bluuebunny/crossref_metadata_2025 huggingface dataset:
Sonic hedgehog-dependent recruitment of GABAergic interneurons into the developing visual thalamus
Authors: Somaiya Rachana Deven , Stebbins Katelyn , Gingrich Ellen C , Xie Hehuang , Campbell John N , Garcia A Denise R , Fox Michael A
Published: November 2022

Axons of retinal ganglion cells (RGCs) play critical roles in the development of inhibitory circuits in visual thalamus. We previously reported that RGC axons signal astrocytes to induce the expression of fibroblast growth factor 15 (FGF15), a motogen required for GABAergic interneuron migration into visual thalamus. However, how retinal axons induce thalamic astrocytes to generate Fgf15 and influence interneuron migration remains unknown. Here, we demonstrate that impairing RGC activity had little impact on interneuron recruitment into mouse visual thalamus. Instead, our data show that retinal-derived sonic hedgehog (SHH) is essential for interneuron recruitment. Specifically, we show that thalamus-projecting RGCs express SHH and thalamic astrocytes generate downstream components of SHH signaling. Deletion of RGC-derived SHH leads to a significant decrease in Fgf15 expression, as well as in the percentage of interneurons recruited into visual thalamus. Overall, our findings identify a morphogen-dependent neuron–astrocyte signaling mechanism essential for the migration of thalamic interneurons.
Contrasting effects of Ksr2, an obesity gene, on trabecular bone volume and bone marrow adiposity
Authors: Gomez Gustavo A , Rundle Charles H , Xing Weirong , Kesavan Chandrasekhar , Pourteymoor Sheila , Lewis Robert E , Powell David R , Mohan Subburaman
Published: November 2022

Pathological obesity and its complications are associated with an increased propensity for bone fractures. Humans with certain genetic polymorphisms at the kinase suppressor of ras2 (KSR2) locus develop severe early-onset obesity and type 2 diabetes. Both conditions are phenocopied in mice with Ksr2 deleted, but whether this affects bone health remains unknown. Here we studied the bones of global Ksr2 null mice and found that Ksr2 negatively regulates femoral, but not vertebral, bone mass in two genetic backgrounds, while the paralogous gene, Ksr1, was dispensable for bone homeostasis. Mechanistically, KSR2 regulates bone formation by influencing adipocyte differentiation at the expense of osteoblasts in the bone marrow. Compared with Ksr2’s known role as a regulator of feeding by its function in the hypothalamus, pair-feeding and osteoblast-specific conditional deletion of Ksr2 reveals that Ksr2 can regulate bone formation autonomously. Despite the gains in appendicular bone mass observed in the absence of Ksr2, bone strength, as well as fracture healing response, remains compromised in these mice. This study highlights the interrelationship between adiposity and bone health and provides mechanistic insights into how Ksr2, an adiposity and diabetic gene, regulates bone metabolism.
Supply forecasting and profiling of urban supermarket chains based on tensor quantization exponential regression for social governance
Authors: Li Dazhou , Zhou Bo , Lin Chuan , Gao Jian , Gao Wei , Gao Aimin
Published: November 2022

Background

During the COVID-19 pandemic, the accurate forecasting and profiling of the supply of fresh commodities in urban supermarket chains may help the city government make better economic decisions, support activities of daily living, and optimize transportation to support social governance. In urban supermarket chains, the large variety of fresh commodities and the short shelf life of fresh commodities lead to the poor performance of the traditional fresh commodity supply forecasting algorithm.

Methods

Unlike the classic method of forecasting a single type of fresh commodity, we proposed a third-order exponential regression algorithm incorporating the block Hankle tensor. First, a multi-way delay embedding transform was used to fuse multiple fresh commodities sales to a Hankle tensor, for aggregating the correlation and mutual information of the whole category of fresh commodities. Second, high-order orthogonal iterations were performed for tensor decomposition, which effectively extracted the high-dimensional features of multiple related fresh commodities sales time series. Finally, a tensor quantization third-order exponential regression algorithm was employed to simultaneously predict the sales of multiple correlated fresh produce items.

Results

The experiment result showed that the provided tensor quantization exponential regression method reduced the normalized root mean square error by 24% and the symmetric mean absolute percentage error by 22%, compared with the state-of-the-art approaches.
CT Perfusion in Evaluation of Cervical Lymph Node Metastasis in Head and Neck Malignancies: A Cross-sectional Study
Authors: . Rekha , Mittal MK
Published: November 2022

Introduction: Though, many cross-sectional modalities are available for evaluation of cervical lymph node metastasis but their results are highly variable. There is paucity of the literature in India, regarding lymph nodal assessment using Computed Tomography (CT) perfusion in head and neck malignancies even though, there is high incidence of oral cancer in India. Aim: To assess the role of Computed Tomography Perfusion (CTP) in evaluation of cervical lymph nodes in head and neck malignancies, by using CT perfusion parameters as compared to histopathology. Materials and Methods: This cross-sectional study was conducted in the Department of Radiodiagnosis in collaboration with the Department of Surgery and Pathology at Vardhman Mahavir Medical College and Safdarjung Hospital, Delhi, India, from October 2017 to April 2019. The study included 30 newly diagnosed head and neck cancer patients, with 46 nodes planned for surgical neck dissection. Computed tomography scan of neck was acquired using Siemens Somatom Definition Flash 256CT scanner. Reconstruction and post processing was performed on workstation and perfusion parameters were obtained to generate the CT perfusion maps. Differentiation between benign and malignant lymph nodes was done, on the basis of CT perfusion parameters such as Blood Flow (BF), Blood Volume (BV), Mean Transit Time (MTT) and Permeability Surface (PS), which were compared with histopathological findings of resected lymph nodes. McNemar’s test was applied for comparison and statistical analysis. Receiver Operating Characteristic (ROC) curve of quantitative parameters were obtained, for the detection of sensitivity, specificity, Positive Predictive Value (PPV), Negative Predictive Value (NPP) and diagnostic accuracy. Results: Out of 46 nodes, 23 were metastatic and 23 were non metastatic. The average value of BF in metastatic nodes was 174.61±71.76 mL/100g/min, BV was 16.32±11.9 mL/100g, MTT was 4.83±2.54 seconds and PS was 49.3±28.59 mL/100g/min. The average values for non metastatic nodes were: BF 88.06±34.4 mL/100g/min, BV: 9.89±7.63 mL/100g, MTT: 13.11±18.58 seconds and PS: 37.07±29.26 mL/100g/ min. The differences between the parameters like blood flow (p-value <0.0001), blood volume (p-value=0.005) and MTT (p-value=0.002) in malignant and benign nodes were significant. In case of blood flow, sensitivity was 82.61% and diagnostic accuracy was 84.78%. In case of blood volume, sensitivity was 91.30% and diagnostic accuracy was 73.91%. In case of mean transit time, sensitivity was 56.52% and diagnostic accuracy was 73.91%. In case of permeability surface, sensitivity was 91.30% and diagnostic accuracy was 67.39%. Conclusion: Blood flow and blood volume values were significantly increased in metastatic cervical lymph nodes as compared to non metastatic nodes, whereas MTT was significantly low. Permeability surface showed equivocal results.
Comparative Efficacy of Addition of Fentanyl and Neostigmine to Isobaric 0.75% Ropivacaine in Elderly Patients undergoing Transurethral Resection of Prostate under Spinal Anaesthesia: A Double-blinded Randomised Clinical Study
Authors: Shukla Pragya , Loha Sandeep , Verma Ravi Kumar
Published: November 2022

Introduction: Ropivacaine is a well-accepted local anaesthetic drug used in spinal and peripheral nerve blocks.It provides a better central nervous system and cardiovascular stability in comparison to bupivacaine. Adjuvants are also added with these drugs as they provide haemodynamic stability, increase efficacy, and decrease the dose requirement of the local anaesthetic drugs. Aim: To determine the efficacy of isobaric 0.75% ropivacaine in combination with normal saline, fentanyl, and neostigmine in Elderly Patients Undergoing Transurethral Resection of Prostate Under Spinal Anaesthesia. Materials and Methods: This double-blinded randomised clinical study conducted in the Department of Anaesthesiology and Critical Care, Institute of Medical Sciences-Banaras Hindu University, Varanasi, Uttar Pradesh, India, from July 2018 to June 2019. The study included 90 patients who were divided into three groups of 30 subjects each. Ropivacaine combined with normal saline in group RNS, ropivacaine combined with fentanyl in group RFE and ropivacaine combined with neostigmine in group RNE, was administered intrathecally.Heart rate, respiratory rate, blood pressure, onset and duration of sensory and motor block, along with the duration of analgesia, were recorded at different time points. The Analysis Of Variance (ANOVA) was used to compare the variables among the three study groups, and Student-Newman-Keuls was used to compare the data between the groups. Results: Patients in group RNE showed a significant fall in heart rate in comparison to groups RNS and RFE at all the time points. Respiratory rate followed a similar pattern in group RNE (p-value <0.001), except at 5 min and 90 min. Mean systolic blood pressure showed a significant rise in group RNE compared to the other two groups, while diastolic blood pressure followed a similar trend at 10, 20, 30, and 60 min. The SpO2 (%) was comparable among groups. The onset of sensory loss was significantly earlier in group RNE (239.6±28.8 sec) than in groups RNS (298.1±27.8 sec), and RFE (261.9±32.2 sec). The duration of the sensory block was significantly longer in group RFE (227.8±30.5 min). The mean time to the onset of motor block (480.7±30.2 sec) and analgesia (582.33±30.2 min) was longer in group RNE than in other groups. Conclusion: The addition of neostigmine to ropivacaine intrathecally is a reliable method to prolong spinal anaesthesia but close monitoring of vitals is desirable.
Prevalence of Dry Eye Disease in Glass Industry Workers of Firozabad, Uttar Pradesh, India: A Cross-sectional Observational Study
Authors: Mazumdar Shefali , Singh Ravindra Kumar , Satsangi SK , Agarwal Renu
Published: November 2022

Introduction: Dry eye disease is a rising occupational hazard in India. Firozabad, is known for its glass manufacturing work worldwide. As there is limited evidence available on dry eye prevalence in glass industry workers, the present study was undertaken. Aim: To find out the prevalence and severity of Dry Eye Disease (DED) in glass industry workers of Firozabad, Uttar Pradesh, India. Materials and Methods: The cross-sectional observational study was conducted in Department of Ophthalmology at Sarojini Naidu Medical College, Agra, Uttar Pradesh, India, from March 2020 to September 2021 among the glass industry workers at Firozabad. It was a field survey, conducted in the factories only. Ocular Surface Disease Index (OSDI) Questionnaire was presented to 500 randomly selected glass industry workers to screen for dry eye disease. Based on subject’s response to OSDI questionnaire, score was calculated and then evaluated with an OSDI chart to assess the magnitude of dry eye symptoms. The final diagnosis and grading of dry eye was done on the basis of Schirmer’s test. Chi-square test was used to detect the association between variables. Statistical Package for Social Sciences (SPSS) software (version 28.0) was used for analysis. A p-value <0.05 was regarded as significant. Results : In the present study, the overall dry eye prevalence in glass industry workers was 28%. Mild dry eye was present in 3.2% of workers, moderate dry eye in 16.8%, and severe dry eye was present in 8% of glass workers. There was significant association between dry eye and working hours (per day) in two different study groups (<8 hrs and >8 hrs) (χ2 =20.9, p-value <0.001). There was also a significant association between the prevalence of dry eye and the years of exposure (in years) in the glass industry (χ2 =51.4, p-value <0.001). Conclusion: Prevalence of dry eye in glass industry workers is significantly high. Glass industry workers should undergo regular eye checkups to pick up DED as the earliest.
Prevalence of Genomic Resistance to Macrolide in Mycoplasma Isolates among Children with Community Acquired Pneumonia
Authors: Kalpana Sivasambo , Selvakumar Shanmugam , Lakshmi Velmurugan , Dhandapani Prabhu , Suruliraj Prem
Published: November 2022

Introduction: Mycoplasma pneumonia is traditionally susceptible to macrolides, tetracycline and fluroquinolones. Since, tetracycline and fluoroquinolone are used cautiously in children, macrolides remain the antibiotic of choice for treating Mycoplasma pneumonia. But, resistance to macrolides has been reported in mycoplasma since the 2000s especially from Asia. Currently, there is no evidence on macrolide resistance of mycoplasma pneumoniae from India. Aim: To identify the prevalence of genomic resistance to macrolides in mycoplasma isolates among children hospitalised with community acquired pneumonia. Materials and Methods: This descriptive cross-sectional study was conducted in Department of Paediatrics at Institute of Child Health and Hospital for Children, Chennai, Tamil Nadu, India (tertiary care centre for children in South India), from September 2019 to August 2020. Children between 2 months to 12 years of age, who were hospitalised with community acquired pneumonia were included in the study. The sampling procedure used was induced sputum or mini bronchoalveolar lavage (in intubated children). The samples were processed for culture in Pleuropneumonia Like Organisms (PPLO) agar. The culture isolates showing the typical fried egg colonies were subjected to polymerase chain reaction to detect the presence of resistance conferring mutation in the P1 adhesin gene MPN141. Chi-square test was used to test statistical significance. Results: Among the 268 children included in the study, mycoplasma pneumonia was positive in 33 (12.3%) cases. Mycoplasma pneumoniae was most common in children aged 1-5 years old (51.5%), followed by infants (36.4%) and children aged 5-12 (12.1%). There was no significant difference in distribution among males (39.4%) and females (60.6%) (p-value=0.08). None of the mycoplasma isolates in the study showed mutation for resistance conferring genes. Conclusion: Macrolide resistance conferring genes were not identified in the study population, which may indicate that the mycoplasma strains from this part of India are still susceptible to macrolides.
Utilization of Fish Bone Waste for Food
Authors: Junianto . , Brainerd Eliazer , Maghfira Riezka , Suyono Muhamad Lanang Ariel , Rizki Azka Fathir , Pratama Rifqi Listyawan , Barkah Syahrul Muhamad
Published: November 2022

This article study aims to explain the use of fishbone waste in its processed form on the daily food product, to analyze to an extent it’s strength and potential in Indonesia and identify its usage. The method used in this study is a qualitative descriptive method using secondary data sources. This research is done by reviewing and analyzing various literature data correlated to fish bone usage. From the associated data, it’s discovered that fish bone mainly used as a calcium source in wich has different value depending on the sources. The utillization of fish bone as food ingredients is mainly used as a substitute or an additive used for product improvements and still being developed to this day.
Impact of Therapeutic Foods on Hematological Parameters of Malnourished Children Admitted at the Intensive Nutritional Recovery Center of Tessaoua, Maradi, Niger Republic
Authors: Lawaly Maman Manzo , Hannanou Katinka Ibrahim , Mansour Abdou Maman , Mansour Moussa Haladou , Mayaki Zoubeirou Alzouma
Published: November 2022

Aims: To evaluate the impact of the therapeutic foods on the hematological parameters of malnourished children below 5 years of age.

Study Design:  This is a Prospective cross-sectional study.

Place and Duration of Study: Intensive Nutritional Recovery Center of Tessaoua, Maradi, Niger republic, from June 15 to September 10, 2022.

Methodology: We included 60 malnourished children (36 males, 24 females; age range 6-59 months) who are severely acutely malnourished. Standard survey forms that were developed for the purpose have permitted to obtain sociodemographic and hematological data (hemoglobin level and leukocyte count) concerning malnourished children.

Results: The results showed that among the 60 children that were surveyed, at entry, 50 (83.33%) were recorded to have a low hemoglobin level and 36 (60%) with highly elevated leucocyte count. At the end of treatment with therapeutic foods, it was observed an overall normalization of these parameters during an average duration of hospitalization.

Conclusion: Therapeutic foods have shown a significant positive influence on hematological parameters of malnourished children. Their use should therefore be encouraged in line with other strategies in order to ensure good and rapid recovery in malnourished children under five years of age.
sources of flint and chert in northern Britain
Authors: Wickham-Jones Caroline R , Collins G H
Published: November 2022

This article lists the potential sources of flint and chert in prehistoric Scotland using information from the publications of the Geographical Survey of Great Britain. The authors highlight the quality of local stone nodules in determining the development of stone knapping and the status of large stone items in Scotland.
Built to last
Authors: Robertson Alistair , Lochrie Julie , Timpany Scott , Bailey Laura , Mynett Abbey , Shillito Lisa-Marie , Smith Catherine
Published: November 2022

A programme of archaeological investigations undertaken in advance of the Forth Replacement Crossing, at Castlandhill on Rosyth and Echline Fields in South Queensferry, revealed archaeological features dating from the Mesolithic to medieval periods, including Neolithic Grooved Ware and Impressed Ware pottery dating to the 4th to 3rd millennium BC. The principal focus of this paper is the Mesolithic remains, which are of major significance for the study of Mesolithic habitation in Scotland; particularly given the finding of a sunken-floored structure at Echline Fields that has returned a date of approximately 8300 cal BC. It is the second such structure discovered in Scotland and also the earliest yet. The sites add to a growing group of Mesolithic settlements characterised by semi-permanence and value of place in a period that has often been more associated with high mobility and temporary camps. Lithic information from these sites further acknowledges the presence of narrow-blade technology in northern Britain during this pioneering period.
Seeds, fruits and nuts in the Scottish Mesolithic
Authors: Bishop Rosie R , Church Mike J , Rowley-Conwy Peter A
Published: November 2022

Over the past few decades, the potential importance of plants within European Mesolithic economies has frequently been discussed, but there has been little systematic consideration of the archaeobotanical evidence for Mesolithic plant consumption in Scotland. This paper assesses the use of plants in the Scottish Mesolithic economy using the archaeobotanical evidence from 48 sites. It is argued that plants were systematically, and, in some cases, intensively, exploited in Mesolithic Scotland. Though plan remains were extremely sparse at most sites, it is suggested that uneven archaeological sampling and taphonomic factors, together with the relatively short duration of occupation of many sites, may be responsible for the restricted range and frequency of edible tax in most assemblages.
Gamma‐interferon causes a selective induction of the lysosomal proteases, cathepsins B and L, in macrophages
Authors: Laha Tamara T. , Hawley Marguerite , Rock Kenneth L. , Goldberg Alfred L.
Published: July 2002

Previous studies have indicated that acid‐optimal cysteine proteinase(s) in the endosomal‐lysosomal compartments, cathepsins, play a critical role in the proteolytic processing of endocytosed proteins to generate the antigenic peptides presented to the immune system on major histocompatibility complex (MHC) class II molecules. The presentation of these peptides and the expression of MHC class II molecules by macrophages and lymphocytes are stimulated by γ‐interferon (γ‐IFN). We found that treatment of human U‐937 monocytes with γ‐IFN increased the activities and the content of the two major lysosomal cysteine proteinases, cathepsins B and L. Assays of protease activity, enzyme‐linked immunosorbant assays (ELISA) and immunoblotting showed that this cytokine increased the amount of cathepsin B 5‐fold and cathepsin L 3‐fold in the lysosomal fraction. By contrast, the aspartic proteinase, cathepsin D, in this fraction was not significantly altered by γ‐IFN treatment. An induction of cathepsins B and L was also observed in mouse macrophages, but not in HeLa cells. These results suggest coordinate regulation in monocytes of the expression of cathepsins B and L and MHC class II molecules. Presumably, this induction of cysteine proteases contributes to the enhancement of antigen presentation by γ‐IFN.
Analysis of recombination junctions in extrachromosomal circular DNA obtained by in‐gel competitive reassociation
Authors: Iwasaki Toshiyasu , Ohki Rieko , Kiyama Ryoiti , Oishi Michio
Published: July 2002

Essentially all eukaryotic cells contain circular extrachromosomal DNA as a result of excision from the chromosomes. To obtain insight into the nature of recombination associated with the occurrence of such DNA species and its biological significance, we analyzed a library enriched in recombination junctions which was constructed by a novel DNA subtraction technique; in‐gel competitive reassociation (IGCR). Furthermore, we also introduced inverse PCR to characterize chromosomal DNA fragments containing the recombination junctions. At least 45% of the clones in the library constructed by the IGCR procedure comprised DNA with recombination junctions. Nucleotide sequence analysis of the recombination junctions indicated that three of four extrachromosomal DNAs thus analyzed were produced through recombination between sequences with a 3–5 bp homology in the chromosomes. One extrachromosomal DNA was apparently generated through non‐homologous recombination, possibly by end‐to‐end joining. These results have demonstrated the usefulness of IGCR in concentrating recombination junctions, which provide the most direct evidence for the mechanism of the recombinational events involved, from highly complex genomes.
The importance of the GTP‐binding protein tissue transglutaminase in the regulation of cell cycle progression
Authors: Mian S. , El Alaoui S. , Lawry J. , Gentile V. , Davies P.J.A. , Griffin M.
Published: July 2002

Tissue transglutaminase (tTgase) is a GTP‐binding Ca2+‐dependent enzyme which catalyses the post‐translational modification of proteins via ϵ(γ‐glutamyl) lysine bridges. Recent evidence suggests that the GTP‐binding activity of tTgase may be important in intracellular signalling thus explaining some of the diverse suggested roles for the enzyme. In the following work a malignant hamster fibrosarcoma (Met B) has been stably transfected with both the full length tTgase cDNA (wild type) and a mutant form of the cDNA whereby the active site cysteine (Cys 277) has been replaced by serine. Expression of this mutant cDNA leads to a protein with GTP binding activity which is deficient of protein crosslinking activity. When synchronised into S‐phase and allowed to progress through the cell cycle tTgase transfected clones (both mutant and wild type), when compared to transfected controls, show a delayed progression from S‐phase to G2/M when analysed by flow cytometry which appears to be elicited by the G‐protein activity of the tTgase.
Identification of a member of mouse semaphorin family
Authors: S. Inagaki  , T. Furuyama  , Y. Iwahashi
Published: July 2002

Grasshopper semaphorin I (Sema I) and its related proteins, chick collapsin and mouse Sema III contribute to the axon guidance by their repellent actions [5,9,12]. We have identified a member of semaphorin gene family from the mouse brain and named it M‐Sema F. The N‐terminal encodes a semaphorin domain that is similar between Sema I–III [6] followed by a single putative immunoglobulin‐like domain, a transmembrane domain, and a proline‐rich intracellular domain. M‐Sema F mRNA is expressed widely in the nervous tissues during development. These suggest that M‐Sema F is a transmembrane member of the semaphorin family of the vertebrate which may function in the developing neuronal network.
Parity and the prevalence of overweight
Authors: Arroyo P. , Avila‐Rosas H. , Fernández V. , Casanueva E. , Galván D.
Published: July 2002

Objective: To assess the association between previous term pregnancies and the prevalence of overweight in a group of urban women, controlling for the influence of age. Methods: One thousand twelve women, living in middle and low socioeconomic areas of Mexico City, were interviewed at home and their reproductive histories studied. Height and weight were measured in a clinical setting using controlled procedures. Overweight (BMI > 25) was the dependent variable used to calculate odds ratios and to perform a multiple logistic regression analysis. Results: Age and parity were significantly associated with the prevalence of overweight. Controlling for age, two or more previous pregnancies significantly increased the magnitude of the association. Conclusion: During the reproductive years parity seems to increase the risk of overweight in low and middle socioeconomic level urban women.
Early expression of the BM88 antigen during neuronal differentiation of P19 embryonal carcinoma cells
Authors: Boutou E. , Hurel C. , Matsas R.
Published: July 2002

Previous studies have shown that the BM88 antigen, a neuron‐specific molecule, promotes the differentiation of mouse neuroblastoma cells [23] (Mamalaki A., Boutou E., Hurel C., Patsavoudi E., Tzartos S. and Matsas R. (1995) The BM88 antigen, a novel neuron‐specific molecule, enhances the differentiation of mouse neuroblastoma cells. J. Biol. Chem. 270, 14201–14208). In particular, stably transfected with the BM88 cDNA, Neuro 2a cells over‐expressing the BM88 antigen are morphologically distinct from their non‐transfected counterparts; they exhibit enhanced process outgrowth and a slower rate of division. Moreover, they respond differentially to growth factors [10] (Gomez J., Boutou E., Hurel C., Mamalaki A., Kentroti S., Vernadakis A. and Matsas R. (1998) Overexpression of the neuron‐specific molecule BM88 in mouse neuroblastoma cells: Altered responsiveness to growth factors. J. Neurosci. Res. 51, 119–128). In order to further elucidate the role of the BM88 antigen in the differentiation of developing neurons we used the in vitro system of differentiating P19 cells which closely resembles early murine development in vivo. In this study, P19 cells were driven to the neuronal pathway with retinoic acid. We examined by immunofluorescence studies the expression of the BM88 antigen in these cells and we found that it correlates well with the expression of the polysialylated form of the neural cell adhesion molecule (PSA‐NCAM) which characterizes early differentiating post‐mitotic neurons. In contrast, very few of the BM88 antigen‐positive/PSA‐NCAM‐positive cells expressed neurofilament protein, a marker of more mature neurons. Our findings, in accordance with previously reported data, strongly suggest that the BM88 antigen is involved in the early stages of differentiation of neuronal cells.
Optimization of High Molecular Weight Pullulan Production by Aureobasidium pullulans in Batch Fermentations
Authors: Gibson Larry H. , Coughlin Robert W.
Published: July 2002

Of five strains of Aureobasidium pullulans studied, NRRL Y‐2311–1 yielded the highest titer (26.2 g/L) of pullulan and formed the lowest amount of melanin‐like pigment. Sucrose was superior to glucose as the carbon and energy source on the basis of yield and titer of pullulan produced. Pullulan titer was higher (26.2 vs 5.1 g/L), biomass concentration was lower (6.9 vs 12.7 g/L), and DO was lower (0 vs 60% of saturation) when the fermenter was agitated by a marine propeller compared to Rushton impellers. Pullulan produced by strain NRRL Y‐2311–1 ranged in weight‐average molar mass ( Mw) from 486 KDa and number‐average molar mass ( Mn) from 220 Da on day 1 of growth to 390 KDa and 690 Da on day 6; Mw declined by about 35% from day 1 to day 3, the day of maximum pullulan titer. For the other strains, the ranges of molar mass on the day of maximum pullulan titer were 338–614 KDa ( Mw) and 100–6820 Da ( Mn).
Homologous Human Blood Protein Separation Using Immobilized Metal Affinity Chromatography: Protein C Separation from Prothrombin with Application to the Separation of Factor IX and Prothrombin
Authors: Wu Huiping , Bruley Duane F.
Published: July 2002

Protein C (PC) is a natural anticoagulant and antithrombotic present in human blood at a concentration of 4 μg/mL. Its deficiency can result in excessive clotting and thrombosis. Protein C can be obtained from human blood plasma; however, there are other coagulant proteins in blood, including prothrombin (factor II), which is present in relatively large amounts and is one of the most active components. Protein C and prothrombin are homologous proteins with similar biochemical features; therefore, immunoaffinity chromatography is used for their separation. However, this technology is very expensive, protein C recovery and activity is low, and contamination problems with mouse antibody are likely. Immobilized metal affinity chromatography (IMAC) utilizes the protein metal‐binding properties for protein separation. Protein C has twelve surface‐accessible histidines, which are the major metal‐binding groups for IMAC separation. After investigating metal ion‐binding properties of protein C, we used an IDA‐Cu column to separate protein C and prothrombin. Following protein adsorption to the column, prothrombin was washed out using a sodium phosphate buffer containing 2 mM imidazole and protein C was recovered with 15 mM imidazole in the buffer. The mild elution condition allows a high protein C activity and a high recovery. Also, this technology introduces no immunoglobulins, and it is relatively inexpensive. IMAC could replace the immunoaffinity technology for the large‐scale separation of protein C from blood plasma Cohn Fraction IV‐1. In addition, this work demonstrates a significant application of this technology for the separation of factor IX from prothrombin. Prothrombin has proven to be a harmful contaminant in factor IX cocktails that have been administered to humans in the treatment of hemophilia B.
Selective and facile cyclization of N‐chloroacetylated peptides from the C4 domain of HIV Gp120 in LiCl/DMF solvent systems
Authors: Robey F.A.
Published: July 2002

Lithium salts have been reported to mediate the solubilization of peptides in organic solvents in 1989 (Seebach, D., Thaler, A. & Beck, A. K. Helv. Chim. Acta 1989; 72, 857–867). The use of Li salts in an organic solvent to influence cyclization of a reactive peptide that only polymerizes in an aqueous solvent, has not been reported. Here, the selective and facile cyclization of N‐chloroacetylated, C‐cysteine amide peptides from the C4 domain of HIV‐1 gp120 in LiCl/DMF solvent systems is demonstrated. The addition of stoichiometric amounts of Tris base to 1 mg/mL peptide in LiCl/DMF solutions was sufficient to drive the cyclization to completion within 3 h at ambient temperatures. Cyclic peptides were the only detectable reaction products and these were confirmed using reversed‐phase HPLC and mass spectrometric analyses of the final products. In aqueous solutions at pH 7.4, only polymers were obtained as judged by HPLC and SDS–PAGE. The method of using Li salts in an organic solvent to enhance the cyclization of unprotected amphipathic peptides may be useful in many situations beyond those described here.
Cow's milk protein intolerance and chronic constipation in children
Authors: Daher Silvia , Tahan Soraia , Solé Dirceu , Naspitz Charles K. , Da Silva Patrício Francy Reis , Neto Ulysses Fagundes , De Morais Mauro Batista
Published: July 2002

Cow's milk protein (CMP) allergy was investigated in 25 children (age‐range 3 months to 11 years) with chronic constipation. A diagnosis of constipation was made on the basis of a history of painful elimination of hard stools for at least 1 month, whether or not associated with a reduced frequency of stools or soiling. The children were evaluated using clinical parameters and the following laboratory tests: total serum immunoglobulin E (IgE); specific IgE (radioallergosorbent test [RAST]) for whole cow's milk, α‐lactoalbumin, β‐lactoglobulin, and a food group; and skin‐prick tests with whole milk, α‐lactoalbumin, β‐lactoglobulin, and casein. Following the evaluation, the children were submitted to a CMP‐free diet for a period of 4 weeks. In seven patients (28%), constipation disappeared during the CMP‐free diet and reappeared within 48–72 h following challenge with cow's milk. In two infants a rectal biopsy revealed allergic colitis and they therefore did not undergo the challenge. High serum levels of total IgE were observed in five of the children who showed a clinical improvement (71%), a positive skin‐test in two (29%), and detectable specific IgE in two (29%). These results suggest that CMP allergy or intolerance should be considered as a cause of chronic refractory constipation in children, although the underlying mechanism still require further investigation.
Gingival fine needle aspiration cytology in acute leukemia
Authors: Abdullah Bashar H. , Yahya Hassan I. , Kummoona Raja K. , Hilmi Ferial A. , Mirza Khalid B.
Published: July 2002

Background:  Oral findings in acute leukemia (AL) are common and could be the presenting feature of the disease, namely, gingival enlargement, ulceration, bleeding, and infection. Gingival enlargement in AL is either due to leukemic infiltration, or due to reactive hyperplasia. To differentiate between them a biopsy is required, but being highly contraindicated, biopsy has been substituted in this study by fine needle aspiration cytology (FNAC).

Method:  Gingival FNAC was performed on different sites in the upper and lower gingiva. Each site represents an interdental papilla, which was selected according to the clinical presentation, i.e., being enlarged or not. Seventy‐two adult AL patients received a cytological and clinical examination in this study, and the cases were classified and categorized according to the French‐American‐British (FAB) criteria.

Results:  Twenty‐one cases were diagnosed as being infiltrated, 16 with gingival enlargement, 4 with no evidence of enlargement. In one case the infiltration affected the alveolar mucosa of an edentulous patient. In six cases the ginigva was enlarged without being infiltrated (reactive hyperplasia). Leukemic gingival enlargement was seen mostly in patients with acute myeloid leukemia, particularly M4 and M5 subtypes; however, two patients with acute mixed lineage leukemia (AMLL) were both affected with leukemic infiltration.

Conclusion:  FNAC was shown to be a simple, non‐traumatic and useful diagnostic procedure for screening leukemic infiltration in gingival tissues in AL patients.
Effects and distribution of the enamel matrix derivative Emdogain® in the periodontal tissues of rat molars transplanted to the abdominal wall
Authors: Hamamoto Yoshioki , Kawasaki Naoko , Jarnbring Fredrik , Hammarström Lars
Published: July 2002

Abstract – The enamel matrix derivative Emdogain® (EMD) has been found to promote regeneration of lost periodontal tissues. We have studied the effects and distribution of EMD in the periodontal tissues of maxillary rat molars transplanted to a subcutaneous position in the abdominal wall. The molars were transplanted with or without EMD either immediately after extraction or after drying for 30 min. After 2 days, 1, 2 or 4 weeks the rats were killed and the teeth were examined by means of light microscopy and immunohistochemistry with anti‐amelogenin antibodies. Teeth transplanted immediately after extraction showed formation of alveolar bone separated from the dental roots by a periodontal space, regardless of the use of EMD. Among the teeth that were transplanted with EMD after drying for 30 min, new alveolar bone was formed in five out of eight teeth after 2 and 4 weeks. None of the teeth that were dried for 30 min and transplanted without EMD showed alveolar bone formation. Only one tooth transplanted with EMD showed root resorption after drying, while resorption was noted in all teeth transplanted without EMD. All teeth that were transplanted with EMD and none of the teeth that were transplanted without EMD showed an immunohistochemical reaction for amelogenin. After 2 days, amelogenin was precipitated on all surfaces exposed at the transplantation procedure. Later, the immunoreactive material was redistributed to cells at the root surface, where it was still demonstrable after 4 weeks. In conclusion, EMD is accumulated in cells at the root surface and promotes regeneration of the periodontal tissues of the transplanted teeth. It also seems to promote healing of root resorption.
Events in the immortalizing process of primary human mammary epithelial cells by the catalytic subunit of human telomerase
Authors: KIM Hyunggee , FARRIS James , CHRISTMAN Shelly A. , KONG Byung-Whi , FOSTER Linda K. , O'GRADY Scott M. , FOSTER Douglas N.
Published: July 2002

The in vitro immortalization of primary human mammary epithelial (HME) cells solely by the exogenous introduction of the catalytic subunit of human telomerase (hTERT) has been achieved. Early passage hTERT-transfected HME (T-HME) cells continuously decreased the length and density of telomeres even in the presence of telomerase activity, with a significant number of cells staining positive for senescence-associated β-galactosidase (SA-β-gal). Subsequently, with the increase in cell passages, the copy number of the exogenously transfected hTERT gene and the percentage of SA-β-gal positive cells were found to decrease. Eventually, a single copy of the exogenous hTERT gene was observed in the relatively later passage T-HME cells in which telomere length was elongated and stabilized without obvious activation of endogenous hTERT and c-Myc expression. In T-HME cells, the expression of two p53 regulated genes p21WAF and HDM2 increased (as in primary senescent HME cells), and was found to be further elevated as the function of p53 was activated by treatment with DNA-damaging agents. p16INK4a was shown to be significantly higher in the primary senescent HME and the early passage T-HME cells when compared with the primary presenescent HME cells, with a dramatic repression of p16INK4a observed in the later passage T-HME cells. In addition, the expression of E2F1 and its transcription factor activity were found to be significantly higher in the later passage T-HME cells when compared with the earlier passage T-HME cells. Together, our results indicate that in vitro immortalization in HME cells may require the activation of the function of telomerase and other genetic alterations such as the spontaneous loss of p16INK4a expression.
Antithyroid drugs and Graves' disease — prospective randomized assessment of long‐term treatment
Authors: Maugendre D. , Gatel A. , Campion L. , Massart C. , Guilhem I. , Lorcy Y. , Lescouarch J. , Herry J. Y. , Allannic H.
Published: July 2002

OBJECTIVE

Although antithyroid drugs (ATD) are widely used in the treatment of Graves' disease, management protocols, especially treatment duration, remain a subject of debate. The rate of relapse after short‐term regimens of less than 6 months with ATD at decreasing doses is higher than after longer treatments from 12 to 24 months. As no prospective study has provided data on even longer protocols exceeding 2 years, we conducted a prospective trial to determine potential benefits of a 42‐month treatment compared with an 18‐month treatment.

DESIGN, PATIENTS AND MEASUREMENTS

The aim of this prospective randomized trial was to compare relapse rates achieved two years after treatment withdrawal in patients who received carbimazole at decreasing doses for 18 months (n = 62) vs 42 months (n = 72). In addition to clinical relapse rate, the percentage of patients who normalized antithyroperoxidase (TPO) antibody and anti‐TSH receptor stimulating antibody (TSAb) levels and early iodine uptake at the end of treatment were assessed as outcome criteria.

RESULTS

The relapse rate two years after discontinuation of treatment did not differ significantly in patients treated for 18 months from those treated for 42 months (36% vs 29%, NS). At the end of treatment, there was no significant difference between the two groups in the percentage of anti‐TPO positive patients (53% vs 46%, NS) or early iodine uptake (27% vs 21%, NS). Although the percentage of patients with TSAb was significantly lower in the 42‐month treatment group (18% vs 42%, P = 0.004) at treatment withdrawal, the percentage of TSAb‐positive patients did not significantly decrease between 18 and 42 months in this group (27% vs 18%, NS).

CONCLUSION

Treatment duration greater than 18 months did not improve remission rate determined 2 years after treatment withdrawal or immunological variables or early iodine uptake measured at the time of discontinuation of treatment. These findings would indicate that, when a defined duration treatment is planned, prolonging treatment beyond 18 months does not provide any additional benefit.
Responses of tropical forest tree seedlings to irradiance and the derivation of a light response index
Authors: Agyeman V. K. , Swaine M. D. , Thompson J.
Published: July 2002

1  Seedlings of 16 West African timber tree species were grown in six neutral shadehouses with irradiances 2%, 6%, 10%, 28%, 44% and 66% of unshaded values.

2  Tested species included well‐known pioneers, which showed negative growth in 2% irradiance but which responded strongly to increased irradiance, and very shade‐tolerant non‐pioneers, which had positive growth in 2% irradiance but responded only slightly to increased irradiance.

3  For all species, maximum growth occurred at an irradiance varying between 10% and 44%. The inhibition of growth at higher irradiance was greater in the more shade‐tolerant species.

4  Data from growth analyses were used in a principal components analysis to quantify the species’ positions on a light response gradient. The growth variables most strongly correlated with this gradient were relative growth rate (RGR) and relative diameter growth in 2% irradiance, and also apparent quantum efficiency and leaf mortality rate in both 2% and 66% irradiance. These results draw attention to the importance of performance in deep forest shade in defining differences among tropical forest tree species.

5  The second (independent) axis of the principal components analysis separated three species that are known from other experiments to be drought tolerant. Other species’ traits of functional significance (adult deciduousness, seed size, seed dispersal) were only weakly, if at all, associated with the gradient of light response.

6  Possible methods for efficient placement of other species on the gradient of light response are discussed. Formulation of standardized conditions simulating deep shade would allow diameter growth to be used as a simple non‐destructive measure of a species’ position on the gradient of light response, which could then be applied to any tropical forest flora.
A single‐chain antibody fragment is functionally expressed in the cytoplasm of bothEscherichia coliand transgenic plants
Authors: Tavladoraki Paraskevi , Girotti Alessandra , Donini Marcello , Arias Francisco Javier , Mancini Camillo , Morea Veronica , Chiaraluce Roberta , Consalvi Valerio , Benvenuto Eugenio
Published: July 2002

Despite the well‐known crucial role of intradomain disulfide bridges for immunoglobulin folding and stability, the single‐chain variable fragment of the anti‐viral antibody F8 is functionally expressed when targeted to the reducing environment of the plant cytoplasm. We show here that this antibody fragment is also functionally expressed in the cytoplasm ofEscherichia coli. A gel shift assay revealed that the single‐chain variable fragment (scFv) accumulating in the plant and bacterial cytoplasm bears free sulfhydryl groups. Guanidinium chloride denaturation/renaturation studies indicated that refolding occurs even in a reducing environment, producing a functional molecule with the same spectral properties of the native scFv(F8). Taken together, these results suggest that folding and functionality of this antibody fragment are not prevented in a reducing environment. This antibody fragment could therefore represent a suitable framework for engineering recombinant antibodies to be targeted to the cytoplasm.
Signalling by CD95 and TNF receptors: Not only life and death
Authors: Magnusson Carina , Vaux David L
Published: July 2002

Members of the TNF family of receptors play important roles in normal physiology and in defence. The recent rapid progress in the understanding of the mechanisms of apoptosis has been accompanied by assumptions that TNF family receptors such as CD95(Fas/APO‐1) only have a role in regulating cell survival. While regulation of cell death is one important function of TNF family receptors, they are capable of activating signal transduction pathways that have many other effects. The present review will focus on signalling of some TNF family receptors in the immune system, not only for apoptosis, but also for survival or activation.
Vaccination of children following a previous hypotonic–hyporesponsive episode
Authors: Goodwin H , Nash M , Gold M , Heath TC , Burgess MA
Published: July 2002

Background: Hypotonic–hyporesponsive episodes (HHE) were once considered a contraindication to pertussis vaccination.

Methodology: To evaluate the safety of further vaccination, chart review was conducted of 66 children who had experienced an HHE, and who were assessed in one of three special clinics in three capital city tertiary paediatric hospitals. Immediately prior to the HHE, 63 children had received whole‐cell pertussis vaccine, three had received acellular pertussis vaccine.

Results: Sixty‐four of the 66 children were vaccinated in the special clinic, 59 with pertussis‐containing vaccines (55 acellular, four whole‐cell), without serious adverse events. There were no HHE after these further vaccinations.

Conclusions: Previously healthy children who experience HHE reactions can safely continue standard vaccination schedules.