudomonas aeruginosa infection can cause a wide array of skin manifestations. While some infections are mild, as are the cases with hot tub folliculitis and toe web or nail infection, others are a result of sepsis and can be fatal without prompt treatment. The classic skin finding of P. aeruginosa sepsis is Ecthyma gangrenosum, but other signs such as papules, petechiae, and hemorrhagic bullae can also be seen. Suppurative panniculitis can also be caused by P. aeruginosa sepsis and clinically manifests as solitary or multiple subcutaneous nodules. Reports in the literature describe these nodules in the setting of clinical sepsis or with positive blood cultures. We report a case of localized subcutaneous nodules on the leg caused by P. aeruginosa in a patient without sepsis or positive blood cultures. The source of the infection was thought to be from a traumatic inoculation. This raises the possibility that P. aeruginosa can cause subcutaneous nodules from a localized infection, perhaps via lymphangitic spread without the manifestations of sepsis.


Yuck, This Biscuit Looks Lumpy! Neophobic Levels and Cultural Differences Drive Children's Check-All-That-Apply (CATA) Descriptions and Preferences for High-Fibre Biscuits.
Food neophobia influences food choice in school-aged children. However, little is known about how children with different degrees of food neophobia perceive food and to what extent different sensory attributes drive their liking. This paper explores liking and sensory perception of fibre-rich biscuits in school-aged children (n = 509, age 9-12 years) with different degrees of food neophobia and from five different European countries (Finland, Italy, Spain, Sweden and United Kingdom). Children tasted and rated their liking of eight commercial biscuits and performed a Check-All-That-Apply task to describe the samples and further completed a Food Neophobia Scale. Children with a higher degree of neophobia displayed a lower liking for all tasted biscuits (p < 0.001). Cross-cultural differences in liking also appeared (p < 0.001). A negative correlation was found between degree of neophobia and the number of CATA-terms used to describe the samples (r = -0.116, p = 0.009). Penalty analysis showed that degree of food neophobia also affected drivers of biscuit liking, where particularly appearance terms were drivers of disliking for neophobic children. Cross-cultural differences in drivers of liking and disliking were particularly salient for texture attributes. Further research should explore if optimizing appearance attributes could be a way to increase liking of fibre-rich foods in neophobic children.


The Short-Chain Fatty Acid Butyrate Attenuates Pulmonary Vascular Remodeling and Inflammation in Hypoxia-Induced Pulmonary Hypertension.
Pulmonary hypertension (PH) is a progressive cardiovascular disorder in which local vascular inflammation leads to increased pulmonary vascular remodeling and ultimately to right heart failure. The HDAC inhibitor butyrate, a product of microbial fermentation, is protective in inflammatory intestinal diseases, but little is known regarding its effect on extraintestinal diseases, such as PH. In this study, we tested the hypothesis that butyrate is protective in a Sprague-Dawley (SD) rat model of hypoxic PH. Treatment with butyrate (220 mg/kg intake) prevented hypoxia-induced right ventricular hypertrophy (RVH), hypoxia-induced increases in right ventricular systolic pressure (RVSP), pulmonary vascular remodeling, and permeability. A reversal effect of butyrate (2200 mg/kg intake) was observed on elevated RVH. Butyrate treatment also increased the acetylation of histone H3, 25-34 kDa, and 34-50 kDa proteins in the total lung lysates of butyrate-treated animals. In addition, butyrate decreased hypoxia-induced accumulation of alveolar (mostly CD68+) and interstitial (CD68+ and CD163+) lung macrophages. Analysis of cytokine profiles in lung tissue lysates showed a hypoxia-induced upregulation of TIMP-1, CINC-1, and Fractalkine and downregulation of soluble ICAM (sICAM). The expression of Fractalkine and VEGFα, but not CINC-1, TIMP-1, and sICAM was downregulated by butyrate. In rat microvascular endothelial cells (RMVEC), butyrate (1 mM, 2 and 24 h) exhibited a protective effect against TNFα- and LPS-induced barrier disruption. Butyrate (1 mM, 24 h) also upregulated tight junctional proteins (occludin, cingulin, claudin-1) and increased the acetylation of histone H3 but not α-tubulin. These findings provide evidence of the protective effect of butyrate on hypoxic PH and suggest its potential use as a complementary treatment for PH and other cardiovascular diseases.


Spermatogonial stem cells: mouse and human comparisons.
Spermatogonial stem cells (SSCs) have unique characteristics in that they produce sperm that transmit genetic information from generation to generation and they can be reprogrammed spontaneously to form embryonic stem (ES)-like cells to acquire pluripotency. In rodents, it is generally believed that the A-single (A(s)) is the stem cell population, whereas the A-paired (A(pr)) and A-aligned (A(al)) represent the progenitor spermatogonial population. The A(1) to A(4) cells, intermediate, and type B spermatogonia are considered differentiated spermatogonia. In human, very little information is available about SSCs, except for the earlier work of Clermont and colleagues who demonstrated that there are two different types of A spermatogonia, the A(dark) and A(pale) spermatogonia. The A(dark) spermatogonia were referred to as the reserve stem cells, whereas the A(pale) were considered the renewing stem cells. In this review, we outline several spermatogonial renewal schemes for both rodents and primates, including man. We also compare phenotypic markers for spermatogonia/spermatogonial stem cells in rodents and humans and address SSC potential and therapeutic application.


Clonal characterization of the human IgG antibody repertoire to Haemophilus influenzae type b polysaccharide. II. IgG antibodies contain VH genes from a single VH family and VL genes from at least four VL families.
To define the V gene family repertoire of human IgG anti-Haemophilus influenzae type b polysaccharide antibodies, we purified six IgG1 and nine IgG2 anti-Hib-PS antibodies to monoclonality from immune serum of six individuals and performed N-terminal amino acid sequence analysis. Of the 15 clonal antibodies we examined, all H chain V regions were of the VHIII family. In contrast, the L chains of these antibodies were clearly from at least four different VL families; VKI, VKII, VKIII, and V lambda. Interestingly. VL family expression correlated with the cross-reactivity of these antibodies to the capsular carbohydrate of Escherichia coli K100. VKII antibodies did not cross-react, whereas antibodies expressing V lambda, VKI, or VKIII generally cross-reacted. We conclude that L chain V regions are very important contributors to the limited heterogeneity in this antibody repertoire.


Predator-by-Environment Interactions Mediate Bacterial Competition in the Dictyostelium discoideum Microbiome.
Interactions between species and their environment play a key role in the evolution of diverse communities, and numerous studies have emphasized that interactions among microbes and among trophic levels play an important role in maintaining microbial diversity and ecosystem functioning. In this study, we investigate how two of these types of interactions, public goods cooperation through the production of iron scavenging siderophores and predation by the social amoeba Dictyostelium discoideum, mediate competition between two strains of Pseudomonas fluorescens that were co-isolated from D. discoideum. We find that although we are able to generally predict the competitive outcomes between strains based on the presence and absence of either D. discoideum or iron, predator-by-environment interactions result in unexpected competitive outcomes. This suggests that while both cooperation and predation can mediate the competitive abilities and potentially the coexistence of these strains, predicting how combinations of different environments affect even the relatively simple microbiome of D. discoideum remains challenging.


Prevalence and causes of blood donor deferrals among clients presenting for blood donation in northern Tanzania.
Blood is an important requirement in different medical and surgical conditions with half of all donations are from developing countries. Lack of eligibility among blood donors who present for blood transfusion, called blood donor deferral is associated with the unsustainable and inadequate amount of blood collected by blood banks worldwide. However, the prevalence and causes of blood donor deferrals are not well known in Tanzania where less than one-third of actual needs of blood is collected, leading to unmet demand of blood for transfusion, and causing unwanted morbidity and mortality. This was a retrospective analysis of blood donors at northern zone blood transfusion center, Tanzania from January to December. 2016. Donor's data were transferred to Statistical Package for Social Studies (SPSS) program version 20.0 for analysis. Descriptive statistics was used to summarize data and comparisons made by type of donor and deferrals using Chi-square test. A total of 14377 participants were studied whereby 12775 (88.9%) were voluntary non-remunerated blood donors. The blood donor deferral rate was 12.7% and deferral was significantly more likely in females, with increasing age above 31 years, who came from nearby regions from where the blood bank is located and/or a family replacement donor (P value <0.01). Overall, infections contributed to 62% of all deferrals and low hemoglobin was the leading cause of temporary deferrals while Hepatitis B lead the permanent deferral causes. Blood donor deferral is a significant problem in northern Tanzania and accounts for more than one-tenth of all prospective blood donors. Latent and active infections are the leading cause of blood donor deferrals, a picture that mirrors other low income countries especially those located in sub-Saharan Africa. Results of this study calls for appropriate preventive interventions to address prevalent causes of deferrals such as infections with HIV and HBV to tackle low hemoglobin.


Production of short-chain fructooligosaccharides (scFOS) using extracellular β-D-fructofuranosidase produced by Aspergillus thermomutatus.
Aspergillus thermomutatus produces an extracellular β-D-fructofuranosidase when cultured in Khanna medium with sucrose as additional carbon source at 30°C under agitation for 72 hr. Addition of glucose and fructose in the culture medium affected the production of the enzyme negatively. The optimum hydrolytic activity was achieved at 60°C and pH 5.0, with half-life (T50) of 30 hr at 50°C and 62% of its activity maintained at pH 5.0 for 48 hr. The extracellular extract containing β-D-fructofuranosidase was effective in producing fructooligosaccharides (FOS), mainly 1-kestose. The highest concentration of FOS was obtained at 30°C and 60°C, indicating the existence of at least two enzymes with transfructosylating activity. At 30°C, the maximal FOS concentration was obtained from 48 to 72 hr, while at 60°C, it was achieved only at 72 hr. The best production of FOS (86.7 g/L) was obtained using 500 g/L sucrose as substrate. PRACTICAL APPLICATION: Fructooligosaccharides (FOS) are linear oligomers of fructose units with important applications in the food industry as sweetening agents and biopreservatives. Due to the presence of β-glycosidic bonds, they cannot be hydrolyzed by human enzymes, allowing the use of FOS-containing products by diabetics. FOS used in the preparation of dairy products imparts humectancy to soft baked products, lowers the freezing point of frozen desserts, provides crispness to low-fat cookies, and provides many other advantages. Diets containing FOS can reduce the levels of triglycerides and cholesterol and improve the absorption of ions, such as Ca2+ and Mg2+ . FOS also exhibit bifidogenic effect on Bifidobacterium and Lactobacillus strains in the colon. Industrially, FOS is produced during the transfructosylation reaction of sucrose catalyzed by β-D-fructofuranosidase. Identifying new sources of β-D-fructofuranosidase is an important challenge to meet its industrial demand.


Reactivity of NK Cells Against Ovarian Cancer Cells Is Maintained in the Presence of Calcium Phosphate Nanoparticles.
Calcium phosphate nanoparticles (CaP-NPs) are biodegradable carriers that can be functionalized with biologically active molecules. As such, they are potential candidates for delivery of therapeutic molecules in cancer therapies. In this context, it is important to explore whether CaP-NPs impair the natural or therapy-induced immune cell activity against cancer cells. Therefore, in this study, we have investigated the effects of different CaP-NPs on the anti-tumor activity of natural killer (NK) cells using different ovarian cancer (OC) cell line models. We explored these interactions in coculture systems consisting of NK cells, OC cells, CaP-NPs, and therapeutic Cetuximab antibodies (anti-EGFR, ADCC-inducing antibody). Our experiments revealed that aggregated CaP-NPs can serve as artificial targets, which activate NK cell degranulation and impair ADCC directed against tumor targets. However, when CaP-NPs were properly dissolved by sonication, they did not cause substantial activation. CaP-NPs with SiO2-SH-shell induced some activation of NK cells that was not observed with polyethyleneimine-coated CaP-NPs. Addition of CaP-NPs to NK killing assays did not impair conjugation of NK with OC and subsequent tumor cytolytic NK degranulation. Therapeutic antibody coupled to functionalized CaP-NPs maintained substantial levels of antibody-dependent cellular cytotoxic activity. Our study provides a cell biological basis for the application of functionalized CaP-NPs in immunologic anti-cancer therapies.


Serum dioxins and polychlorinated biphenyls are associated with growth among Russian boys.
We evaluated the associations of serum dioxins and polychlorinated biphenyls (PCBs) with longitudinally assessed growth measurements among peripubertal Russian boys. A total of 499 boys from Chapaevsk, Russia, aged 8 to 9 years were enrolled in the study from 2003 to 2005 and were followed prospectively for 3 years. Blood samples were collected and physical examinations were conducted at entry and repeated at annual study visits. Multivariate mixed-effects regression models for repeated measures were used to examine the associations of serum dioxins and PCBs with longitudinal measurements of BMI, height, and height velocity. Serum dioxin (total 2005 toxic equivalency [TEQ] median: 21.1 pg/g lipid) and PCBs (median sum of PCBs: 250 ng/g lipid) were measured in 468 boys. At study entry and during 3 years of follow-up, >50% of the boys had age-adjusted BMI and height z scores within 1 SD of World Health Organization-standardized mean values for age. Boys in the highest exposure quintile of the sum of dioxin and PCB concentrations and total TEQs had a significant decrease in mean BMI z scores of 0.67 for dioxins and TEQs and 1.04 for PCBs, compared with boys in the lowest exposure quintile. Comparison of the highest versus the lowest quintile revealed that higher serum PCB concentrations were associated with significantly lower height z scores (mean z-score decrease: 0.41) and height velocity (mean decrease: 0.19 cm/year) after 3 years of follow-up. Our findings suggest that exposures to dioxins and PCBs are associated with reduced growth during the peripubertal period and may compromise adult body mass, stature, and health.


Approaches to family planning in Muslim communities.
Addressing the cultural and religious beliefs around the issue of family planning has been a big challenge for the international development community. The concept of family planning has raised some concerns regarding its acceptability within Muslim populations. While some Muslim states and organisations have adopted a rather cautious approach to the issue, others have gone to the extent of inviting religious leaders to present religiously sound interpretations (fatwa) on the subject. Alongside these deliberations are some alarming statistics on maternal health. The World Health Organization estimates that worldwide 211 million women become pregnant each year and that about two-thirds of them deliver live infants. The remaining one-third of pregnancies end in miscarriage, stillbirth or induced abortion. Some 200 million women in developing countries have an unmet need for effective contraception. These statistics and the ongoing discussion surrounding family planning in the Muslim communities raise legitimate questions. How is family planning perceived within the Muslim community? Does Islam address the issue of family planning? Is it permissible? How should appropriate family planning programmes within Muslim settings be developed and applied? This article seeks to present the ongoing debate on family planning within the Muslim community and offer recommendations to organisations for effective strategy implementation of family planning programmes within Muslim settings. The article provides a brief background on the historical development of family planning in the Muslim community, and outlines Muslim perceptions on this issue. It concludes with recommendations for non-governmental organisations on how to effectively implement acceptable family planning programmes within Muslim settings.


Plasticity of substance P in mature and aged sympathetic neurons in culture.
The effect of age on the plasticity of the putative peptide neurotransmitter substance P (SP) was examined in the rat superior cervical sympathetic ganglion. Explantation of ganglia from 6-month-old rats to serum-supplemented culture resulted in a tenfold increase in SP concentration, reproducing results previously obtained for ganglia from neonatal rats. Veratridine prevented the increase in SP concentration in adult ganglia, and tetrodotoxin blocked the veratridine effect, suggesting that membrane depolarization and sodium influx prevented the rise in the SP content of adult ganglia as well as of neonatal ganglia. However, the time courses of the increase in the amount of the peptide differed in neonatal and mature ganglia, suggesting that some aspects of regulation may differ in the two. The effects of aging on neural plasticity were further analyzed by explanting ganglia from 2-year-old rats. No significant increase in SP concentration was observed in these ganglia. Remarkable plasticity thus seems to persist in mature neurons but may be deficient in aged sympathetic neurons.


MicroRNA-100 Suppresses Chronic Vascular Inflammation by Stimulation of Endothelial Autophagy.
The interaction of circulating cells within the vascular wall is a critical event in chronic inflammatory processes, such as atherosclerosis, but the control of the vascular inflammatory state is still largely unclear. This study was undertaken to characterize the function of the endothelial-enriched microRNA miR-100 during vascular inflammation and atherogenesis. Based on a transcriptome analysis of endothelial cells after miR-100 overexpression, we identified miR-100 as a potent suppressor of endothelial adhesion molecule expression, resulting in attenuated leukocyte-endothelial interaction in vitro and in vivo as shown by flow cytometry and intravital imaging. Mechanistically, miR-100 directly repressed several components of mammalian target of rapamycin complex 1-signaling, including mammalian target of rapamycin and raptor, which resulted in a stimulation of endothelial autophagy and attenuated nuclear factor κB signaling in vitro and in vivo. In a low-density lipoprotein receptor-deficient atherosclerotic mouse model, pharmacological inhibition of miR-100 resulted in enhanced plaque lesion formation and a higher macrophage content of the plaque, whereas a systemic miR-100 replacement therapy had protective effects and attenuated atherogenesis, resulting in a decrease of plaque area by 45%. Finally, analysis of miR-100 expression in >70 samples obtained during carotid endarterectomy revealed that local miR-100 expression was inversely correlated with inflammatory cell content in patients. In summary, we describe an anti-inflammatory function of miR-100 in the vascular response to injury and inflammation and identify an important novel modulator of mammalian target of rapamycin signaling and autophagy in the vascular system. Our findings of miR-100 as a potential protective anti-athero-miR suggest that the therapeutic replacement of this microRNA could be a potential strategy for the treatment of chronic inflammatory diseases, such as atherosclerosis, in the future.


The Association of Ambient Air Pollution With Cataract Surgery in UK Biobank Participants: Prospective Cohort Study.
Air pollution is associated with chronic diseases of later life. Cataract is the most common cause of blindess globally. It is biologically plausible that cataract risk is increased by pollution exposure. Therefore, the relationship between air pollution and incident cataract surgery was examined. This was a prospective, observational study involving 433,727 UK Biobank participants. Ambient air pollution measures included particulates, nitrogen dioxide (NO2) and nitrogen oxides (NOx). Outdoor air pollution was estimated based on land use regression models. Participants undergoing cataract surgery in either eye were ascertained via data linkage to the National Health Service procedure statistics. Those undergoing cataract surgery within 1 year of baseline assessment and those reporting cataract at baseline were excluded. Cox proportional hazards models were used to examine the associations between air pollutants and incident cataract surgery, adjusting for sociodemographic and lifestyle factors. There were 16,307 incident cases of cataract surgery. Higher exposure to PM2.5 was associated with a 5% increased risk of incident cataract surgery (per interquartile range [IQR] increase). Compared to the lowest quartile, participants with exposures to PM2.5, NO2, and NOx in the highest quartile were 14%, 11%, and 9% more likely to undergo cataract surgery, respectively. A continuous exposure-response relationship was observed, with the likelihood of undergoing cataract surgery being progressively higher with greater levels of PM2.5, NO2, and NOx (P for trend P < 0.001). Although the results of our study showed a 5% increased risk of future cataract surgery following an exposure to PM2.5, NO2, and NOx, the effect estimates were relatively small. Further research is required to determine if the associations identified are causal.


Crystallographic characterization of CCG repeats.
CCG repeats are highly over-represented in exons of the human genome. Usually they are located in the 5' UTR but are also abundant in translated sequences. The CCG repeats are associated with three tri-nucleotide repeat disorders: Huntington's disease, myotonic dystrophy type 1 and chromosome X-linked mental retardation (FRAXE). In this study, we present two crystal structures containing double-stranded CCG repeats: one of an RNA in the native form, and one containing LNA nucleotides. Both duplexes form A-helices but with strands slipped in the 5' (native structure) or the 3' direction (LNA-containing structure). As a result, one of two expected C-C pairs is eliminated from the duplex. Each of the three observed C-C pairs interacts differently, forming either one weak H-bond or none. LNA nucleotides have no apparent effect on the helical parameters but the base stacking is increased compared to the native duplex and the distribution of electrostatic potential in the major groove is changed. The CCG crystal structures explain the thermodynamic fragility of CCG runs and throw light on the observation that the MBNL1 protein recognises CCG runs, as well as CUG and CAG, but not the relatively stable CGG repeats.


Development and evaluation of a 3-D virtual pronunciation tutor for children with autism spectrum disorders.
The deficit in speech sound production in some children with autism spectrum disorder (ASD) adds to their communication barriers. The 3-D virtual environments have been implemented to improve their communication abilities. However, there were no previous studies on the use of a 3-D virtual pronunciation tutor designed specifically to train pronunciation for children with ASD. To fill this research gap, the current study developed and evaluated a 3-D virtual tutor which served as a multimodal and real-data-driven speech production tutor to present both places and manners of Mandarin articulation. Using an eye-tracking technique (RED 5 Eye Tracker), Experiment 1 objectively investigated children's gauged attention distribution online while learning with our computer-assisted 3-D virtual tutor in comparison to a real human face (HF) tutor. Eye-tracking results indicated most participants showed more interests in the visual speech cues of the 3-D tutor, and paid some degree of absolute attention to the additional visual speech information of both articulatory movements and airflow changes. To further compare treatment outcomes, training performance was evaluated in Experiment 2 with the ASD learners divided into two groups, with one group learning from the HF tutor and the other from the 3-D tutor (HF group vs. 3-D group). Both groups showed improvement with the help of computer-based training in the post-intervention test based on the calculation of a 5-point Likert scale. However, the 3-D group showed much higher gains in producing Mandarin stop and affricate consonants, and apical vowels. We conclude that our 3-D virtual imitation intervention system provides an effective approach of audiovisual pronunciation training for children with ASD.


Interpretation of the transpulmonary pressure in the critically ill patient.
Mechanical ventilation is a life-saving procedure, which takes over the function of the respiratory muscles while buying time for healing to take place. However, it can also promote or worsen lung injury, so that careful monitoring of respiratory mechanics is suggested to titrate the level of support and avoid injurious pressures and volumes to develop. Standard monitoring includes flow, volume and airway pressure (Paw). However, Paw represents the pressure acting on the respiratory system as a whole, and does not allow to differentiate the part of pressure that is spent di distend the chest wall. Moreover, if spontaneous breathing efforts are allowed, the Paw is the sum of that applied by the ventilator and that generated by the patient. As a consequence, monitoring of Paw has significant shortcomings. Assessment of esophageal pressure (Pes), as a surrogate for pleural pressure (Ppl), may allow the clinicians to discriminate between the elastic behaviour of the lung and the chest wall, and to calculate the degree of spontaneous respiratory effort. In the present review, the characteristics and limitations of airway and transpulmonary pressure monitoring will be presented; we will highlight the different assumptions underlying the various methods for measuring transpulmonary pressure (i.e., the elastance-derived and the release-derived method, and the direct measurement), as well as the potential application of transpulmonary pressure assessment during both controlled and spontaneous/assisted mechanical ventilation in critically ill patients.


Impact of End-Stage Renal Disease and Acute Kidney Injury on ICU Outcomes in Patients With Sepsis.
To report the characteristics and outcomes of patients with sepsis in the intensive care unit (ICU) with end-stage renal disease (ESRD) and acute kidney injury (AKI) compared to patients with nonkidney injury (non-KI). Retrospective study of all patients with sepsis admitted to the ICU of a university hospital within a 12-month time period. Data were obtained from the University Health Consortium database and a chart review of the electronic medical records. We identified 39 cases of ESRD, 106 cases of AKI, and 103 cases of non-KI. Intensive care unit mortality was 15.4% for ESRD, 30.2% for AKI, and 13.6% for non-KI ( P < .01). Hospital mortality was 20.5% for ESRD, 32.1% for AKI, and 13.6% for non-KI ( P < .01). Early AKI and late AKI had an ICU mortality of 24.4% versus 50% ( P <.01), hospital mortality of 26.8% versus 50% ( P = .03), ICU length of stay (LOS) of 3 and 6 days ( P = .04), and hospital LOS of 7 and 12.5 days ( P <.01), respectively. Patients with sepsis having AKI have a higher mortality rate than those with ESRD and non-KI. Hospital and ICU mortality rates for patients with ESRD were similar to non-KI patients. Late AKI compared to early AKI had a higher mortality and longer LOS.


Exploring activity landscapes with extended similarity: is Tanimoto enough?
Understanding structure-activity landscapes is essential in drug discovery. Similarly, it has been shown that the presence of activity cliffs in compound data sets can have a substantial impact not only on the design progress but also can influence the predictive ability of machine learning models. With the continued expansion of the chemical space and the currently available large and ultra-large libraries, it is imperative to implement efficient tools to analyze the activity landscape of compound data sets rapidly. The goal of this study is to show the applicability of the n-ary indices to quantify the structure-activity landscapes of large compound data sets using different types of structural representation rapidly and efficiently. We also discuss how a recently introduced medoid algorithm provides the foundation to finding optimum correlations between similarity measures and structure-activity rankings. The applicability of the n-ary indices and the medoid algorithm is shown by analyzing the activity landscape of 10 compound data sets with pharmaceutical relevance using three fingerprints of different designs, 16 extended similarity indices, and 11 coincidence thresholds.


Cosmetic oral and maxillofacial surgery.
The dental specialty of oral and maxillofacial surgery over the last fifteen years has been intimately involved in not only altering the bony facial skeleton, but also the esthetic defects of the facial soft tissue drape. These procedures are currently being taught, in many of our residency programs, along side the art and science of dentoalveolar surgery. When one combines the modification of both the skeletal and soft tissues, the resultant effect can be greater than the sum of its parts. Should one take this a step further and add to the list, proper arch form as prepared by the orthodontist along with cosmetic shape and appearance of the dentition by the restorative dentist using the latest in cosmetic dental techniques, what transforms is a result that has never been seen heretofore in the annals of dentofacial care. To follow are a number of case reports illustrating some salient points.


Losing sleep over speaking up at work: A daily study of voice and insomnia.
Organizational scholars have examined a number of antecedents of insomnia in a search for ways to prevent insomnia and its negative implications for the workplace. However, most studies have focused on the antecedents that are beyond employee control. Therefore, our collective understanding of how employees can modify their workplace behaviors to reduce the symptoms of insomnia and prevent its adverse consequences has remained limited. In this study, we examined whether the expression of voice, as a prosocial yet psychologically costly behavior that is under employee control, affects employee sleep quality, and whether sleep quality affects the expression of voice on the next workday. Having surveyed 113 full-time employees twice a day for 10 workdays, we found that employees who express promotive voice at work experience higher positive affect at the end of the workday, more effectively detach from work in the evening, and are less likely to suffer from insomnia at night. We also found that employees who express prohibitive voice at work experience higher negative affect at the end of the workday, less effectively detach from work in the evening, and are more likely to experience insomnia at night. Our study further demonstrates that, while insomnia is not related to the expression of prohibitive voice on the next day, sleep-deprived employees are less likely to engage in promotive voice because of being psychologically depleted. The results of our study suggest that sleep problems might be mitigated if employees regulate their engagement in costly workplace behaviors, such as voice. (PsycInfo Database Record (c) 2023 APA, all rights reserved).


Hepatic biomarker responses to organic contaminants in feral chub (Leuciscus cephalus)--laboratory characterization and field study in the Sava River, Croatia.
As a widely spread cyprinid fish species, the European chub (Leuciscus cephalus) has been used extensively in biomonitoring programs. However, no laboratory dose-response and/or time course studies related to applied biomarkers have been reported on chub yet. In order to address this issue, specimens of juvenile chub caught in September 2005 in the Sava River, Croatia, were laboratory exposed to various (0.25-50 mg/kg) doses of either model polycyclic aromatic hydrocarbons (PAH) premutagen benzo[a]pyrene (BaP), or beta-naphthoflavone (beta-NF), a well-known model cytochrome 1A (CYP1A) inducer, for 3 (BaP) or 5 d (beta-NF). The responses of several hepatic biomarkers were determined in the exposed fish: The hepatic 7-ethoxyresorufin-O-deethylase activity, CYP1A content, glutathione S-transferase (GST) activity, liver bioactivation potential, and the amount of hydroxylated polycyclic aromatic hydrocarbon bile metabolites determined by the fixed wavelength fluorescence and the high-performance liquid chromatography technique. The relevance of determined biomarker responses has been analyzed further and crosscorrelated with the same set of biomarkers, as well as with tissue concentrations of polycyclic aromatic hydrocarbons and polychlorinated biphenyls, determined in chub specimens collected in September 2005 at five different polluted locations along the Sava River. The species-specific upper and lower limits in responses of studied biomarkers were determined and the obtained ranges successfully evaluated in real field situation. With the exception of the GST activity, all other biomarkers determined in chub proved to be valuable indicators of environmental pollution. Finally, the results of the present study demonstrated that the same strategy of laboratory characterization in combination with field evaluation should be used regularly in the selection of optimal biomarkers and indicator species.


[Chester-Erdheim's disease. A case].
We report the 32nd case of a multivisceral form of Erdheim-Chester disease. This exceptional pathology is a diffuse xanthogranulomatosis which comes within the scope of histiocytosis. The originality of this case is due to cerebral localizations and to the fact that some symptoms have been observed for a long time: diabetes insipidus, exophthalmos and stubborn intertrigo.


CD34+ peripheral blood progenitors as a target for genetic correction of the two flavocytochrome b558 defective forms of chronic granulomatous disease.
Chronic granulomatous disease (CGD) can result from any of four single gene defects involving components of the superoxide (O2-.)-generating phagocyte NADPH oxidase (phox). The phox transmembrane flavocytochrome b558 is composed of two peptides, gp91phox and p22phox. Mutations of gp91phox cause X-linked CGD, whereas mutations of p22phox cause one of the three autosomal recessive forms of CGD. We used the Maloney leukemia virus-based MFG retrovirus vector to produce replication defective retroviruses encoding gp91phox or p22phox. To maximize viral titer MFG retroviruses do not contain internal promoter or resistance elements. Epstein-Barr virus transformed B-lymphocyte cell lines (EBV-B) derived from normal individuals contain phox components and produce O2-., whereas those derived from CGD patients show the CGD defect. Transduction of gp91phox or p22phox-deficient CGD EBV-B lines resulted in correction of O2-. production from a barely detectable baseline to an average 7.2% and 13.8% of normal control, respectively, without any selective regimen to enrich for transduced cells. CD34+ hematopoietic progenitor cells, the therapeutic target for gene therapy of CGD, were isolated from peripheral blood of CGD patients, transduced with MFG-phox retroviruses, and differentiated in culture to mature phagocytes. Transduction of progenitors corrected the gp91phox (seven patients) and p22phox (two patients) CGD phagocyte oxidase defect to 2.5% and 4.9% of normal O2-. production, respectively, representing an 87-fold and 161-fold increase. These studies show correction of flavocytochrome b558-deficient CGD in primary hematopoietic progenitors, providing a basis for development of gene therapy for the X-linked gp91phox and autosomal p22phox-deficient forms of CGD.


Maintenance of mitochondrial oxygen homeostasis by cosubstrate compensation.
Mitochondria maintain a constant rate of aerobic respiration over a wide range of oxygen levels. However, the control strategies underlying oxygen homeostasis are still unclear. Using mathematical modeling, we found that the mitochondrial electron transport chain (ETC) responds to oxygen level changes by undergoing compensatory changes in reduced electron carrier levels. This emergent behavior, which we named cosubstrate compensation (CSC), enables the ETC to maintain homeostasis over a wide of oxygen levels. When performing CSC, our ETC models recapitulated a classic scaling relationship discovered by Chance [Chance B (1965) J. Gen. Physiol. 49:163-165] relating the extent of oxygen homeostasis to the kinetics of mitochondrial electron transport. Analysis of an in silico mitochondrial respiratory system further showed evidence that CSC constitutes the dominant control strategy for mitochondrial oxygen homeostasis during active respiration. Our findings indicate that CSC constitutes a robust control strategy for homeostasis and adaptation in cellular biochemical networks.


Particulate Study on NeoProfen, a Neonatal Injectable Product.
NeoProfen or sterile ibuprofen L-lysine at 10 mg/mL ibuprofen, in 2 mL single-use Type I glass vials is often a first choice medication used to close a patent ductus arteriosus in neonatal patients from 500 to 1500 g body weight. Visible particulate matter was found in vials that were placed on a commercial stability program prior to the approved expiration date of 2 years. A combination of instrumental techniques including inductively coupled plasma-mass spectrometry, x-ray photoelectron spectroscopy, scanning electron microscopy energy dispersive x-ray spectrometry, and Raman and Fourier transform infrared microspectroscopy was used to evaluate stability, pilot batch and packaging samples in a root cause investigation. The particulate matter was shown to consist largely of ibuprofen aluminum salts of various stoichiometries. It developed over time by a substitution mechanism, in which the ibuprofen anion in solution reacts with the aluminum oxide network of the borosilicate glass giving the ibuprofen aluminum salt with =Al-OH remaining in the network. For corrective action an alternate Type I borosilicate glass vial with interior coating, not found in the original vial, was chosen for the product to prevent this occurrence. NeoProfen (sterile preservative-free ibuprofin L-lysine at 17 mg/mL in a single-use glass vial) is used to close a clinically significant patent ductus arteriosus in premature infants no more than 32 weeks gestational age. The neonatal population is especially sensitive to outside chemical, physical and environmental conditions because of incompletely developed organ systems, low birth weight and other underlying conditions. Two batches of this product were voluntarily recalled by the manufacturer, Lundbeck, and investigated for the source of particulate matter observed during a commercial stability testing program. This was found to result from an interaction between the product and the Type I borosilicate glass vial where ibuprofen substitutes for the aluminum oxide network in the glass, forming an ibuprofen aluminum hydroxide salt as particulate. In order to prevent this salt formation an alternate glass vial was chosen which had interiors treated using a chemical vapor deposition technique. These vials were found to preserve NeoProfen quality properties during short term stress and medium term stability studies.


Impact of Charlson comorbidity index varies by age in patients with prostate cancer treated by radical prostatectomy: a competing risk regression analysis.
To investigate the prognostic impact of the Charlson comorbidity index (CCI) on either cancer-specific mortality (CSM) or other-cause mortality (OCM) according to age in patients with prostate cancer (PC) who underwent radical prostatectomy (RP). Data from 336 patients who underwent RP for PC between 1992 and 2005 were analyzed. Variables, including the preoperative prostate-specific antigen (PSA), prostate volume, clinical stage, and pathologic stage, were compared across age groups (<65 or ≥65 years old). Preexisting comorbidities were evaluated by the CCI, and patients were classified into two CCI score categories (0 or ≥1). The median (interquartile range) follow-up period was 96 (85-121) months. Subjects were divided into two subgroups according to age: <65 years (n = 151) or ≥65 years (n = 185). There was no significant difference in PSA, biopsy Gleason sum, body mass index, pathologic stage, or CCI between the two age groups. OCM was significantly associated with the CCI score (P = 0.011). Cumulative incidence estimates obtained from competing risk regression analysis indicated that CCI was not associated with CSM (P = 0.795) or OCM (P = 0.123) in the ≥65-year group. However, in men <65 years, cumulative incidence estimates for OCM were significantly associated with CCI (P = 0.036). CCI was independently associated with OCM after RP, but only in men <65 years old. CCI was not associated with CSM in either age group. Accordingly, a thorough evaluation of patient's comorbidities is mandatory when considering aggressive surgical treatment, especially in relatively young patients.


[Carcinoma of the cervix stages Ib, II, III. Results of treatment and prognostic factors. Retrospective analysis of 396 cases].
Between 1976 and 1986 we have treated 396 patients with carcinoma of the uterine cervix, stages Ib, II and III. The mean age was 54.2 years. The staging system used was the FIGO classification, modified according to the Institut Gustave-Roussy criteria. There were in stage Ib 117 cases, in early stage II 124 cases, in late stage II 63 cases and in stage III 92 cases. Treatment methods used were radiotherapy and surgery combined for localised pelvic lesions (stage Ib and stage II) and radiotherapy alone for those cases that were central and going out to the side wall of the pelvis (late stage II and stage III). The actuarial five-year survival rate was 91% for stage Ib, 67% for early stage II, 48% for late stage II and 28% for stage III. The main causes of failure were recurrence in the pelvis in 15% of treated areas and distant metastases in 13%. The state of the lymph nodes was another prognostic factor but the histological subtype and age were not. 7% of cases had severe complications (urinary, digestive and lymphatic).


Murine cytotoxic T cells specific to respiratory syncytial virus recognize different antigenic subtypes of the virus.
The aim of this study was to establish whether cytotoxic T cells (Tc), raised against respiratory syncytial virus (RSV) in the mouse, are specific to the strain of immunizing virus, or cross-reactive between virus strains. Several recent studies using monoclonal antibodies have begun to define the antigenic variation among strains of RSV. It is likely that a successful RSV vaccine will need to contain antigenic determinants from more than one subtype, but since only the highest levels of neutralizing antibody are able to give complete protection against RSV infection, a vaccine may also need to elicit a cellular immune response. We have recently described H-2-restricted, RSV-specific Tc following RSV infection in the mouse; we present here evidence that polyclonal RSV-specific Tc in the mouse recognize syngeneic target cells infected with every human strain of RSV tested, whatever the subtype. The only RSV strain that appeared not to be recognized was bovine RSV, which seems unable to infect mouse cells; however, bovine cells, infected with bovine RSV and fixed with glutaraldehyde, primed mice for Tc which recognized human strains of RSV.


Down syndrome screening in assisted conception twins: an iatrogenic medical challenge.
The objective of this study was to provide a critical analysis of the impact of assisted conception on prenatal screening for Down syndrome (DS) in twin pregnancies and the value of various screening modalities for early detection of anomalies. The literature was searched using PubMed and the Cochrane Library focusing on prenatal screening and antenatal care of assisted-conception twin pregnancies. Serum screening alone is of limited value in detecting aneuploid twins, because the unaffected cotwin can "mask" the abnormal serum results of an affected one. In addition, this test can designate the pregnancy as at high risk but not identify the affected fetus. Nuchal translucency (NT) screening is the best available modality and a highly effective screening method for twin pregnancies. Among twins, NT alone has a 69% DS detection rate, first-trimester combined NT and serum biochemistry has a 72% DS detection rate, and an integrated screen will have an 80% DS detection rate at a 5% FPR. The data in the literature concerning the effect of assisted conception on maternal serum screening markers in twin pregnancies are scarce. Down syndrome screening in assisted-conception twins presents clinical and technical challenges. Therefore, assisted-conception twins need close monitoring from conception to delivery, by a practitioner familiar with the available screening modalities and their relative accuracy.


Patterns of contraceptive use within teenagers' first sexual relationships.
Teenagers have a high unintended pregnancy rate, in part because of inconsistent use or nonuse of contraceptives. It is important to determine how partner and relationship characteristics are related to contraceptive use and consistency within adolescents' first sexual relationships. Logistic and multinomial logistic regression analyses of data from 1,027 participants in the first two waves of the National Longitudinal Study of Adolescent Health examined the influence of relationship and partner characteristics on ever-use and consistent use of contraceptive methods during teenagers' first sexual relationships. Teenagers who had waited a longer time between the start of a relationship and first sex with that partner, discussed contraception before first having sex or used dual contraceptive methods had significantly increased odds of ever or always using contraceptives. Adolescents who had taken a virginity pledge, had an older partner, had a greater number of close friends who knew their first partner, or reported having a relationship that was not romantic but that involved holding hands, kissing and telling their partners they liked or loved them had decreased odds of contraceptive use or consistency. As relationship length increased, teenagers were more likely to ever have used a method, but less likely to always have used a method. Parents and programs should encourage teenagers to delay sexual intercourse, discuss contraception with partners before initiating sex and be vigilant about contraceptive use, particularly in long-term sexual relationships and in relationships with older partners.


The Role of Abnormal Hemostasis and Fibrinolysis in Morbidity and Mortality of Acute Promyelocytic Leukemia.
Despite the improved therapeutic advances in the management of acute promyelocytic leukemia (APL), a significant early mortality during induction, also referred to as early death (ED), remains an obstacle for further improvement in outcome. Hemorrhagic complications are the most common cause of morbidity and mortality. Perturbed hemostatic dysfunction is present as the result of abnormalities in both the coagulation and the fibrinolytic systems. The activation of coagulation is distinct from the classical disseminated intravascular coagulation. Multiple abnormalities in the fibrinolytic system have recently been identified. The most significant change is increased production of tissue plasminogen activator (tPA) and its receptor annexin A2 by the APL promyelocytes. Among the hemorrhagic complications, intracranial hemorrhage predominates. The pathogenesis of this catastrophic event is elucidated by new evidence of adverse effect of tissue plasminogen activator (tPA) on the brain, including both the plasmin-dependent and plasmin-independent pathways. In order to address the hemorrhagic complications, a thorough understanding of the hemostatic dysfunction is essential. In this article, our current concept of the abnormal hemostasis in APL is reviewed. The failure to reduce the early death rate, despite the introduction of effective therapy, will also be discussed.


Intratumoral molecular heterogeneity in a BRAF-mutant, BRAF inhibitor-resistant melanoma: a case illustrating the challenges for personalized medicine.
Targeted therapies are increasingly being used to treat a variety of cancers. Their efficacy depends upon the accurate detection and targeting of a specific mutation or aberration in the tumor. All cancers, such as melanoma, are molecularly heterogeneous, with drug-resistant subclones present before the treatment or emerging as a result of targeted therapies. Here, we show intralesional molecular heterogeneity in a progressing V600E BRAF-mutant melanoma metastasis from a patient treated for 7 months with the BRAF inhibitor vemurafenib. In the single metastasis, two distinct subclones were observed, both V600E BRAF-mutant and only one with an additional G13R NRAS mutation. Molecular heterogeneity even at the intralesional level shows that personalizing or adjusting therapies based on genotyping of a portion of a single lesion may not accurately depict the molecular profile or drivers of oncogenesis across the entire patient's melanoma.


Differential uptake of alkylating agents by normal and leukemic lymphocytes.
When 14C-labeled cyclophosphamide and nitrogen mustard were incubated separately with normal lymphocytes and lymphocytes from patients with chronic lymphocytic leukemia, the amount of radioactivity associated with the normal cells far exceeded that detected on the leukemic lymphocytes. This comparative diminution may be analogous to the impaired PHA response and excess surface immunoglobulin which serve as identifying markers of the malignant B cell. Cytotoxicity and neuraminidase experiments indicated that drug uptake by lymphocytes is not capricious and may occur in an optimum, predetermined fashion. Although surface uptake and therapeutic response are not necessarily directly interrelated, initial peripheral contact with an antineoplastic agent may be an essential step which modifies tumor sensitivity or resistance.


Human plague system associated with rodent diversity and other environmental factors.
Plague remains a threat to public health and is considered as a re-emerging infectious disease today. Rodents play an important role as major hosts in plague persistence and driving plague outbreaks in natural foci; however, few studies have tested the association between host diversity in ecosystems and human plague risk. Here we use zero-inflated generalized additive models to examine the association of species richness with human plague presence (where plague outbreaks could occur) and intensity (the average number of annual human cases when they occurred) in China during the Third Pandemic. We also account for transportation network density, annual precipitation levels and human population size. We found rodent species richness, particularly of rodent plague hosts, is positively associated with the presence of human plague. Further investigation shows that species richness of both wild and commensal rodent plague hosts are positively correlated with the presence, but only the latter correlated with the intensity. Our results indicated a positive relationship between rodent diversity and human plague, which may provide suggestions for the plague surveillance system.


Fabrication and characterization of PDMS microlenses based on elastomeric molding technology.
Elastomeric molding technology is adopted to fabricate polydimethilsiloxane (PDMS)-based solid microlenses, as they can have different focal lengths and achieve different lens types (such as concave and convex) using the same mold structure. The only parameter that needs to be controlled is the pressure applied during the molding process. These PDMS lenses can also be transformed from the original spherical ones into microlenses having elliptic surface profiles simply by stretching their substrates in one direction. This provides the process with additional tunability of astigmatism.


Usefulness of biochemical markers of bone turnover in rats after clodronate administration.
Dichloromethylene bisphosphonate (clodronate), an inhibitor of bone resorption, has been administered subcutaneously (1.25 mg/day, 3 consecutive days) to control male albino Wistar rats (n = 8). Variations in the levels of serum alkaline phosphatase (AP) and osteocalcin (BGP), biochemical markers of bone formation, and in urinary hydroxyproline (OH-Prol) and serum tartrate-resistant acid phosphatase (TRAP), markers of bone resorption, have been analyzed. A significant decrease was observed in OH-Prol/creatinine and TRAP. We found a positive correlation between the percentage of decrease, with respect to basal values, of TRAP and OH-Prol/creatinine (p < 0.05). These results suggest that TRAP could be a useful marker to evaluate the changes of bone resorption induced by bisphosphonates in the rat together with the classical marker OH-Prol. A significant decrease was observed in AP. BGP also decreased significantly, but the decrease was relatively small compared to the coefficient of variation of the method. We suggest the preferential use of AP determination, instead of BGP, in the study of the effects produced by bisphosphonates on bone formation in rats, if hepatic function is maintained.


Immunohistochemistry of Pneumocystis carinii infection.
Pneumocystis carinii is the pre-eminent pulmonary pathogen and leading cause of death in patients with acquired immunodeficiency syndrome (AIDS). The diagnosis of this organism depends upon the morphologic demonstration of the cyst wall, trophozoite, or sporozoite in specimens from the lower respiratory tract. A variety of histochemical stains have been used to identify P. carinii, each with considerable limitations in specificity. Extrapulmonary spread and unusually destructive pulmonary patterns associated with P. carinii, although once considered rare, are seen occasionally in patients with AIDS. Traditional stains have proven to be less reliable in extrapulmonary sites. In 13 patients with AIDS, we stained formalin-fixed paraffin-embedded autopsy lung and other visceral organ sections using monoclonal antibody 3F6 (Dako, Santa Barbara, CA) to P. carinii. The antibody stained P. carinii in the lungs of seven patients with P. carinii pneumonia by Gomori methenamine silver stain (GMS). Numerous aggregates of P. carinii cysts were marked within alveoli, as is usually seen with other stains. No antibody staining was present in autopsy lung sections from non-AIDS patients with viral, fungal, or bacterial pneumonia. Clinically occult extrapulmonary P. carinii infection was seen in 4/7 (57%) patients with P. carinii pneumonia at autopsy. Monoclonal antibody to P. carinii stained organisms in all four (100%) patients with disseminated disease, as compared with 1/4 (25%) staining with GMS. Antibody staining of P. carinii was demonstrated in sections of thyroid, adrenal, and carinal lymph node (one patient), esophagus (one patient), kidney (one patient), and heart, thyroid, kidney, adrenal, liver, stomach, pancreas, spleen, and bone marrow (one patient).(ABSTRACT TRUNCATED AT 250 WORDS)


Retinoblastoma protein in non-small cell lung carcinoma, cells arrested for growth by retinoic acid.
The growth of non-small cell lung carcinoma (NSCLC) cells was inhibited by retinoic acid after 16 h of treatment. However, the growth of all cell lines except one became refractory to retinoic acid after 48 h of exposure. The expression of the hyperphosphorylated retinoblastoma protein species (RB p110) was observed to be increased fivefold to tenfold in NSCLC cells within 16 h of exposure to retinoic acid. In the H460a and H226b cell lines, p110 showed some conversion to the underphosphorylated p105 form after 24 h of retinoic acid treatment. After 48 h, p105 became the predominant form, along with a 60K M(r) species. After 72 h, expression of all RB protein species became almost undetectable in H460a and H226b cells, concomitant with increases in cell growth rates. A different pattern of RB expression was observed in the H322j and H358 cell lines. In these cells, both p110 and p105 were induced within 8 h of retinoic acid treatment. Furthermore, the elevated levels and the phosphorylation state of RB in retinoic acid-treated H322j and H358 cells remained essentially unchanged for up to 72 h and only low amounts of the 60K M(r) species were detected. I believe that a post-translational mechanism is responsible for the retinoic acid-mediated induction of RB, since levels of RB mRNA remained relatively unchanged during the time course of retinoic acid exposure. I conclude that retinoic acid inhibited the growth of NSCLC cells by inducing high levels of RB and that increases in RB levels occurred as a result of either an increase in stability of the protein or by downregulation of an RB-specific protease. The inhibition of growth by retinoic acid must involve other molecular events, since the H596b cell line, which lacks RB protein, exhibited growth properties in the presence of retinoic acid similar to those of cells in which RB expression was induced.