Evidence of a functional reorganization in the auditory dorsal stream following unilateral hearing loss.
Unilateral hearing loss (UHL) generates a disruption of binaural hearing mechanisms, which impairs sound localization and speech understanding in noisy environments. We conducted an original study using fMRI and psychoacoustic assessments to investigate the relationships between the extent of cortical reorganization across the auditory areas for UHL patients, the severity of unilateral hearing loss, and the deficit in binaural abilities. Twenty-eight volunteers (14 UHL patients) were recruited (twenty-two females and six males). The brain imaging analysis demonstrated that UHL induces a shift in aural dominance favoring the better ear, with a cortical reorganization located in the non-primary auditory areas, ipsilateral (same side) to the better ear. This reorganization is correlated not only to the hearing loss severity but also to spatial localization abilities. A regression analysis between brain activity and patient's performance clearly showed that the spatial hearing deficit was linked to a functional alteration of the posterior auditory areas known to process spatial hearing. Altogether, our study reveals that UHL alters the dorsal auditory stream, which is deleterious to spatial hearing.


Moyamoya disease in a Hispanic child: a case report.
Moyamoya disease was initially described by Suzuki and Takaku in 1963 as a radiographic phenomenon relating to the tiny collateral vessels characteristic of the disease that resemble a cloud or puff of smoke. The disease is rare and initially it was believed that the disease was confined to the Japanese population. It consists of occlusive vascular disease at the Circle of Willis with a tendency toward multiple ischemic neurological events and small strokes. In older populations it can often be associated with further vascular degeneration and intracerebral hemorrhage. This paper discusses the diagnosis, treatment, and management of moyamoya disease in a Hispanic child. The case of a Hispanic child who presented with transient ischemic attacks over a period of 1 year is reported. Magnetic resonance imaging (MRI) revealed occlusive vascular disease in the posterior Circle of Willis. Digital subtraction cerebral angiography showed vascular occlusion at the base of the skull with collateral leptomeningeal and posterior circulation contribution in a pattern typical of moyamoya disease. Technetium was injected for a SPECT study demonstrating less uptake in the left frontal and left parieto-occipital regions. The patient underwent a left superficial temporal-to-middle cerebral artery anastamosis followed by a right-sided anastamosis in a second operation. The patient tolerated the cerebral revascularization and was symptom-free at 6-month follow-up. Cerebral angiography demonstrated improved perfusion in both cerebral hemispheres postoperatively. This article reports the occurrence of moyamoya in a Hispanic child. It illustrates the improved perfusion postoperatively as seen on digital subtraction cerebral angiography. Direct revascularization is felt to be difficult in children and alternatives such as encephaloduroarteriosynangiosis have been advocated. Direct revascularization was effective in treating moyamoya disease in this instance. Most of the discussion of moyamoya disease has been focused on the Japanese and Far East population. This report confirms the entity as occurring in a Hispanic individual in the United States with no known Japanese ancestry.


Propofol Protects Rats and Human Alveolar Epithelial Cells Against Lipopolysaccharide-Induced Acute Lung Injury via Inhibiting HMGB1 Expression.
High-mobility group box 1 (HMGB1) plays a key role in the development of acute lung injury (ALI). Propofol, a general anesthetic with anti-inflammatory properties, has been suggested to be able to modulate lipopolysaccharide (LPS)-induced ALI. In this study, we investigated the effects of propofol on the expression of HMGB1 in a rat model of LPS-induced ALI. Rats underwent intraperitoneal injection of LPS to mimic sepsis-induced ALI. Propofol bolus (1, 5, or 10 mg/kg) was infused continuously 30 min after LPS administration, followed by infusion at 5 mg/(kg · h) through the left femoral vein cannula. LPS increased wet to dry weight ratio and myeloperoxidase activity in lung tissues and caused the elevation of total protein and cells, neutrophils, macrophages, and neutrophils in bronchoalveolar lavage fluid (BALF). Moreover, HMGB1 and other cytokine levels were increased in BALF and lung tissues and pathological changes of lung tissues were excessively aggravated in rats after LPS administration. Propofol inhibited all the above effects. It also inhibited LPS-induced toll-like receptor (TLR)2/4 protein upexpression and NF-κB activation in lung tissues and human alveolar epithelial cells. Propofol protects rats and human alveolar epithelial cells against HMGB1 expression in a rat model of LPS-induced ALI. These effects may partially result from reductions in TLR2/4 and NF-κB activation.


Clear cell carcinoma of cervix and vagina in progeny of women who received diethylstilbestrol: three cases with scanning and transmission electron microscopy.
Three cases of clear cell carcinoma of the cervix and vagina in progeny of women administered diethylstilbestrol (DES) were studied ultrastructurally. The patients were young and had a positive history of maternal ingestion of DES during pregnancy. Similar ultrastructural features were observed in all cases. The neoplastic cells were arranged in solid nests or glands. Ultrastructural findings in these cases were similar to those of previously reported clear cell carcinomas of the ovary and endometrium, which seems to indicate a common Müllerian origin11, 16 as opposed to a mesonephric origin. 10 A transition between vaginal adenosis and the neoplasm was not observed.


Substances released during the decomposition of Vallisneria natans and Thalia dealbata.
Two types of aquatic plants commonly used for the ecological restoration of rivers and lakes, Vallisneria natans (Lour.) Hara and Thalia dealbata Fraser ex Roscoe, were selected and grouped by plant parts (root, stem and foliage), and decomposing release experiments were conducted. The influence of the released substances on the water quality was analyzed, as well as the amount of nutrients released by each part of these two plants. The calculated maximum chemical oxygen demand releases from the foliage of V. natans and the foliage of T. dealbata were approximately 5.4 g/kg and 22.65 g/kg, respectively. Through three-dimensional fluorescence spectrum and parallel factor analyses, the different material compositions of the decomposing liquids from the plants were determined, and the main dissolved organic components of the decomposing liquid of V. natans were amino-acid-like and microbially derived humics, and those T. dealbata were soluble microbial by-product-like substances. The carbon-to-nitrogen ratio and humification index of each experimental group were compared. The experimental results showed that different parts of V. natans and T. dealbata had different rates of nutrient release. The dissolved organic matter in the decomposed solution can be utilized by microorganisms, which have the potential to become additional carbon sources. This study provides a new method for the treatment of aquatic plant litter. Different plant species can be used in combination according to their characteristics to ensure that better results are achieved during water treatment processes that use plant decomposing liquids as additional carbon sources.


Rapid clearance of HBe antigen and development of anti-HBe antibody in acute viral hepatitis.
The behaviour of the HBe/anti-HBe system in AVH was evaluated, using Magnius technique, by testing serum samples from 47 patients; 29 of them were followed during the clinical evolution of the disease until complete remission was achieved. HBe was more frequent in samples taken from patients in the first 7 days after the onset of jaundice (18/33 = 54%) than in samples collected later (3/14 = 21%). During the clinical evolution of the disease we could always demonstrate the disappearance of HBe not later than 12 days after the onset of jaundice. In one patient studied from the incubation period HBe disappeared before any clinical or laboratory evidence of disease. In 8/29 cases (27%) anti-HBe developed starting from the 15th day of illness, but 4 of these had had no detectable HBe during the acute phase. No significant difference could be demonstrated between HBe +ve and -ve cases in the maximum values of SGPT and bilirubin and the duration of the disease. The changing pattern of the HBe/anti-HBe system could account for the different incidences of these markers reported by many authors in AVH. Our findings support the hypothesis that HBe develops in every HBsAg +ve AVH case. Therefore, it is not the presence of HBe in the early stage, but the persistence of this marker that might be important in predicting progression to chronicity.


Effects of cryo-injury on progesterone receptor(s) of canine spermatozoa and its response to progesterone.
The integrity of sperm progesterone (P4) receptor(s) and its response to steroid stimulation might be crucial for the maintenance of sperm fertilizing ability after cryopreservation. The aim of the current investigation was to study the effect of cryo-procedures on canine sperm P4 receptor(s). In addition, alteration of P4 receptor(s) at the molecular level and their functional integrity following cryo-procedures was evaluated. Fresh and frozen-thawed semen samples (n=6 same dogs) after capacitation were treated with 10 microg/mL P4 to induce the acrosome reaction (AR, FITC-PNA staining). Parallel samples were treated with 50% canine seminal plasma (SP) prior to AR induction with P4. The percentages of AR in capacitated fresh and frozen-thawed semen samples after treatment with P4 were 31.0+/-6.7 and 21.6+/-4.1% (P<0.05), respectively. The percentage of AR in fresh and frozen-thawed semen samples pretreated with SP and incubated with P4 was; 11.5+/-4.8 and 16.5+/-2.0% (P<0.05), respectively. The incidence of the spontaneous AR (P>0.05) in fresh and frozen-thawed semen samples at the onset (5.5+/-2.2 and 6.1+/-1.8%; respectively) and after a 2h (9.6+/-5.1 and 10.4+/-2.7%; respectively) capacitation, avoiding P4 stimulation, were not different. The percentage of progesterone-BSA-FITC staining over the acrosomal region was 18.3+/-10.3% in fresh semen, 36.0+/-11.9% in capacitated (P<0.05) and less than 5% in SP treated spermatozoa. This staining was barely visible in frozen-thawed spermatozoa regardless of capacitation status. In western blot analysis, mAb C262 recognized two bands (54 and 65 kDa). Digitonin treated fresh and frozen-thawed spermatozoa, labeled with [3H]-progesterone, revealed that the P4 binding capacity decreased from 6.0+/-4.4 in fresh to 3.0+/-2.1 nM in frozen-thawed spermatozoa. In nearly all samples tested (except one) 65 kDa protein band decreased significantly after freeze-thaw procedures while the 54kDa protein was increased. These results indicate that the reduced incidence of AR in response to P4 in frozen spermatozoa is possibly due to the conformational changes of P4 receptor(s) and/or reduced P4 receptor density derived from freezing injury.


[Action of cortical extracts of the left and right hemispheres on the recovery of conditioned reflex activity in rats after unilateral removal of the frontal cortex].
The recovery of active avoidance conditioned reflex (AACR) was investigated after unilateral frontal cortex extirpation. Intraperitoneal injection of extracts from left or right brain cortex (1 mg/kg) of healthy rats (LE or RE) stimulated AACR recovery in animals with lobectomy on the same side. If RE was extracted 9 days after left side brain extirpation, i.e. during the period of the development of compensatory processes, its effect on AACR recovery was stronger in left-operated animals, while in right-operated animals it remained unchanged.


The efficacy of long-term captopril treatment on micro- and macroalbuminuria in hypertensive diabetics.
Microalbuminuric [16] and macroalbuminuric [17] hypertensive insulin dependent diabetics were followed up for 4 years after the initiation of captopril therapy to assess the efficacy of ACE inhibitor therapy on albuminuria and blood pressure normalisation. Within the first six months of captopril therapy mean systolic blood pressure decreased in microalbuminuric and macroalbuminuric patients from 168.1 +/- 17.6 mmHg to 134.4 +/- 12.1 mmHg (19.2 +/- 7.1%) and from 177.6 +/- 16.8 mmHg to 143.5 +/- 12.7 (18.9 +/- 6.7%) mmHg, respectively. Mean diastolic blood pressure, similarly, showed a decrease from 91.9 +/- 9.1 mmHg to 74.4 +/- 10.3 mmHg (19.0 +/- 9.4%) in the microalbuminuric and from 95.3 +/- 13.7 mmHg to 78.2 +/- 7.3 (16.9 +/- 9.5%) mmHg in the macroalbuminuric group. After six months of captopril administration albumin excretion rates decreased as well, from 97.4 +/- 35.9 micrograms/min to 51.9 +/- 19.9 micrograms/min (46.9 +/- 7.6%) and from 766.7 +/- 577.9 micrograms/min to 365.1 +/- 298.4 micrograms/min (50.4 +/- 8.4%) in the micro- and macroalbuminuric groups, respectively. Thereafter, mean albumin excretion rates and blood pressure rose significantly, but at the end of the fourth year they were still significantly lower compared to that of the pretreatment period. After four years, albumin excretion rates were 71.3 +/- 29.6 micrograms/min in the microalbuminuric and 391.2 +/- 204.7 micrograms/min in the macroalbuminuric group. We conclude that ACE inhibitor therapy results in a rapid decrease of albuminuria and blood pressure, and despite a slow gradual increase, the albumin excretion rates and blood pressure values remain significantly lower than the initial values after four years.


Digital mammography: are there advantages in screening for breast cancer?
Digital mammography separates the processes of image acquisition, processing, and display, which allows for the optimization of each process. The result addresses some of the limitations of screen film mammography. This work reviews the advantages of the decoupling of the processes and the clinical trials comparing digital mammography with film-screen mammography in the screening setting. Advanced applications of digital mammography, such as contrast-enhanced digital mammography and tomosynthesis, are also discussed.


Infrared Thermography for Estimating Supraclavicular Skin Temperature and BAT Activity in Humans: A Systematic Review.
Brown adipose tissue (BAT) is a thermogenic tissue with potential as a therapeutic target in the treatment of obesity and related metabolic disorders. The most used technique for quantifying human BAT activity is the measurement of 18 F-fluorodeoxyglucose uptake via a positron emission tomography/computed tomography scan following exposure to cold. However, several studies have indicated the measurement of the supraclavicular skin temperature (SST) by infrared thermography (IRT) to be a less invasive alternative. This work reviews the state of the art of this latter method as a means of determining BAT activity in humans. The data sources for this review were PubMed, Web of Science, and EBSCOhost (SPORTdiscus), and eligible studies were those conducted in humans. In most studies in which participants were first cooled, an increase in IRT-measured SST was noted. However, only 5 of 24 such studies also involved a nuclear technique that confirmed increased activity in BAT, and only 2 took into account the thickness of the fat layer when measuring SST by IRT. More work is needed to understand the involvement of tissues other than BAT in determining IRT-measured SST; at present, IRT cannot determine whether any increase in SST is due to increased BAT activity.


Mitigating the pro-oxidant state and melanogenesis of Retinitis pigmentosa: by counteracting mitochondrial dysfunction.
Retinitis pigmentosa (RP) is a group of mitochondrial diseases characterized by progressive degeneration of rods and cones leading to retinal loss of light sensitivity and, consequently, to blindness. To date, no cure is available according to the clinical literature. As a disease associated with pigmentation-related, pro-oxidant state, and mitochondrial dysfunction, RP may be viewed at the crossroads of different pathogenetic pathways involved in adverse health outcomes, where mitochondria play a preeminent role. RP has been investigated in a number of experimental and clinical studies aimed at delaying retinal hyperpigmentation by means of a number of natural and synthetic antioxidants, as well as mitochondrial cofactors, also termed mitochondrial nutrients (MNs), such as alpha-lipoic acid, coenzyme Q10 and carnitine. One should consider that each MN plays distinct-and indispensable-roles in mitochondrial function. Thus, a logical choice would imply the administration of MN combinations, instead of individual MNs, as performed in previous studies, and with limited, if any, positive outcomes. A rational study design aimed at comparing the protective effects of MNs, separately or in combinations, and in association with other antioxidants, might foresee the utilization of animal RP models. The results should verify a comparative optimization in preventing or effectively contrasting retinal oxidative stress in mouse RP models and, in prospect, in human RP cases.


Cutaneous and venous complications of pentazocine abuse.
A 55-year-old man with a two-year history of intramuscular pentazocine abuse had classical cutaneous findings as well as secondary venous thrombosis, which, to our knowledge, is previously undescribed. Histologic evaluation, including direct immunofluorescent microscopy, did not establish the mechanism of tissue destruction.


The transport mechanism of monocarboxylate transporter on spinosin in Caco-2 cells.
The aim of this study was to determine the uptake mechanism of spinosin (SPI) by the monocarboxylic acid transporters (MCTs) in Caco-2 cells. The Caco-2 cells were pretreated with various monocarboxylic acids, and the uptake of spinosin from Caco-2 cells was measured by High Performance Liquid Chromatography (HPLC). Preloading of various monocarboxylic acids enhanced the uptake of SPI, especially salicylic acid (a substrate of MCTs) had a 23.4 times increase in SPI uptake, indicating that the monocarboxylic acid transporters had an efflux effect on SPI uptake and salicylic acid had a strong inhibition on SPI efflux in Caco-2 cells. At the same time, the uptake of SPI through Caco-2 cells was Na(+)- and temperature-dependent, pretreatment without Na(+) significantly increased the uptake of SPI by 1.85 times and incubated at low temperature (4 °C) SPI uptake increased 20% than that of 37 °C. Furthermore, SPI was transported mainly via a carrier-mediated transport: [Vmax = 5.364 μg/mg protein, Km = 657.0 μg/mL]. The uptake of spinosin (SPI) in Caco-2 cells was mainly regulated by the monocarboxylic acid transporters along with Salicylic acid.


Windpipe controls Drosophila intestinal homeostasis by regulating JAK/STAT pathway via promoting receptor endocytosis and lysosomal degradation.
The adult intestinal homeostasis is tightly controlled by proper proliferation and differentiation of intestinal stem cells. The JAK/STAT (Janus Kinase/Signal Transducer and Activator of Transcription) signaling pathway is essential for the regulation of adult stem cell activities and maintenance of intestinal homeostasis. Currently, it remains largely unknown how JAK/STAT signaling activities are regulated in these processes. Here we have identified windpipe (wdp) as a novel component of the JAK/STAT pathway. We demonstrate that Wdp is positively regulated by JAK/STAT signaling in Drosophila adult intestines. Loss of wdp activity results in the disruption of midgut homeostasis under normal and regenerative conditions. Conversely, ectopic expression of Wdp inhibits JAK/STAT signaling activity. Importantly, we show that Wdp interacts with the receptor Domeless (Dome), and promotes its internalization for subsequent lysosomal degradation. Together, these data led us to propose that Wdp acts as a novel negative feedback regulator of the JAK/STAT pathway in regulating intestinal homeostasis.


Angiotensin II-mediated vascular changes in aged offspring rats exposed to perinatal nicotine.
This study determined the long-term influence of prenatal nicotine exposure (PN) on blood pressure and vascular functions in the aged offspring rats. PN did not affect body weight and plasma adrenocorticotropic hormone level; however, it significantly reduced plasma angiotensin I and angiotensin II in both sexes. Systolic pressure in the male aged PN offspring was significantly higher. Angiotensin II-increased mean arterial pressure was higher in the aged PN offspring than that in the control regardless of sex. AT1 receptor blocker losartan, not AT2 receptor antagonist PD123319, reduced blood pressure in the aged PN rats more than that in the control. In the aged PN offspring, angiotensin II-increased vessel contraction and intracellular calcium level were higher in small mesenteric arteries. Acetylcholine-mediated vascular relaxation was weaker, and nitric oxide-related endothelial functions were damaged in aortic rings of PN offspring. Thickness of the wall of mesenteric arteries was increased in the male aged PN offspring. Ratio of AT1/AT2 receptors was significantly increased in the vessel of the PN group regardless of sex. These data provide new information on the very long term influence of PN on vascular structures and functions in the aged offspring, demonstrate that the aged PN female rats were not free of vascular risks after menopause, and suggest that multiple pathways may be involved in the detrimental alterations of the cardiovascular system of the PN rats.


Trans-ethnic polygenic risk scores for body mass index: An international hundred K+ cohorts consortium study.
While polygenic risk scores hold significant promise in estimating an individual's risk of developing a complex trait such as obesity, their application in the clinic has, to date, been limited by a lack of data from non-European populations. As a collaboration model of the International Hundred K+ Cohorts Consortium (IHCC), we endeavored to develop a globally applicable trans-ethnic PRS for body mass index (BMI) through this relatively new international effort. The polygenic risk score (PRS) model was developed, trained and tested at the Center for Applied Genomics (CAG) of The Children's Hospital of Philadelphia (CHOP) based on a BMI meta-analysis from the GIANT consortium. The validated PRS models were subsequently disseminated to the participating sites. Scores were generated by each site locally on their cohorts and summary statistics returned to CAG for final analysis. We show that in the absence of a well powered trans-ethnic GWAS from which to derive marker SNPs and effect estimates for PRS, trans-ethnic scores can be generated from European ancestry GWAS using Bayesian approaches such as LDpred, by adjusting the summary statistics using trans-ethnic linkage disequilibrium reference panels. The ported trans-ethnic scores outperform population specific-PRS across all non-European ancestry populations investigated including East Asians and three-way admixed Brazilian cohort. Here we show that for a truly polygenic trait such as BMI adjusting the summary statistics of a well powered European ancestry study using trans-ethnic LD reference results in a score that is predictive across a range of ancestries including East Asians and three-way admixed Brazilians.


[Effect of different culture media on viability and tau protein expression in rat hippocampal slices].
To explore the effect of different culture media on viability and expression of tau protein in organotypic hippocampal slice. Brain slices (400 microm) from 1, 2, 4, and 8 week-old Wistar rats were prepared and cultured in minimum essential medium (MEM) or Dulbecco's modified eagle medium: nutrient mixture (DMEM/F12) medium respectively for 21 days. Viability of the slices was measured by lactate dehydrogenase (LDH) assay and expression of tau protein was detected by Western blot. The viability of the slices was not influenced significantly by the two different culture media, while the expression level of tau protein was significantly higher in DMEM/F12 than in MEM (P < 0.05), especially in the slices from 2 and 4 week-old rats. The slices from 2 or 4 week-old rat hippocampi and DMEM/F12 medium may be the preferred choice for tau associated researches. An ideal Alzheimer's disease model may be established based on the results of these researches.


Cloning and characterization of the tandemly arranged bovine lymphotoxin and tumour necrosis factor-alpha genes.
The screening of a bovine genomic library with a human tumour necrosis factor-alpha (TNF-alpha) cDNA probe resulted in the isolation of a 7.2 kb DNA fragment containing the entire bovine TNF-alpha gene. Analysis of this genomic clone showed that it also contains the bovine lymphotoxin (LT, TNF-beta) gene. Comparison to published sequences of human, murine, ovine and rabbit counterparts allowed us to delineate the coding sequences, the promoters and the enhancers of these two genes. Sequences involved in the regulation of translation and in the mRNA stability were found in the 3' untranslated regions.


Selective feticide reverses intrahepatic cholestasis of pregnancy in twins discordant for growth: A case report.
Twin gestations are associated with an increased risk of intrahepatic cholestasis in pregnancy (ICP), probably attributed to the elevated pregnancy hormones. We report a case of a dichorionic diamniotic twin pregnancy, at the third trimester, complicated with ICP and severe, selective fetal growth restriction (sFGR). A 32-year-old primiparous woman with a dichorionic, diamniotic twin gestation conceived via in vitro fertilization (IVF) presented with pruritus at the maternity care unit at 26+4 weeks of pregnancy. Following a detailed assessment, she was diagnosed with severe sFGR and ICP. During her hospitalization, selective feticide of the FGR fetus was decided and a remarkable improvement in the symptoms and the laboratory findings of ICP was noticed. The incidence of ICP is reported to be higher in twin pregnancies, especially those conceived via IVF, compared with singletons. The optimal timing of delivery and management of twin pregnancies complicated with ICP remain unclear. In our case, selective reduction of the FGR fetus led to the resolution of ICP.


Study in the chemical constituents of plantago cynops L. and antibacterial evaluation of verbascoside.
Oleanolic acid, verbascoside, plantarenaloside and four sterolic derivatives were isolated from the aerial parts of Plantago cynops L. These constituents in this species were determined and described for the first time. An antibacterial evaluation was carried out for verbascoside.


Characterization of bone diagenesis by histology in forensic contexts: a human taphonomic study.
The diagenesis of a bone in the postmortem period causes an identifiable deterioration in histology. This degradation is characterized by a collagenous alteration, which can be observed very early. In order to develop a method for determining a postmortem interval for medico-legal use, two ribs collected from six human bodies were studied prospectively over 2 years. Each bone was studied after staining with Sirius red to demonstrate the degradation of collagen as a function of time. This study demonstrated a time-based bone alteration characterized by the architectural degradation of the lamellar bone, without any microbial influence in this postmortem period. The staining was carried out by using Sirius red and correlated this alteration with a collagenic degradation by chemical hydrolysis owing to the affinity of this dye to the amino acids lysine, hydroxylysine, and arginine. Our work asserts that human bone samples that were studied in a controlled environment and analyzed for 24 months underwent a diagenetic trajectory whose main element was collagen hydrolysis.


Recurrent Mutations in APC and CTNNB1 and Activated Wnt/β-catenin Signaling in Intraductal Papillary Neoplasms of the Bile Duct: A Whole Exome Sequencing Study.
This study aimed to elucidate the genetic landscape of biliary papillary neoplasms. Of 28 cases examined, 7 underwent whole exome sequencing, while the remaining 21 were used for validation studies with targeted sequencing. In the whole exome sequencing study, 4/7 cases had mutations in either APC or CTNNB1, both of which belong to the Wnt/β-catenin pathway. Somatic mutations were also identified in genes involved in RAS signaling (KRAS, BRAF), a cell cycle regulator (CDC27), histone methyltransferase (KMT2C, KMT2D), and DNA mismatch repair (MSH3, MSH6, PMS1). Combined with discovery and validation cohorts, mutations in APC or CTNNB1 were observed in 6/28 subjects (21%) and were mutually exclusive. When the cases were classified into intraductal papillary neoplasms of the bile duct (IPNBs, n=14) and papillary cholangiocarcinomas (n=14) based on the recently proposed classification criteria, mutations in APC and CTNNB1 appeared to be entirely restricted to IPNBs with 6/14 cases (43%) harboring mutations in either gene. These genetic alterations were detected across the 3 nonintestinal histologic types. In immunohistochemistry, the aberrant cytoplasmic and/or nuclear expression of β-catenin was found in not only 5/6 IPNBs with APC or CTNNB1 mutations, but also 6/8 cases with wild-type APC and CTNNB1 (total 79%). In addition, APC and CTNNB1 alterations were exceptional in nonpapillary cholangiocarcinomas (n=29) with a single case harboring CTNNB1 mutation (3%). This study demonstrated recurrent mutations in APC and CTNNB1 in nonintestinal-type IPNBs, suggesting that activation of the Wnt/β-catenin signaling pathway is relevant to the development and progression of IPNBs.


[Clinical observation of mesenchymal stem cell as salvage treatment for refractory acute graft-versus-host disease].
To explore the effect of mesenchymal stem cells (MSCs) on refractory acute graft-versus-host disease (GVHD) failed to second-line immunosuppressive therapy. Twenty-two patients with refractory aGVHD received the treatment of first- and/or second-line immunosuppressive agents in combination with MSCs. The MSCs from bone marrow (BM) of HLA-unrelated third-party donors, were used at the median time of 19 (11 - 49) days after aGVHD onset, at a dose of 1×10(6)/kg once with an interval of 14 days. If the symptoms of aGVHD did not improve after continuous infusion four times, MSCs would be discontinued. Meanwhile the proportion of CD3(+)CD4(+), CD3(+)CD8(+) and CD4(+)CD25(+) was detected by flow cytometry (FCM) before and 4 weeks after the MSCs infusion. The median dose of MSC was 4.8 (2.5 - 6.3)×10(6) cell×kg(-1) with a median infusion of 2.5 (1 - 7) times per case. Twelve patients achieved complete response (CR), four partial response (PR) after treatment. The total effective rate was 72.7% (16/22). With a median follow-up of 246.5 (36 - 1116) days post-transplantation, 11 patients survived and 11 died. The causes of death included GVHD(n = 6), infections (n = 3), leukemia relapse (n = 1) and post-transplant lymphoproliferative diseases (n = 1), respectively. The proportion of CD3(+)CD4(+)/CD3(+)CD8(+) was significantly higher at 4th week after MSCs infusion compared to before infusion (1.58 ± 0.54 vs 0.49 ± 0.19, \%t\% = 0.628, P = 0.04). The number of CD4(+)CD25(+) Treg cells had not changed much compared to before infusion (P = 0.606). MSCs derived from the BM of a third-party donor are effective to treat aGVHD failed to second-line immunosuppressive therapy after allo-HSCT. MSCs might play a role in aGVHD by regulating the rate of CD3(+)CD4(+)/CD3(+)CD8(+).


Adsorption of nitrophenol onto activated carbon: isotherms and breakthrough curves.
The adsorption isotherm of p-nitrophenol onto granular activated carbon in 25 degrees C aqueous solution was experimentally determined by batch tests. Both the Freundlich and the Redlich-Peterson models were found to fit the adsorption isotherm data well. A series of column tests were performed to determine the breakthrough curves with varying bed depths (3-6 cm) and water flow rates (21.6-86.4 cm3/h). Explicit equations for the breakthrough curves of the fixed-bed adsorption processes with the Langmuir and the Freundlich adsorption isotherms were developed by the constant-pattern wave approach using a constant driving force model in the liquid phase. The results show that the half breakthrough time increases proportionally with increasing bed depth but decreases inverse proportionally with increasing water flow rate. The constant-pattern wave approach using the Freundlich isotherm model fits the experimental breakthrough curves quite satisfactorily. A correlation was proposed to predict the volumetric mass-transfer coefficient in the liquid phase successfully. The effects of solution temperature and pH on the adsorption isotherm were also studied and the Tóth model was found to fit the isotherm data well at varying solution temperatures and pHs.


Rapid estimation of Salmonella enterica contamination level in ground beef - Application of the time-to-positivity method using a combination of molecular detection and direct plating.
Little progress has been made in decreasing the incidence rate of salmonellosis in the US over the past decade. Mitigating the contribution of contaminated raw meat to the salmonellosis incidence rate requires rapid methods for quantifying Salmonella, so that highly contaminated products can be removed before entering the food chain. Here we evaluated the use of Time-to-Positivity (TTP) as a rapid, semi-quantitative approach for estimating Salmonella contamination levels in ground beef. Growth rates of 14 Salmonella strains (inoculated at log 1 to -2 CFU/g) were characterized in lean ground beef mTSB enrichments and time-to-detection was determined using culture and molecular detection methods. Enrichments were sampled at five timepoints and results were used to construct a prediction model of estimated contamination level by TTP (superscript indicates time in hours) defined as TTP4: ≥5 CFU/g; TTP6: ≤5, ≥1 CFU/g; TTP8: ≤1, ≥0.01 CFU/g; with samples negative at 8 h estimated ≤0.01 CFU/g. Model performance measures showed high sensitivity (100%) and specificity (83% and 93% for two detection methods) for samples with a TTP4, with false negative rates of 0%.


Bacterial infections of the lower urinary tract treated with ciprofloxacin or cefalexin--a comparative study.
30/30 patients with lower UTI were enrolled in a randomized study to compare ciprofloxacin (250 mg/b.i.d.) and cefalexin (1 g/t.i.d.). In addition, 59 patients with cefalexin-resistant bacterial strains were treated in an open study arm. Ciprofloxacin showed convincingly better clinical and bacteriological efficacy than cefalexin. Both antibiotics were well tolerated. Ciprofloxacin has proved to be superior to cefalexin in the treatment of lower UTI.


Differential regulation of behavioral, genomic, and neuroendocrine responses by CRF infusions in rats.
Studies suggest that behavioral, genomic, and endocrine functions mediated by central corticotropin-releasing factor (CRF)-containing circuits may be differentially regulated. However, this hypothesis has never been tested directly by simultaneous assessment of distinct CRF-mediated responses within the same animal. The present study addressed this issue by concurrently examining the effects of central CRF infusions on anxiety responses, plasma corticosterone release, and c-fos mRNA induction within limbic brain circuits. Bilateral intracerebroventricular (icv) infusions of CRF (0.1-10 microg total) dose-dependently reduced exploratory behavior in a novel open field, increased circulating corticosterone (CORT) levels and augmented c-fos mRNA expression in the central nucleus of the amygdala (CeA) and the hypothalamic paraventricular nucleus (PVN). Plasma CORT levels increased significantly after 0.1 microg CRF, whereas behavioral and genomic responses required at least 1 microg CRF, suggesting that the distinct responses mediated by CRF are differentially regulated. Further characterization of intracerebroventricular CRF at 1 microg also demonstrated a disruption of social interaction behavior. The majority of behavioral effects and the elevated c-fos mRNA expression were attenuated by 10 mg/kg DMP696, a CRF(1) antagonist. However, plasma CORT elevation required 30 mg/kg DMP696 for attenuation. Thus, our studies demonstrate a greater sensitivity of the hypothalamic-pituitary-adrenal axis to intracerebroventricular CRF compared with the induction of innate fear-like responses and associated genomic changes.


The artificial neural network approach based on uniform design to optimize the fed-batch fermentation condition: application to the production of iturin A.
Iturin A is a potential lipopeptide antibiotic produced by Bacillus subtilis. Optimization of iturin A yield by adding various concentrations of asparagine (Asn), glutamic acid (Glu) and proline (Pro) during the fed-batch fermentation process was studied using an artificial neural network-genetic algorithm (ANN-GA) and uniform design (UD). Here, ANN-GA based on the UD data was used for the first time to analyze the fed-batch fermentation process. The ANN-GA and UD methodologies were compared based on their fitting ability, prediction and generalization capacity and sensitivity analysis. The ANN model based on the UD data performed well on minimal statistical designed experimental number and the optimum iturin A yield was 13364.5 ± 271.3 U/mL compared with a yield of 9929.0 ± 280.9 U/mL for the control (batch fermentation without adding the amino acids). The root-mean-square-error for the ANN model with the training set and test set was 4.84 and 273.58 respectively, which was more than two times better than that for the UD model (32.21 and 483.12). The correlation coefficient for the ANN model with training and test sets was 100% and 92.62%, respectively (compared with 99.86% and 78.58% for UD). The error% for ANN with the training and test sets was 0.093 and 2.19 respectively (compared with 0.26 and 4.15 for UD). The sensitivity analysis of both methods showed the comparable results. The predictive error of the optimal iturin A yield for ANN-GA and UD was 0.8% and 2.17%, respectively. The satisfactory fitting and predicting accuracy of ANN indicated that ANN worked well with the UD data. Through ANN-GA, the iturin A yield was significantly increased by 34.6%. The fitness, prediction, and generalization capacities of the ANN model were better than those of the UD model. Further, although UD could get the insight information between variables directly, ANN was also demonstrated to be efficient in the sensitivity analysis. The results of these comparisons indicated that ANN could be a better alternative way for fermentation optimization with limited number of experiments.


Effect of endothelin-1 in man: pretreatment with nifedipine, with indomethacin and with cyclosporine A.
The rise in blood pressure following the intravenous administration of endothelin-1 remained unchanged in healthy male volunteers pretreated with either the calcium-channel antagonist nifedipine (10 mg orally), the cyclo-oxygenase inhibitor indomethacin (150 mg day-1 for three days) or the immunosuppressive agent cyclosporine (5 mg kg-1 body weight for five days). Following administration of nifedipine the rise in plasma concentrations of endothelin-1 during the infusion of the peptide was markedly higher (P less than 0.01) than during control experiments without nifedipine. It is concluded that, in healthy men, nifedipine, indomethacin and cyclosporine do not exert a major influence on the pressor action of endothelin-1. However, nifedipine apparently influences the elimination of endothelin-1 from the circulation in healthy men.


Clinical models of chemoprevention for colon cancer.
Colon cancer is a common malignancy in the westernized world and is incurable in its advanced stages. This article summarizes the currently available information on colorectal cancer chemoprevention. A brief outline of the incidence and etiologic factors is followed by a discussion of the evidence on which chemopreventive strategies for colon cancer are modeled. This includes a description of the development of surrogate endpoint biomarkers and experimental models to study colorectal cancer chemopreventives, a review of the promising colorectal cancer chemopreventives, and a discussion of the issues to be addressed in the design of future chemoprevention trials. The article concludes with an emphasis on the development and validation of biomarkers and selection of high-risk cohorts using genetic and epidemiologic tools as the main goals of future colon cancer chemoprevention trials before large-scale, risk-reduction trials are conducted.


[Risk factors for cytomegalovirus in solid organ transplant recipients].
Cytomegalovirus (CMV) is the most important opportunistic pathogen in patients undergoing solid organ transplantation and increases mortality due to both direct and indirect effects. The most important risk factor for the development of CMV disease is discordant donor-recipient CMV serology (positive donor and negative recipient), which confers more than 50% risk of developing CMV disease if no prophylaxis is given. The use of highly potent antiviral agents for CMV prophylaxis in high-risk patients has changed the characteristics of CMV disease in this population. Other classical risk factors for CMV disease include acute graft rejection, the type of organ transplanted, coinfection with other herpesviruses and the type of immunosuppressive agents employed. New risk factors for this complication have recently been described, including variations in the CMV genotype between donor and recipient and genetic alterations in the recipient's innate immunity. The present review discusses classical risk factors and the latest findings reported on the development of CMV in organ transplant recipients.


Comparison of clinical presentation and outcome of childhood-onset and adulthood-onset of systemic lupus erythematosus among Indonesian patients.
The clinical presentation of childhood-onset systemic lupus erythematosus (SLE) is generally perceived to differ from that of adult-onset SLE. We aimed to compare the demographic and clinical manifestation between childhood-onset vs. adult-onset SLE in a cohort of Indonesian patients at tertiary care centers. This retrospective study included patients in the Hasan Sadikin Lupus Registry from 2008 until December 2017. The demographics, clinical presentations, and outcomes were compared between childhood-onset SLE (<18 years old) (Group 1) and adult-onset SLE (≥18 years old) (Group 2). Eight hundred seventy patients were involved into this study. The proportion of childhood-onset SLE was 20% (174 patients). The mean age of group 1 versus group 2 was 13.56 ± 3.04 vs 30.41 ± 8.54 years. The following clinical manifestations at SLE diagnosis were significantly more common in childhood-onset than in adult-onset SLE patients: hematological disorder (p = 0.033) and arthritis (p = 0.006). While discoid rash (p = 0.036) and photosensitivity (p < 0.001) were significantly found higher in adult-onset SLE. Cyclophosphamide therapy was significantly more common to be used in childhood-onset (38.5% vs 21.0%, p = <0.001). However, frequency of mortality on follow-up tended to be higher in childhood-onset group (11.5% vs 7.0%, p = 0.208). Arthritis and hematologic involvements at SLE diagnosis were more prominent in childhood-onset compared to adult-onset patients, and mortality in childhood-onset SLE during follow-up relatively higher. This data may suggest the need for more aggressive management approach to childhood-onset patients with SLE.


Efficacy of Telephone-Based Cognitive Behavioral Therapy for Weight Loss, Disordered Eating, and Psychological Distress After Bariatric Surgery: A Randomized Clinical Trial.
Weight regain after bariatric surgery is associated with recurrence of obesity-related medical comorbidities and deterioration in quality of life. Developing efficacious psychosocial interventions that target risk factors, prevent weight regain, and improve mental health is imperative. To determine the efficacy of a telephone-based cognitive behavioral therapy (tele-CBT) intervention at 1 year after bariatric surgery in improving weight loss, disordered eating, and psychological distress. This multisite randomized clinical trial was conducted at 3 hospital-based bariatric surgery programs, with recruitment between February 2018 and December 2021. Eligibility for participation was assessed among 314 adults at 1 year after bariatric surgery who were fluent in English and had access to a telephone and the internet. Patients with active suicidal ideation or poorly controlled severe psychiatric illness were excluded. Primary and secondary outcome measures were assessed at baseline (1 year after surgery), after the intervention (approximately 15 months after surgery), and at 3-month follow-up (approximately 18 months after surgery). Data were analyzed from January to February 2023. The tele-CBT intervention consisted of 6 weekly 1-hour sessions and a seventh booster session 1 month later. The control group received standard postoperative bariatric care. The primary outcome was postoperative percentage total weight loss. Secondary outcomes were disordered eating (Binge Eating Scale [BES] and Emotional Eating Scale [EES]) and psychological distress (Patient Health Questionnaire-9 item scale [PHQ-9] and Generalized Anxiety Disorder-7 item scale [GAD-7]). The hypotheses and data-analytic plan were developed prior to data collection. Among 306 patients 1 year after bariatric surgery (255 females [83.3%]; mean [SD] age, 47.55 [9.98] years), there were 152 patients in the tele-CBT group and 154 patients in the control group. The group by time interaction for percentage total weight loss was not significant (F1,160.61 = 2.09; P = .15). However, there were significant interactions for mean BES (F2,527.32 = 18.73; P < .001), EES total (F2,530.67 = 10.83; P < .001), PHQ-9 (F2,529.93 = 17.74; P < .001), and GAD-7 (F2,535.16 = 15.29; P < .001) scores between the tele-CBT group and control group across all times. This study found that tele-CBT delivered at 1 year after surgery resulted in no change in short-term weight outcomes but improved disordered eating and psychological distress. The impact of these psychosocial improvements on longer-term weight outcomes is currently being examined as part of this longitudinal multisite randomized clinical trial. ClinicalTrials.gov Identifier: NCT03315247.


[>Femoral bone infarction revealing compound heterozygous SC sickle cell disease in a Moroccan patient].
Double heterozygosity SC is a major sickle cell syndrome. Its course may be marked by severe or irreversible complications, such as bone infarction. We here report the case of a 17-year-old patient presenting with compound heterozygous SC sickle cell disease following severe knee pain (gonalgia) in order to highlight the risk of delayed diagnosis as well as the need for prevention policies guiding early screening, thus improving patients' management and prognosis.


Symptom-oriented versus syndrome approaches to resolving heterogeneity of neuropsychological functioning in schizophrenia.
Schizophrenia is a heterogeneous disorder. The syndrome and the symptom-oriented approaches are two methods that have been used to examine differences in psychopathological processes across different patients with schizophrenia. The authors indirectly compared these two approaches in their examination of associations between positive symptoms and neuropsychological performance. Positive syndrome and delusion and hallucination severity scores were compared in respective associations to functioning on 12 neuropsychological variables in 73 stable outpatients with schizophrenia. The individual symptoms of the positive syndrome were associated with relatively distinct patterns of neuropsychological performance, suggesting that the symptom-oriented approach was more sensitive.


Expanding and Enhancing Food and Nutrition Education in New York City Public Schools: An Examination of Program Characteristics and Distribution.
To expand their capacity, many schools partner with food and nutrition education programs (FNPs). Public policies and funding can support FNPs, but comprehensive data on the organizations that run FNPs, their program characteristics, or distribution across schools did not exist in NYC. This study aims to help local education and health agencies assess the characteristics of food and nutrition education in schools, as well as to measure progress implementing school policies and practices. A cross-sectional study on NYC FNPs was conducted during the 2016-2017 school year. Survey data on organizations and the FNPs they operate were collected. Data on schools in which FNPs operate were gathered. To determine distribution of FNPs across schools and by school demographics, the database of FNPs in schools was combined with a publicly available database of NYC schools. In 2016-2017, 40 organizations operated 101 FNPs in 56% of NYC public schools. These FNPs varied by goals, content, activities, location, and populations served. Information on these variations can help policymakers, advocates, funders, and schools expand school-based food and nutrition education. To ensure equitable access, more coordination, investment, and collaboration are needed.


Differential expression of HOX genes upon activation of leukocyte sub-populations.
The HOX genes are key determinants of cellular identity both in early development and in the renewal and differentiation of adult blood cells. Although a number of studies have examined the expression of individual HOX genes in defined blood cell lineages, we have undertaken a comprehensive analysis of HOX gene expression in resting and activated lymphocytic and monocytic subpopulations. This has revealed distinct patterns of expression between different cell types and resting and activated states. (Main category A: Erythrocytes, Leukocytes and Hematopoiesis, subcategory: 8: Lymphocytes).


High-risk human papilloma virus infection of the foreskin in asymptomatic boys.
To evaluate the prevalence and genotypes of high-risk human papilloma virus (HPV) infection of the foreskin in asymptomatic boys before first sexual intercourse. We collected 50 consecutive foreskin specimens after radical circumcision. Indication for surgery was phimosis. High-risk HPV status was determined by real-time polymerase chain reaction for the genotypes 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, and 59. Immunohistochemistry for p16(INK4a) was performed. The median age at the time of surgery was 5.5 years (range, 5 months-15 years). High-risk HPV was detected in 6 of 50 foreskins (12%). All positive samples showed HPV16. No association with age or grade of phimosis was observed. Two samples were focally positive for p16(INK4a), both of which were HPV-negative. In a significant proportion of boys, subclinical high-risk HPV infections are found in the foreskin, which could be a reservoir for HPV-associated diseases. Our study generates the hypothesis that nonsexual routes play significant roles in HPV transmission. Because the human foreskin represents a high-risk HPV reservoir, vaccination may be also advised in boys.


Hospitalization and mortality trends among patients with confirmed COVID-19 in the United States, April through August 2020.
The United States has experienced high COVID-19 case counts, hospitalizations, and death rates. This retrospective analysis reports changing trends in the demographics and clinical outcomes of hospitalized US COVID-19 patients between April and August 2020. The Premier Healthcare Database Special Release was used to examine patient demographics of hospitalized COVID-19 patients from all US Census Bureau divisions. Demographics included age, sex, race, and ethnicity. Clinical outcomes included in-hospital mortality, intensive care unit (ICU) admission, and receipt of invasive mechanical ventilation. Overall, 146,491 hospitalized COVID-19 patients were included (mean [SD] age, 61.0 [18.4] years; 51.7% male; 29.6% White non-Hispanic). Monthly total hospitalizations decreased from 44,854 in April to 18,533 in August; ICU admissions increased from 19.8% to 23.6%, and ventilator use and inpatient mortality decreased from 18.6% to 14.5% and 21.0% to 11.4%, respectively. Inpatient mortality was highest in the Middle Atlantic division (20.3%), followed by the New England (19.0%), East North Central (14.2%), and Mountain (13.7%) divisions. Black non-Hispanic patients were overrepresented among hospitalizations (19.0%); this group comprises 12.2% of the US population. Patients aged <65 years made up 53% of hospitalizations and had lower inpatient mortality than those aged ≥65 years. Hospitalizations, ventilator use, and mortality decreased, while ICU admission rates increased from April to August 2020. Older individuals and Black non-Hispanics were found to be at elevated risk of severe outcomes. These trends could inform ongoing patient care and US public health policies to limit the further spread of SARS-CoV-2.


Adductor longus: An anatomical study to better understand groin pain.
We investigated the shear modulus-passive force relationship in the hip adductor longus (AL) muscles of human cadavers and explored the effect of muscle architecture on the elastic properties of the AL muscle using shear wave elastography (SWE). Nine AL muscles were harvested from a soft, embalmed cadaver. The AL muscles were affixed to a custom-built device comprising two clamps, a pulley, and a cable to provide passive loads, which were increased from 0 to 600 g in 60-g increments. The shear modulus of the AL muscle was measured in the proximal (Pro), middle (Mid), and distal (Dis) regions. The masses and anatomical cross-sectional areas (ACSAs) of the AL muscles were measured. The shear modulus-passive load relationship of each tested muscle region was analyzed by fitting a least-squares regression line. Moreover, the rate of increase in the shear modulus per unit load (s) was calculated. The shear modulus and passive force were linearly correlated for all AL muscles in each region (p < 0.01). The mean coefficients of determination (R2 ) for Pro, Mid, and Dis were 0.989, 0.986, and 0.982, respectively. The rate of increase in the shear modulus per unit load significantly correlated with the reciprocal of the muscle mass (r = 0.77, p = 0.02) and ACSA (r = 0.43, p = 0.03). Shear wave elastography can be used as an indirect measure of passive force in any region of the AL muscle. Additionally, the rate of increase in the shear modulus per unit load could be associated with muscle architectural parameters.


Microbiome-wide association studies between phyllosphere microbiota and ionome highlight the beneficial symbiosis of Lactococcus lactis in alleviating aluminium in cassava.
The phyllosphere is one of the most abundant habitats for global microbiota. The ionome is the composition of mineral elements in plants. The correlation between phyllosphere microbiota and the ionome remains elusive in plants, especially in the most important tropical crop cassava. In this study, microbiome-wide association studies (MWASs) of thirty varieties were performed to reveal the association between phyllosphere microbiota and ionomic variations in cassava. Annotation of metagenomic species identified some species that were significantly correlated with ionomic variations in cassava. Among them, Lactococcus lactis abundance was negatively associated with leaf aluminium (Al) levels but positively related to leaf potassium (K) levels. Notably, both the reference and isolated L. lactis showed strong binding capacity to Al. Further bacterial transplantation of isolated L. lactis could significantly decrease endogenous Al levels but increase K levels in cassava, and it can also lead to increased citric acid and lactic acid levels as well as higher transcript levels of K uptake-related genes. Taken together, this study reveals the involvement of phyllosphere microbiota in ionomic variation in cassava, and the correlation between L. lactis abundance and Al and K levels provides novel insights into alleviating Al accumulation and promoting K uptake simultaneously.


Decoding herbal materials of TCM preparations with the multi-barcode sequencing approach.
With the rapid development of high-throughput sequencing technology, approaches for assessing biological ingredients in Traditional Chinese Medicine (TCM) preparations have also advanced. Using a multi-barcode sequencing approach, all biological ingredients could be identified from TCM preparations in theory, as long as their DNA is present. The biological ingredients of several classical TCM preparations were analyzed successfully based on this approach in previous studies. However, the universality, sensitivity and reliability of this approach on a diverse set of TCM preparations remain unclear. In this study, we selected four representative TCM preparations, namely Bazhen Yimu Wan, Da Huoluo Wan, Niuhuang Jiangya Wan, and You Gui Wan, for concrete assessment of the multi-barcode sequencing approach. Based on ITS2 and trnL biomarkers, we have successfully detected the prescribed herbal materials (PHMs) in these representative TCM preparations (minimum sensitivity: 77.8%, maximum sensitivity: 100%). The results based on ITS2 have also shown higher reliability than trnL at species level, while their combination could provide higher sensitivity and reliability. The multi-barcode sequencing approach has shown good universality, sensitivity and reliability in decoding these four representative TCM preparations. In the omics big-data era, this work has undoubtedly made one step forward for applying multi-barcode sequencing approach in PHMs analysis of TCM preparation, towards better digitization and modernization of drug quality control.


[Malignant histiocytosis. Histiocytic medullary reticulosis].
The Authors report a case of malignant histiocytosis apparently localized only in the spleen. The diagnosis of the disease, characterized at the onset by moderate leukopenia and thrombocytopenia, was made possible by the histopathologic examination of the spleen following splenectomy. The disease shows a chronic course. The Authors discuss the clinical and histological features of the case.


Predicting Positive ELISA Results in Dairy Herds with a Preferred Status in a Paratuberculosis Control Program.
Dairy herds participating in the Dutch milk quality assurance program for paratuberculosis are assigned a herd status on the basis of herd examinations by ELISA of individual serum or milk samples, followed by an optional confirmatory fecal PCR. Test-negative herds are assigned Status A; the surveillance of these herds consists of biennial herd examinations. Farmers falsely believing that their Status A herds are Map-free may inadvertently refrain from preventive measures. Therefore, we aimed to develop a predictive model to alert Status A farmers at increased risk of future positive ELISA results. Using data of 8566 dairy herds with Status A in January 2016, two logistic regression models were built, with the probabilities of ≥1 or ≥2 positive samples from January 2017-June 2019 as dependent variables, and province, soil type, herd size, proportion of cattle born elsewhere, time since previous positive ELISA results, and the 95th percentile of the S/P ratios in 2015-2016, as explanatory variables. As internal validation, both models were applied to predict positive ELISA results from January 2019-June 2021, in 8026 herds with Status A in January 2019. The model predicting ≥1 positive sample had an area under the receiver operating characteristics curve of 0.76 (95% CI: 0.75, 0.77). At a cut-off predicted probability πc = 0.40, 25% of Status A herds would be alerted with positive and negative predictive values of 0.52 and 0.83, respectively. The model predicting ≥2 positive samples had lower positive, but higher negative, predictive values. This study indicates that discrimination of Status A herds with high and low risks of future positive ELISA results is feasible. This might stimulate farmers with the highest risks to take additional measures to control any undetected Map infections.


P-glycoprotein: a role in the export of amyloid-β in Alzheimer's disease?
The accumulation of amyloid-β (Aβ) peptides is a key histopathological feature of the Alzheimer's brain. Defective clearance mechanisms result in toxic levels of soluble Aβ40 and Aβ42 oligomers, leading to impaired synaptic function, neurodegeneration and cognitive decline. Growing evidence points to the involvement of P-glycoprotein (P-gp or ABCB1), an ATP-binding cassette transporter highly expressed on the luminal side of the blood-brain barrier, in facilitating the clearance of Aβ from the brain. In this review, we summarise evidence from human, animal and in vitro studies examining the contribution of P-gp to Aβ clearance, and discuss the potential for P-gp as a novel pharmacological target in Alzheimer's disease (AD). P-gp expression and activity in the brain are inversely correlated with ageing, Aβ deposition and AD. Moreover, Aβ itself has been found to compromise the expression of P-gp, thereby exacerbating Aβ deposition and disease. Despite decades of research, the pathophysiology of AD remains elusive. Understanding the normal versus impaired processing and clearance mechanisms affecting Aβ peptides will assist the development of more effective therapeutic agents to combat this progressive neurodegenerative condition that continues to devastate millions of patients globally.


Complement your knowledge with a rare cause of pauci-immune glomerulonephritis.
RPGN can be subdivided into three categories on an immunopathologic basis: pauci-immune glomerulonephritis (PIGN), anti-glomerular basement membrane glomerulonephritis (anti-GBM disease), or immune complex-mediated glomerulonephritis (GN). PIGN is the most common cause of RPGN (80% of cases). The most common etiology of PIGN is anti-neutrophil cytoplasmic antibody (ANCA)-associated GN, which accounts for up to 90% of PIGN. PIGN is unique in that few to no immunoglobulin deposits are seen on glomerular immunofluorescence (IF) and electron microscopy (EM), but it is important to remember that dysregulation of the alternative pathway may result in the deposition of complements leading to inflammatory injury even in PIGN. Membranoproliferative glomerulonephritis (MPGN) is a rare, primary glomerular disorder. Both ANCA-associated GN and complement-mediated MPGN will lack immunoglobulin staining on immunofluorescence (IF) and thus present as PIGN on pathologic examination. This may lead to occurrences where these entities mimic one another, therefore necessitating heightened suspicion and close pathologic examination of a renal biopsy with electron microscopy to differentiate the diagnoses. This case highlights a rare case of C3 GN mimicking ANCA-associated GN.


Activation of p38MAPK contributes to expanded polyglutamine-induced cytotoxicity.
The signaling pathways that may modulate the pathogenesis of diseases induced by expanded polyglutamine proteins are not well understood. Herein we demonstrate that expanded polyglutamine protein cytotoxicity is mediated primarily through activation of p38MAPK and that the atypical PKC iota (PKCiota) enzyme antagonizes polyglutamine-induced cell death through induction of the ERK signaling pathway. We show that pharmacological blockade of p38MAPK rescues cells from polyglutamine-induced cell death whereas inhibition of ERK recapitulates the sensitivity observed in cells depleted of PKCiota by RNA interference. We provide evidence that two unrelated proteins with expanded polyglutamine repeats induce p38MAPK in cultured cells, and demonstrate induction of p38MAPK in an in vivo model of neurodegeneration (spinocerebellar ataxia 1, or SCA-1). Taken together, our data implicate activated p38MAPK in disease progression and suggest that its inhibition may represent a rational strategy for therapeutic intervention in the polyglutamine disorders.


The characterization of Ia+ cells in human parathyroid and islets.
Recent studies in animals have shown prolonged survival of endocrine allografts after treatment of the graft with Ia antibody prior to transplant. In this study, we documented the presence of Ia bearing passenger cells in human parathyroid and islets of Langerhans. Furthermore, we characterized these cells with regard to their expression of HLA-DQ and HLA-DR molecules using indirect immunofluorescence. We found that passenger cells in parathyroid express both HLA-DQ and HLA-DR. In contrast, islets express only HLA-DR. Parathyroid tissue treated first with polyclonal anti-DR or anti-DQ and complement eliminated cells that reacted with monoclonal anti-DR or -DQ probes, respectively. Similarly, islets treated with polyclonal anti-DR and complement eliminated cells that reacted with monoclonal anti-DR reagents. We believe that further characterization of Ia+ cells in human endocrine tissue will provide information that could be used to enhance allograft survival.


Plasma 17 alpha, 20 beta-dihydroxy-4-pregnen-3-one levels during sexual maturation of amago salmon (Oncorhynchus rhodurus): correlation with plasma gonadotropin and in vitro production by ovarian follicles.
Plasma gonadotropin (GtH) and 17 alpha, 20 beta-dihydroxy-4-pregnen-3-one (17 alpha, 20 beta-diOH-prog) levels were assessed by radioimmunoassay during the sexual maturation of female amago salmon (Oncorhynchus rhodurus). Both GtH and 17 alpha, 20 beta-diOHprog levels were low in vitellogenic females (June to September) and in those with full-grown immature oocytes and were elevated in mature and ovulated females to 40 ng/ml for GtH and 50-70 ng/ml for 17 alpha, 20 beta-diOHprog. In vitro production of 17 alpha, 20 beta-diOHprog by ovarian follicles and its stimulation by partially purified chinook salmon gonadotropin (SG-G100) was examined monthly using 18 hr incubations. In June and July, levels were too low to detect (less than 30 pg/ml) in media from all treatment groups. In August and September, SG-G100 at a concentration of 1 microgram/ml stimulated low levels of production (0.2-0.3 ng/ml). Full-grown, immature follicles in October showed a dose-response production of 17 alpha, 20 beta-diOHprog, averaging over 10 ng/ml when incubated with 1 microgram/ml SG-G100. Postovulatory follicles (1-2 days after ovulation) produced large amounts of 17 alpha, 20 beta-diOHprog, averaging over 100 ng/ml with 1 microgram/ml SG-G100. These results are discussed in relation to previous work on amago salmon and other species and together indicate that 17 alpha, 20 beta-diOHprog is the major steroid responsible for oocyte maturation in amago salmon, produced as the follicular mediator of gonadotropin.


The effects of AER and eGFR on outcomes of CVD in patients with T2DM in an urban community over 8 years of multifactorial treatment: the Beijing Communities Diabetes Study 18.
It is well known that diabetic kidney disease is a risk factor for cardiovascular diseases (CVD) in patients with type 2 diabetes mellitus (T2DM). In this study, the effects of urine albumin excretion rate (AER) and estimated glomerular filtration rate (eGFR) on CVD outcomes were analyzed in a population of T2DM. The study was carried out using recorded information of a cohort study. A total of 1,914 patients with T2DM with no prevalent CVD were enrolled in an 8 years prospective study and received multifactorial intervention. The risk of CVD outcomes was assessed according to chronic kidney disease staging, which was categorized using AER (mg/d) and eGFR (mL/min/1.73 m2). The effects of AER and eGFR on risk of CVD onset were also analyzed. During the follow-up period (median 6.8 years), 71 CVD events occurred. At baseline, those with AER ≥300 mg/d and coexisting eGFR 60-89 mL/min/1.73 m2 or <60 mL/min/1.73 m2 showed increased risk for CVD outcomes when compared with "no chronic kidney disease" (AER <30 mg/d and eGFR ≥90 mL/min/1.73 m2). The increased CVD risk was observed in patients who progressed to AER ≥30 mg/d during the follow-up period, whereas patients who progressed to eGFR <90 mL/min/1.73 m2 alone showed no increased CVD risk. During the follow-up period, after multifactorial intervention, 8.7% patients with microalbuminuria and 1.8% patients with overt nephropathy reversed to normoalbuminuria or microalbuminuria. AER is a more sensitive predictor than eGFR for CVD outcomes in T2DM patients. Overt nephropathy can be reversed after multifactorial intervention.


Catalytic control over supramolecular gel formation.
Low-molecular-weight gels show great potential for application in fields ranging from the petrochemical industry to healthcare and tissue engineering. These supramolecular gels are often metastable materials, which implies that their properties are, at least partially, kinetically controlled. Here we show how the mechanical properties and structure of these materials can be controlled directly by catalytic action. We show how in situ catalysis of the formation of gelator molecules can be used to accelerate the formation of supramolecular hydrogels, which drastically enhances their resulting mechanical properties. Using acid or nucleophilic aniline catalysis, it is possible to make supramolecular hydrogels with tunable gel-strength in a matter of minutes, under ambient conditions, starting from simple soluble building blocks. By changing the rate of formation of the gelator molecules using a catalyst, the overall rate of gelation and the resulting gel morphology are affected, which provides access to metastable gel states with improved mechanical strength and appearance despite an identical gelator composition.


[Management of carcinoma of the kidney with metastasis (author's transl)].
The discovery of pulmonary metastasis is not a contraindication to operation. In the light of 21 personal cases and a review of the literature, the authors give their arguments in favour of surgery. No pronostic criterion is certain except for two facts: 1) the course is capricious; 2) spontaneous remissions do not occur. The results of surgery alone do not seem to be influenced by radiotherapy nor chemotherapy.


Developmental Programming: Priming Disease Susceptibility for Subsequent Generations.
Racial and/or ethnic minorities carry the highest burden of many adverse health outcomes intergenerationally We propose a paradigm in which developmental programming exacerbates the effects of racial patterning of adverse environmental conditions, thereby contributing to health disparity persistence. Evidence that developmental programming induces a heightened response to adverse exposures ("second hits") encountered later in life is considered. We evaluated the evidence for the second hit phenomenon reported in animal and human studies from three domains (air, stress, nutrition). Original research including a gestational exposure and a childhood or adulthood second hit exposure was reviewed. Evidence from animal studies suggest that prenatal exposure to air pollutants is associated with an exaggerated reaction to postnatal air pollution exposure, which results in worse health outcomes. It also indicates offspring exposed to prenatal maternal stress produce an exaggerated response to subsequent stressors, including anxiety and hyper-responsiveness of the hypothalamic-pituitary-adrenal axis. Similarly, prenatal and postnatal Western-style diets induce synergistic effects on weight gain, metabolic dysfunction, and atherosclerotic risk. Cross-domain second hits (e.g., gestational air pollution followed by childhood stressor) were also considered. Suboptimal gestational environments induce exaggerated offspring responses to subsequent environmental and social exposures. These developmental programming effects may result in enhanced sensitivity of ongoing, racially patterned, adverse exposures in race/ethnic minorities, thereby exacerbating health disparities from one generation to the next. Empirical assessment of the hypothesized role of priming processes in the propagation of health disparities is needed. Future social epidemiology research must explicitly consider synergistic relationships among social environmental conditions to which gestating females are exposed and offspring exposures when assessing causes for persistent health disparities.


The road to hell is paved with good intentions: a look back at the PANTHER-IPF trial.
The PANTHER-IPF trial was a turning point in treatment of idiopathic pulmonary fibrosis (#IPF) highlighting the importance of randomised controlled trials in determining treatment strategies, even for rare diseases and/or potentially fatal acute events https://bit.ly/3Oi0KwD.


Effects of Salts and Surface Charge on the Biophysical Stability of a Low pI Monoclonal Antibody.
The impact of five representative Hofmeister salts (NaCl, KCl, MgCl2, Na2SO4, and NaSCN) on the thermal stability and aggregation kinetics of a slightly acidic monoclonal antibody (mAb) were investigated under different pH conditions. The thermal stability of the mAb was assessed by measuring the lowest unfolding transition temperature, Tm, with differential scanning fluorimetry. MgCl2 and NaSCN significantly decreased Tm at all three charged states of the mAb, but to the greatest extent when the mAb surface charge was net positive. Non-native aggregation kinetics was monitored by measuring Rayleigh light scattering. When the mAb surface charge was net positive or net neutral, the nucleation rate increased non-monotonically with MgCl2 and NaSCN but decreased monotonically with NaCl, KCl, and Na2SO4. By contrast, when the mAb surface was negatively charged, there were only minor changes in the nucleation rate with all salts tested. Furthermore, there was less structural perturbation and slower aggregation rates when the mAb was net negatively charged than when it was net neutrally or positively charged. The observed salt effects on thermal unfolding are consistent with ion-specific mechanisms dominated by short-range amide backbone binding. On the other hand, the salt effects on nucleation rates appear to be influenced by both amide backbone binding and long-range electrostatic binding of ions to charged amino acid side chains.


Basics of laser application to dermatology.
Q-switched lasers, with a pulse of light sufficiently short (nanosecond-domain) is demonstrated to be useful for treatment of dermal melanocytosis, blue-black tattoos, melanocytic nevi, and solar lentigines, although transient postinflammatory hyperpigmentation usually developed in the irradiated area during the following 3-4 months. If the postinflammatory pigmentation does not disappear after 1 year, incontinentia pigmenti histologica is a possibility. However, the pigment in café-au-lait macules responds variably to treatment. Melasma shows no response to laser. Therefore, accurate diagnosis is the key to success in the laser treatment. Laser treatment of vascular lesions is based on selective absorption by blood with thermal injury to the vessel wall. Therefore, the pulse-width of the vascular-specific lasers must be longer (microsecond-domain) than that of pigment-specific lasers. Because the wavelength of the lasers for vascular lesions, however, cannot penetrate into the deep areas of the skin, not all vascular lesions can be treated. Laser or light-assisted hair removal offers an efficient way to permanently reduce excessive hair growth. Skin rejuvenation is possible by laser or pulsed light with millisecond-domain pulse-width. Because these light sources, however, cause severe damage to the skin surface, the exposure energy must be reduced and the treatment must be combined with cooling devices. Therefore, the clinical results of light-assisted skin rejuvenation are not prominent. In conclusion, the pulse-width and wavelength of the laser light are critical parameters for laser treatments. If we obtain information about these parameters for specific lasers, we can expect the results of the treatment to be positive.


Draft Genome Sequence of Stenotrophomonas bentonitica BII-R7T, a Selenite-Reducing Bacterium Isolated from Spanish Bentonites.
The Gram-negative bacterium Stenotrophomonas bentonitica BII-R7T was isolated from bentonite formations. Like other species within the genus Stenotrophomonas, strain BII-R7T possesses high tolerance to numerous heavy metals, suggesting potential for bioremediation purposes. The draft genome sequence reported here comprises 4.37 Mb with a G+C content of 66.5% and 3,796 predicted protein-coding sequences.


The effects of intensive dietary weight loss and exercise on gait in overweight and obese adults with knee osteoarthritis. The Intensive Diet and Exercise for Arthritis (IDEA) trial.
The Intensive Diet and Exercise for Arthritis (IDEA) trial was an 18-month randomized controlled trial that enrolled 454 overweight and obese older adults with symptomatic and radiographic knee osteoarthritis (OA). Participants were randomized to either exercise (E), intensive diet-induced weight loss (D), or intensive diet-induced weight loss plus exercise (D + E) interventions. We previously reported that the clinical benefits of D + E were significantly greater than with either intervention alone (e.g., greater pain reduction, and better function, mobility, and health-related quality of life). We now test the hypothesis that D + E has greater overall benefit on gait mechanics compared to either intervention alone. Knee joint loading was analyzed using inverse dynamics and musculoskeletal modeling. Analysis of covariance determined the interventions' effects on gait. The D + E group walked significantly faster at 18-month follow-up (1.35 m s-1) than E (1.29 m s-1, p = 0.0004) and D (1.31 m s-1, p = 0.0007). Tibiofemoral compressive impulse was significantly lower (p = 0.0007) in D (1069 N s) and D + E (1054 N s) compared to E (1130 N s). D had significantly lower peak hip external rotation moment (p = 0.01), hip abduction moment (p = 0.0003), and peak hip power production (p = 0.016) compared with E. Peak ankle plantar flexion moment was significantly less (p < 0.0001) in the two diet groups compared with E. There also was a significant dose-response to weight loss; participants that lost >10% of baseline body weight had significantly (p = 0.0001) lower resultant knee forces and lower muscle (quadriceps, hamstring, and gastrocnemius) forces than participants that had less weight loss. Compared to E, D produces significant load reductions at the hip, knee, and ankle; combining D with E attenuates these reductions, but most remain significantly better than with E alone.


Development of reconstructive blepharoplasty: the temporofacial rotation flap. Was it first conceived on the Spree, on the Seine, or on the Manzanares?
The development of reconstructive blepharoplasty in the 19th century is analyzed and the techniques classified according to their principles. Besides the main contributions of French, German, and Italian plastic surgeons, those made by two Spanish surgeons are summarized. Argumosa, who published an article on the tempororotation flap two years before Dieffenbach, and Hysern, who developed the first musculocutaneous transposition flaps for eyelid reconstruction, are usually omitted in the literature. After Dieffenbach's publication, German and French surgeons argued about who contributed to the development of plastic surgery in general and to blepharoplasty in particular, while in Spain controversies arose about the importance of Hysern's and Argumosa's contributions. The background and causes of these disputes are analyzed and compared with the present day dissemination and publication of new techniques.


Relationship of cataract symptoms of preoperative patients and vision-related quality of life.
This study was conducted in an attempt to describe the status of cataract symptoms of preoperative cataract patients, as well as to determine the relationship between cataract symptom and vision-related quality of life measures. A cross-sectional study design was used. Study subjects were selected using non-probabilistic methods. The study sample consisted of 132 patients scheduled for cataract surgery in one eye. Information was obtained from study subjects by conducting face-to-face interviews. Study subjects were adult cataract patients at Samsung and Kunyang General Hospitals in Seoul, Korea, and Tulane University Hospital and Clinics in New Orleans, Louisiana, United States of America. Degree to which study subjects were "bothered" by cataract symptoms was measured by using the 5-item Cataract Symptom Scores (CSS). The number of symptoms, highest scored symptom among the five, number of symptoms extremely bothering, and the mean of cataract symptom score were created based on the CSS measure. Vision-related quality of life was assessed by using the Visual Function 14 Items (VF-14) and Global Measure of Vision (GMV) for trouble and satisfaction. The symptoms "blurry vision" and "worsening of vision" were most frequently reported. The symptom "colors looking different" was rarely reported among preoperative cataract patients. The degree to which study subjects were bothered by "blurry vision" and "seeing glare, halo, or rings" was highly associated with visual function, visual trouble, and visual satisfaction. Multiple regression analysis revealed that the newly developed cataract symptom-related scales, as well as total amount of degree to which study subjects were "bothered" by symptoms were highly related to vision-related quality of life measures. These relationships were persistent after controlling for major socio-demographic variables. The results indicate that cataract symptoms are highly associated with vision-related quality of life. The results also suggest that the number of symptoms, highest scored symptom among the five, number of symptoms extremely bothering, and the mean cataract symptom scores should be considered an important cataract symptom-related scale when analyzing quality of life, including cataract symptom.


Modality difference in the physiological properties and morphological characteristics of the trigeminal sensory neurons.
A-delta nociceptive neurons divided according to fast-conducting (n = 21) and slow-conducting (n = 13) types were found to be different in electrophysiology and morphology, while between warm temperature (n = 23) and warm temperature and touch (n = 38) neurons, and between touch (n = 26) and vibrotactile (n = 59) neurons their characteristics were found to be similar.


Minimizing sperm oxidative stress using nanotechnology for breeding programs in rams.
Artificial insemination (AI) is a routine breeding technology in animal reproduction. Nevertheless, the temperature-sensitive nature and short fertile lifespan of ram sperm samples hamper its use in AI. In this sense, nanotechnology is an interesting tool to improve sperm protection due to the development of nanomaterials for AI, which could be used as delivery vehicles. In this work, we explored the feasibility of vitamin E nanoemulsion (NE) for improving sperm quality during transport. With the aim of evaluating this proposal, ejaculates of 7 mature rams of Manchega breed were collected by artificial vagina and extended to 60 × 106 spz/mL in Andromed®. Samples containing control and NE (12 mmol/L) with and without exogenous oxidative stress (100 µmol/L Fe2+/ascorbate) were stored at 22 and 15 ºC and motility (CASA), viability (YO-PRO/PI), acrosomal integrity (PNA-FITC/PI), mitochondrial membrane potential (Mitotracker Deep Red 633), lipoperoxidation (C11 BODIPY 581/591), intracellular reactive oxygen species (ROS) production and DNA status (SCSA®) monitored during 96 h. Our results show that NE could be used to maintain ram spermatozoa during transport at 15 and 22 ºC for up to 96 h, with no appreciable loss of kinematic and physiological characteristics of freshly collected samples. The storage of ram spermatozoa in liquid form for 2-5 d with vitamin E nanoemulsions may lead more flexibility to breeders in AI programs. In view of the potential and high versatility of these nanodevices, further studies are being carried out to assess the proposed sperm preservation medium on fertility after artificial insemination.


Cell-Sized confinement in microspheres accelerates the reaction of gene expression.
Cell-sized water-in-oil droplet covered by a lipid layer was used to understand how lipid membranes affect biochemical systems in living cells. Here, we report a remarkable acceleration of gene expression in a cell-sized water-in-oil droplet entrapping a cell-free translation system to synthesize GFP (green fluorescent protein). The production rate of GFP (V(GFP)) in each droplet remained almost constant at least for on the order of a day, which implies 0(th)-order reaction kinetics. Interestingly, V(GFP) was inversely proportional to radius of droplets (R) when R is under 50 μm, and V(GFP) in droplets with R ∼ 10 μm was more than 10 times higher than that in the bulk. The acceleration rates of GFP production in cell-sized droplets strongly depended on the lipid types. These results demonstrate that the membrane surface has the significant effect to facilitate protein production, especially when the scale of confinement is on the order of cell-size.


Phylogenetic relationships among the Ibero-African cobitids (Cobitis, cobitidae) based on cytochrome b sequence data.
We estimated the phylogenetic relationships of all Ibero-African spined loaches of the genus Cobitis using the complete mitochondrial cytochrome b gene (1140bp). We analysed 93 individuals of seven cobitid species found in all the principal drainages of the Iberian Peninsula and North Africa. A molecular phylogeny was used to revise current systematics of the Ibero-African Cobitis species and to infer a biogeographical model for Cobitis in the Western Mediterranean area during the Cenozoic period. Phylogenetic analysis provided support for the monophyly of two mtDNA clades: Clade A or Italian Clade with the Italian species (C. bilineata, C. zanandreai), and Clade B or the Ibero-African Clade. The Ibero-African Clade included all species endemic for the Iberian Peninsula (C. vettonica, C. calderoni, and C. paludica) and North Africa (C. maroccana). The species C. paludica does not constitute a natural group, and could be divided into at least four monophyletic mtDNA lineages with moderate to high bootstrap values and posterior probability support. Phylogenetic relationships of the Ibero-African species were not resolved satisfactorily, but in all analyses C. calderoni from Northern Iberian Peninsula was basal. We have re-calibrated a molecular clock for the genus Cobitis (0.68% per million year by pairwise) using populations inhabiting both sides of the Gibraltar Strait. Application of this Cobitis mtDNA clock provides evidence that the Messinian salinity crisis played a primary role in the diversification of some Ibero-African cobitid species. The basal polytomies of the Ibero-African Clade might suggest that all mtDNA lineages diversified rapidly.


Neighborhood effects on nonword visual processing in a language with shallow orthography.
Neighborhood size and neighborhood frequency were orthogonally varied in two experiments on Italian nonwords. In Experiment 1, an inhibitory effect of neighborhood frequency on visual lexical decision was found: The presence of one high-frequency neighbor increased response latencies and error rates to nonwords. By contrast, no effect of neighborhood size and no neighborhood size x neighborhood frequency interaction were found. In Experiment 2, a facilitatory effect of neighborhood size on nonword naming was shown: Naming latencies were faster when nonwords had a large neighborhood. In the naming experiment, there was no effect of neighbors' frequency and no neighborhood size x neighborhood frequency interaction. An additional role for bigram frequency was found whereas syllable frequency did not give any independent contribution. These results further corroborate the view that, in a language with transparent orthography like Italian, despite a substantial contribution of sublexical print-to-sound mapping due to the language's high regularity/consistency, reading aloud of nonlexical material may benefit from the contribution of the lexical component.


Prevalence and perinatal risk factors of parent-reported colic, abdominal pain and other pain or discomforts in infants until 3 months of age - A prospective cohort study in PreventADALL.
To estimate the prevalence and perinatal risk factors associated with parent reported colic, abdominal pain and pain or other discomforts in infants until 3 months of age. Infant colic is a common concern for parents and clinicians. The prevalence varies in different studies and its symptoms overlap with other conditions like abdominal pain and discomfort. Diagnosis criteria are challenging, pathogenesis unclear and risk factors are conflicting. This was a prospective cohort study. The 1852 mother-child pairs from the PreventADALL prospective birth-cohort answering the 3 months questionnaire were included. Information on perinatal risk factors was collected from the inclusion visit and questionnaires during pregnancy at 18 and 34 weeks, as well as birth charts. STROBE checklist was followed. The reported prevalence of colic was 3% (59/1852), abdominal pain 22% (415/1852) and pain or other discomfort 6% (119/1852), with a total of 26% (478/1852) infants. Mothers on sick leave in pregnancy and reporting any allergic diseases had a significantly higher odds of reporting infant colic, abdominal pain and pain or other discomforts. Mothers with higher perceived stress in pregnancy exhibited a trend towards higher odds for reporting infant pain. Mothers coming from Sweden were less likely to report infant abdominal pain compared to mothers from Norway. The prevalence of abdominal pain and pain or other discomforts was higher than the prevalence of colic. Perinatal risk factors connected to maternal health were associated with all three symptoms. Colic and abdominal pain are stressful, symptoms overlap and risk factors for both can be identified in pregnancy. Our study suggests that it is difficult for parents to distinguish among infant colic, abdominal pain and other pain or discomfort and some report two or all three symptoms. Identifying the perinatal risk factors associated with infant pain may help target and support parents.


Postnatal development of transmural gradients in expression of ion channels and Ca²⁺-handling proteins in the ventricle.
Transmural gradients in myocyte action potential duration (APD) and Ca(2+)-handling proteins are argued to be important for both the normal functioning of the ventricle and arrhythmogenesis. In rabbit, the transmural gradient in APD (left ventricular wedge preparation) is minimal in the neonate. During postnatal development, APD increases both in the epicardium and the endocardium, but the prolongation is more substantial in the endocardium leading to a significant transmural gradient. We have investigated changes in the expression of ion channels and also Ca(2+)-handling proteins in the subepicardial and subendocardial layers of the left ventricular free wall in neonatal (2-7 days of age) and adult male (~6 months of age) New Zealand White rabbits using quantitative PCR and also, when possible, in situ hybridisation and immunohistochemistry. In the adult, there were significant and substantial transmural gradients in Ca(v)1.2, KChIP2, ERG, K(v)LQT1, K(ir)2.1, NCX1, SERCA2a and RyR2 at the mRNA and, in some cases, protein level-in every case the mRNA or protein was more abundant in the epicardium than the endocardium. Of the eight transmural gradients seen in the adult, only three were observed in the neonate and, in two of these cases, the gradients were smaller than those in the adult. However, in the neonate there were also transmural gradients not observed in the adult: in HCN4, Na(v)1.5, minK, K(ir)3.1 and Cx40 mRNAs - in every case the mRNA was more abundant in the endocardium than the epicardium. If the postnatal changes in ion channel mRNAs are used to predict changes in ionic conductances, mathematical modelling predicts the changes in APD observed experimentally. It is concluded that many of the well known transmural gradients in the ventricle develop postnatally.


Difference between calculated and direct-measured low-density lipoprotein cholesterol in subjects with diabetes mellitus or taking lipid-lowering medications.
We evaluated factors that caused differences between calculated low-density lipoprotein cholesterol (C-LDL-C) and direct-measured LDL-C (D-LDL-C) and compared them in subjects with diabetes mellitus (DM) or taking lipid-lowering medications. 21,452 subjects (9,177 women, 12,275 men; 8.1% with DM and 8.5% on lipid-lowering medications) were included in the analysis. Participants were classified into 3 groups, i.e., group 1: the subjects without DM and not on lipid-modifying drugs (n = 18,287), group 2: without DM and on lipid-modifying drugs (n = 1,423), and group 3: with DM (n = 1,742). LDL-C concentrations were either directly measured by a homogenous method or calculated by Friedewald formula. There was a significant correlation between C-LDL-C and D-LDL-C (r = 0.966, P < .001). The absolute values of the differences between two LDL-C values were 7.0 ± 6.2 mg/dl and 6.6 ± 7.3% (6.6 ± 5.9 mg/dl and 6.0 ± 6.5%, 8.8 ± 6.7 mg/dl and 9.1 ± 9.7%, and 10.1 ± 7.3 mg/dl and 10.7 ± 10.1% in group 1, 2, and 3 respectively, P < .001). The subjects with the absolute value of the differences of LDL-C ≥10% was 20.2% (17.3%, 31.3%, and 41.1% in group 1, 2, and 3 respectively, P < .001). In the multiple logistic regression analysis, high triglyceride (≥150 mg/dl), low high-density lipoprotein cholesterol (HDL-C) (<40 mg/dl), male gender, obesity (body mass index ≥25 kg/m(2)), DM and taking lipid-lowering drugs were significant associated with high LDL-differences (the absolute value of the differences ≥10% or ≥10 mg/dl). D-LDL-C was generally higher by 5 mg/dl or 5% than C-LDL-C. The differences C-LDL-C and D-LDL-C were higher in subjects with DM and on lipid-lowering medications. Male gender, high triglyceride, low HDL-C, and obesity were also associated with the greater differences between C-LDL-C and D-LDL-C.


Chemical and Structural Differences in Cell Wall Polysaccharides of Two Monokaryotic Strains and Their Resulting Dikaryon of Agaricus bisporus
Compatible monokaryotic strains of Agaricus bisporus ATCC 36975 and 36976 and the resulting dikaryon of their mating were grown in a liquid medium, and their respective cell walls were prepared. Significant differences were not found in the gross chemical composition of the three hyphal types. However, the neutral carbohydrate composition of the complete walls and their fractions was found to be somewhat different in each strain. More consistent differences were encountered in the chemical structure of the distinct polysaccharidic wall fractions in the three types of organisms. Some of these structural wall differences can be considered as characteristic markers for differentiating the mono- and dikaryotic types of A. bisporus.


Directed induction of alveolar type I cells derived from pluripotent stem cells via Wnt signaling inhibition.
Alveologenesis is a developmental step involving the expansion of the lung surface area which is essential for gas exchange. The gas exchange process is mediated by alveolar type I (AT1) cells, which are known to be differentiated from alveolar type II (AT2) or bipotent cells. Due to the difficulty of isolating and culturing primary AT1 cells, the mechanism underlying their differentiation is not completely understood. We performed single-cell RNA sequencing (scRNA-seq) of fibroblast-dependent alveolar organoids (FD-AOs), including human induced pluripotent stem cell (hiPSC)-derived epithelial cells and fetal lung fibroblasts, and identified hiPSC-derived AT1 (iAT1) cells. A comparison of the FD-AOs and fibroblast-free alveolar organoids showed that iAT1 cells were mainly present in the FD-AOs. Importantly, the transcriptomes of iAT1 cells were remarkably similar to those of primary AT1 cells. Additionally, XAV-939, a tankyrase inhibitor, increased iAT1 cells in passaged FD-AOs, suggesting that these cells were differentiated from hiPSC-derived AT2 (iAT2) cells through the inhibition of canonical Wnt signaling. Consequently, our scRNA-seq data allowed us to define iAT1 cells and identify FD-AOs as a useful model for investigating the mechanism underlying human AT1 cell differentiation from AT2 cells in vitro.


Mitochondrial dysfunction and its role in motor neuron degeneration in ALS.
Mitochondria play a pivotal role in many metabolic and apoptotic pathways that regulate the life and death of cells. Accumulating evidence suggests that mitochondrial dysfunction is involved in the pathogenesis of amyotrophic lateral sclerosis (ALS). Mitochondrial dysfunction may cause motor neuron death by predisposing them to calcium-mediated excitotoxicity, by increasing generation of reactive oxygen species, and by initiating the intrinsic apoptotic pathway. Morphological and biochemical mitochondrial abnormalities have been described in sporadic human ALS cases, but the implications of these findings in terminally ill individuals or in post-mortem tissues are difficult to decipher. However, remarkable mitochondrial abnormalities have also been identified in transgenic mouse models of familial ALS expressing mutant Cu, Zn superoxide dismutase (SOD1). Detailed studies in these mouse models indicate that mitochondrial abnormalities begin prior to the clinical and pathological onset of the disease, suggesting that mitochondrial dysfunction may be causally involved in the pathogenesis of ALS. Although the mechanisms whereby mutant SOD1 damages mitochondria remain to be fully understood, the finding that a portion of mutant SOD1 is localized in mitochondria, where it forms aberrant aggregates and protein interactions, has opened a number of avenues of investigation. The future challenges are to devise models to better understand the effects of mutant SOD1 in mitochondria and the relative contribution of mitochondrial dysfunction to the pathogenesis of ALS, as well as to identify therapeutic approaches that target mitochondrial dysfunction and its consequences.