Tuning the response of osteoblast-like cells to the porous-alumina-assisted mixed-oxide nano-mound arrays.
Nanostructuring of biomaterials is used to create an appropriate interfacial layer that promotes stable cellular adhesion, proliferation, and differentiation on orthopedic and dental implants. Here, we synthesize self-organized arrays of composite-oxide nano-mounds through anodizing Al/Ta bilayers sputtered on substrates to cover the "missing" smallest size range of 10-40 nm for structuring an advanced inorganic biomaterial-Al2 O3 -doped Ta2 O5 films. The osteoblast-like cells appear to be able to recognize the finest differences in the film nano-morphologies. In the absence of serum proteins, the adhesion and cell growth are substantially enhanced on the 20 and 40 nm nanoarrays while in complete medium the cells show better initial adhesion on the 10 nm nanoarrays. The proliferation assay reveals a significant rise in cell number on the 20 and 40 nm nanoarrays during the first 7 days. A remarkable increase in the alkaline phosphatase activity is noticed on the 40 nm nanoarray. Immunostaining of cells adhered to the nano-mound surfaces shows that the cells are well spread over all the nanostructured films with organized actin fibers. The larger surface areas and improved focal contacts are again associated with the 20 and 40 nm nanoarrays. The findings help improve compatibility of living cells with the metal-oxide nanostructured surfaces developed for tissue engineering. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 1645-1654, 2018.


Molecular characterization of fetal alcohol syndrome using mRNA differential display.
The molecular pathogenesis of fetal alcohol syndrome (FAS) has not been well elucidated. The technique of mRNA differential display was used to characterize the etiology and to identify potential markers for FAS. Out of approximately 1,080 mRNA transcripts in mouse embryos that were analyzed, the levels of three mRNAs were altered by ethanol. Two of these mRNAs (one novel and one encoding heat shock protein 47) were also modulated by another teratogen, 3-methylcholanthrene. The third mRNA, encoding alpha-tropomyosin, was specifically up-regulated by ethanol. Consistent with the Northern blot data, immunoblot analysis demonstrated that the level of alpha-tropomyosin protein (31 kDa, most likely a brain specific isoform) was elevated in the embryos exposed to ethanol.


Quantitative real-time polymerase chain reaction (RQ-PCR) for the rapid detection of SHOX haploinsufficiency in Leri-Weill Syndrome.
The short stature homeobox-containing (SHOX) gene, found on the human sex chromosomes, has a role in bone growth and height determination. Haploinsufficiency of the SHOX gene is believed to be responsible for poor growth such as that observed in ther Leri-Weill syndrome (LWS). This is the first report of the study of SHOX gene copy number by the technique of quantitative real-time polymerase chain reaction (RQ-PCR) in 9 patients with LWS. Only 7 patients (78%) of LWS had one copy of the SHOX gene deleted, but 2 patients (12%) have neither a single copy gene deletion nor point mutation after direct sequencing of all 7 exons. Although the majority of patients with LWS in this study have SHOX gene haploinsufficiency, there are some patients with both copies of the SHOX gene intact with absence of any point mutations in the coding region. This may be due to abnormalities in the upstream promoter, or to the effect of other candidate gene mutations. RQ-PCR is a faster and cheaper method of studying SHOX sing-copy deletions compared with the conventional fluorescence in situ hybridization (FISH), and is recommended for the detection of SHOX gene haploinsufficiency.


Acute torsion of the renal transplant after combined kidney-pancreas transplant.
Surgical complications after combined kidney and pancreas transplantation are a major source of morbidity and mortality. Complications related to the pancreas occur with greater frequency as compared to renal complications. The occurrence in our practice of two cases of renal infarction resulting from torsion about the vascular pedicle led to our retrospective review of similar vascular complications after combined kidney and pancreas transplantation. Charts were reviewed retrospectively, and two patients were identified who experienced torsion about the vascular pedicle of an intra-abdominally placed renal allograft. Two patients who had received combined intraperitoneal kidney and pancreas transplantation presented at 16 and 11 months after transplant, respectively, with abdominal pain and decreased urine output. One patient had radiological documentation of abnormal rotation before the graft loss; unfortunately, the significance of this finding was missed. Diagnosis was made in both patients at laparotomy, where the kidneys were infarcted secondary to torsion of the vascular pedicle. Both patients underwent transplant nephrectomy and subsequently received a successful second cadaveric renal transplant. The mechanism of this complication is a result of the intra-abdominal placement of the kidney, length of the vascular pedicle, excess ureteral length, and paucity of adhesions secondary to steroid administration. These factors contribute to abnormal mobility of the kidney. Technical modifications such as minimizing excess ureteral length and nephropexy may help to avoid this complication.


Goodness of fit assessment of an alcohol intervention program and the underlying theories of change.
The authors conducted an investigation of The Pennsylvania State University's Alcohol Intervention Program Level 2 (AIP2) to determine goodness of fit of the program components and its underpinning theories. They determined that the Health Belief Model, Social Norms Theory, Social Learning Theory, and the Transtheoretical Model Stages of Change and Process of Change are associated with AIP2's program elements. Although elements of AIP2 fit with these theories, gaps did exist within the program. The authors put forth recommendations for additional program elements that would strengthen the fit with these theories and enhance the program. In addition, upon examining the Transtheoretical Model Stages of Change and Process of Change in terms of sequence, the authors found that information from the readiness to change assessment was not being used to tailor the program elements to individual participant needs. Moreover, the postreadiness-to-change measure adapted by a prereadiness-to-change assessment was not useful in determining behavior change.


Comparative studies on Aeromonas strains isolated from Lakes Balaton (Hungary) and Fertó/Neusiedlersee (Hungary).
Ecological and comparative taxonomic investigations were carried out on 49 Aeromonas strains isolated from water samples of two moderately alkaline lakes of Hungary, Lake Balaton and Lake Fertó/Neusiedlersee together with 3 authentic strains of Aeromonas hydrophila. Five phena were created at greater than 92% similarity value using the UPGMA method with the Jaccard coefficient. Strains isolated from Lake Balaton were determined as A. hydrophila, while strains originated from Lake Fertó were identified as A. hydrophila and A. sobria. The Fertó isolates of A. hydrophila grew only at higher salt concentration (5% NaCl). This might be an adaptation to the higher salt contents in the water of Lake Fertó. However, no specific differences were detected in their behaviour against alkaline pH values. The wide range of their degradative enzymes indicate that aeromonads can play an important role in nutrient cycling.


Does neighborhood belonging matter? Examining school and neighborhood belonging as protective factors for Latino adolescents.
Across many investigations, school belonging has been linked to several positive outcomes among adolescents, including academic success and psychological well-being. Based on an ecological framework of child development, this study expands on existing research to explore factors that contribute to adolescents' sense of neighborhood as well as school belonging and investigates how belonging in both contexts is related to Latino adolescents' academic and psychological functioning. Participants consisted of 202 Latino adolescents residing in low-income, urban neighborhoods. Structural equation modeling demonstrated that neighborhood peer support was significantly associated with adolescents' sense of neighborhood belonging, whereas teacher support and school peer support were related to school belonging. Although school belonging was positively associated with higher scores on all academic indicators, neighborhood belonging was negatively related to academic aspirations and expectations, grades, and educational values. Yet, both school and neighborhood belonging were associated with better psychological functioning as manifested by lower reported levels of depression. The importance of investigating Latino adolescents' development within multiple contexts is discussed.


Germline promoter hypermethylation of tumor suppressor genes in gastric cancer.
To explore germline hypermethylation of the tumor suppressor genes MLH1, CDH1 and P16(INK4a) in suspected cases of hereditary gastric cancer (GC). A group of 140 Chinese GC patients in whom the primary cancer had developed before the age of 60 or who had a familial history of cancer were screened for germline hypermethylation of the MLH1, CDH1 and P16(INK4a) tumor suppressor genes. Genomic DNA was extracted from peripheral blood leukocytes and modified by sodium bisulfite. The treated DNA was then subjected to bisulfite DNA sequencing for a specific region of the MLH1 promoter. The methylation status of CDH1 or P16(INK4a) was assayed using methylation-specific PCR. Clonal bisulfite allelic sequencing in positive samples was performed to obtain a comprehensive analysis of the CpG island methylation status of these promoter regions. Methylation of the MLH1 gene promoter was detected in the peripheral blood DNA of only 1/140 (0.7%) of the GC patient group. However, this methylation pattern was mosaic rather than the allelic pattern which has previously been reported for MLH1 in hereditary non-polyposis colorectal cancer (HNPCC) patients. We found that 10% of the MLH1 alleles in the peripheral blood DNA of this patient were methylated, consistent with 20% of cells having one methylated allele. No germline promoter methylation of the CDH1 or P16(INK4a) genes was detected. Mosaic germline epimutation of the MLH1 gene is present in suspected hereditary GC patients in China but at a very low level. Germline epimutation of the CDH1 or P16(INK4a) gene is not a frequent event.


Choosing the Ideal Implant-Supported Prosthesis for an Edentulous Maxilla: Three Key Prosthetic Parameters.
Assessment of patient- and prosthetic-specific factors in identifying the ideal implant-supported prosthesis helps enable the delivery of a dental restoration that can meet the esthetic, functional, physiologic, psychologic, financial, and time goals of the patient. For practitioners, determining the relative risks and benefits of removable and fixed implant-supported prostheses for restoration of an edentulous maxilla can be challenging. Proper treatment selection requires thorough examination, diagnosis, and a systematic approach to treatment planning. Consideration of both prosthetic and surgical outcomes is necessary to ensure the restoration of facial structures and oral function. This article discusses three prosthetic parameters essential to determining whether a patient's needs can be best met with a fixed or removable prosthesis: prosthetic space, lip support, and transition line. Standardized treatment planning considerations are presented to facilitate the clinician's step-by-step diagnostic and decision-making processes.


Trends and Reported Complications in Ankle Arthroplasty and Ankle Arthrodesis in the State of New York, 2009-2018.
Ankle arthroplasty has emerged as a viable alternative to ankle arthrodesis due in large part to recent advancements in both surgical technique and implant design. This study seeks to document trends of arthroplasty and arthrodesis for ankle osteoarthritis in New York State from 2009-2018 in order to determine if patient demographics play a role in procedure selection and to ascertain the utilization of each procedure and rates of complications. Patients 40 years and older from 2009-2018 were identified using International Classification of Diseases, Ninth and Tenth Revisions (ICD-9 and ICD-10), Clinical Modification (CM) diagnosis and procedure codes for ankle osteoarthritis, ankle arthrodesis, and ankle arthroplasty in the New York statewide planning and research cooperative system database. A trend analysis for both inpatient and outpatient procedures was performed to evaluate the changing trends in utilization of ankle arthrodesis and ankle arthroplasty over time. A multivariable logistic regression was used to assess the odds of receiving ankle arthrodesis relative to ankle arthroplasty. Complications were compared between inpatient ankle arthrodesis and arthroplasty using multivariable Cox proportional hazards regression. A total of 3735 cases were included. Ankle arthrodesis increased by 25%, whereas arthroplasty increased by 757%. African American race, federal insurance, workers compensation, presence of comorbidities, and higher social deprivation were associated with increased odds of having an ankle arthrodesis vs an ankle arthroplasty. Compared with ankle arthroplasty, ankle arthrodesis was associated with increased rates of readmission, surgical site infection, acute renal failure, cellulitis, urinary tract infection, and deep vein thrombosis. Ankle arthroplasty volume has grown substantially without a decrease in ankle arthrodesis volume, suggesting that ankle arthroplasty may be selectively used for a different population of patients than ankle arthrodesis patients. Despite the increased growth of ankle arthroplasty, certain patient demographics including patients from minority populations, federal insurance, and from areas of high social deprivation have higher odds of receiving arthrodesis. Level III, retrospective cohort.


Synchronous direct gradient layer and indirect room calorimetry.
A dual direct/indirect room-sized calorimeter is used at the Beltsville Human Nutrition Research Center to measure heat emission and energy expenditure in humans. Because the response times of a gradient layer direct calorimeter and an indirect calorimeter are not equivalent, the respective rate of heat emission and energy expenditure cannot be directly compared. A system of equations has been developed and tested that can correct the respective outputs of the direct gradient layer calorimeter and indirect calorimeter for delays due to the response times of the measurement systems. Performance tests using alcohol combustion to simulate a human subject indicate accurate measurements of heat production from indirect (99.9 +/- 0.4%), indirect corrected for response time (99.9 +/- 0.5%), direct (99.9 +/- 0.8%), and direct corrected for response time (99.9 +/- 0.8%) calorimetry systems. Results from 24-h measurements in 10 subjects indicate that corrected heat emission is equivalent to (99.8 +/- 2.0%) corrected energy expenditure. However, heat emission measured during sleep was significantly greater (14%) than energy expenditure, suggesting a change in the energy stored as heat in the body. This difference was reversed during the day. These results illustrate how the simultaneous measurement of heat emission and energy expenditure provides insights into heat regulation.


Major colorectal cancer resection should not be denied to the elderly.
Adjuvant therapy after surgery for colorectal cancer is often denied to the elderly for various reasons. This study was to determine morbidity and mortality risk after surgery in the elderly and whether this is affected by adjuvant therapy. Data were collected prospectively and entered on a database for all patients undergoing resection of colorectal cancer between January 1994 and July 2000. A total of 304 patients were included, 65 aged 80 years and over. There were 84 deaths, 21 (30%) in the over 80s, and 63 (26%) in the under 80s (P=0.51). The 'in-hospital' mortality was 10.1% in the over 80s and 3.8% in the under 80s (P=0.056). In the over 80s the colon was more affected than the rectum (P=0.002). The over 80s were less likely to be offered adjuvant therapy, 7.2% vs 42.1% (P<0.001). The 5 year survival (all-cause mortality) in the over 80s was 58.5% and 47.6% in the under 80s (P=0.25). Cox's regression analysis of all patients identified the following factors to be independently related to overall survival: age>80 years, post-operative leak, increasing Dukes stage and distant recurrence of disease. This study has demonstrated that surgery should not be denied to elderly patients with colorectal cancer as despite a higher post-operative morbidity and mortality rate and with the absence of adjuvant therapy, favourable long-term outcome can be achieved by resectional surgery alone.


Rocaglamide overcomes tumor necrosis factor-related apoptosis-inducing ligand resistance in hepatocellular carcinoma cells by attenuating the inhibition of caspase-8 through cellular FLICE-like-inhibitory protein downregulation.
The enhancement of apoptosis is a therapeutic strategy used in the treatment of cancer. Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a promising antitumor agent. However, hepatocellular carcinoma (HCC) cells exhibit marked resistance to the induction of cell death by TRAIL. The present study investigated whether rocaglamide, a naturally occurring product isolated from the genus Aglaia, is able to sensitize resistant HCC cells to TRAIL-mediated apoptosis. Two HCC cell lines, HepG2 and Huh-7, were treated with rocaglamide and/or TRAIL and the induction of apoptosis and effects on the TRAIL signaling pathway were investigated. The in vivo efficacy of rocaglamide was determined in TRAIL-resistant Huh-7-derived tumor xenografts. Rocaglamide significantly sensitized the TRAIL-resistant HCC cells to apoptosis by TRAIL, which resulted from the rocaglamide-mediated downregulation of cellular FLICE-like inhibitory protein and subsequent caspase-8 activation. Furthermore, rocaglamide markedly inhibited tumor growth from Huh-7 cells propagated in severe combined immunodeficient mice, suggesting that chemosentization also occurred in vivo. These data suggest that rocaglamide acted synergistically with TRAIL against the TRAIL-resistant HCC cells. Thus, it is concluded that rocaglamide as an adjuvant to TRAIL-based therapy may present a promising therapeutic approach for the treatment of HCC.


Separation and characterization of late endosomal membrane domains.
Very little is known about the biophysical properties and the lipid or protein composition of membrane domains presumably present in endocytic and biosynthetic organelles. Here we analyzed the membrane composition of late endosomes by suborganellar fractionation in the absence of detergent. We found that the internal membranes of this multivesicular organelle can be separated from the limiting membrane and that each membrane population exhibited a defined composition. Our data also indicated that internal membranes may consist of at least two populations, containing primarily phosphatidylcholine or lysobisphosphatidic acid as major phospholipid, arguing for the existence of significant microheterogeneity within late endosomal membranes. We also found that lysobisphosphatidic acid exhibited unique pH-dependent fusogenic properties, and we speculated that this lipid is an ideal candidate to regulate the dynamic properties of this internal membrane mosaic.


[Splenic injury during left nephrectomy for renal cell cancer].
Left radical nephrectomy is the second most common cause of splenic injury during transabdominal oncological surgery in the upper left quadrant of the abdomen. The incidence of iatrogenic splenectomy during left nephrectomy is estimated to be between 4.3% and 13.2%. The spleen may be injured in three ways: traction, application of retractors or directly by the surgeon's instruments. Capsular tears, lacerations, avulsions and subcapsular haematomas are the injuries most frequently encountered. The aim of this study was to assess frequency of splenic injury and evaluate methods and results of management of splenic injury during left nephrectomy for renal cell carcinoma. Left radical nephrectomy was performed in 768 consecutive patients for renal cell carcinoma. The mean patient age was 52 years (range 34 to 88 years). The mean tumor size was 58 mm (range 28 to 230 mm). Depending on tumor size or surgeon's preferences transabdominal or retroperitoneal approach was performed. Of the 768 left nephrectomies 34 (4.4%) resulted in splenic injury. Splenectomy was required in 18 cases (2.3%). Splenic injuries were more common during transperitoneal approach. The incidence of iatrogenic spleen trauma was 7.5% during transperitoneal approach and 0.8% during retroperitoneal approach. Surgical treatment of splenic injury was performed in 41% of all splenic injuries. In 3 cases splenorrhaphy was performed, in 2 cases we used tissue glue, in 11 cases TachoComb was used. In 2 patients autotransplantation of splenic parts was performed. Splenic injury during left nephrectomy for renal cell carcinoma is an uncommon complication. The use of haemostatic agent seems to be an effective method of treatment of superficial splenic injuries. Proper management of splenic injury enables preservation of this important organ.


The glycan-specific sulfotransferase (R77W)GalNAc-4-ST1 putatively responsible for peeling skin syndrome has normal properties consistent with a simple sequence polymorphisim.
Expanded access to DNA sequencing now fosters ready detection of site-specific human genome alterations whose actual significance requires in-depth functional study to rule in or out disease-causing mutations. This is a particular concern for genomic sequence differences in glycosyltransferases, whose implications are often difficult to assess. A recent whole-exome sequencing study identifies (c.229 C > T) in the GalNAc-4-ST1 glycosyltransferase (CHST8) as a disease-causing missense R77W mutation yielding the genodermatosis peeling skin syndrome (PSS) when homozygous. Cabral et al. (Genomics. 2012;99:202-208) cite this sequence change as reducing keratinocyte GalNAc-4-ST1 activity, thus decreasing glycosaminoglycan sulfation, as the mechanism for this blistering disorder. Such an identification could point toward potential clinical and/or prenatal diagnosis of a harmful medical condition. However, GalNAc-4-ST1 has minimal activity toward glycosaminoglycans, instead modifying terminal β1,4-linked GalNAc on N- and O-linked oligosaccharides on specific glycoproteins. We find expression, processing and catalytic activity of GalNAc-4-ST1 completely equivalent between wild type and (R77W) sulfotransferases. Moreover, keratinocytes have little or no GalNAc-4-ST1 mRNA, indicating that they do not express GalNAc-4-ST1. In addition, loss-of-function of GalNAc-4-ST1 primarily presents as reproductive system aberrations rather than skin effects. These findings, an allele frequency of 0.004357, and a 10-fold difference in prevalence of CHST8 (c.299 C > T, R77W) across different ethnic groups, suggest that this sequence represents a "passenger" distributed polymorphism, a simple sequence variant form of the enzyme having normal activity, rather than a "driver" disease-causing mutation that accounts for PSS. This study presents an example for guiding biomedical research initiatives, as well as medical and personal/family perspectives, regarding newly-identified genomic sequence differences.


Dermatology and HIV/AIDS in Africa.
Human immunodeficiency virus and the acquired immunodeficiency syndrome (HIV/AIDS) have greatly complicated dermatologic disease and the required care in most regions of Africa. Opportunistic infections, ectoparasites, Kaposi sarcoma, and skin manifestations of systemic infections are exceedingly common in patients with HIV/AIDS. Dermatologists have contributed significantly to our knowledge base about HIV/AIDS and have played an important educational role regarding the clinical manifestations historically. Because of the increased burden of skin disease in Africa due to the HIV/AIDS epidemic we must redouble our efforts to provide dermatology education to care providers in Africa. We review the burden of skin disease in Africa, how it relates to HIV/AIDS and global infectious disease, current educational strategies in Africa to address this need, and suggest potential solutions to move these efforts forward.


Water-assisted one-pot synthesis of N-doped carbon supported Ru catalysts for heterogeneous catalysis.
For the first time, a simple yet efficient water-assisted one-pot pyrolysis (WAOP) strategy was developed to in situ liberate the inaccessible Ru active sites confined inside N-doped carbon. The liberated Ru/CN catalysts exhibit a 9-fold improvement in catalytic activity for quinoline hydrogenation compared with catalysts obtained from the water-free pyrolysis process, and high tolerance for selective hydrogenation of various quinolines substituted with different functional groups. We anticipate that WAOP addresses a key issue that currently plagues carbon-based catalyst synthesis and should lead to improvements in fields as diverse as chemical production and environmental protection.


ERS International Congress 2022: highlights from the Interstitial Lung Diseases Assembly.
This article contains a selection of scientific highlights in the field of interstitial lung diseases (ILDs) presented at the hybrid European Respiratory Society International Congress 2022. Early Career Members of Assembly 12 summarise recent advances in translational and clinical research in idiopathic interstitial pneumonias, ILDs of known origin, sarcoidosis and other granulomatous diseases, and rare ILDs. Many studies focused on evaluation of diagnostic and prognostic (bio)markers, and novel pharmacological and nonpharmacological treatment options for different ILDs. In addition, new insights in clinical, physiological and radiological features of various rare ILDs were presented.


A Discovery of Relevant Hepatoprotective Effects and Underlying Mechanisms of Dietary Clostridium butyricum Against Corticosterone-Induced Liver Injury in Pekin Ducks.
Clostridium butyricum (C. butyricum) can attenuate oxidative stress, inflammation, and hepatic fatty deposition in poultry, however, the underlying mechanisms for this in Pekin ducks remain unclear. This study evaluated these hepatoprotective effects and the underlying mechanisms in a corticosterone (CORT)-induced liver injury model in Pekin ducks fed a C. butyricum intervention diet. A total of 500 Pekin ducks were randomly divided into five groups: one group (CON group) was only provided with a basal diet, three groups were provided a basal diet with 200 mg/kg (LCB group), 400 mg/kg (MCB group), or 600 mg/kg (HCB group) C. butyricum, respectively, and one group was provided a basal diet with 150 mg/kg aureomycin (ANT group) for 42 d. At 37 days-old, all ducks received daily intraperitoneal injections of CORT for five days to establish a liver injury model. C. butyricum intervention alleviated liver injury by decreasing the liver organ indices, hepatic steatosis and hepatocyte necrosis, and improving liver function, antioxidant capacity, and inflammatory factors. Hepatic RNA-seq revealed 365 differentially expressed genes (DEGs) between the MCB and CON groups, with 229 up- and 136 down-regulated DEGs in the MCB group. Between the MCB and ANT groups, 407 DEGs were identified, including 299 up- and 108 down-regulated genes in MCB group. Some DEGs in the MCB group related to oxidative stress and inflammatory responses such as Sod3, Tlr2a/b, and Il10, which were up-regulated, while Apoa1, Cyp7a1, Acsl1/5, Fasn, Ppar-γ, and Scd, which are involved in lipid metabolism, were down-regulated, indicating that these genes were responsive to dietary C. butyricum for the alleviation of corticosterone-induced hepatic injury. Toll-like receptor signaling, PI3K-Akt signaling pathway, cytokine-cytokine receptor interaction, peroxisome proliferator-activated receptor (PPAR) signaling pathway, adipocytokine and glycerophospholipid metabolism signaling pathway were significantly enriched in the MCB group. These findings indicate that C. butyricum intervention can protect Pekin ducks from corticosterone-induced liver injury by the modulation of immunoregulatory- and lipid metabolism-related genes and pathways.


Formulation and in vitro evaluation of upconversion nanoparticle-loaded liposomes for brain cancer.
Aim: This work focused on the development of transferrin-conjugated theranostic liposomes consisting of docetaxel (DXL) and upconversion nanoparticles for the diagnosis and treatment of gliomas. Materials & methods: Upconversion nanoparticles and docetaxel-loaded theranostic liposomes were prepared by a solvent injection method. Formulations were analyzed for physicochemical properties, encapsulation efficiency, drug release, elemental analysis, cytotoxicity and fluorescence. Results: The particle size was around 200 nm with spherical morphology and an encapsulation efficiency of up to 75.93%, was achieved for liposomes with an in vitro drug release of 71.10%. The IC50 values demonstrated enhanced cytotoxicity on C6 glioma cells with targeted liposomes in comparison with nontargeted liposomes. Conclusion: Prepared theranostic liposomes may be promising for clinical validation after an in vitro and in vivo evaluation on cell lines and animals, respectively.


Modulation of glycosaminoglycan biosynthesis by retinoids.
In conclusion, retinoids modulate phenotypic changes such as morphology, adhesion, and growth rate. These changes also result in specific alterations of glycosaminoglycans. We have observed increases in the degree of sulfation in fibroblast matrix heparan sulfate and, a change in the ratio of sulfamido to ester sulfate in matrix heparan sulfate in 407 cell surface. Matrix glycosaminoglycans have been functionally implicated in cellular interactions, and have a specific role in adhesion and growth rates. Our results are consistent with the proposed role for heparan sulfate. It is possible that some of the modulation in cellular behavior resulting from retinoid treatment may be mediated by cell surface and cellular changes in heparan sulfate and other glycosaminoglycans.


Evidence for at least eight Fanconi anemia genes.
Fanconi anemia (FA) is an autosomal recessive chromosomal breakage disorder with diverse clinical symptoms including progressive bone marrow failure and increased cancer risk. FA cells are hypersensitive to crosslinking agents, which has been exploited to assess genetic heterogeneity through complementation analysis. Five complementation groups (FA-A through FA-E) have so far been distinguished among the first 20 FA patients analyzed. Complementation groups in FA are likely to represent distinct disease genes, two of which (FAC and FAA) have been cloned. Following the identification of the first FA-E patient, additional patients were identified whose cell lines complemented groups A-D. To assess their possible assignment to the E group, we introduced selection markers into the original FA-E cell line and analyzed fusion hybrids with three cell lines classified as non-ABCD. All hybrids were complemented for cross-linker sensitivity, indicating nonidentity with group E. We then marked the three non-ABCDE cell lines and examined all possible hybrid combinations for complementation, which indicated that each individual cell line represented a separate complementation group. These results thus define three new groups, FA-F, FA-G, and FA-H, providing evidence for a minimum of eight distinct FA genes.


Persistent Malnutrition and Associated Factors among Children under Five Years Attending Primary Health Care Facilities in Limpopo Province, South Africa.
Despite years of interventions intended to reduce child malnutrition in South Africa, its negative effects, stunting in particular, persist mainly among children under five years old living in under-resourced regions. A cross-sectional study was conducted to determine the prevalence of malnutrition and associated factors among 404 children under age five attending childcare services with their mothers in selected healthcare facilities of Limpopo Province, South Africa. Anthropometry, socio-demographics and obstetric history were collected. Height-for-age, weight-for-age and body mass index-for-age Z-scores were used to determine stunting, underweight and thinness among children, respectively. Logistic regression analyses were performed to generate the factors associated with malnutrition. Stunting (45.3%) was the prevalent form of malnutrition among children under age five, affecting boys (51.7%) more than girls (38.8%) and children aged 12-23 months (62.4%) more than those <11 months old (40.1%), in addition to the overall prevalence of underweight (29.0%) and thinness (12.6%). Boys had increased odds of stunting (adjusted odds ratio, AOR = 2.07, 95% CI: 1.26-3.41, p = 0.004) and underweight (AOR = 2.17, 95% CI: 1.32-3.57, p = 0.002) than girls. Children aged 12-23 months were more likely to be stunted (AOR = 4.79, 95% CI: 2.36-9.75, p ≤ 0.0001) than children aged ≤11 months. Delayed introduction of solid foods increased the odds of stunting (AOR = 5.77, 95% CI: 2.63-12.64, p ≤ 0.0001) and underweight (AOR = 2.05, 95% CI: 1.08-3.89, p = 0.028). Children with normal birth weight were less likely to be thin (AOR = 0.42, 95% CI: 0.19-0.92, p = 0.029) and underweight (AOR = 0.34, 95% CI: 0.17-0.68, p = 0.003) than children who had low birth weight. Children whose mothers had obtained secondary school education (AOR = 0.39, 95% CI: 0.16-0.97, p = 0.044), and Grade 12 or post-Grade 12 education (AOR = 0.32, 95% CI: 0.12-0.83, p = 0.020) were less likely to be stunted than were children of mothers who had only primary school education. Suboptimal complementary feeding predisposed children to stunting and underweight. National nutrition programs should be context-specific to improve the introduction of complementary foods among children, especially in the remote and poor areas.


Interaction of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) with owl monkey kidney cells in enhancing the yields of Herpesvirus saimiri (HVS) and its antigens.
Pre- and posttreatment with N-methyl-N'-nitro-nitrosoguanidine (MNNG) of owl monkey kidney (OMK) cells infected with Herpesvirus saimiri (HVS) resulted in one to three logs higher yields of virus, depending upon the dose of MNNG. A higher percentage of cells also showed HVS early antigen (EA) and late antigen (LA) by immunofluorescence when OMK cells infected with HVS were fed with medium containing MNNG. The high yields of HVS were also observed by electron microscopy. MNNG did not induce HVS-EA in HVS nonproducer lymphoblastoid T cells, nor did it enhance TPA-induced EA to LA. The data suggest that MNNG could be useful in obtaining high yields of virus and/or antigen-producing cells for immunofluorescence or other biochemical experiments, especially from those strains of HVS which grow poorly in vitro. The interaction of MNNG and HVS could also be useful for in vitro transformation or in vivo enhancement of the malignant process.


Interpretation of the effect of an oscillating electric field on membrane enzymes.
Theoretical expressions for the frequency and amplitude dependence of the rate of a catalyzed reaction are fitted to the data of Graziana et al. (1990) [Graziana, A., Ranjeva, R., & Teissié, J. (1990) Biochemistry 29, 8313-8318] for Ca2+ uptake by carrot protoplasts in an oscillating electric field. This uptake is a direct (linear) measure of the rate of increase of ATP caused by a plasma membrane enzyme in the oscillating field. The fit gives 20 ms and 33 microseconds for the relaxation times of the enzyme and roughly 3 for the effective number of elementary changes displaced across the membrane by a conformational change of the enzyme in its catalytic cycle. Additional experiments are suggested to define further the mechanism of the enzymatic reaction.


[Death certificates of women in childbearing age: search for maternal deaths].
In Brazil, there is a lack of complete records on death certificates, and its reliability is questioned, especially for causes attributed to pregnancy and childbirth. We investigated, based on death certificates of women in reproductive age, any fields for identifying maternal deaths. Documentary research, conducted in hospital records. We analyzed in death certificates, maternal and no maternal deaths, inconclusive deaths and hidden deaths. To analyze the underlying causes of death we used ICD 10th Revision. Of the 301 death certificates reviewed, 60% had the fields 43/44 completed, and 40% had these fields blank and/or ignored. We found 58.5% of no maternal deaths, 2% of maternal deaths and 39.5% inconclusive. The analysis of inconclusive deaths allowed us to classify 4.3% as hidden deaths. To overcome the incompletitudes of civil registries, it is necessary that all health professionals be committed to the reliability of the information, so the priority target could be reached.


Differential maturation of beta-adrenoceptor-mediated responses in the lamb fetus.
To study the functional maturity of beta-receptor-mediated responses, seven chronically catheterized lamb fetuses, 93-107 days of gestation, and seven fetuses, 116-134 days of gestation, received intravenous randomly sequenced infusions of isoproterenol (ISO) 0.03, 0.06, and 0.125 micrograms.kg-1.min-1 for 10 min separated by 45-min intervals or two saline infusions followed by 0.125 micrograms.kg-1.min-1 ISO after treatment with 0.5 mg/kg propranolol (PRO). Each fetus received the two treatments 24-48 h apart. In immature fetuses, plasma renin activity (PRA) of 2.0 +/- 0.7 ng.ml-1.h-1 did not change with either protocol. In mature fetuses, PRA of 7.5 +/- 2.5 ng.ml-1.h-1 increased two- to three-fold after the infusion of the highest two doses of ISO (P less than 0.003). Propranolol blocked this response. No significant changes were observed after the infusions of the lowest dose of ISO or saline. Both groups showed significant heart rate increases with all doses of ISO. Propranolol injection decreased heart rate significantly and blocked responses to ISO. We conclude that although a cardiac beta-receptor-mediated response is present by 93 days of gestation in the lamb fetus, a renal beta-receptor-mediated response, renin secretion, is absent.


Attention, intention, and strategy in preparatory control.
The neural mechanisms underlying different forms of preparatory control were examined using event-related fMRI. Preparatory brain activation was monitored in relation to different types of advance information: (1) random task cues indicating which of two possible tasks to perform upon subsequent target presentation; (2) task-ambiguous target stimuli; or (3) targets for which the correct response could be pre-determined. Three types of activation pattern were observed in different brain regions. First, more posterior regions of lateral prefrontal cortex (LPFC) and parietal cortex were activated by both advance task cues and advance targets, but with increased and more sustained activation for the latter. Second, more anterior regions of LPFC and parietal cortex were selectively activated by advance targets. Importantly, in these regions preparatory activation was not further modulated by the availability of advance response information. In contrast, preparatory activation in a third set of brain regions, including medial frontal cortex, reflected the utilization of advance response information, but by only a subset of participants. These results suggest three types of preparatory control: attentional (stimulus-oriented), intentional (action-oriented), and a possibly strategic component that might determine inter-individual differences in response readiness. Notably, the absence of regions selectively or even preferentially activated during cue-based preparation argues against certain conceptualizations of task-selective attention under cued task-switching conditions.


[Poly-traumatic cases. Skills and tricks].
From a personal perspective based on her experience, the author explains various "tricks" and skills which can be used when facing different emergency situations. In this article, the author's recommendations deal with poly-traumatic cases.


Two cases of "hrB-like" autoantibodies appearing as alloantibodies.
Two cases are described in which autoantibodies mimicked alloantibodies. The direct antiglobulin test (DAT) on the red cells (RBCs) from both patients was negative when routine manual hexadimethrine bromide (Polybrene) and enzyme-linked antiglobulin techniques were used. The RBCs also did not react on direct bromelin and direct Polybrene tests. However, an "hrB-like" antibody was eluted from the RBCs of both patients. The sera from these patients reacted with all e+ hrB+ RBCs but not with e+ hrB-, e-, or their own RBCs. The antibody in the serum of one patient was not adsorbed by R2R2 RBCs. Serologic tests initially suggested (by direct testing and adsorption studies) that the serum antibodies were alloantibodies rather than autoantibodies. RBCs taken from one patient, 8 months after her sample was first referred to our laboratory, reacted with a serum sample from her first admission. An RBC sample taken from the other patient, initially typed e+ and hrB- but 1 month later typed e+ and hrB+ by using the same anti-hrB sera, was used to test the earlier samples.


[Effect of using icodextrin as a starting therapy for peritoneal permeability].
Peritoneal permeability differs between patients at the time of starting peritoneal dialysis (PD) and it can increase along with time on the technique. This fact is related to peritonitis, the biocompatibility of the dialysis fluids and the use of glucose as osmotic agent. The aim of the present study was to evaluate if the use of one exchange a day of icodextrine from the time of DP initiation affects the evolution of peritoneal permeability. 56 patients starting PD (mean age: 48.3 +/- 14.0; 62.5% males; 17.9% diabetics) that used one exchange a day with icodextrine from the time of starting PD. We performed a peritoneal transport kinetic study at the time of starting PD and then every 6 months during two years. We calculated the peritoneal mass transfer area coefficient of creatinine (Cr-MTAC) and urea (U-MTAC) as well as the D/P creatinine relationship (D/P Cr). As a control group we used the results of Cr-MTC of 249 patients that had used glucose as the only osmotic agent from the time of starting PD. The peritoneal transport, calculated using Cr-MTC, U-MTC and D/P Cr, diminished at 12 months (11.7+/-5.7 vs. 8.1+/-3.1; 23.5+/-7.3 vs. 18.9+/-3.8; 0.72+/-0.09 vs. 0.67+/-0.08; respectively), staying stable afterwards.We found that high transporters (HA) patients (higher quatril Cr-MTC ) showed a higher diminution of Cr-MTAC along the first year of treatment. The diminution of Cr-MTAC after 12 months using icodextrine was significantly higher (p<0.001) that the one observed in the control group that only used glucose as osmotic agent (10.5+/-5.3 vs. 10.1+/-4.6). We found that high transporters (HA) patients (higher quatril Cr-MTC) showed a higher decrease of Cr-MTAC along the first year of treatment. the use of icodextrine at the time of starting PD might help to correct the high transport status observed in some patients during the first months of treatment. The peritoneal transport kinetic studies performed at 6 and 12 months after starting PD are more representative of the long term peritoneal transport characteristics of the patients than those performed at the time of starting PD.


Effects of naloxone on limited-access ethanol drinking in rats.
The hypothesis that naloxone (NAL) decreases oral ethanol intake in rats by inducing a conditioned taste aversion (CTA) to ethanol was investigated. Rats were trained to drink 8% ethanol (v/v) on a 1-hr limited-access schedule. They received 4 days of intraperitoneal injections of 10 mg/kg of NAL, 10 min before limited-access (-10MIN group), immediately after limited-access (1HR group), or 3 hr after limited-access (3HR). Ethanol intake decreased in the -10MIN and 1HR groups during the injection period and on the postinjection day. In experiment 2, rats received 4 days of NAL injections when ethanol was not available (pre-exposure), and then the paradigm was repeated. In this experiment, there was no suppression of ethanol intake for any group on the postinjection day. The decrease in ethanol intake during injections observed for the 1HR in experiment 1 and the sustained suppression postinjection was interpreted as a CTA. Pre-exposure in experiment 2 abolished the CTA. Differences in the pattern ethanol intake for the -10MIN and 3HR groups during the experiments, however, suggest that a CTA is not the sole mechanism underlying NAL's suppressant effects on ethanol intake. In conclusion, in rats both the dose of NAL and the relative timing of NAL injections and ethanol drinking effect subsequent NAL suppression of ethanol intake.


MR Arthrography of the Hip.
Magnetic resonance imaging (MRI) of the hip has been valuable in the diagnosis of occult osseous abnormalities and of periarticular soft tissue disorders. MRI has been less useful in the evaluation of acetabular labral tears and other intra-articular abnormalities. Image optimization is more technically challenging in the hip than in smaller joints because the overlying soft tissues are thicker, resulting in decreased spatial and contrast resolutions that may not be adequate to distinguish the acetabular labrum from subchondral bone, articular cartilage, and joint capsule. MR arthrography (MRA) extends the capabilities of conventional MRI because contrast solution separates intra-articular structures and outlines abnormalities. In hips with suspected acetabular labral injury, arthrographic MR images demonstrate the location and length of tears and the presence of associated capsular defects. Arthrographic MR images may also enable the accurate diagnosis of cartilage lesions and intra-articular loose bodies. This article illustrates normal arthrographic MR features of the hip as well as pathologic disorders of the acetabular labrum, capsule, and articular cartilage


Abnormal alkaline phosphatase isoenzymes detected in the serum of elderly patients.
Alkaline phosphatase (ALP) isoenzyme analysis of 101,832 serum samples was performed by electrophoresis using cellulose acetate membrane, and abnormal bands at the alpha(1) and alpha(2) globulin positions were detected in 23 samples. The physicochemical properties of these abnormal ALP isoenzyme fractions were examined. In brief, the abnormal fractions were heat-sensitive and inhibited by L-phenylalanine, and neither sialic acid in their polysaccharide chains nor the glycosyl-phosphatidylinositol anchor was detected by the enzyme treatment. The physicochemical properties of abnormal ALP isoenzyme fractions detected in Patients 1 - 22 were similar to those of the adult small intestine type. However, the molecular weight of the adult small intestine type abnormal fractions was smaller than that of the normal fractions. These adult small intestine type abnormal bands at the alpha(1)- to alpha(2)-globulin positions, which were identified in the serum of Patients 1 - 22, were detected in elderly patients. Most of them had various basal diseases such as renal insufficiency, fracture, interstitial pneumonia, and chronic pancreatitis. Some of them had severe diseases such as rectal cancer, descending colon cancer, and septic shock. In Patient 23, the polysaccharide chain had sialic acid and was heat sensitive physicochemical properties that were similar to those of the Kasahara ALP variant.


Depot neuroleptics: side effects and safety.
Long-acting depot neuroleptics are likely to receive increasing consideration in the maintenance treatment of schizophrenia because of the growing recognition of noncompliance with daily oral medication in this patient population. The impending availability of haloperidol decanoate in the United States may also increase the use of long-acting neuroleptics. The long-term safety of these drugs, therefore, deserves close scrutiny. Based on experiences with other depot neuroleptics, the safety profile of haloperidol decanoate is unlikely to differ from short-acting formulations of haloperidol. In general, tardive dyskinesia remains the most troubling problem associated with long-term neuroleptic treatment. The available information indicates that all neuroleptics, including long-acting preparations, share the same propensity for causing tardive dyskinesia. Furthermore, the use of the lowest effective dose of a neuroleptic appears to minimize the severity of tardive dyskinesia should it develop. Extrapyramidal symptoms, notably akinesia and akathisia, are also potential concerns of maintenance treatment with oral and depot neuroleptics; however, the occurrence of these side effects appears dose related. Thus, the clinician can favorably influence the benefit-to-risk ratio of long-acting depot neuroleptics by minimizing potential side effects through the use of a low dose strategy.


Staphylococcus aureus Bacteremia: Contemporary Management.
Staphylococcus aureus bacteremia (SAB) is a serious cause of bloodstream infection associated with significant morbidity and mortality. Complications include deep-seated foci of infection including infective endocarditis, device-associated infection, osteoarticular metastases, pleuropulmonary involvement, and recurrent infection. With the 30-day all-cause mortality being around 20%, a collaborative effort of early Infectious Diseases (ID) consultation and Antimicrobial Stewardship Program (ASP) involvement will show improved SAB outcomes and therapy optimization.1.


The time course of attentional disengagement from angry faces in social anxiety.
While impaired attentional disengagement from threatening stimuli is thought to enhance social anxiety, it is unclear when the impaired disengagement occurs accurately. We used a gap task (Experiment 1) and an overlap task (Experiment 2) to reveal the impaired attentional disengagement from angry faces in socially anxious people with non-treatment seeking undergraduates. High (N = 17 in Experiments 1 and 2) and low socially anxious people (N = 17 in Experiment 1 and 19 in Experiment 2) were asked to fixate on an angry or neutral face presented at the center of a screen. Then, they discriminated the peripheral target stimuli. When there was a temporal gap between the face and target in Experiment 1 (gap task), the reaction times (RTs) for angry and neutral faces did not differ for all participants. However, when there was no gap and the face continued to appear in Experiment 2 (overlap task), the RTs for angry faces in high socially anxious people were longer than those for neutral faces after presentation times of 300 ms or longer. In low socially anxious people, the RTs following the angry and neutral faces did not differ. These results suggest that high socially anxious people face difficulty in disengaging attention from angry faces after recognizing them.


Insight of Genus Corynebacterium: Ascertaining the Role of Pathogenic and Non-pathogenic Species.
This review gathers recent information about genomic and transcriptomic studies in the Corynebacterium genus, exploring, for example, prediction of pathogenicity islands and stress response in different pathogenic and non-pathogenic species. In addition, is described several phylogeny studies to Corynebacterium, exploring since the identification of species until biological speciation in one species belonging to the genus Corynebacterium. Important concepts associated with virulence highlighting the role of Pld protein and Tox gene. The adhesion, characteristic of virulence factor, was described using the sortase mechanism that is associated to anchorage to the cell wall. In addition, survival inside the host cell and some diseases, were too addressed for pathogenic corynebacteria, while important biochemical pathways and biotechnological applications retain the focus of this review for non-pathogenic corynebacteria. Concluding, this review broadly explores characteristics in genus Corynebacterium showing to have strong relevance inside the medical, veterinary, and biotechnology field.


Developments in the Catalytic Asymmetric Synthesis of Agrochemicals and Their Synthetic Importance.
Catalytic asymmetric synthesis has become an essential tool for the enantioselective synthesis of pharmaceuticals, natural products, and agrochemicals (mainly fungicides, herbicides, insecticides, and pheromones). With continuous growing interest in both modern agricultural chemistry and catalytic asymmetric synthesis chemistry, this review provides a comprehensive overview of some earlier reports as well as the recent successful applications of various catalytic asymmetric syntheses methodologies, such as enantioselective hydroformylation, enantioselective hydrogenation, asymmetric Sharpless epoxidation and dihydroxylation, asymmetric cyclopropanation or isomerization, organocatalyzed asymmetric synthesis, and so forth, which have been used as key steps in the preparation of chiral agrochemicals (on R&D, piloting, and commercial scales). Chiral agrochemicals can also lead the new generation of such chemicals having specific and novel modes of action for achieving sustainable crop protection and production. Some perspectives and challenges for these catalytic asymmetric methodologies in the synthesis of chiral agrochemicals are also briefly discussed in the final section of the manuscript. This review will provide the insight regarding understanding, development, and evaluation of catalytic asymmetric systems for the synthesis of chiral agrochemicals among the agrochemists.


A phase I/II study of siltuximab (CNTO 328), an anti-interleukin-6 monoclonal antibody, in metastatic renal cell cancer.
Serum interleukin (IL)-6 levels correlate with disease outcomes in renal cell carcinoma (RCC) patients. Siltuximab, a chimeric, murine-human mAb against IL-6, was evaluated in a three-part phase I/II study in patients with progressive metastatic RCC. In part 1, 11 patients received 1, 3, 6, or 12mgkg-¹ at weeks 1, 4 and q2w × 2 thereafter; in part 2, 37 patients randomly received 3 or 6 mgkg-¹ q3w × 4; in part 3, 20 low-risk patients received 6mgkg-¹ q2w × 6. Modified WHO response criteria were assessed at weeks 7, 11, the 6-week follow-up, and when clinically indicated. Siltuximab was well tolerated overall, with no maximum tolerated dose or immune response observed. In all, 5 out of 11, 17 out of 37, and 9 out of 20 patients in parts 1, 2, and 3, respectively, received extended treatment beyond 4-6 initial infusions. In part 2, stable disease (SD) (≥11weeks) or better was achieved by 11 out of 17 (65%) 3 mgkg-¹ treated patients (one partial response (PR) ~8 months, 10 SD) and 10 out of 20 (50%) 6mgkg-¹ treated patients (10 SD). In part 3, documented complete or PR was not observed, but 13 out of 20 (65%) patients achieved SD. Siltuximab stabilised disease in >50% of progressive metastatic RCC patients. One PR was observed. Given the favourable safety profile of siltuximab and poor correlation of tumour shrinkage with clinical benefit demonstrated for other non-cytotoxic therapies, further evaluation of dose-escalation strategies and/or combination therapy may be considered for patients with RCC.


Infectious complications of human T cell leukemia/lymphoma virus type I infection.
Infection with human T cell leukemia/lymphoma virus type I (HTLV-I) has been etiologically associated with two diseases: adult T cell leukemia and HTLV-I-associated myelopathy/tropical spastic paraparesis. Increasing evidence suggests that HTLV-I infection may be associated with immunosuppression and, as a consequence, affect the risk and expression of several other infectious diseases, of which the best studied are strongyloidiasis, tuberculosis, and leprosy. In strongyloidiasis, coinfection with HTLV-I appears to result in a higher rate of chronic carriage, an increased parasite load, and a risk of more severe infection. In tuberculosis, a decrease in delayed-type hypersensitivity to Mycobacterium tuberculosis has been established, but whether this decrease is clinically significant has yet to be determined. In leprosy, an increased risk of disease is suggested, but the published studies are all too poorly controlled to draw definite conclusions.


Prognostic factors in first-time care seekers due to acute low back pain.
There is limited knowledge on prognostic factors for developing chronic low back pain (LBP) at an early stage of LBP. The objectives of this study were to investigate the clinical course of pain and disability, and prognostic factors for non-recovery after 1-year, in patients seeking help for the first time due to acute LBP. An inception cohort study included 123 patients with acute LBP lasting less than 3 weeks and consulting primary care for the first time. Main outcome measures were pain intensity, Roland-Morris disability questionnaire (RMQ), and sickness absence. Eleven patients (9%) did not return for the 12-month follow-up. There were large and significant reductions in pain intensity (P<0.001) and RMQ scores (P<0.001) during follow-up. Patients with neurological signs showed significantly less improvement in pain (P=0.001) and RMQ (P=0.004) compared with those without neurological signs. The proportions with sickness absence due to LBP at 6, 9, and 12 months were 7%, 8%, and 9%, respectively. At 12 months, 17% of patients had not fully recovered. Multivariate logistic regression analyses showed that high scores on a psychosocial screening (acute low back pain screening questionnaire) and emotional distress (Hopkin's symptom check list) were significantly associated with non-recovery at 12 months, with odds ratios of 4.4 (95% confidence interval 1.1-17.4) and 3.3 (1.1-10.2), respectively.


Structure-activity relationships and human relevance for perfluoroalkyl acid-induced transcriptional activation of peroxisome proliferation in liver cell cultures.
Perfluoroalkyl acids (PFAAs) are widely distributed and environmentally persistent agents whose potential toxicity is not yet fully characterized. Perfluorooctanoic acid (PFOA) and perfluorooctane sulfonic acid elicit a number of potential toxicities in rodents, the most prevalent of which are governed by activation of the peroxisome proliferator-activated receptor alpha (PPARalpha). The purpose of this investigation was twofold: (1) To conduct a structure-activity relationship study of the transcriptional activation of peroxisome proliferation in primary rat liver cell cultures for PFAA-related carboxylic and sulfonic acids of varying carbon chain length and (2) to explore whether this activity can be translated to human liver cells in culture. Exposure to PFOA caused a dose-dependent stimulation of the expression of acyl-CoA oxidase (Acox), Cte/Acot1, and Cyp4a1/11 transcripts that are indicative of peroxisome proliferation in primary rat hepatocytes. PFOA concentrations of 30 microM and above caused cell injury characterized by the expression of Ddit3. Perfluorobutanoic acid (PFBA), on the other hand, stimulated Acox, Cte/Acot1, and Cyp4a1/11 gene expression in primary rat hepatocytes only at concentrations of 100 microM and above. Neither PFOA nor PFBA at concentrations up to 200 microM stimulated PPARalpha-related gene expression in either primary or HepG2 human liver cells. These data demonstrate that (1) PFFAs cause a concentration- and chain length-dependent increase in expression of gene targets related to cell injury and PPARalpha activation in primary rat hepatocytes, (2) the sulfonates are less potent than the corresponding carboxylates in stimulating PPARalpha-related gene expression in rat hepatocytes, and (3) stimulation of PPARalpha-mediated gene transcription is a mechanism that is not shared by human liver cells, adding further substantiation that PPARalpha-dependent liver toxicity in rodents does not extrapolate to assessing human health concerns.


Phospholipase D hydrolysis of choline phosphoglycerides is selective for the alkyl-linked subclass of Madin-Darby canine kidney cells.
Madin-Darby canine kidney (MDCK) cells were used to study the synthesis of diglycerides from choline phospholipids (PC) in response to 12-O-tetradecanoylphorbol-13-acetate (TPA). In this system, diglyceride formation was blocked in the presence of ethanol (0.5%), and a corresponding amount of phosphatidylethanol (PEt) was formed, indicating that phospholipase D is responsible for the diglyceride production. Analysis of the subclasses of phosphatidylethanol revealed 1-O-alkyl-(alkyl), 1-O-alk-1'-enyl-(alkenyl), and 1-acyl species of PEt (38.0, 8.3, and 53.7%, respectively). The molecular species of the alkyl-PEt most closely matched the alkyl-PC. No change in the relative amounts of alkyl- versus acyl-PEt was observed with time after stimulation. Comparison of the alkyl content of PEt (38.0%) and the parent PC (15.2%) indicated a marked selectivity for the alkyl subclass of PC. A cell-free assay (Huang, C., Wykle, R. L., Daniel, L. W., and Cabot, M. C. (1992) J. Biol. Chem. 267, 16859-16865) for phospholipase D was also used to confirm the selectivity of the enzyme for alkyl-PC versus acyl-PC. The predominant molecular species of PEt contained saturated acyl or alkyl chains in position-1 and monounsaturated residues in position-2 accounting for approximately 50% of the total PEt. 1-O-Octadecyl-2-oleoyl-sn-glycerol, a representative alkyl molecular species, was synthesized and tested for its effect upon protein kinase C derived from MDCK cells. This alkyl-diglyceride (DG) neither stimulated protein kinase C nor inhibited its activation by diacylglycerol. In summary, TPA-stimulated phospholipase D is selective for the alkyl-PC subclass in MDCK cells. The alkyl-DG subsequently formed does not appear to function as a second-messenger in activating protein kinase C.


Weaning practices in children up to 19 months of age in Madrid.
The weaning practices of 344 children in Madrid, aged 3-19 months, were investigated by controlled interview of their mothers or caretakers. The results were compared with national and international guidelines for the introduction of complementary foods; there are no Spanish guidelines. We found only minor differences between the practices in three different socioeconomic groups. Generally, mothers in Madrid followed national and international guidelines well. However, more attention should be paid to aspects such as the timing of the introduction of the first complementary food, the nature of gluten, the addition of salt and sugar to baby meals and the introduction of potentially allergenic foods. Also, manufacturers of baby foods that are sold in Spain could play a useful role by reducing the number of ingredients used in products advised for the initial phase of weaning and by avoiding the use of salt and sugar in their products.


Detection of Plasmodium falciparum DNA in saliva samples stored at room temperature: potential for a non-invasive saliva-based diagnostic test for malaria.
Current malaria diagnostic methods require blood collection, that may be associated with pain and the risk of transmitting blood-borne pathogens, and often create poor compliance when repeated sampling is needed. On the other hand, the collection of saliva is minimally invasive; but saliva has not been widely used for the diagnosis of malaria. The aim of this study was to evaluate the diagnostic performance of saliva collected and stored at room temperature using the OMNIgene®•ORAL kit for diagnosing Plasmodium falciparum malaria. Paired blood and saliva samples were collected from 222 febrile patients in Cameroon. Saliva samples were collected using the OMNIgene®•ORAL (OM-501) kit and stored at room temperature for up to 13 months. Thick blood film microscopy (TFM) was used to detect P. falciparum blood-stage parasites in blood. Detection of P. falciparum DNA in blood and saliva was based on amplification of the multi-copy 18 s rRNA gene using the nested-polymerase chain reaction (nPCR). Prevalence of malaria detected by TFM, nPCR-saliva and nPCR-blood was 22, 29, and 35%, respectively. Using TFM as the gold standard, the sensitivity of nPCR-saliva and nPCR-blood in detecting P. falciparum was 95 and 100%, respectively; with corresponding specificities of 93 and 87%. When nPCR-blood was used as gold standard, the sensitivity of nPCR-saliva and microscopy was 82 and 68%, respectively; whereas, the specificity was 99 and 100%, respectively. Nested PCR-saliva had a very good agreement with both TFM (kappa value 0.8) and blood PCR (kappa value 0.8). At parasitaemia > 10,000 parasites/µl of blood, the sensitivity of nPCR-saliva was 100%. Nested PCR-saliva detected 16 sub-microscopic malaria infections. One year after sample collection, P. falciparum DNA was detected in 80% of saliva samples stored at room temperature. Saliva can potentially be used as an alternative non-invasive sample for the diagnosis of malaria and the OMNIgene®•ORAL kit is effective at transporting and preserving malaria parasite DNA in saliva at room temperature. The technology described in this study for diagnosis of malaria in resource-limited countries adds on to the armamentarium needed for elimination of malaria.


The 'rejuvenating factor' TIMP-2 is detectable in human blood components for transfusion.
Tissue inhibitor of metalloproteinases 2 (TIMP-2) is a protein suspected to be crucial in numerous physiological and pathological processes such as morphogenesis, tissue remodelling and metastasis suppression. In animal models, the administration of TIMP-2 to aged mice improved their cognitive functions. Therefore, one can hypothesize that differences in TIMP-2 levels between blood donors and recipients might influence cognitive functions also in humans. However, the stability of TIMP-2 during processing and storage of blood components for transfusion has not been intensively investigated so far. This study determined TIMP-2 concentrations in fresh-frozen plasma (FFP), erythrocyte concentrate (EC) and pathogen-inactivated platelet concentrate (PI-PC) depending on the donor's demographic factors age and gender. Tissue inhibitor of metalloproteinases 2 was measured in FFP (n = 30), EC (n = 12) and PI-PC (n = 12) using a Q-Plex single-plex immunoassay for chemiluminescence-based detection. Absolute quantification of TIMP-2 was performed with Q-view software. Fresh umbilical cord plasma was used as a positive control. Tissue inhibitor of metalloproteinases 2 was detected in FFP (30/30 samples), EC (11/12 samples) and PI-PC (12/12 samples). The median TIMP-2 concentration in EC (17·2 ng/ml; range: 0-26·5 ng/ml) was significantly lower compared with FFP (63·4 ng/ml; range: 44·4-87·3 ng/ml) or PI-PC (69·9 ng/ml; range: 39·9-83·6 ng/ml). Across all blood components, TIMP-2 levels are comparable in male and female donors and independent of age. Tissue inhibitor of metalloproteinases 2 is detectable and stable in FFP, PI-PC and, in low concentration, EC. It can be hypothesized that TIMP-2 will be transmitted to recipients during transfusion.


Flavonoids from Tephrosia major. A new prenyl-beta-hydroxychalcone.
The roots and aerial parts of Tephrosia major Micheli, afforded a new prenylated-beta-hydroxychalcone, characterized as 2',6'-dihydroxy-3'-prenyl-4'-methoxy-beta-hydroxychalcone. In addition, seven prenylated flavonoids, two rotenoids, beta-sitosterol, stigmasterol, lupeol and quercetin were isolated. The structure of the new beta-hydroxy chalcone was established by spectroscopic methods, including 2D NMR experiments.


Notch signaling in mammalian hair cell regeneration.
In the inner ear, Notch signaling has been shown to have two key developmental roles. The first occurs early in otic development and defines the prosensory domains that will develop into the six sensory organs of the inner ear. The second role occurs later in development and establishes the mosaic-like pattern of the mechanosensory hair cells and their surrounding support cells through the more well-characterized process of lateral inhibition. These dual developmental roles have inspired several different strategies to regenerate hair cells in the mature inner ear organs. These strategies include (1) modulation of Notch signaling in inner ear stem cells in order to increase hair cell yield, (2) activation of Notch signaling in order to promote the formation of ectopic sensory regions in normally non-sensory regions within the inner ear, and (3) inhibition of Notch signaling to disrupt lateral inhibition and allow support cells to transdifferentiate into hair cells. In this review, we summarize some of the promising studies that have used these various strategies for hair cell regeneration through modulation of Notch signaling and some of the challenges that remain in developing therapies based on hair cell regeneration.


Molecular genetics of signal transduction in Dictyostelium.
In conditions of starvation, the free living amoebae of Dictyostelium enter a developmental program: The cells aggregate by chemotaxis to form a multicellular structure that undergoes morphogenesis and cell-type differentiation. These processes are mediated by a family of cell surface cAMP receptors (cARs) that act on a specific heterotrimeric G protein to stimulate actin polymerization, activation of adenylyl and guanylyl cyclases, and a host of other responses. Most of the components in these pathways have mammalian counterparts. The accessible genetics of this unicellular organism facilitate structure-function analysis and enable the discovery of novel genes involved in the regulation of these important pathways.


Characterizations of the Interactions between Escherichia coli Periplasmic Chaperone HdeA and Its Native Substrates during Acid Stress.
The bacterial acid-resistant chaperone HdeA is a "conditionally disordered" protein that functions at low pH when it undergoes a transition from a well-folded dimer to an unfolded monomer. The dimer dissociation and unfolding processes result in exposure of hydrophobic surfaces that allows binding to a broad range of client proteins. To fully elucidate the chaperone mechanism of HdeA, it is crucial to understand how the activated HdeA interacts with its native substrates during acid stress. Herein, we present a nuclear magnetic resonance study of the pH-dependent HdeA-substrate interactions. Our results show that the activation of HdeA is not only induced by acidification but also regulated by the presence of unfolded substrates. The variable extent of unfolding of substrates differentially regulates the HdeA-substrate interaction, and the binding further affects the HdeA conformation. Finally, we show that HdeA binds its substrates heterogeneously, and the "amphiphilic" model for HdeA-substrate interaction is discussed.


Ethnobotany of food plants in the high river Ter valley (Pyrenees, Catalonia, Iberian Peninsula): non-crop food vascular plants and crop food plants with medicinal properties.
The present study reports a part of the findings of an ethnobotanical research project conducted in the Catalan region of the high river Ter valley (Iberian Peninsula), concerning the use of wild vascular plants as food and the medicinal uses of both wild and cultivated food plants. We have detected 100 species which are or have been consumed in this region, 83 of which are treated here (the remaining are the cultivated food plants without additional medicinal uses). Some of them, such as Achillea ptarmica subsp. pyrenaica, Convolvulus arvensis, Leontodon hispidus, Molopospermum peloponnesiacum and Taraxacum dissectum, have not been previously reported, or have only very rarely been cited or indicated as plant foods in very restricted geographical areas. Several of these edible wild plants have a therapeutic use attributed to them by local people, making them a kind of functional food. They are usually eaten raw, dressed in salads or cooked; the elaboration of products from these species such as liquors or marmalades is a common practice in the region. The consumption of these resources is still fairly alive in popular practice, as is the existence of homegardens, where many of these plants are cultivated for private consumption.


Human B-1 and B-2 B Cells Develop from Lin-CD34+CD38lo Stem Cells.
The B-1 B cell population is an important bridge between innate and adaptive immunity primarily because B-1 cells produce natural Ab. Murine B-1 and B-2 cells arise from distinct progenitors; however, in humans, in part because it has been difficult to discriminate between them phenotypically, efforts to pinpoint the developmental origins of human B-1 and B-2 cells have lagged. To characterize progenitors of human B-1 and B-2 cells, we separated cord blood and bone marrow Lin-CD34+ hematopoietic stem cells into Lin-CD34+CD38lo and Lin-CD34+CD38hi populations. We found that transplanted Lin-CD34+CD38lo cells, but not Lin-CD34+CD38hi cells, generated a CD19+ B cell population after transfer into immunodeficient NOD.Cg-Prkdcscid Il2rgtm1wjl/SxJ neonates. The emergent CD19+ B cell population was found in spleen, bone marrow, and peritoneal cavity of humanized mice and included distinct populations displaying the B-1 or the B-2 cell phenotype. Engrafted splenic B-1 cells exhibited a mature phenotype, as evidenced by low-to-intermediate expression levels of CD24 and CD38. The engrafted B-1 cell population expressed a VH-DH-JH composition similar to cord blood B-1 cells, including frequent use of VH4-34 (8 versus 10%, respectively). Among patients with hematologic malignancies who underwent hematopoietic stem cell transplantation, B-1 cells were found in the circulation as early as 8 wk posttransplantation. Altogether, our data demonstrate that human B-1 and B-2 cells develop from a Lin-CD34+CD38lo stem cell population, and engrafted B-1 cells in humanized mice exhibit an Ig-usage pattern comparable to B-1 cells in cord blood.


Secretory co-factors in next-generation cellular therapies for cancer.
Since chimeric antigen receptor (CAR) T-cell therapies for hematologic malignancies were approved by the U.S. Food and Drug Administration, numerous "next-generation" CAR T cells have been developed to improve their safety, efficacy, and applicability. Although some of these novel therapeutic strategies are promising, it remains difficult to apply these therapies to solid tumors and to control adverse effects, such as cytokine release syndrome and neurotoxicity. CAR T cells are generated using highly scalable genetic engineering techniques. One of the major strategies for producing next-generation CAR T cells involves the integration of useful co-factor(s) into the artificial genetic design of the CAR gene, resulting in next-generation CAR T cells that express both CAR and the co-factor(s). Many soluble co-factors have been reported for CAR T cells and their therapeutic effects and toxicity have been tested by systemic injection; therefore, CAR T cells harnessing secretory co-factors could be close to clinical application. Here, we review the various secretory co-factors that have been reported to improve the therapeutic efficacy of CAR T cells and ameliorate adverse events. In addition, we discuss the different co-factor expression systems that have been used to optimize their beneficial effects. Altogether, we demonstrate that combining CAR T cells with secretory co-factors will lead to next-generation CAR T-cell therapies that can be used against broader types of cancers and might provide advanced tools for more complicated synthetic immunotherapies.


The structure of hepatitis B envelope and molecular variants of hepatitis B virus.
Accumulated evidence in recent years has shown that the variation of hepatitis B virus (HBV) genomes may have profound implications for our understanding of hepatitis B pathogenesis and prevention. Attention has focused on areas of the outer envelope coded by the S gene which are involved in the induction of a protective neutralising antibody response, and mutations which directly affect the production of C gene products, one of which is considered as a target for immune T cells involved in virus clearance. This review highlights recent experimental data which emphasizes the role of such mutations in the establishment and maintenance of chronic HBV infections and focuses attention on the significance of HBV variants with respect to the expanding use of HBV vaccines for mass immunization.


Comparison between the story recall test and the word-list learning test in Korean patients with mild cognitive impairment and early stage of Alzheimer's disease.
Among verbal memory tests, two that are commonly used to measure the ability of verbal memory function in cognitive impairment are story recall tests and word-list learning tests. However, research is limited regarding which test might be more sensitive in discriminating between normal cognitive aging and patients with Alzheimer's disease (AD) in the Korean population. The purpose of the current study was to compare the word-list learning test (Seoul Verbal Learning Test; SVLT) and the story recall test (Korean Story Recall Test; KSRT) to determine which test is more sensitive in discriminating between individuals with normal cognitive aging and patients with mild cognitive impairment (MCI) and early stage of AD in Korea. A total of 53 healthy adults, 127 patients with MCI, and 72 patients with early stage of AD participated in this study. The receiver-operating characteristic (ROC) curve and area under the curve (AUC) were evaluated to compare these two tests. The results showed that the AUC of the SVLT was significantly larger than the AUC of the KSRT in all three groups (healthy adults vs. MCI and early stage of AD; healthy adults vs. MCI; healthy adults vs. early stage of AD). However, in comparison of patients with MCI and early stage of AD, the AUC of SVLT and the AUC of KSRT were not significant. The word-list learning test is a more useful tool for examining verbal memory function for older adults in Korea than the story recall test.


Identification of residues involved in neurotensin binding and modeling of the agonist binding site in neurotensin receptor 1.
The neurotensin receptor 1 (NTR1) subtype belongs to the family of G protein-coupled receptors and mediates most of the known effects of the neuropeptide including modulation of central dopaminergic transmission. This suggested that nonpeptide agonist mimetics acting at the NTR1 might be helpful in the treatment of Parkinson's disease and schizophrenia. Here, we attempted to define the molecular interactions between neurotensin-(8-13), the pharmacophore of neurotensin, and the rat NTR1. Mutagenesis of the NTR1 identified residues that interact with neurotensin. Structure-activity studies with neurotensin-(8-13) analogs identified the peptide residues that interact with the mutated amino acids in the receptor. By taking these data into account, computer-assisted modeling techniques were used to build a tridimensional model of the neurotensin-(8-13)-binding site in which the N-terminal tetrapeptide of neurotensin-(8-13) fits in the third extracellular loop and the C-terminal dipeptide binds to residues at the junction between the extracellular and transmembrane domains of the receptor. Interestingly, the agonist binding site lies on top of the previously described NTR1-binding site for the nonpeptide neurotensin antagonist SR 48692. Our data provide a basis for understanding at the molecular level the agonist and antagonist binding modes and may help design nonpeptide agonist mimetics of the NTR1.


A multitarget assay for inhibitors of membrane-associated steps of peptidoglycan biosynthesis.
Peptidoglycan synthesis begins in the cytoplasm with the condensation of UDP-N-acetyl glucosamine (UDP-GlcNAc) and phosphoenolpyruvate catalyzed by UDP-N-acetylglucosamine enolpyruvoyl transferase. UDP-GlcNAc is also utilized as substrate for the glycosyltransferase MurG, a membrane-bound enzyme that catalyzes the production of lipid II. Membranes from Escherichia coli cells overproducing MurG support peptidoglycan formation at a rate approximately fivefold faster than membranes containing wild-type levels of MurG. Conditions have been optimized for the production of large amounts of membranes with increased levels of MurG, allowing the development of an assay suitable for high-throughput screening of large compound libraries. The quality of the purified membranes was assessed by electron microscopy and also by testing cross-linked peptidoglycan production. Moreover, kinetic studies allowed the determination of optimal concentrations of the substrates and membranes to be utilized for maximum sensitivity of the assay. Using a 96-well assay format, the IC50 values for vancomycin, tunicamycin, flavomycin, and bacitracin were 1.1 microM, 0.01 microg/ml, 0.03 microg/ml, and 0.7 microg/ml, respectively.


Cariology in Japanese dental schools.
The aim of this study was to evaluate the teaching of cariology in Japanese dental schools. Postal questionnaires were sent to all Japanese dental schools. Twenty-five of the 29 Japanese dental schools (response rate: 86%). The results were in many areas similar to those reported from North America. The non-surgical approach to the management of primary caries predominated, but no schools used bacteriological tests in caries diagnosis and only one school recommended antibacterial treatment. Four of the 25 responding schools advocated the treatment of primary enamel lesions by surgical intervention while three awaited operative intervention until the lesions had reached the middle third of the dentine. Recommendation of fluoride treatment was less common than in North America. The use of electronic devices to diagnose occlusal caries lesions was more common than in North America, but 'explorer catch' was the most frequently used method to diagnose primary and secondary caries in dental schools in both regions. Research in cariology over the years has led to altered views on how and when to treat caries lesions. The interpretation of research findings and the transfer of knowledge by teachers in dental schools have an impact on the practice of operative dentistry, but the diversity in the teaching of cariology indicates that the interpretation and implementation of modern principles in the treatment of caries differ quite markedly in Japanese dental schools.


Outpatient treatment and early discharge of symptomatic pulmonary embolism: a systematic review.
The published literature regarding the safety of outpatient treatment of symptomatic pulmonary embolism (PE) was systematically summarised. A literature search was performed using the PubMed and EMBASE databases. Studies in which patients had symptomatic PE and the antithrombotic treatment was administered completely at home or the patients were discharged early were selected. A scoring system was used to divide studies into three quality categories. Short- and long-term outcomes were extracted: all-cause mortality, death from PE or from major haemorrhage, recurrent venous thromboembolism, and major bleeding. Eleven observational studies were included. No randomised controlled studies were identified. No study fulfilled the criteria for high quality. A total of 928 patients with symptomatic PE were treated completely as outpatients or discharged early; haemodynamic instability and hypoxia were the main exclusion criteria. No patient died during the first 7 days of antithrombotic treatment. Outpatient treatment of symptomatic PE is not based on high-quality evidence. Although the published data suggest that certain subgroups of haemodynamically and respiratorily stable patients may be safely treated at home when a well-defined management programme is applied, further studies are warranted for a short-term prognostic risk stratification of this PE subgroup.


Transcriptomic Changes Associated with Pregnancy in a Marsupial, the Gray Short-Tailed Opossum Monodelphis domestica.
Live birth has emerged as a reproductive strategy many times across vertebrate evolution; however, mammals account for the majority of viviparous vertebrates. Marsupials are a mammalian lineage that last shared a common ancestor with eutherians (placental mammals) over 148 million years ago. Marsupials are noted for giving birth to highly altricial young after a short gestation, and represent humans' most distant viviparous mammalian relatives. Here we ask what insight can be gained into the evolution of viviparity in mammals specifically and vertebrates in general by analyzing the global uterine transcriptome in a marsupial. Transcriptome analyses were performed using NextGen sequencing of uterine RNA samples from the gray short-tailed opossum, Monodelphis domestica. Samples were collected from late stage pregnant, virgin, and non-pregnant experienced breeders. Three different algorithms were used to determine differential expression, and results were confirmed by quantitative PCR. Over 900 opossum gene transcripts were found to be significantly more abundant in the pregnant uterus than non-pregnant, and over 1400 less so. Most with increased abundance were genes related to metabolism, immune systems processes, and transport. This is the first study to characterize the transcriptomic differences between pregnant, non-pregnant breeders, and virgin marsupial uteruses and helps to establish a set of pregnancy-associated genes in the opossum. These observations allowed for comparative analyses of the differentially transcribed genes with other mammalian and non-mammalian viviparous species, revealing similarities in pregnancy related gene expression over 300 million years of amniote evolution.


Arsenic-enriched groundwaters of India, Bangladesh and Taiwan--comparison of hydrochemical characteristics and mobility constraints.
Arsenic (As) enrichment in groundwater has become a major global environmental disaster. Groundwater samples were collected from 64 sites located in the districts of 24-Parganas (S), and Nadia in West Bengal, India (Bhagirathi sub-basin), and 51 sites located in the districts of Comilla, Noakhali, Magura, Brahman baria, Laxmipur, Munshiganj, Faridpur and Jhenaida in Bangladesh (Padma-Meghna sub-basin). Groundwater samples were also collected from two As-affected areas (Chianan and Lanyang plains) of Taiwan (n = 26). The concentrations of major solutes in groundwater of the Padma-Meghna sub-basin are more variable than the Bhagirathi sub-basin, suggesting variations in the depositional and hydrological settings. Arsenic concentrations in groundwaters of the studied areas showed large variations, with mean As concentrations of 125 μg/L (range: 0.20 to 1,301 μg/L) in Bhagirathi sub-basin, 145 μg/L (range: 0.20 to 891 μg/L) in Padma-Meghna sub-basin, 209 μg/L (range: 1.3 to 575 μg/L) in Chianan plain, and 102 μg/L (range: 2.5 to 348 μg/L) in Lanyang plain groundwater. The concentrations of Fe, and Mn are also highly variable, and are mostly above the WHO-recommended guideline values and local (Indian and Bangladeshi) drinking water standard. Piper plot shows that groundwaters of both Bhagirathi and Padma-Meghna sub-basins are of Ca-HCO(3) type. The Chianan plain groundwaters are of Na-Cl type, suggesting seawater intrusion, whereas Lanyang plain groundwaters are mostly of Na-HCO(3) type. The study shows that reductive dissolution of Fe(III)-oxyhydroxides is the dominant geochemical process releasing As from sediment to groundwater in all studied areas.


Functional interactions of nucleocapsid protein of feline immunodeficiency virus and cellular prion protein with the viral RNA.
All lentiviruses and oncoretroviruses examined so far encode a major nucleic-acid binding protein (nucleocapsid or NC* protein), approximately 2500 molecules of which coat the dimeric RNA genome. Studies on HIV-1 and MoMuLV using in vitro model systems and in vivo have shown that NC protein is required to chaperone viral RNA dimerization and packaging during virus assembly, and proviral DNA synthesis by reverse transcriptase (RT) during infection. The human cellular prion protein (PrP), thought to be the major component of the agent causing transmissible spongiform encephalopathies (TSE), was recently found to possess a strong affinity for nucleic acids and to exhibit chaperone properties very similar to HIV-1 NC protein in the HIV-1 context in vitro. Tight binding of PrP to nucleic acids is proposed to participate directly in the prion disease process. To extend our understanding of lentiviruses and of the unexpected nucleic acid chaperone properties of the human prion protein, we set up an in vitro system to investigate replication of the feline immunodeficiency virus (FIV), which is functionally and phylogenetically distant from HIV-1. The results show that in the FIV model system, NC protein chaperones viral RNA dimerization, primer tRNA(Lys,3) annealing to the genomic primer-binding site (PBS) and minus strand DNA synthesis by the homologous FIV RT. FIV NC protein is able to trigger specific viral DNA synthesis by inhibiting self-priming of reverse transcription. The human prion protein was found to mimic the properties of FIV NC with respect to primer tRNA annealing to the viral RNA and chaperoning minus strand DNA synthesis.


The Visible Emission Spectrum of the HgGa(2) Excimer.
A spectroscopic study of the Ga-Hg excimer has been carried out for the first time. The vapor mixture excited by a RF discharge shows emission bands with an intensity maximum at 5020 Å. The density of Hg in this study was in the range of 10(18)-10(19) atoms/cc and that of Ga about 10(11) atoms/cc. High-resolution spectra with a dispersion of 1.2 Å/mm were obtained, and vibrational structures were resolved and analyzed on the basis of a triatomic species HgGa(2). Copyright 2001 Academic Press.


[Target blood pressure values from the cardiologic viewpoint].
Recent large intervention trials in hypertension have shown that diastolic blood pressure--even in the presence of ischemic heart disease--can be reduced below 90 mm Hg with further benefit for the patient. This has lead to discussions about a new definition of target blood pressure in hypertension treatment. From a cardiological point of view, the total cardiovascular risk of the patient has to be calculated according to a coronary risk profile. This risk stratification of the patient helps to make a decision about pharmacological treatment of blood pressure and target blood pressure. The higher the risk of the individual patient, the more aggressive blood pressure lowering is indicated.


Fat Grafting to the Breast: Clinical Applications and Outcomes for Reconstructive Surgery.
This article is a review of fat grafting for breast reconstruction. The use of small volume fat grafting for the correction of step-off deformities, intrinsic deformities, and extrinsic deformities of the breast, and the uses of large volume fat grafting for total breast reconstruction, correction of implant complications with simultaneous implant exchange with fat, and correction of noncancer chest wall deformities is reviewed. Cancer monitoring and the risks of cancer recurrence following fat-grafting to the breast is also reviewed.


Treatment of severe steroid-dependent nephrotic syndrome (SDNS) in children with tacrolimus.
Severe steroid-dependent nephrotic syndrome (SDNS) is a common type of nephrotic syndrome (NS) observed in childhood. Steroid-sparing agents such as calcineurin inhibitors (CNIs) are used to avoid steroid toxicity in SDNS. Tacrolimus (TAC) has been prescribed for maintaining remission of NS in patients who have developed treatment resistance or adverse effects with cyclosporin A (CYA) at our institution since 1995. The aim of this study was to compare the efficacy and complications of TAC with CYA in the management of severe SDNS. We report a retrospective longitudinal clinical series of patients with SDNS, all of whom have been treated with TAC. Ten SDNS children (eight males) were reviewed quarterly from time of initial referral to the present day during 93 completed treatment patient years. Nine patients had minimal change disease and one had focal segmental glomerulosclerosis on their first biopsy. The median age at diagnosis was 2.9 years (range 1.6-12.9). The median age at initial referral was 3.9 years (range 2.2-12.9). All patients initially responded to prednisolone at 60 mg/m2/day, and subsequent frequent relapses were treated sequentially with oral cyclophosphamide 168 mg/kg over 8-12 weeks (n = 10), CYA (n = 10), intravenous mustine or a second course of cyclophosphamide (n = 7) and then TAC (n = 10). The initial daily treatment of CYA and TAC in two divided doses was 5 and 0.1 mg/kg/day, respectively; targeted 12 h blood drug levels were of 50-100 microg/l for CYA and 5-10 microg/l for TAC. Patients underwent renal biopsy and the formal glomerular filtration rate (GFR) was measured using plasma clearance of the Inutest method every 2-3 years while receiving CNIs. Six patients continued with TAC; in four patients, TAC was discontinued because of poor response (n = 2), hypertension (n = 1) and glucose intolerance (n = 1). For CYA and TAC treatment periods, the median NS relapse rate was two and one relapses per year, respectively, and cumulative steroid dosage was 73.9 and 105.2 mg/kg/day, respectively (P = 0.54). The reduction in GFR was 5.8 and 11.7 ml/min/1.73 m2 during these periods. Three of the 10 patients showed histological evidence of mild CNI nephrotoxicity over the whole of the CNI treatment period despite achieving target therapeutic drug levels; no significant change in measured or calculated GFR over this prolonged CNI therapy was observed. Antihypertensive medication was prescribed for 11 of 31 CYA and 22 of 40 TAC treatment years. Growth was maintained during the entire CNI therapy period with median change in height SD scores (SDS) of +0.37 and -0.03 over CYA and TAC, respectively (P = 0.13). In conclusion, we observed that the replacement of CYA by TAC does not lead to a better management of severe SDNS.


Conformations of the Huntingtin N-term in aqueous solution from atomistic simulations.
The first 17 amino acids of Huntingtin protein (N17) play a crucial role in the protein's aggregation. Here we predict its free energy landscape in aqueous solution by using bias exchange metadynamics. All our findings are consistent with experimental data. N17 populates four main kinetic basins, which interconvert on the microsecond time-scale. The most populated basin (about 75%) is a random coil, with an extended flat exposed hydrophobic surface. This might create a hydrophobic seed promoting Huntingtin aggregation. The other main populated basins contain helical conformations, which could facilitate N17 binding on its cellular targets.


Cell wall-associated effectors of plant-colonizing fungi.
Plant-colonizing fungi secrete a cocktail of effector proteins during colonization. After secretion, some of these effectors are delivered into plant cells to directly dampen the plant immune system or redirect host processes benefitting fungal growth. Other effectors function in the apoplastic space either as released proteins modulating the activity of plant enzymes associated with plant defense or as proteins bound to the fungal cell wall. For such fungal cell wall-bound effectors, we know particularly little about their molecular function. In this review, we describe effectors that are associated with the fungal cell wall and discuss how they contribute to colonization.


Acetylcholinesterase inhibition in the crayfish Procambarus clarkii exposed to fenitrothion.
Laboratory and field studies were conducted to evaluate the effects of fenitrothion (O,O-dimethyl O-4-nitro-m-tolyl phosphorothioate) on the crayfish Procambarus clarkii. Acetyl- and butyryl-cholinesterase activities were measured in the muscle of P. clarkii exposed to different doses of fenitrothion (4, 20, and 100 microg/liter) for different times (up to 48 hr). A positive correlation was found between both cholinesterases, and acetylcholinesterase (AChE) was selected as a biomarker of exposure to this compound. Significant AChE depression (20%) was detected 2 hr after exposure to 20 microg/liter of fenitrothion, reaching a maximum at 48 hr (47%), followed by a slow recovery. Reactivation techniques using the nucleophilic reagent pyridine 2-aldoxime methiodide were assayed in fenitrothion-poisoned specimens, and the results suggested the utility of this method to diagnose exposure, particularly when control animals are not available. Finally, AChE inhibition was used to test a field population of P. clarkii potentially exposed to high concentrations of the organophosphorus pesticide fenitrothion, and a 55% inhibition was detected.


Zingiberene targets the miR-16/cyclin-B1 axis to regulate the growth, migration and invasion of human liver cancer cells.
Liver cancer or hepatocellular carcinoma (HCC) is considered as one of the most frequent malignancies with significantly high morbidity and mortality across the globe. MicroRNAs (miRs) are regarded as important regulators of liver cancer formation and its development. However, the full biochemical mechanism of their role is still very less understood. The main objective of the current research work was to examine the role of miR-16/cyclin-B1 axis in liver cancer regulation and how this pathway along with liver cancer migration and invasion are targeted by zingiberene molecule. Quantitative reverse transcriptase polymerase chain reaction was used to evaluate miR-16 expression in HCC cell lines. Western blotting was performed to evaluate the expression of the miR-16 target genes. Effects on cell migration and invasion were evaluated by in vitro wound healing assay and transwell Matrigel assay, respectively. Effects of zingiberene on HCC cell viability were evaluated by MTT assay. Zingiberene treatment led to downregulation of miR-16 in HepG2 human hepatocellular carcinoma cells, accompanied by induction of G0/G1 cell cycle arrest targeting cyclin B1 as direct target. These effects were also accompanied by inhibition of cell migration and invasion, indicating that miR-16 can have a significant role as liver cancer suppressor after zingiberene treatment. Luciferase reporter assay confirmed that miR-16, which was one of HCC downregulated miRs, directly targeted Cyclin B1 in HCC cells. The current study indicates miR-16/cyclin B1 axis might have significant applications as a therapeutic target for patients with liver cancer.