Examining inequalities in the uptake of the school-based HPV vaccination programme in England: a retrospective cohort study.
Although uptake of Human Papillomavirus (HPV) vaccine is high in the United Kingdom, it is unknown whether the programme has been delivered equitably by ethnicity or deprivation. This study aimed to investigate factors associated with HPV vaccine initiation and completion within the routine HPV vaccination programme in the South West of England. Data were retrieved for young women eligible for routine vaccination from 2008/09 to 2010/11 from three Primary Care Trusts (PCTs)/local authorities. Multivariable logistic regression models were developed to examine factors associated with uptake of HPV vaccination. Of 14 282 eligible young women, 12 658 (88.6%) initiated, of whom 11 725 (92.6%) completed the course. Initiation varied by programme year (86.5-89.6%) and PCTs/local authorities (84.8-91.6%). There was strong evidence for an overall difference of initiation by ethnicity (P < 0.001), but not deprivation quintile (P = 0.48). Young women educated in non-mainstream educational settings were less likely to initiate and, if initiated, less likely to complete (both P < 0.001). HPV vaccination uptake did not vary markedly by social deprivation. However, associations with ethnicity and substantially lower uptake in non-mainstream educational settings were observed. Research to identify reasons for low vaccine uptake in these population groups is required.


In vivo fluorescent labeling and tracking of extracellular matrix.
Connective tissues are essential building blocks for organ development, repair and regeneration. However, we are at the early stages of understanding connective tissue dynamics. Here, we detail a method that enables in vivo fate mapping of organ extracellular matrix (ECM) by taking advantage of a crosslinking chemical reaction between amine groups and N-hydroxysuccinimide esters. This methodology enables robust labeling of ECM proteins, which complement previous affinity-based single-protein methods. This protocol is intended for entry-level scientists and the labeling step takes between 5 and 10 min. ECM 'tagging' with fluorophores using N-hydroxysuccinimide esters enables visualization of ECM spatial modifications and is particularly useful to study connective tissue dynamics in organ fibrosis, tumor stroma formation, wound healing and regeneration. This in vivo chemical fate mapping methodology is highly versatile, regardless of the tissue/organ system, and complements cellular fate-mapping techniques. Furthermore, as the basic chemistry of proteins is highly conserved between species, this method is also suitable for cross-species comparative studies of ECM dynamics.


Cutaneous tumors in pregnancy.
Pregnancy alters the frequency and natural course of certain skin tumors. Pregnancy-associated changes in melanocytic nevi are transient, and there is no substantiated evidence of increased risk of malignant transformation of melanocytic nevi in gestation. Characteristic vascular and pigment-related dermatoscopic features are helpful in evaluating pigmented lesions, but a biopsy should be performed for significant change or other worrisome features in a lesion. Outcomes for pregnancy-associated melanoma do not appear to be poorer compared with nonpregnancy melanoma; however, data are limited for advanced (stage III/IV) melanoma. Some studies suggest increased propensity for lymphovascular spread, but more data are needed for definitive conclusions and guidelines on prognostication, workup, and treatment of pregnancy-associated melanoma. Vascular tumors, particularly pyogenic granuloma (granuloma gravidarum), occur with increased frequency and are associated with pro-angiogenic hormonal influences. Dermatofibrosarcoma protuberans has a more aggressive course during pregnancy with both prompt surgical treatment and close monitoring for recurrence being indicated.


Endogenous estrogen, testosterone and progesterone levels in relation to breast cancer risk.
Multiple lines of evidence support a central role of hormones in the etiology of breast cancer. In epidemiologic studies, considerable effort has focused on delineating the role of endogenous hormones in risk of breast cancer among postmenopausal women. Recently, substantial additional data has accrued from prospective studies where endogenous hormones are measured in study subjects prior to disease diagnosis. In this review, the epidemiologic evidence linking sex steroids--estrogens, testosterone, and progesterone, specifically--with subsequent risk of breast cancer in both premenopausal and postmenopausal women is summarized. Overall, a strong positive association between breast cancer risk and circulating levels of both estrogens and testosterone has now been well confirmed among postmenopausal women; women with hormone levels in the top 20% of the distribution (versus bottom 20%) have a two- to three-fold higher risk of breast cancer. Evidence among premenopausal women is more limited, though increased risk associated with higher levels of testosterone is consistent. However, both positive and null associations have been observed with estrogens and progesterone and clearly more evaluation is needed.


Comparison of the cytotoxic responses of Escherichia coli (E. coli) AMC 198 to different fullerene suspensions (nC60).
Fullerenes are set to be produced on an industrial scale in anticipation of their wide applications. This calls for research on their environmental and health impacts. This study investigates and compares the cell toxicity of different aqueous fullerene aggregates. Popular C(60) dispersal methods were used to prepare four types of nC(60) aggregates. These aggregates were tested against the indicator species Escherichia coli (E. coli) AMC 198. With aggregates of around 150 nm in diameter, the THF/nC(60) suspension was very toxic and gave rise to a half maximal effective concentration (EC(50)) of 0.54 mg L(-1) in E. coli. By contrast, the Tol/nC(60) suspension exhibited a cytoprotective role while the Aqu-N(2)/nC(60) and Aqu-O(2)/nC(60) suspensions enhanced the metabolism of E. coli. Although some toxicants, such as THF and THF-peroxide, were introduced into the THF/nC(60) suspension during the dispersion, the toxicity of nC(60) itself cannot be neglected.


Earthworms, as ecosystem engineers, influence multiple aspects of a salamander's ecology.
Ecosystem engineers create habitat that can be used by other species in multiple ways, such as refugees from predators, places to breed, or areas with increased prey resources. I conducted a series of enclosure experiments to: (1) determine if salamanders use earthworm burrows, and (2) examine the potential influence of earthworm burrow use and indirect effects on salamander intra- and interspecific competition, predator avoidance, and seasonal performance. I found that one species of woodland salamander, Plethodon cinereus, used earthworm burrows 50% of the time when burrows were present. Neither adults nor juveniles of the congeneric P. glutinosus used earthworm burrows. Intraspecific, but not interspecific, competition by P. cinereus affected salamander behavior when earthworms were absent, with P. cinereus found under cover objects >70% of the time when alone or with a P. glutinosus, but only 40% of the time when with another P. cinereus. When earthworms were present, the behavior of P. cinereus was similar across salamander treatments. Earthworms decreased the amount of leaf litter and microinvertebrates, although this did not affect salamander mass. In subsequent experiments using only P. cinereus, the refuge provided by earthworm burrows increased the survival of P. cinereus over the winter and allowed P. cinereus to avoid being consumed by the common garter snake (Thamnophis sirtalis). Because earthworm burrows provide a refuge for P. cinereus during intraspecific encounters, in the presence of a predator and over the winter, they may serve as an important belowground-aboveground linkage in eastern forests where salamanders are common.


Can a selective PPARγ modulator improve glycemic control in patients with type 2 diabetes with fewer side effects compared with pioglitazone?
INT131 besylate is a potent, nonthiazolidinedione, selective peroxisome proliferator-activated receptor γ (PPARγ) modulator (SPPARM) designed to improve glucose metabolism while minimizing the side effects of full PPARγ agonists. This placebo-controlled study compared the efficacy and side effects of INT131 besylate versus 45 mg pioglitazone HCl in subjects with type 2 diabetes (T2D). This was a 24-week randomized, double-blind, placebo- and active-controlled study of 0.5-3.0 mg INT131 versus 45 mg pioglitazone or placebo daily in 367 subjects with T2D on sulfonylurea or sulfonylurea plus metformin. The primary efficacy analysis was the comparison of change from baseline to week 24 in hemoglobin A1c (HbA1c) across treatment groups. Fluid status was assessed with a prospective scoring system for lower-extremity pitting edema. INT131 had a steep dose response for efficacy as measured by changes in HbA1c. After 24 weeks' treatment, the 0.5-mg dose demonstrated minimal efficacy (HbA1c -0.3 ± 0.12%) and the 2-mg dose demonstrated near-maximal efficacy (HbA1c -1.1 ± 0.12%), which was not statistically different from the efficacy of 45 mg pioglitazone (HbA1c -0.9 ± 0.12%; P < 0.01 for noninferiority). With the 1-mg dose, INT131 provided significant improvements in glycemic control (HbA1c 0.8 ± 0.12; P < 0.001 vs. placebo) but with less edema, weight gain, and hemodilution than observed with 45 mg pioglitazone. INT131 demonstrated dose-dependent reductions in HbA1c, equivalent to 45 mg pioglitazone, but with less fluid accumulation and weight gain, consistent with its SPPARM design.


Encrusted cystitis.
We report a case of a patient who developed encrusted cystitis after transurethral resection of the prostate. This rare urological condition is characterized by intramucosal calcifications and is commonly preceded by urological instrumentation. Urea-splitting bacteria, most commonly Corynebacterium urealyticum, are the causative pathogen. Treatment is a combination of antibiotics, urine acidification, and endoscopic removal of encrustations.


Effect of calcium-antagonist and calmodulin-antagonist drugs on calmodulin-dependent contractions of chemically skinned vascular smooth muscle from rabbit renal arteries.
1. Renal arteries from rabbits were chemically skinned by incubation with Triton X-100, and subsequently stored in buffered glycerol. 2. In the presence of Mg-ATP, of EGTA-buffered calcium, and of calmodulin, miniature strips of the skinned arteries developed tension the strength of which was approx. 15-20% of that of viable renal arteries. 3. Tension development was dependent on the concentration of both calcium and calmodulin. 4. The effect of eight vasodilator drugs, the majority of them being "calmodulin antagonists" or "calcium antagonists", on the skinned arteries was assessed. In concentrations up to 10(-3) M, verapamil, D-600, and hydralazine proved to be ineffective, and the same was found with the dihydropyridine derivatives, nifedipine and felodipine, at 0.6 X 10(-3) M and 0.8 X 10(-4) M, respectively, i.e. at saturation in a 9:1 contracting buffer/ethanol mixture (v/v). 5. In a concentration-dependent manner, trifluoperazine, W-7, and fendiline relaxed Ca-calmodulin-induced tension or prevented tension development when given prior to the activation by Ca-calmodulin. However, considerably higher concentrations of the drugs were necessary for half-maximal relaxation than the reported concentrations for half-maximal saturation of hydrophobic binding sites at the calmodulin molecule. 6. These findings suggest that at therapeutic blood levels, the vasodilator properties of calcium antagonists and other direct vasodilators cannot be explained by interference with the binding of myosin light chain kinase to calmodulin.


Evidence for the major contribution of remodeling-based bone formation in sclerostin-deficient mice.
Bone formation by osteoblasts is achieved through remodeling-based bone formation (RBBF) and modeling-based bone formation (MBBF). The former is when bone formation occurs after osteoclastic bone resorption to maintain bone mass and calcium homeostasis. The latter is when new bone matrices are added on the quiescent bone surfaces. Administration of anti-sclerostin neutralizing antibody promotes MBBF in ovariectomized rats and postmenopausal women. However, it remains to be elucidated which mode of bone formation mainly occurs in Sost-deficient mice under physiological conditions. Here, we show that two-thirds of bone formation involves RBBF in 12-week-old Sost-deficient mice (C57BL/6 background). Micro-computed tomography and histomorphometric analyses showed that the trabecular bone mass in Sost-KO mice was higher than that in Sost+/- mice. In contrast, the osteoclast number remained unchanged in Sost-KO mice, but the bone resorption marker TRAP5b in serum was slightly higher in those mice. Treatment with anti-RANKL antibody increased the trabecular bone mass of Sost+/- or Sost-KO mice. Bone formation markers such as osteoid surfaces, the mineral apposition rate, and bone formation rate were almost completely suppressed in Sost+/- mice treated with anti-RANKL antibody compared with vehicle-treated Sost+/- mice. In Sost-KO mice, treatment with anti-RANKL antibody suppressed those parameters by more than half. These findings indicate that RBBF accounts for most of the bone formation in Sost+/- mice, whereas approximately two-thirds of bone formation is estimated to be remodeling-based in 12-week-old Sost-deficient mice. Furthermore, anti-RANKL antibody may be useful for detecting MBBF on trabecular bone.


NBIGV-DB: A dedicated database of non-B cell derived immunoglobulin variable region.
Immunoglobulins (Ig) are important immune molecules that possess highly diverse variable region sequences enabling antigen recognition. According to classical immune theory, B lymphocytes have been considered the only source of Ig production (B-Igs). However, accumulating evidence have suggested that Igs are also produced by many non-B cells (non-B Igs), including epithelial cells, neurons, germ cells, as well as myeloid cells of hemopoietic system. Besides acting as bona fide antibodies, Non-B Igs have alternative cellular functions, such as promotion of cell survival, adhesion and migration. More importantly, Unlike the unlimited sequence diversity of B-Igs, the non-B Igs exhibit conserved V(D)J patterns across the same lineages. To support the analysis and comparison of variable region sequences from Igs, produced by B and non-B cells, we established a database (NBIGV) constituted by a non-B Ig variable region repertoire, which includes 727,989 VHDJH and VκJκ recombination sequences of non-B Igs sequenced from mouse samples. Upon database search, users can view, browse and investigate the variable region sequences of non-B Igs according to respective mice strains and tissues as well as Ig classes. Moreover, users can easily download selected sequences and/or compare sequences of interest with known non-B Ig sequences present in the database using NCBI-BLAST algorithms. Additionally, our database integrates a submission page and supplementary sample information. The NBIGV database may serve as a valuable resource for sequence analyses of Non-B Igs. NBIGV database is freely available at http://nbigv.org.


Functional Outcomes in Cochleostomy and Round Window Insertion Technique: Difference or No Difference?
With the introduction and rapid development of Cochlear Implants since the 1970s, there has been marked improvement in the speech recognition and spoken language skills of the implanted profoundly deaf children. The cochlear implant can be done by means of different techniques, traditionally by Cochleostomy method and round window membrane (RWM) insertion technique. Post operatively, the functional outcomes are measured by many scores more commonly by Categories of Auditory Performance (CAP) and Speech Intelligibility Rating (SIR) scores. To study the speech and hearing perception skills in pediatric cases of Congenital non syndromic bilateral profound sensorineural hearing loss after Cochleostomy and Round Window Insertion technique of Cochlear Implantation. 31 patients clinically diagnosed as congenital non syndromic bilateral profound sensorineural hearing loss who had undergone Cochlear implantation either by Cochleostomy or by RWM insertion technique and fulfilling the eligibility criteria were enrolled for study. Post operatively functional outcomes were assessed subjectively by measuring CAP and SIR scores. All the patients showed increase in their CAP and SIR scores post-operatively, measured at 03 months, 06 months and 01 year after Cochlear Implantation. The mean CAP and SIR scores in the two groups were comparable at 03 months, 06 months and 1 year after surgery. There was no significant difference in the speech and hearing perception skills of post implantees in the two groups (p value < 0.05). There is no difference in functional outcomes of Cochlear implantation by Cochleostomy and round window membrane insertion technique.


Improvement of sinonasal mucociliary function by endoscopic sinus surgery in patients with chronic rhinosinusitis.
Mucociliary clearance is an important defense mechanism for upper and lower airway. Chronic rhinosinusitis has been frequently associated with mucociliary dysfunction. Endoscopic sinus surgery (ESS) is recommended for treatment-resistant sinusitis in order to improve mucociliary function. The present study investigated the effect of ESS on the saccharin time (mucociliary clearance time) in relation to symptom profile assessed by the Sino-nasal Outcome Test (SNOT)-22, and disease severity based on the Lund-Kennedy endoscopic scores and Lund-Mackay computed tomographic (CT) scores. The present cohort study included 22 patients with chronic rhinosinusitis who were considered a candidate for ESS due to lack of response to medical treatment. Saccharin test was used before and 3 months after ESS to evaluate changes in mucociliary function. The CT scan was used for preoperative assessment of sinonasal anatomy. In addition, diagnostic nasal endoscopy was performed before and 3 months after ESS. The CT scan and nasal endoscopy findings were respectively analyzed based on the Lund-Mackay and the Lund-Kennedy staging systems recommended for chronic rhinosinusitis. Symptom severity was assessed from before to 3 months after ESS using SNOT-22. The present study showed decreased saccharin time in patients with chronic rhinosinusitis three months after ESS. The mean pre-operative and post-operative saccharin time were 23.4 and 16.9 min, respectively. There was also a trend toward significance for the effect of changes in the SNOT-22 scores on changes in saccharin time. By contrast, no effects of age, gender, pre-operative Lund-Mackay CT scores, and changes in Lund-Kennedy endoscopy scores were observed on post-ESS saccharin time. The study confirms that ESS in patients with treatment-resistant chronic rhinosinusitis confers improvement in performance on the saccharin test of mucociliary function.


Is eye padding routinely necessary after uncomplicated phacoemulsification?
To investigate the value of eye padding following uncomplicated phacoemulsification under peribulbar anaesthesia. A prospective randomised controlled study was conducted to compare the effect of a conventional eye pad and shield with that of a clear eye shield applied without a pad in 83 patients undergoing routine phacoemulsification under peribulbar anaesthesia without lid block. The primary outcome measures were corneal fluorescein staining, discomfort, diplopia and mobility. Moderate or severe corneal fluorescein staining on the first post-operative day was significantly more common in the pad and shield group (39%) than in the clear shield group (19%) (p < 0.01). There was no significant difference in post-operative pain as measured either by visual analogue scale or by categorical pain scale. Forty per cent of the clear shield group reported transient post-operative diplopia during the immediate post-operative period compared with 7% of the pad and shield group (p < 0.001). There was no significant difference in reported mobility between the two groups. Following phacoemulsification under peribulbar anaesthesia, the use of a gauze eye pad is associated with greater corneal fluorescein staining than a clear plastic shield without pad and offers no reduction in discomfort. A clear shield protects the globe against direct trauma, is associated with reduced moderate to severe corneal staining and facilitates vision in the early post-operative period. Transient diplopia reported by some patients given a clear shield is not disabling and would not be expected to occur in patients with one seeing eye. The use of a clear shield alone is a safe alternative to eye padding and offers important advantages in patients with one seeing eye.


Metformin suppresses vascular smooth muscle cell senescence by promoting autophagic flux.
Vascular smooth muscle cell (VSMC) senescence in the vasculature results in vascular aging as well as age-related diseases, while metformin improves the inflamm-aging profile by enhancing autophagy. However, metformin's impact on VSMC senescence is largely undefined. To test the hypothesis that metformin exerts an anti-senescence role by restoring autophagic activity in VSMCs and vascular tissues. Animal models established by angiotensin II (Ang II) induction and physiological aging and senescent primary VSMCs from the aortas of elderly patients were treated with metformin. Cellular and vascular senescence were assessed by measuring the amounts of senescence-associated β-galactosidase and senescence markers, including p21 and p53. Autophagy levels were assessed by autophagy-related protein expression, transmission electron microscope, and autolysosome staining. In order to explore the underlying mechanism of the anti-senescence effects of metformin, 4D label-free quantitative proteomics and bioinformatic analyses were conducted, with subsequent experiments validating these findings. Ang II-dependent senescence was suppressed by metformin in VSMCs and vascular tissues. Metformin also significantly improved arterial stiffness and alleviated structural changes in aged arteries, reduced senescence-associated secretory phenotype (SASP), and improved proliferation and migration of senescent VSMCs. Mechanistically, the proteomic analysis indicated that autophagy might contribute to metformin's anti-senescence effects. Reduced autophagic flux was observed in Ang II-induced cellular and vascular senescence; this reduction was reversed by metformin. Specifically, metformin enhanced the autophagic flux at the autophagosome-lysosome fusion level, whereas blockade of autophagosome-lysosome fusion inhibited the anti-senescence effects of metformin. Metformin prevents VSMC and vascular senescence by promoting autolysosome formation.


Phase II trial of irinotecan plus docetaxel in cisplatin-pretreated relapsed or refractory oesophageal cancer.
This phase II trial assessed the toxicity and efficacy of irinotecan plus docetaxel in cisplatin-pretreated oesophageal cancer. Irinotecan 160 mg m(-2) plus docetaxel 65 mg m(-2) once every 3 weeks led to severe myelosuppression in four patients, all of whom experienced neutropenic fever. After amendment of this regimen, 24 patients (male/female=18/6; median age=58.5 years; ECOG performance status 0/1/2=9/11/4) with advanced oesophageal cancer (adenocarcinoma/epidermoid carcinoma=13/11) received irinotecan 55 mg m(-2) plus docetaxel 25 mg m(-2) on days 1, 8 and 15 of a 28-day cycle. Serious adverse events occurred in five patients, one with lethal outcome (pneumonia). Haematological toxicity >or=3 degrees was rare, whereas nonhaematological toxicity >or=3 degrees was noted in nine out of 24 patients (asthenia in five patients, diarrhoea in three patients, nausea/emesis in two patients, constipation in one patient). Median survival time was 26 (range 2-70) weeks. Response rate, assessed according to the WHO criteria, was 12.5% (95% CI 2.7-32.4%); rate of disease stabilisation (partial remission and stable disease) was 33.3% (95% CI 15.6-55.3%) with a median duration of 18.5 (range 16-51) weeks. Although the nonhaematological toxicity proved to be considerable, weekly irinotecan plus docetaxel is feasible and shows some activity in extensively pretreated patients with oesophageal cancer.


Low temperature thermal desorption-chemical oxidation hybrid process for the remediation of organic contaminated model soil: A case study.
The efficiencies of thermal desorption and oxidation process in engineering soil remediation are to some extent limited due to the huge loss of heat to the soil or underground water or mass transfer resistance in the soil. To enhance the oxidation and improve the energy utilization. Herein, the thermal desorption process and the oxidation process (by ozone) are combined together to remediate the organic contaminated model soil (take the 2,4-dimethylaniline (2,4-DMA) as pollutant). Results show that this hybrid process could not only reduce the thermal desorption temperature (as low as 50-90 °C), but also improve the oxidation efficiency significantly, even achieving 100% degradation of 2,4-DMA in less than 10 min in the soil. It is found that the remediation efficiency by the hybrid process is also highly dependent on different operational parameters, including the heating temperature, ozone concentration, especially the moisture content in the soil. The results suggest that a proper content of water in the soil is beneficial for the degradation of pollutants by the hybrid process. Finally, the degradation kinetics and mechanisms of 2,4-DMA in the soil by the hybrid process has been primarily discussed. These findings suggest that the low temperature enhanced chemical oxidation process would be a promising method for future remediation of organic contaminated soil due to its relatively low energy consuming and high removal efficiency in shorter time.


Internet-based monitoring and benchmarking in ambulatory surgery centers.
Each year the number of surgical procedures performed on an outpatient basis increases, yet relatively little is known about assessing and improving quality of care in ambulatory surgery. Conventional methods for evaluating outcomes, which are based on assessment of inpatient services, are inadequate in the rapidly changing, geographically dispersed field of ambulatory surgery. Internet-based systems for improving outcomes and establishing benchmarks may be feasible and timely. Eleven freestanding ambulatory surgery centers (ASCs) reported process and outcome data for 3,966 outpatient surgical procedures to an outcomes monitoring system (OMS), during a demonstration period from April 1997 to April 1999. ASCs downloaded software and protocol manuals from the OMS Web site. Centers securely submitted clinical information on perioperative process and outcome measures and postoperative patient telephone interviews. Feedback to centers ranged from current and historical rates of surgical and postsurgical complications to patient satisfaction and the adequacy of postsurgical pain relief. ASCs were able to successfully implement the data collection protocols and transmit data to the OMS. Data security efforts were successful in preventing the transmission of patient identifiers. Feedback reports to ASCs were used to institute changes in ASC staffing, patient care, and patient education, as well as for accreditation and marketing. The demonstration also pointed out shortcomings in the OMS, such as the need to simplify hardware and software installation as well as data collection and transfer methods, which have been addressed in subsequent OMS versions. Internet-based benchmarking for geographically dispersed outpatient health care facilities, such as ASCs, is feasible and likely to play a major role in this effort.


Clinical impact of type 2 diabetes mellitus on outcomes after one-lung ventilation during thoracic surgery: a retrospective cohort study.
The preoperative presence of diabetes mellitus (DM) has been recently demonstrated to be a risk factor for adverse events after thoracic surgery. However, the specific effects of presence of DM preoperatively on thoracic surgery is not known. This study aimed to investigate the association between preoperative DM and clinical outcomes and the short-term survival rates after thoracic surgery. In this retrospective, observational, and matched-pair analysis study, patients receiving thoracic surgery from a tertiary university hospital in 2 consecutive years were grouped as either type 2 DM (T2DM) or controlled within the first 24 hours after surgery. Multivariate Cox regression was conducted to investigate the impact of T2DM within the first 24 hours of admission on in-intensive care unit (ICU) and hospital survival. Among the included thoracic patients, 41 (8.4%) had T2DM and 450 (91.6%) did not have T2DM. In the single-factor analyses, T2DM patients were shown to have a higher preoperative white blood cells (WBCs) count, increased release of immunoglobulin A, complement C3 and C4, impaired kidney function with high level of urea, and low expression of alanine aminotransferase (ALT) and monoamine oxidase (MAO). In multivariate analyses, the preoperative urea level was associated with a low-grade risk of dying for the ICU survival time. In contrast, preoperative complement C3 level favored a positive contribution in-ICU survival. Besides the complement C3 level, immunoglobulin A level remained a positive contribution in regression models of hospital survival. Pre-admission T2DM was not associated with an increased in-ICU and hospital mortality among patients with thoracic surgery. Furthermore, they were accompanied by impaired kidney function, activated inflammation and liver function.


Phage-display immunoprecipitation sequencing of the antibody epitope repertoire in inflammatory bowel disease reveals distinct antibody signatures.
Inflammatory bowel diseases (IBDs), e.g., Crohn's disease (CD) and ulcerative colitis (UC), are chronic immune-mediated inflammatory diseases. A comprehensive overview of an IBD-specific antibody epitope repertoire is, however, lacking. Using high-throughput phage-display immunoprecipitation sequencing (PhIP-Seq), we identified antibodies against 344,000 antimicrobial, immune, and food antigens in 497 individuals with IBD compared with 1,326 controls. IBD was characterized by 373 differentially abundant antibody responses (202 overrepresented and 171 underrepresented), with 17% shared by both IBDs, 55% unique to CD, and 28% unique to UC. Antibody reactivities against bacterial flagellins dominated in CD and were associated with ileal involvement, fibrostenotic disease, and anti-Saccharomyces cerevisiae antibody positivity, but not with fecal microbiome composition. Antibody epitope repertoires accurately discriminated CD from controls (area under the curve [AUC] = 0.89), and similar discrimination was achieved when using only ten antibodies (AUC = 0.87). Individuals with IBD thus show a distinct antibody repertoire against selected peptides, allowing clinical stratification and discovery of immunological targets.


Development and characterization of ferrihydrite-modified diatomite as a phosphorus adsorbent.
A novel phosphorus adsorbent, ferrihydrite-modified diatomite was developed and characterized in this study. The ferrihydrite-modified diatomite was made through surface modification treatments including NaOH treatment and ferrihydrite deposition on raw diatomite. In the NaOH treatment, surface SiO(2) of diatomite was partially dissolved in the NaOH solution. The dissolved Si contributed to form the stable 2-line ferrihydrite which deposited into the macropores and mesopores of diatomite. Blocking macropores and larger mesopores of diatomite with 0.24g Fe/g of 2-line ferrihydrite resulted in a specific surface area of 211.1m(2)/g for the ferrihydrite-modified diatomite, which is 8.5-fold increase than the raw diatomite (24.77m(2)/g). The surface modification also increased the point of zero charge (pH(PZC)) values to 10 for the ferrihydrite-modified diatomite from 5.8 for the raw diatomite. Because of the increased surface area and surface charge, the maximum adsorption capacity of ferrihydrite-modified diatomite at pH 4 and pH 8.5 was increased from 10.2mgP/g and 1.7mgP/g of raw diatomite to 37.3mgP/g and 13.6mgP/g, respectively.


How to diagnose and manage neurological toxicities of immune checkpoint inhibitors: an update.
As the use of cancer immunotherapy with immune checkpoint inhibitors (ICIs) is expanding rapidly for the treatment of many tumor types, it is crucial that both neurologists and oncologists become familiar with the diagnosis and treatment of neurological immune-related adverse events (n-irAEs). These are rare complications, developing in their severe forms in only 1-3% of the patients, but are highly relevant due to their mortality and morbidity burden. The diagnosis of n-irAEs is-however-challenging, as many alternative diagnoses need to be considered in the complex scenario of a patient with advanced cancer developing neurological problems. A tailored diagnostic approach is advisable according to the presentation, clinical history, and known specificities of n-irAEs. Several patterns characterized by distinct clinical, immunological, and prognostic characteristics are beginning to emerge. For example, myasthenia gravis is more likely to develop after anti-programmed cell death protein 1 (PD-1) or anti-programmed cell death ligand 1 (PD-L1) treatment, while meningitis appears more frequently after anti-cytotoxic T lymphocyte antigen-4 (CTLA-4) therapy. Also, peripheral neuropathy and Guillain-Barré syndrome seem to be more common in patients with an underlying melanoma. Central nervous system disorders (CNS) are less frequent and are more often associated with lung cancer, and some of them (especially those with limbic encephalitis and positive onconeural antibodies) have a poor prognosis. Herein, we provide an update of the recent advances in the diagnosis and treatment of neurological toxicities related to ICI use, focusing on the exclusion of alternative diagnoses, diagnostic specificities, and treatment of n-irAEs.


Bacterial phagocytosis activates extracellular signal-regulated kinase and p38 mitogen-activated protein kinase cascades in human neutrophils.
The hypothesis that bacterial phagocytosis by human polymorphonuclear neutrophils (PMNs) stimulates MAPK cascades that regulate respiratory burst activation was tested. Extracellular response kinase (ERK) and p38 kinase, but not c-Jun NH2-terminal kinase, activities were increased within 5 min of phagocytosis of plasma-opsonized Staphylococcus aureus (S-SA), reached maximum at 20-30 min, and remained elevated through 60 min. The role of Fcy receptors was examined using gamma globulin-opsonized SA (IgG-SA), whereas CR3 receptors were activated by particulate beta-glucan. IgG-SA stimulated a maximal ERK activity at 30 min, whereas p38 activity was maximal at 5 min. Beta-glucan stimulated maximal ERK activity at 5 min and maximal p38 activity at 2 min. Non-opsonized bacteria were ingested at 10% of the level of S-SA and stimulated a minimal increase in ERK and p38 activity at 60 min. S-SA stimulation of ERK was inhibited by wortmannin, LY294002, and genistein, but not calphostin C; whereas p38 stimulation was inhibited by calphostin C and genistein, but not wortmannin and LY294002. Simultaneous measurement of phagocytosis and H2O2 production by flow cytometry was used to assess the role of ERKs and p38 kinase in phagocytosis. The MEK inhibitor PD098059 had no significant effect on phagocytosis or H2O2 production. The p38 kinase inhibitor SB203580 significantly attenuated H2O2 production, whereas phagocytosis was unaffected. In conclusion, bacterial phagocytosis stimulates ERK and p38 activation by distinct signal transduction pathways. Phagocytosis-stimulated p38 kinase activity is necessary for optimal H2O2 production.


Primary Follicular Lymphoma of Anterior Maxilla Rectifying the Importance of Diagnostic Algorithm: A Case Report.
Primary Follicular Lymphoma of the oral cavity is one of the rarest variants of non-Hodgkin's lymphoma b-cell subtype. Now a days, increased incidence of extranodal occurrence in oral cavity and its conjoining behavior with epithelial malignancies possess the need for precise and timely diagnosis of the entity to prevent abrupt over-treatment. In this string, we report a case of primary follicular lymphoma of oral cavity which initially masqueraded oral squamous cell carcinoma but later its diagnosis as follicular lymphoma led to early treatment of the patient which led to good prognosis. The online version contains supplementary material available at 10.1007/s12070-023-03774-6.


[Impact of the Core Training Law on preventive medicine and public health training and other common medical specialties].
The purpose of the Core Training Law (CTL) is to amend specialised medical training to include 24 months of common training. The aim of this study is to assess its potential impact on the Preventive Medicine and Public Health (PM&PH) training programme and other medical specialties. The programmes of the 21 common medical specialties were analysed and the recommended training periods for each specialty collected, before the information was agreed upon by three observers. The training impact was calculated as the percentage of months that should be amended per specialty to adapt to the common training schedule. The Preventive Medicine and Public Health training programme is the specialty most affected by the Core Training Law (100%, 24 months). Intensive medicine (0%, 0 months) and medical oncology (17%, 4 months) is the least affected. The CTL affects the common medical specialties in different ways and requires a complete reorganisation of the activities and competencies of PM&PH professionals.


Development and Evaluation of Sustained Oral Ketorolac Tromethamine Particulate Matrix via Bioadhesive Chitosan Based Freeze-Dried Solid Dispersions.
This study aimed to develop and evaluate chitosan (CTS) solid dispersion particulate matrix (SDPM) for sustained oral delivery of ketorolac tromethamine (KT). SDPM formulations were prepared by freeze drying method and characterized for their effectiveness and biological activities via in vitro and in vivo assessment. Powder's flowability and bioadhesion of SDPM increased compared to KT-CTS physical mixtures and the raw materials. DSC analysis proved that the extent of drug crystallinity in matrix particles reduced as the amount of CTS content increased. FT-IR spectroscopy suggested drug-polymer interaction that was prominent in SDPM (1:7). In vitro drug release and simulated plasma profiles showed the superiority of SDPM (1:7) in sustaining drug release up to 12h. The optimized formula was stable during the storage time whereas the similarity factor (f2) for in vitro release data before and at the end of the study was 92%. Furthermore, in vivo bioactivity studies confirmed that the ulcerogenic property of SDPM (1:7) remarkably decreased compared to the standard drug while the analgesic and anti-inflammatory properties were maintained. Results suggested freeze-dried chitosan based SDPM (1:7) as a potential candidate for sustained oral administration of KT.


A patient with stage IIIB advanced breast cancer who is still alive 24 years after surgery: a case report and remarks on the treatment strategies.
In recent years, a number of agents possessing novel mechanisms, such as cyclin-dependent kinase 4/6 (CDK 4/6) inhibitors and PIK3CA inhibitors, have been developed for the treatment of hormone receptor-positive (HR+) human epidermal growth factor receptor type negative (HER2-) advanced or recurrent breast cancer. As a result, the treatment strategies for advanced or recurrent breast cancer have changed significantly. The combination of CDK 4/6 inhibitors administration and endocrine therapy is now widely used in the treatment of HR+ HER2- recurrent breast cancer with improved outcomes. In 2021, abemaciclib was approved as post-operative adjuvant combination therapy with endocrine therapy for HR+ HER2- advanced breast cancer and is expected to suppress postoperative recurrence. A range of new agents are being developed in addition to CDK4/6 inhibitors that provided more options of treatment strategies for advanced or recurrent breast cancer, which in turn could improve outcomes. However, the prognosis for the recurrent HR+ HER2- breast cancer remains poor, overall survival (OS) is still very low and a complete cure is difficult even with the treatments. In 1998, 24 years ago, neoadjuvant chemotherapy (NAC) and the concept of subtypes were not even widespread, the number of available drugs was far fewer than today, the clinical treatment guidelines had not been established. Nevertheless, we experienced a case of HR+ HER2- advanced breast cancer, stage IIIB at the initial diagnosis, which was consistently treated with the aim of complete cure and with the various treatments available at the time, resulting in long-term survival. 24 years have passed since the initial surgery, the patient has continued to do well despite repeated recurrences and remissions. We report here a case of long-term survival in advanced breast cancer of 24 years after surgery, and remark for future treatment strategies that not bound by the conventional treatment policy that emphasizes quality of life without aiming for complete cure.


Rimonabant, a selective cannabinoid1 receptor antagonist, protects against light-induced retinal degeneration in vitro and in vivo.
The endocannabinoid system is involved in some neurodegenerative diseases such as Alzheimer's disease. An endogenous constellation of proteins related to cannabinoid1 receptor signaling, including free fatty acids, diacylglycerol lipase, and N-acylethanolamine-hydrolyzing acid amidase, are localized in the murine retina. Moreover, the expression levels of endogenous agonists of cannabinoid receptors are changed in the vitreous fluid. However, the role of the endocannabinoid system in the retina, particularly in the light-induced photoreceptor degeneration, remains unknown. Therefore, we investigated involvement of the cannabinoid1 receptor in light-induced retinal degeneration using in vitro and in vivo models. To evaluate the effect of cannabinoid1 receptors in light irradiation-induced cell death, the mouse retinal cone-cell line (661W) was treated with a cannabinoid1 receptor antagonist, rimonabant. Time-dependent changes of expression and localization of retinal cannabinoid1 receptors were measured using Western blot and immunostaining. Retinal damage was induced in mice by exposure to light, followed by intravitreal injection of rimonabant. Electroretinograms and histologic analyses were performed. Rimonabant suppressed light-induced photoreceptor cell death. Cannabinoid1 receptor expression was upregulated by light exposure. Treatment with rimonabant improved both a- and b-wave amplitudes and the thickness of the outer nuclear layer. These results suggest that the cannabinoid1 receptor is involved in light-induced retinal degeneration and it may represent a therapeutic target in the light-induced photoreceptor degeneration related diseases.


Dynamic interaction of obesity, age, MCP-1 Level, and ACE-1 gene with the severity of knee osteoarthritis: a cross-sectional study.
The risk factors most strongly associated with knee osteoarthritis (OA) are old age and obesity. However, few studies have evaluated the interaction between aging and obesity in conjunction with inflammatory markers and knee OA severity as part of a complete assessment of knee OA management. Therefore, this study aims to evaluate the interaction between obesity, age, inflammation [including the I/D polymorphism of angiotensin converting enzyme-1 (ACE-1)], and the severity of knee OA. A total of 80 knee OA patients were included in this cross-sectional study. The severity of knee OA was determined based on the Kellgren-Lawrence system. All patients underwent physical and radiological examination; monocyte chemoattractant protein 1 (MCP-1) markers were measured. The parameters of the ACE-1 gene were examined with sequencing DNA. There was a significant relationship between age and severity of knee OA (P=0.007), with subjects aged greater than or equal to 65 having a 3.56-fold higher risk of developing moderate to severe OA than subjects aged less than 65. There was a significant difference between body weight and knee OA severity (P=0.026), in which subjects weighing greater than or equal to 60 kg had 3.14 times the risk of experiencing severe knee OA. Multivariate regression analysis indicated that age was the strongest independent variable for knee OA severity compared with body weight. MCP-1 levels were significantly higher in mild knee OA than in moderate to severe knee OA. The DD genotype of the ACE-1 gene increases the risk of severe knee OA by four times in subjects aged greater than or equal to 65 compared to subjects aged less than 65. However, the DD genotype of the ACE-1 gene does not increase the risk of severe knee OA in subjects weighing greater than or equal to 60 kg. While obesity and age were found to be associated with the severity of knee OA, age emerged as the independent risk factor for knee OA severity. Furthermore, MCP-1 levels were significantly higher in cases of mild knee OA compared to severe knee OA. It was observed that the DD genotype of the ACE-1 gene increases the risk of severe knee OA in individuals aged 65 years or older.


Organizational Readiness and Response During COVID-19: Reflections From a Sexual Assault Agency Serving a Predominately African American Community.
The coronavirus disease 2019 (COVID-19) pandemic forced victim service organizations to establish new service provision protocols to include remote/telehealth services. We conducted N = 12 qualitative interviews with sexual assault advocates working in an urban agency in a predominately African American U.S. city to understand how they adapted services to meet the needs of their community. A thematic analysis revealed this organization was under-prepared for prolonged interruption of in-person services. Even though this agency was able to create telehealth options, many clients did not have the financial and technological resources to utilize these services. Advocates reported that survivors expressed a strong preference for in-person services, which afford more privacy and confidentiality. The pervasive digital divide within this urban community limited survivors' access to comprehensive services and jeopardized their safety.


How tech-savvy employees make the difference in core facilities: Recognizing core facility expertise with dedicated career tracks: Recognizing core facility expertise with dedicated career tracks.
Core facilities have a different mission than academic research labs. Accordingly, they require different career paths and structures.


Synaptic vesicle 2C and its synaptic-related function.
Synaptic vesicle 2 C (SV2C) is a novel isoform belonging to the synaptic vesicle 2 (SV2) protein superfamily; a family of proteins known to have roles in vesicle trafficking, exocytosis and neurotransmission. In humans, SV2C is expressed in evolutionarily older brain regions, and is a known receptor for botulinum neurotoxin/A (BoNT/A), controlling glucose-evoked granule recruitment and regulating dopamine release, thus serving as a potential target molecule in the treatment of psychosis. In addition, recent researches have shown that SV2C regulates hypertension and accelerates venous thromboembolism (VTE) and coagulation pathways and may play roles in several non-nervous system diseases. In terms of regulation, SV2C is positively regulated by both alendronate and statins. As SV2C may provide a potential novel therapeutic target for psychosis and other diseases, this article reviews the progress made thus far in understanding the structure, distribution, function and regulation of SV2C.


Longitudinal associations among maternal depressive symptoms, child emotional caretaking, and anxious/depressed symptoms in pediatric cancer.
Research has shown that children experience increased emotional distress when engaging in emotional caretaking of a parent. The current study is the first to examine this process in families in which the source of the stress is the child's illness. Prospective associations were tested among mothers' depressive symptoms near the time of their child's cancer diagnosis, mothers' expressed distress and their child's emotional caretaking during an interaction task, and child anxious/depressed symptoms at 1 year postdiagnosis. Families (N = 78) were recruited from two pediatric hospitals soon after their child's (Ages 5-18) new diagnosis or relapse of cancer. Mothers reported on their own depressive symptoms and their child's anxious/depressed symptoms near the time of diagnosis or recurrence (Time 1) and 1 year later (Time 3). At Time 2 (4 months after Time 1), mother-child dyads completed a video-recorded discussion of their experience with cancer that was coded for observed maternal expressed distress (anxiety, sadness) and observed child emotional caretaking. Maternal expressed distress during the interaction was significantly related to more emotional caretaking behaviors by both boys and girls. Results of a moderated mediation model showed that child emotional caretaking at Time 2 significantly mediated the relation between maternal depressive symptoms at Time 1 and child anxious/depressed symptoms at Time 3 for girls but not for boys. The findings suggest that children's emotional caretaking behaviors contribute to subsequent anxious/depressed symptoms for girls, but not for boys, with cancer. (PsycINFO Database Record (c) 2018 APA, all rights reserved).


Anterolateral thigh adipofascial flap combined with skin paddle in three-dimension scalp reconstruction: A case report.
Management of cranial and scalp defects is always challenging. Complication rates following cranioplasty are still high (10-40 %), including bone graft resorption, infection, and T-mesh implant exposure due to thinning of soft tissue (Yeap et al., 2019; Alkhaibary et al., 2020; Cho and Kang, 2017 [1-3]). We present a case of a 38-year-old male with autologous bone graft resorption accompanied by thinning of the forehead skin. He underwent cranioplasty with titanium mesh (T-mesh) and scalp reconstruction with an anterolateral thigh (ALT) flap in a single-stage operation. A 2-component ALT flap was used: the skin paddle covered the scalp defect to reduce closure tension, and especially the adipofascial flap lining beneath the forehead skin to increase thickness. Postoperative results were great in terms of coverage function and aesthetics. The phenomenon of soft tissue thinning causing titanium mesh exposure is a common complication with a rate of 10-14 % (Yeap et al., 2019; Dong et al., 2020; Maqbool et al., 2018 [1, 4, 5]). Using adipofascial ALT flap as a sub-scalp filler material has many advantages over the latissimus dorsi (LD) flap to increase scalp thickness but still ensure aesthetics. The results suggest a new direction in using adipofascial ALT flap as filler material to increase scalp thickness to reduce T-mesh exposure following cranioplasty.


Bioactivity of the Protein Hydrolysates Obtained from the Most Abundant Crustacean Bycatch.
The animals from bycatch of the shrimp fisheries can be a source of natural products and bioactive compounds. Thus, the present study aimed to evaluate the bioactivity of protein hydrolysates prepared from the two most abundant crabs from the bycatch of shrimp fisheries in Brazil (Callinectes ornatus and Hepatus pudibundus). Samples of C. ornatus and H. pudibundus were collected in the region of Ubatuba, State of São Paulo, Brazil. Muscles with small pieces of exoskeleton of both species were hydrolyzed using two enzymes, Alcalase 2.4 L® or Protamex®. The in vitro antioxidant capacity was analyzed used three methods: DPPH, sulfhydryl groups, and peroxyl radicals. Additionally, the cytotoxicity of the hydrolysates was investigated using pre-osteoblasts cells. The results showed that the degree of hydrolysis (DH) of H. pudibundus was superior to DH of C. ornatus using both enzymes and was higher when using the enzyme Alcalase 2.4 L® (32.0% ± 1.9). The analysis suggested that the hydrolysates have antioxidant activity. Besides that, no cytotoxic effect was observed on cell viability. Thus, protein hydrolysates of C. ornatus and H. pudibundus have bioactivity, which add value to these bycatch species and suggests their potential use as nutraceutical ingredient in the food industry.


Alfentanil for urgent caesarean section in a patient with severe mitral stenosis and pulmonary hypertension.
We present the case of a parturient with severe mitral stenosis and pulmonary hypertension who received general anaesthesia using alfentanil for urgent Caesarean section. Alfentanil promoted haemodynamic stability and allowed immediate postoperative extubation. Epidural morphine provided postoperative analgesia. This combination permitted early ambulation and prevention of thromboembolism. A disadvantage of this technique, neonatal respiratory depression, was promptly reversed with a single dose of naloxone. The anaesthetic management of mitral stenosis in pregnancy is discussed and the neonatal pharmacokinetics of maternally administered alfentanil are presented.


Slowed Compensation Responses to Altered Auditory Feedback in Post-Stroke Aphasia: Implications for Speech Sensorimotor Integration.
Developing a clearer understanding of impairments that underlie the behavioral characteristics of aphasia is essential for the development of targeted treatments and will help inform theories of speech motor control. Impairments in sensorimotor integration of speech in individuals with conduction aphasia have previously been implicated in their repetition deficits. However, much less is known about the extent to which these integrative deficits occur outside of conduction aphasia and how this manifests behaviorally in areas other than speech repetition. In this study, we aimed to address these issues by examining the behavioral correlates of speech sensorimotor impairment under altered auditory feedback (AAF) and their relationship with the impaired ability to independently correct for online errors during picture naming in people with aphasia. We found that people with aphasia generate slower vocal compensation response to pitch-shift AAF stimuli compared with controls. However, when the timing of responses was controlled for, no significant difference in the magnitude of vocal pitch compensation was observed between aphasia and control groups. Moreover, no relationship was found between self-correction of naming errors and the timing and magnitude of vocal compensation responses to AAF. These findings suggest that slowed compensation is a potential behavioral marker of impaired sensorimotor integration in aphasia.


Spontaneous noncardiac chest pain. Evaluation by 24-hour ambulatory esophageal motility and pH monitoring.
Noncardiac chest pain can be a diagnostic dilemma because patients rarely experience spontaneous chest pain in the laboratory. Therefore, we studied 24 patients with chronic, daily, substernal chest pain with a prototype 24-h ambulatory esophageal motility and pH system. Spontaneous chest pain episodes were correlated with pH less than 4 and abnormal motility changes (mean amplitude and duration, maximum amplitude and duration, or percentage of abnormal peristalsis) defined as exceeding the patient's normal esophageal motility pattern. Twenty-two patients experienced a total of 92 spontaneous chest pain episodes. Eleven chest pain episodes (12%) occurred during abnormal motility, whereas 18 episodes (20%) were associated with pH less than 4 and four episodes (4%) had both abnormalities. The majority of chest pain episodes, 59 events (64%), did not have any association with motility or pH. Abnormal maximum duration and amplitude were the motility changes most frequently associated with chest pain. Overall, 13 of 22 patients (59%) had at least one chest pain episode correlating with abnormal motility or pH (range 33%-100%). Therefore, we conclude that ambulatory esophageal motility and pH monitoring is useful in the evaluation of noncardiac chest pain. pH abnormalities (20%) are more commonly associated with chest pain than motility abnormalities (12%). However, the majority of chest pain episodes (64%) did not correlate with either abnormality and may be the result of lowered esophageal pain threshold for distention, i.e., the "irritable esophagus."


Matrix and serine protease expression during leukemic cell differentiation induced by aclacinomycin and all-trans-retinoic acid.
In myeloid leukemia, immature leukemic cells are able to egress into peripheral blood to infiltrate extra-medullary organs. We therefore analyzed the migrating and invasive potential of human HL-60 and NB4 cell lines, representative of acute myelogenous leukemia, their ability to express matrix metalloproteases (MMPs), tissue inhibitors of metalloproteases (TIMPs) and urokinase plasminogen activator (uPA) in response to differentiating agents. Granulocytic differentiation by all-trans-retinoic acid (ATRA) and aclacinomycin (ACLA) strongly increased HL-60 and NB4 cell migration and invasion. At mRNA and protein levels, these cell lines produced significant amounts of MMP-9 (HL-60<NB4). Granulocytic differentiation by ACLA increased both pro and active forms of MMP-9 whereas ATRA decreased them and stimulated uPA mRNAs. TIMP-1, the physiological MMP inhibitor, increased during granulocytic differentiation whereas TIMP-2 did not significantly vary. Use of Batimastat and aprotinin suggests that ATRA was active by modulating the uPA system while ACLA interfered with MMP expression. In conclusion, our data demonstrate that HL-60 and NB4 cells express MMPs and uPA which are differentially regulated by the differentiating agents ATRA and ACLA and suggest the clinical usefulness of MMPs and serine protease inhibitors in the prophylaxis and treatment of the ATRA syndrome.


Cell cycle status in human corneal endothelium.
Corneal endothelium is the single-cell layer that forms a physical barrier between the corneal stroma and aqueous humour. The barrier and ionic 'pump' functions of corneal endothelium help regulate stromal hydration. Loss of endothelial cells due to increasing age, trauma, disease, dystrophy, or previous corneal transplants can reduce the density of endothelial cells to a critical point below which the stroma becomes edematous and visual acuity is lost. Throughout life, division of endothelial cells either does not occur or occurs at a rate too slow to adequately replace dead cells. Thus, the major means of repairing the monolayer is by cell migration and/or enlargement. The basis for the lack of endothelial cell proliferation is not yet fully understood, although it is clear that cells do retain proliferative capacity. Previous studies from this laboratory have identified certain environmental conditions that may be responsible for maintaining these cells in a non-replicative state in vivo. In addition, corneal endothelial cells exhibit intrinsic, age-related differences in relative proliferative capacity. The studies described below provide evidence to support the hypothesis that, with age, an increasing number of HCEC enter a replicative senescence-like state in which they become increasingly refractive to mitogenic stimulation. This decreasing sensitivity to mitogens appears to be mediated, at least in part, by age-dependent alterations in the relative expression and activity of the cyclin-dependent kinase inhibitors, p27KIP1, p16INK4A, and p21CIP1.


Spatiotemporal Changes in the Bacterial Community of the Meromictic Lake Uchum, Siberia.
Lake Uchum is a newly defined meromictic lake in Siberia with clear seasonal changes in its mixolimnion. This study characterized the temporal dynamics and vertical profile of bacterial communities in oxic and anoxic zones of the lake across all four seasons: October (autumn), March (winter), May (spring), and August (summer). Bacterial richness and diversity in the anoxic zone varied widely between time points. Proteobacteria was the dominant bacterial phylum throughout the oxic and anoxic zones across all four seasons. Alphaproteobacteria (Loktanella) and Gammaproteobacteria (Aliidiomarina) exhibited the highest abundance in the oxic and anoxic zone, respectively. Furthermore, there was a successional shift in sulfate-reducing bacteria (SRB) and sulfur-oxidizing bacteria in the anoxic zone across the seasons. The most dominant SRB, Desulfonatronovibrio sp., is likely one of the main producers of hydrogen sulfide (H2S) and typically accumulates the most H2S in winter. The representative anoxygenic phototrophic bacterial group in Lake Uchum was purple sulfur bacteria (PSB). PSB were dominant (60.76%) in summer, but only had 0.2-1.5% relative abundance from autumn to spring. Multivariate analysis revealed that the abundance of these SRB and PSB correlated to the concentration of H2S in Lake Uchum. Taken together, this study provides insights into the relationships between changes in bacterial community and environmental features in Lake Uchum.


Transfection and promoter analysis in embryonic stem cells.
Embryonic stem (ES) cells are useful for the molecular analysis of developmental pathways when combined with efficient genetic manipulation and in vitro differentiation procedures. To facilitate the genetic modification of ES cells, we describe how to analyze the transcriptional activities of different promoter systems. The DNA segments from a number of promoters are subcloned into the multiple cloning sites of the green fluorescent protein (GFP) reporter plasmid. The resulting recombinant plasmids are used to transfect ES cells by lipofection. Using luciferase-normalized GFP assays, we observed differing promoter activities in transient transfections of ES cells. The judicious selection of the optimum promoter system for specific genetic modification of ES cells will ensure the successful generation of transgenic ES cell lines.


In vivo analysis of recycling endosomes in Caenorhabditis elegans.
The microscopic nematode Caenorhabditis elegans (C. elegans) serves as an excellent animal model for studying membrane traffic. This is due in part to its highly advanced genetics and genomics, and a transparent body that allows the visualization of fluorescently tagged molecules in the physiologically relevant context of the intact organism. Notably, C. elegans oocytes, coelomocytes, and intestinal epithelia have been established as facile cellular models to explore nonpolarized and polarized cell membrane trafficking mechanisms. In this chapter, we describe in vivo C. elegans assays, utilizing fluorescent dyes or proteins, to examine the molecular mechanisms that control endocytosis and endocytic recycling. Tissue-specific, steady-state imaging and associated quantitative analysis allow the identification and interpretation of subcellular events in the intact animal. To better understand the kinetic characteristics of recycling tubules that mediate membrane protein recycling, we describe recently developed dynamic-imaging assays in intestinal epithelial cells. Such methods bring new clarity to the system, helping to elucidate the functional roles of recycling mediators.


Attitudes towards human fetal tissue research: Survey of researchers and the public in Japan.
The rules for human fetal tissue (HFT) research in Japan are unclear. In this paper, we conducted a web survey to examine the attitudes of Japanese researchers (n=535) and the public (n=3,000) toward HFT research. The results demonstrated that 5.8% of researchers and 18.8% of the public explicitly opposed HFT research, and 71.8% of the researchers thought that the rules for HFT research need to be clarified. Even among researchers who intended to consider conducting HFT research, 74.2% responded that the rules should be clarified. Although different from attitudes to make decisions regarding HFT donation, being non-religious and in their reproductive age among women in the public group were factors for accepting attitudes toward HFT research. To establish the rules, it is necessary to develop a system that can adequately protect vulnerable women who are asked to provide HFT.


Doxepin protects cultured neurons against oxidative stress-induced injury.
To investigate protective effects of doxepin (Dox) on cultured neuronal injury induced by oxidative stress. Cytotoxicities of glutamate (Glu 0.5 mmol/L, 15 min), sodium dithionite (0.5 mmol/L, 24 h) or hemoglobin (Hb100 mg/L,24 h)and the protective effects of Dox were observed. Exposure of cultured neurons to Glu, sodium dithionite and Hb developed a neurotoxicity expressed in the thiazol blue tetrazolium bromide (MTT) assay, the increase of lactate dehydrogenase (LDH) leakage, malondialdehyde (MDA) content and intracellular [Ca2+]i accumulation, as well as the decrease of superoxide dismutase (SOD) activity. DOX 1-100 nmol/L significantly inhibited all above changes. Dox protects cultured neurons against oxidative stress-induced injury by suppressing intracellular [Ca2+]i accumulation, decreasing lipid peroxide generation and stimulating antioxidant enzyme.


[Factors influencing the administration of sedatives to infants and their significance].
An epidemiological survey on 1,100 young mothers has enabled the authors to carry out a study on how frequent it is for mothers to give sedatives to their infants under one year of age and on the way it can be linked with demographical, sociological and psychological factors. The longitudinal study---until they are 3---of a small group of babies has shown that practitionners asked to prescribe sedatives or sleeping pills for babies should consider it as a risk indicator.


HIV acquisition during pregnancy and postpartum is associated with genital infections and partnership characteristics.
The objective of this study is to determine the risk and cofactors for HIV acquisition during pregnancy and postpartum. A prospective cohort study : Pregnant women in western Kenya were enrolled if HIV seronegative at that visit or within 3 months. Serial HIV nucleic acid amplification tests (NAATs) were conducted at 1 to 3-month intervals to 9 months postpartum. Genital swabs were collected for detection of chlamydia and gonorrhoea at baseline, and for trichomonas, bacterial vaginosis and yeast at baseline and follow-up. Among 1304 pregnant women, median age was 22 years, 78% were married for a median of 4 years, 66% reported knowing partner HIV status and 8% reported using condoms. Study retention was 98%. During 1235 person-years of follow-up, HIV incidence was 2.31/100 person-years [95% confidence interval (95% CI) 0.71-4.10]. Incident HIV was associated with syphilis (hazard ratio 9.18, 95% CI 2.15-39.3), chlamydia (hazard ratio 4.49, 95% CI 1.34-15.0), bacterial vaginosis (hazard ratio 2.91, 95% CI 1.25-6.76), yeast (hazard ratio 3.46, 95% CI 1.46-8.19), sexually transmitted infection (STI) history (hazard ratio 3.48, 95% CI 1.31-9.27), lifetime number of sex partners (hazard ratio 1.19, 95% CI 1.03-1.37), partner age discordance (hazard ratio 1.07 per year, 95% CI 1.02-1.13) and shorter marriage (hazard ratio 1.19 per year, 95% CI 1.03-1.38). No women with incident HIV reported an HIV-infected partner. In multivariate analyses, chlamydia, older partners and yeast infection remained significant; however, power was limited. Pregnant and lactating women may not perceive HIV risk and rarely used condoms. Prevention and treatment of genital infections and risk stratification to identify women for pre-exposure prophylaxis (PrEP) could decrease HIV acquisition in pregnant/lactating women.


[Cutaneous microdialysis. Use in dermatology].
The determination of concentrations of exogenous or endogenous substances in the dermis, e.g. for pharmacokinetic studies, is technically difficult. Performing skin biopsies or inducing suction blisters results in the disintegration of the tissue and allows only single measurements. Recently, cutaneous microdialysis, a new simple and minimally invasive technique for continuously measuring of substances in the dermis in vivo, has been introduced in dermatological research. According to the principle of dialysis, a semipermeable membrane is inserted in the dermis and due to a concentration gradient between the interstitial space and a perfusate, substances diffuse through the pores of the membrane and can be analyzed in the dialysate. Cutaneous microdialysis represents a useful technique for pharmacokinetic or pharmacodynamic studies as well as for investigations regarding cutaneous physiology and pathophysiology.


[Comparison of two amine-modified chemical platforms for DNA microarray preparation].
To study two amine modification procedures for DNA microarray preparation based on polymeric coatings. One of the proposed approaches utilized poly-amine coating of silanized slides activated by 1,4-phenylene diisothiocyanate, and the other employed acrylic acid-co-acrylamide copolymer and 1-(3-dimethylamino propyl)-3-ethylcarbodiimide hydrochloride/N-hydroxysuccinimide as the coating agents and activator, respectively. The modified slides were used for preparing lambda phage DNA microarrays, whose properties were analyzed by hybridization. Formation of dendrimeric structure and polymer was observed on the surface of the slides. The signal spots in uniform, steady and regular shape, in comparison with the commercial CMT-GAPS slides, indicated successful manufacture of the microarrays. The two platforms are suitable for microarray preparation, and the method of acrylic acid-co-acrylamide copolymer modification is more preferred.


Correlation of intra-tumour heterogeneity on 18F-FDG PET with pathologic features in non-small cell lung cancer: a feasibility study.
We evaluated the feasibility to correlate intra-tumour heterogeneity as visualized on 18F-FDG PET with histology for NSCLC. For this purpose we used an ex-vivo model. The procedure was feasible in all operated patients. We have shown that this method is suitable for correlating intra-tumour heterogeneity in tracer uptake with histology.


[ICAM-1 and immunological cell adhesion in the skin].
The adhesion molecule ICAM-1 mediates cell contact among leukocytes and between leukocytes and various non-lymphoid cells. A substantial body of experimental evidence indicates a crucial role of ICAM-1 in the biology of cutaneous inflammation. The current status of knowledge on ICAM-1 is reviewed and discussed with regard to the pathophysiology of inflammatory and neoplastic disorders of the skin.


Trimetazidine as a potential neuroprotectant in transient global ischemia in gerbils: a behavioral and histological study.
The effect of Trimetazidine (TMZ) as a potential neuroprotectant against stroke was studied in the gerbil model of transient forebrain global ischemia. Animals were subjected to a 5-min period of ischemia and assessed 4 and 21 days later. Gerbils were divided into two groups: in group one, gerbils were treated with TMZ at a dose of 25 mg/kg given by intraperitoneal injection prior to ischemia. In group two, gerbils were treated with TMZ at a dose of 25 mg/kg given intraperitoneally after ischemia. Saline-injected gerbils served as controls. Histological evaluation of neuronal damage was carried out using the silver staining technique in gerbils 4 and 21 days after the start of the experimental protocol. Behavioral functions were assessed in gerbils from the 14th to the 21st day after the start of the experimental protocol using the Morris water maze test. Results obtained from this study showed no significant difference between saline treated TMZ-treated gerbils when TMZ was administered after ischemia. When TMZ was administered prior to ischemia, there was a reduction in neuronal damage although it did not reach statistical significance and a statistically significant improvement in behavior. We conclude that TMZ shows signs of promise as a neuroprotective agent, and further studies should look at pre-treatment with different doses and different times.


Evaluation of lymph node metastases: comparison of gadofluorine M-enhanced MRI and diffusion-weighted MRI in a rabbit VX2 rectal cancer model.
To compare the diagnostic performance of a diffusion-weighted imaging (DWI) dataset and a gadofluorine M-enhanced imaging dataset for identifying lymph node (LN) metastases in a rabbit rectal cancer model. VX2 carcinomas were injected into the rectum of 26 rabbits. Four weeks later, T2-weighted imaging (T2WI), pre-T1WI, DWI, and post-T1WI were performed. Two radiologists independently reviewed the DWI set (T2WI, pre-T1WI, DWI) and the gadofluorine M set (T2WI, pre- and post-T1WI) and recorded their confidence scores for LN metastasis on a per-LN basis. Receiver operating characteristic (ROC) analysis was performed to compare the area under the ROC curve (A(z) ) of the two imaging sets. Histopathologic results were used as the reference standard. The A(z) and sensitivity of the gadofluorine M set were comparable to those of the DWI set (A(z) , for reader 1, 0.849, 0.829, P = 0.571; for reader 2, 0.923, 0.876, P = 0.212; sensitivity, for reader 1, 97%, 97%; for reader 2, 97%, 92%, P = 0.304). The specificity of the former was greater than that of the latter (for reader 1, 65%, 53%, P = 0.0003; for reader 2, 81%, 68%, P = 0.01). Gadofluorine M-enhanced images provided greater specificity than DWI for identifying LN metastases, whereas the A(z) and sensitivity of the former were comparable to those of the latter.


Effect of clenbuterol, a new beta 2 selective adrenoceptor stimulant, on the release of histamine and SRS-A from passively sensitized guinea-pig chopped lungs.
Effect of clenbuterol on the release of histamine and SRS-A from passively sensitized guinea-pig lungs was studied in a superfused-cascade bioassay system, and compared with isoproterenol and salbutamol. One-week-old guinea-pig lungs were used, since histamine and SRS-A were released more in these than in 6- or 12-week old lungs during anaphylaxis. Clenbuterol, isoproterenol and salbutamol (10(-8) - 10(-7) M) were found to inhibit the releases of histamine and SRS-A. These actions were inhibited by propranolol. These results suggest that clenbuterol inhibits the releases of histamine and SRS-A via the beta-adrenoceptor.


PDI augments kainic acid-induced seizure activity and neuronal death by inhibiting PP2A-GluA2-PICK1-mediated AMPA receptor internalization in the mouse hippocampus.
Protein disulfide isomerase (PDI) is a redox-active enzyme and also serves as a nitric oxide donor causing S-nitrosylation of cysteine residues in various proteins. Although PDI knockdown reduces α-amino-3-hydroxy-5-methylisoxazole-4-propionic acid receptor (AMPAR)-mediated neuronal activity, the underlying mechanisms are largely unknown. In the present study, we found that under physiological condition PDI knockdown increased CaMKII activity (phosphorylation) in the mouse hippocampus. However, PDI siRNA inhibited protein phosphatase (PP) 2A-mediated GluA2 S880 dephosphorylation by increasing PP2A oxidation, independent of S-nitrosylation. PDI siRNA also enhanced glutamate ionotropic receptor AMPA type subunit 1 (GluA1) S831 and GluA2 S880, but not GluA1 S845 and GluA2 Y869/Y873/Y876 phosphorylations, concomitant with the enhanced protein interacting with C kinase 1 (PICK1)-mediated AMPAR internalization. Furthermore, PDI knockdown attenuated seizure activity and neuronal damage in response to kainic acid (a non-desensitizing agonist of AMPAR). Therefore, these findings suggest that PDI may regulate surface AMPAR expression through PP2A-GluA2-PICK1 signaling pathway, and that PDI may be one of the therapeutic targets for epilepsy via AMPAR internalization without altering basal neurotransmission.


Renal, cardiovascular and endocrine responses of fetal sheep at 0.8 of gestation to an infusion of amino acids.
Amino acid infusions increase renal blood flow (RBF) and glomerular filtration rate (GFR) and stimulate tubular reabsorption in adults. To characterize the effects of amino acids on fetal renal haemodynamics, tubular sodium reabsorption, acid-base homeostasis and plasma renin levels, 11 chronically catheterized fetal sheep aged 121 +/- 1 days (term ~150 days) were infused I.V. for 4 h with alanine, glycine, proline and serine (0.1, 0.1, 0.06 and 0.06 mmol min(-1), respectively) in 0.15 M saline at 0.165 ml min(-1). Eight control fetuses were given saline. During amino acid infusion, plasma amino acid levels increased up to 20-fold (P < 0.005). GFR increased by 50 +/- 8 % (P < 0.001); there was only a small transient increase in RBF. Proximal fractional sodium reabsorption fell from 74.6 +/- 2.9 to 55.5 +/- 5.4 % (P < 0.005). Distal sodium delivery increased, but a smaller percentage of this distal sodium load was reabsorbed (P < 0.005). Thus fractional sodium reabsorption fell from 95.5 +/- 0.9 to 81.4 +/- 2.0 % (P < 0.005). There was a large diuresis, natriuresis, kaliuresis and increase in osmolar excretion (P < 0.005). Plasma sodium and chloride concentrations fell (P < 0.005). Plasma osmolality did not change. Plasma renin levels fell (P < 0.05), cortisol levels increased (P < 0.05), and there was a compensated metabolic acidosis. Thus the fetal sheep kidney demonstrated a remarkable functional capacity to respond to amino acid infusion. The increase in filtration fraction and the lack of an increase in RBF suggest that efferent arteriolar vasoconstriction occurred, a very different response from the renal vasodilatation seen in adult animals.


Efficacy of seven anthelmintics against Ancylostoma ceylanicum in the golden hamster, Mesocricetus auratus.
In an experimental patent infection of Ancylostoma ceylanicum in golden hamsters the efficacy of seven anthelmintics was tested; bephenium hydroxynaphthoate, tetramisole, tetrachlorethylene, phenylene 1,4-diisothiocyanate, thiabendazole, parbendazole and mebendazole were studied. Mebendazole was the most effective, with parbendazole, phenylene 1,4-diisothiocyanate, thiabendazole and bephenium hydroxynaphthoate also satisfactory. Tetrachlorethylene and tetramisole were the least effective in our screen.


Fermentation of milk permeate by proteolytic bacteria for protease production.
Kinetics of cell growth and protease production by four proteolytic bacterial strains, namely, Bacillus subtilis EMCC 1020, Bacillus megaterium EMCC 1057, Serratia marcescens EMCC 1247, and Pseudomonas fluorescens EMCC 1221, in milk permeate, compared with other fermentation media, were studied. The pH values, lactose utilization, and biochemical oxygen demand (BOD) reduction in milk permeate also were investigated. The four strains were able to grow in milk permeate, and to produce considerable amounts of protease, reaching 289 (EMCC 1020), 252 (EMCC 1057), 263 (EMCC 1247), and 212 (EMCC 1221) U/mL after 30 h of fermentation. The growth and enzyme activity of the four strains were greater in milk permeate than those in other fermentation media. The protease-producing bacteria were able to utilize lactose in milk permeate with values ranging from 37.62 to 54.97% and to reduce the BOD of milk permeate by 50.59-63.65%. Milk permeate proved to be the best medium for enzyme production by all organisms examined.


A rare combination of pheochromocytoma and gastrointestinal stromal tumour in a patient with neurofibromatosis 1 syndrome-a case report.
Neurofibromatosis 1 is a rare inherited autosomal dominant syndrome. It comprises 90 % of neurofibromatosis cases. These patients may develop various types of tumours in early age, especially multiple neurofibromas with a high risk of developing malignant peripheral nerve sheath tumours. Other tumours can also develop like pheochromocytoma, optic nerve and brain stem gliomas, carcinoids and rarely gastrointestinal stromal tumours. A combination of pheochromocytoma and gastrointestinal stromal tumour is very rare. Only a few cases have been reported. Here, we are reporting a case of NF1 syndrome with a combination of pheochromocytoma and gastrointestinal stromal tumour with additional findings of multiple clear cell nodules and brown fat in the periadrenal connective tissue.


Carotid stent fracture and restenosis management.
We report an unusual case of asymptomatic accelerated right carotid artery in-stent restenosis in a patient referred for revascularization of a de novo stenosis of her left internal carotid artery.


Reversed-phase liquid chromatographic separation of 3',5'-cyclic ribonucleotides.
A rapid, reversed-phase "high-performance" liquid-chromatographic separation of the five naturally occurring cyclic ribonucleotides is described. The separation, optimized for the measurement of these compounds in biological samples, is short (25 min), sensitive (50-100 pmol), and requires no sample pre-concentration steps. We also report an alternative isocratic elution mode, optimized for a rapid and selective analysis for adenosine 3',5'-cyclic phosphate. The identity of chromatographic peaks in biological extracts is confirmed by several methods: retention times, co-chromatography with the reference compounds, absorbance ratios, enzymatic peak-shift with cyclic nucleotide phosphodiesterase, and stopped-flow ultraviolet-scanning techniques.


Comparison of the metabolic changes in rats with hypertension secondary to fructose feeding or renal artery stenosis.
Hypertension was induced in rats by either renal artery stenosis or a fructose-enriched diet, and the consequent changes in plasma glucose, insulin, and triglyceride (TG) concentrations, and the steady-state plasma insulin (SSPI) and glucose (SSPG) concentrations in response to a 180-min continuous infusion of glucose and insulin in these two groups of hypertensive rats, were compared to values in a sham-operated group with normal blood pressure. Mean (+/- SEM) blood pressure was significantly higher than the control values (121 +/- 3 mm Hg) at the end of the study in rats with renal artery stenosis (178 +/- 13 mm Hg) and fructose-fed rats (151 +/- 5 mm Hg), whereas left ventricular weight was only significantly (P < .01) higher in rats with renal artery stenosis. Plasma glucose concentration was the same in all three groups, but fructose-fed rats had significantly higher plasma insulin (59 +/- 7 microU/mL) and TG (317 +/- 48 mg/dL) concentration than either sham-operated rats (30 +/- 4 microU/mL and 121 mg/dL) or rats with renal artery stenosis (34 +/- 5 microU/mL and 124 +/- 14 mg/dL). Although SSPI concentrations were similar (approximately 250 microU/mL) in all three groups of rats, SSPG concentrations were significantly higher (P < .01) in the fructose-fed rats (187 +/- 10 mg/dL) than in either sham-operated normotensive rats (120 +/- 6 mg/dL) or hypertensive rats with renal artery stenosis (133 +/- 4 mg/dL). Thus, insulin resistance, hyperinsulinemia, and hypertriglyceridemia developed in rats with fructose-induced hypertension, whereas none of these changes were seen in rats with renal artery stenosis.


An energetics model of an aquatic predator and its application to life-history optima.
A bioenergetics simulation model of the growth and life history of the aquatic predator Nephelopsis obscura Verrill was developed and validated using both experimentation and sensitivity analysis. Sensitivity analysis demonstrated that the model's internal feedbacks resulted in stability similar to homeostatic biological mechanisms. The experimental validation showed the model very accurately predicts growth at 10°C and 15°C but is slightly biased at 20°C. Simulation output was also consistent with the observed data on Nephelopsis from the site from which the simulation input data were obtained and indicated that Nephelopsis growth is more sensitive to prey variation among years than to temperature variation. Although built using data from a population at one extreme of the spectrum observed in life history and growth, the model was able to emulate the growth of Nephelopsis throughout its range. Thus, the variability in size and life history observed in the field can be explained as the result of a plastic phenotype responding to different habitat conditions.


Fatal pulmonary failure attributable to viral pneumonia with human herpes virus 6 (HHV6) in a young immunocompetent woman.
Adult respiratory distress syndrome (ARDS) attributable to viral pneumonia is described mainly in immunodeficient persons. ARDS caused by human herpes virus 6 (HHV6) is extremely rare, and to our knowledge only 1 case has been reported in the literature. We present the case of a young woman who developed fatal pulmonary failure most probably attributable to HHV6 pneumonia.


Acidification in the rat proximal jejunum.
1. Production of hydrogen ions by the rat proximal jejunum was investigated using the everted sac technique. 2. Acidification occurred in the absence of glucose, increasing on addition of glucose to reach a maximal value. An apparent Km of 1-78 mM was derived for the glucose-dependent process. 3. Acidification in the presence of glucose was inhibited by 10 mM-2:4-dinitrophenol, 10 mM phlorrhizin, 10 mM aminophylline and anaerobiosis. 4. Histamine, ethylenediamine tetraacetic acid (EDTA), ouabain and acetazolamide, compounds known to alter acid production in gastric mucosa had no effect on jejunal acidification. 5. Galactose and 3-O-methylglucose failed to increase acidification; in contrast, mannose and fructose did lead to increases, indicating metabolic origin of the hydrogen ions. 6. Serosal and mucosal lactate production were measured and the calculated percentage of hydrogen ions possibly derived from this source was shown to account for only a small proportion of acidification. 7. The greatest increase in acidification with minimal simultaneous production of lactate occurred with ATP which was shown not to enter intestinal tissues. 8. A hypothesis for acidification, that of the break-down at the mucosal surface of ATP from intracellular metabolic sources, is proposed and its relevance to the postulated microclimate is discussed.


Rat embryonic hippocampal neurons express a new class A calcium channel variant.
The aim of this study was to investigate, using a RT-PCR strategy, rat voltage-gated class A calcium channel (alpha1A) splice variants during rat hippocampus development. Results demonstrate the presence of multiple alpha1A mRNAs with the hippocampus formation and revealed a new variant of the rat alpha1A subunit (alpha1A-EFe) that diverges from alpha1A-a in the EF-hand domain. alpha1A-EFe expression in hippocampal neurons is restricted to the embryonic period. This in vivo developmental program is recapitulated in dissociated cultures of E17 embryonic hippocampal neurons. These data demonstrate that rat hippocampus neurons express a unique alpha1A splice variant during the embryonic period and suggest that alternative RNA splicing may modulate neuronal calcium channel properties during development.


The 3'-UTR C>T polymorphism of the oxidized LDL-receptor 1 (OLR1) gene does not associate with coronary artery disease in Italian CAD patients or with the severity of coronary disease.
Oxidized low-density lipoproteins (OxLDLs) play a critical role in endothelial dysfunction, which is implicated in the pathogenesis of atherosclerosis. Vascular endothelial cells internalize and degrade oxLDL through the endothelial lectin-like oxidized LDL receptor 1 (OLR1). OLR1 is up-regulated in several pathological conditions, including hypertension, hyperlipidemia, diabetes, atherosclerosis and inflammation, and represents therefore a good candidate for coronary artery disease (CAD). Recently, a 3'-UTR (188 C>T) SNP in the OLR1 gene has been reported to be associated with coronary artery stenosis and myocardial infarction. In the present study we investigated whether the OLR1 gene 188 C>T SNP is a genetic risk marker for CAD in Italian patients with angiographically defined coronary atherosclerosis, and assessed its relation with clinical and metabolic abnormalities, including severity of disease (classified as restenosis, single- or multiple coronary vessels disease, and MI). The 3'-UTR C>T SNP was detected in real-time PCR in 351 subjects with CAD and in 215 control subjects. The OLR1-T allele frequencies were 48.9% in the CAD subjects and 47.7% in controls, with no significant difference between the two groups. Also, the 3'-UTR C>T SNP did not associate with any of the parameters of severity of disease. Furthermore, none of the other clinical and metabolic parameters were associated with the OLR1 gene SNP. Our observations suggest that, in our population, the 3'-UTR C>T polymorphism of the OLR1 gene is unlikely to play a role in the pathogenesis of coronary artery disease.


Lesion of the ponto-geniculo-occipital pathways in kittens. I. Effects on sleep and on unitary discharge of the lateral geniculate nucleus.
Suppression of geniculate ponto-geniculo-occipital (PGO) waves was achieved in kittens by performing bilateral mesencephalic lesions of the PGO pathways. Sleep-wake parameters and the spontaneous unit activity in the lateral geniculate nucleus (LGN) were then chronically recorded during one month after the lesions. Whereas the states of vigilance remained normal, a significant reduction of the unitary discharge in the LGN during paradoxical sleep (PS) and particularly of its phasic component, was observed concomitantly with the suppression of PGO waves. It is concluded that phasic discharge of LGN neurons during PS is controlled by PGO-related mechanisms and that its suppression during the developmental period provides a model for studying its significance on brain maturation.


Dispersal of yeasts and bacteria by Drosophila in a temperate forest.
The dispersal of yeasts, bacteria and molds by Drosophila and Hymenoptera was examined in a deciduous woods in Bloomington, Indiana. Drosophila were found to be significant vectors of yeasts and bacteria, while Hymenoptera also transmitted molds. Analyses of their microbe contents indicated sap feeding Drosophila carried mainly yeasts, fungus feeding Drosophila carried primarily bacteria and ants carried primarily molds.


Epidemiological and genetic characterization of porcine epidemic diarrhea virus in the Mekong Delta, Vietnam, from 2015 to 2017.
Porcine epidemic diarrhea (PED) virus (PEDV) is a highly contagious virus. PED was first identified in 2008 and has greatly affected the Vietnamese pig production economy. The aim of this study was to investigate the epidemiological and genetic characteristics of PEDV in piglet herds in the Mekong Delta, Vietnam. Diarrheal stool and intestinal samples from 2262 piglets from 191 herds in five provinces were collected to test for the presence of PEDV. Ten PEDV strains were randomly selected for sequencing, and four genes encoding PEDV structural proteins were analyzed. The rates of herds and samples positive for PEDV were 27.23% and 27.72%, respectively. In positive herds, the morbidity and mortality of PEDV-positive piglets were 97.97% and 79.06%, respectively, with most of the infected piglets under 7 days of age. Phylogenetic analysis showed that the 10 PEDV strains from this study clustered with genotype G2 strains from Vietnam and neighboring countries. Many amino acid substitutions were identified in important antigenic regions in the spike protein of the 10 strains when compared to four PEDV vaccine strains. This study provides novel insights into the epidemiology and genetic diversity of circulating PEDV strains, which could facilitate the development of an appropriate and proactive strategy for controlling PED.


Standard biopsy forceps versus large-capacity forceps with and without needle.
Endoscopic biopsy forceps vary in size and design. The purpose of this prospective randomized study was to compare the quality and quantity of gastric tissue obtained by needle and non-needle versions of standard biopsy forceps and newly designed large capacity forceps. Fifty consecutive patients who underwent endoscopy with gastric biopsy forceps were enrolled in the study. There was no significant difference in the presence of crush artifact between the two forceps, both with and without the presence of a needle. Both needle and non-needle versions of the large capacity biopsy forceps were found to obtain significantly larger sized specimens (p = 0.02) than needle and non-needle versions of the standard biopsy forceps. Overall, there was no significant difference in the depth of specimen obtained when comparing the large capacity forceps to standard forceps. Needle versions of each forceps were found to obtain significantly deeper biopsies than non-needle versions of each forceps. In conclusion, our study found that large capacity forceps obtained larger specimens than standard biopsy forceps. Further clinical trials with a larger study population need to be undertaken to determine the impact of these findings on the determination of diagnoses.


The role of preservation in the variability of regenerative medicine products.
Regenerative medicine (RM) has the potential to restore or establish normal function of cells, tissues and organs that have been lost due to age, disease or injury. It is common for the site of raw material collection, site of manufacture and site of clinical use to be different for RM products, and at the same time cells must remain viable and functional during transportation among different sites. Freezing products down to cryogenic temperatures along with cold chain transportation has become an effective method of preserving RM products. The quality of RM products along this supply chain represents the cumulative effects of all of the processing steps and all of the reagents used in the process. A variety of sources of variability in the preservation of RM products can result in both cell losses and greater variability in the quality of RM products. The purpose of this article is to review the sources of variability in the preservation process as well as the methods by which variability can be controlled or avoided.


Characterization of the different polypeptide components and analysis of subunit assembly in ferritin.
Ferritin was dissociated into subunits by various denaturants and the subunits were examined by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Human, horse, rat, and rabbit ferritins all exhibited characteristic patterns of heterogeneity; components with molecular weights of about 19,000, 11,000, and 8,000 were invariably found in these preparations. This result contradicts earlier reports that ferritin consists of 24 identical subunits. These polypeptides were isolated, purified in the presence of low concentrations of detergent, and characterized. Evidence based on amino acid compositions, NH2-terminal analysis and investigation of detergent-induced breakdown products, indicated that the 19,000 molecular weight component is a composite of the 8,000 and 11,000 molecular weight chains. Circular dichroism studies showed that the 19,000 molecular weight polypeptide retained appreciable amounts of ordered secondary structure whereas the two lower molecular weight peptides were unfolded to a much greater extent. If the 8,000 and 11,000 molecular weight polypeptides were recombined in equimolar amounts and the denaturant was completely removed, a substance with electrophoretic mobility and morphological appearance of native apoferritin was obtained.


Clinical and electrophysiological findings and follow-up in tarsal tunnel syndrome.
The authors report clinical and electrophysiological findings in 59 patients with tarsal tunnel syndrome (TTS) and follow-up in 23 of them. The entrapment was prevalent in females; was bilateral in 6 patients and involved medial plantar in 7 and lateral plantar nerves in two cases. Eleven presented with other nerve entrapment syndromes or focal mononeuropathies, due to hereditary neuropathy with liability to pressure palsy or systemic diseases. The other 48 subjects had TTS without any other related entrapment syndromes: 23 were idiopathic cases, 13 had a history of local trauma, 3 had systemic diseases and the others had external or intrinsic compressions. The most frequent symptoms were paraesthesia or dysaesthesia (86% of feet) and pain (55%). Hypoaesthesia of the sole and weakness of toe flexion were evident in 74% and 22% of feet, respectively. Absence of sensory action potential or slowing of sensory conduction velocity (SCV) of the plantar nerves were present in 77% of feet; significant differences of SCV between affected and unaffected plantar nerves and/or between distal sural and plantar nerves were evident in 14%. Abnormalities of plantar SCV were therefore absent in only 9% of feet. Distal motor latency was delayed in 55% and electromyography showed neurogenic changes in 45% of sole muscles. Five cases (6 feet) underwent surgery with excellent or good results in 5, 4 of them also showing improvement in distal conduction of the plantar nerves. Nine were treated with local steroid injections, with good results shown in 6 patients. Nine other patients who did not receive any therapy showed a disappearance of symptoms or good outcome in 6 cases. The subjects with poor therapeutic results had S1 radiculopathy or systemic diseases. The authors underline that patients with connective tissue diseases should not be treated by surgical decompression because they may have subclinical neuropathy. Some subjects with idiopathic or trauma-induced TTS recover spontaneously. Surgical release should be limited to cases with space-occupying lesions and when conservative treatments fail.