Biochemical alterations induced by the mixture were greater than those recorded for each of the individual insecticides. The joint action analysis, based on the obtained biochemical data, revealed the dominance of antagonistic action among MET and ABM. Zinc supplementation achieved noticeable ameliorative effects. It was concluded that zinc may act as a powerful antioxidant, especially in individuals who are occupationally exposed daily to low doses of such pesticides.Exposure to mixtures of toxicants  is common in real life and a subject of current concern. The present investigation was undertaken to assess some toxicological effects in male rats following exposure to methomyl (MET), abamectin (ABM), and their combination (MET+ABM), and to evaluate the ameliorative effect of zinc co-administration. Three groups of rats were designated for MET, ABM, and the mixture treatments. Three other groups were designated for zinc in conjunction with the pesticides. Additionally, one group received water only (control), and the other represented a positive zinc treatment. The obtained results revealed that MET was acutely more toxic than ABM. The tested pesticides induced significant elevation in lipid peroxidation and catalase levels, while declined the levels of the other tested parameters e.g., Superoxide dismutase (SOD), Glutathione-S-transferase (GST), Glutathione peroxidase (GPx), Glutathione reductase (GR), Cytochrome P S-methyl N-(methylcarbamoyloxy) thioacetimidate (C5H10N2OS), is an oximecarbamate insecticide produced by DuPont since 1966. It is used widely for controlling insect pests on fruits, vegetables, vines, grains, soybeans, and cotton all over the world  oxy] ethanimidothioate; IUPAC), was obtained from BASIF Limited Co., Cairo, Egypt, as Nudrin® (90% SP). Abamectin, a biopesticide (molecular weight: 873.1 (avermectin B1a); 860.1 (avermectin B1b); empirical formula: C48H72O14 (80% avermectin B1a); C47H70O14 (20% avermectin B1b), was obtained from Al-Mustafa Triad Co., Cairo, Egypt, asVabcomic® (1.8% EC). Zinc chloride (ZnCl2) powder (molecular weight: 136.29), a product of Oxford Laboratory, was purchased from Neminath Industrial Estate No. 6 Navghar, Vasai East, Thane.Methomyl, an oximecarbamate insecticide (methyl 450 (CYP450), and hormonal biomarkers: testosterone (T) and tetraiodothyronine-thyroxine (T4).Diagnostic kits used in the present study were obtained from Biodiagnostic Co., Giza, Egypt. These were oxidative/anti-oxidative biomarkers: lipid peroxidation (LPO), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), glutatione-S-transferase (GST), cytochrome PRattusnorvegicus), 60 days of age and with average weights of 110 ± 20 g, were obtained from the Animal Breeding House of the National Research Centre (NRC), Dokki, Cairo, Egypt, and maintained in clean plastic cages in the laboratory animal room (23 ± 2 °C) on a standard pellet diet and had free access to water in daily dark/light cycle of 12/12 h. Rats were acclimatized for one week prior to experimentation. The experimental work on rats was performed with the approval of the Animal Care and Experimental Committee, College of Agriculture in Damanhur, Egypt, and according to the guidance for care and use of laboratory animals [Healthy male albino rats of the Wister strain  for commercial formulations of methomyl (MET) and abamectin (ABM) on male rats. For each insecticide, three doses were prepared in water based on active ingredient (a.i.) contents . Four rats were used for each tested dose, in addition to four rats given water only, serving as a control group. Dosing was performed by gavages with 0.5 mL solutions. The 24h-LD50 values were estimated according to Finney [50 values, the equivalent to the 1/10 was used in the present study .Preliminary tests were carried out to determine the median lethal dose (LD2 in drinking water at a concentration of 227 mg/L (as Zn) according to Goel et al. [ad libitum for all groups. The experimental duration was extended up to 42 days, and the doses of insecticides were adjusted weekly according to changes in body weights of the tested animals. Observations on animals were performed one day after another to record any treatment-related clinical signs of toxicity.A total of 64 male rats were divided into eight groups (Gs), each contained eight animals. G1 (Cont) received water free of any pesticide and served as negative control. G2  administered ZnCll et al. . G3 (METg) for 10 min using a BOECO centrifuge model C-28, Germany, to separate the plasma. The plasma samples were kept in a deep freezer (−20 °C) until analysis within one week. Finally, the rats were sacrificed by decapitation. Liver, kidneys, and testes were quickly removed, weighed individually, and preserved in 10% formalin saline for further histological studies in another work.At the end of the experimental duration, blood samples were withdrawn from the animals under ether anesthesia by puncturing the retro orbital venous plexus with a fine sterilized glass capillary. Blood was collected into heparinized glass tubes to separate plasma and left for 20 min at room temperature, then centrifuged at 3000 rpm . The analyses were carried out in accordance to the pamphlet instructions given by the manufacturers, and in light of the published methods.p ≤ 0.05 and p ≤ 0.01 levels.All the obtained data were statistically analyzed using Statistical Analysis (SAS) Software Program . Data were analyzed as factorial arrangements of the kind of emulsifying and storage period in a complete randomized design with three replicates. Comparisons among the means of different treatments were achieved by using the least significant difference procedure (LSD) at 1, and A2 represent the mean values obtained from the biochemical estimation of a studied parameter: M for the mixture value; A1 and A2 for the values of the individual compounds in that mixture; and C for the control value.According to Mansour and Refaie , the typ(i)Case of positive effect ; where I.I. > 1 means potentiation; I.I. = 1 means additive; I.I. < 1 means antagonism.(ii)Case of negative effect ; where I.I. > 1 means antagonism; I.I. = 1 means additive; I.I. < 1 means potentiation.(iii)Case of no observed effect: In such a case, it is supposed that treatment with the mixture and each of its individual compounds do not induce statistically significant differences between the values of the measured parameters; and thus control and treatments have nearly the same values. Here, the interaction index (I.I.), if determined, will equal 1 . The prior statistical examination for the dataset of a given biochemical measurement would assist the differentiation between anadditive case and that of no effect.The rating of the interaction indices is subject to one of three cases as follows:For accuracy, a “safety factor” of ±0.05 is added to the indices values when ranking the joint action. This means that additive effect will be considered as I.I. = 1 ± 0.05 , and the other categories have to be ranked accordingly .According to Mansour and Gamet-Payrastre , alteratOn the other hand, the “Amelioration Index” (AI) could be estimated by comparing the results of a given biochemical parameter in the groups of pesticides + antioxidant agent  with the results of the control groups to assess the ameliorative effect of Zn. As AI approaches 1, the amelioration reaches a high degree of normalization to the control value : (3)Amel50 for methomyl (MET) and abamectin (ABM) against the used male rats indicated that MET was more toxic (17.0 mg/kg b.w.) than ABM (20.3 mg/kg b.w.). These values were used to calculate the doses used in the present study .The estimated oral LDAfterten days of insecticidal treatments, signs of tremors, restless, excitation, dizziness and weakness were observed in MET, ABM and the mixture (MET+ABM)-treated groups. These signs were more pronounced in MET and ABMrat groups. There were no treatment-related clinical signs of toxicity noted inpesticide + Zn treatments. During the last week of experimental duration, most of the signs of toxicity had disappeared.p ≤ 0.01) in the activity of this enzyme. Co-administration of Zn with MET or ABM limited the enzyme decline to some extent (p ≤ 0.05), where their estimated values equaled 0.92 and 0.93 µmol/min/mL, respectively , either with or without zinc administration. Generally, results of control and zinc treatments were of no significant differences. Compared with control result (1.07 µmol/min/mL), the activity of glutathione-S-transferase (GST) recorded 0.85, 0.85, 0.80 and 0.82 µmol/min/mL, respectively for MET, ABM, MET+ABM, and MET+ABM+Zn; indicating highly significant decreases (ectively a.p ≤ 0.01) higher than the values recorded for MET, ABM, and MET+ABM . Co-administration of Zn with the individual pesticides or their mixture limited decline of GPx activity to some extent (p ≤ 0.05).The activity of glutathione peroxidase (GPx) b in the p ≤ 0.01) than the values recorded for MET, ABM, and MET+ABM . Co-administration of Zn with ABM normalized the activity of GR .p ≤ 0.01), accounting to 2.22, 2.36, and 2.71 nmol/mL in the rats treated with MET, ABM, and their mixture, respectively. Such high elevation was limited to some extent (p ≤ 0.05) by co-administration of Zn.The activity of lipid peroxidation (LPO), in terms of malondialdehyde (MDA), in plasma of control rats was found 1.35 nmol/mL a. MDA rep ≤ 0.01) decline in rats treated with MET, ABM, MET+ABM, and MET+ABM+Zn compared with that recorded for the control (104.87 µmol/min/mL). The greatest decline was attributed to MET treatment (88.23 µmol/min/mL). Administration of Zn with MET or ABM limited such a high decline to some extent (p ≤ 0.05).Contrary to MDA, the activity of superoxide dismutase (SOD) b showed p ≤ 0.01) were recorded for MET, ABM, and MET+ABM . Administration of Zn with MET or ABM normalized catalase activity, but failed to achieve a similar result with the mixture (0.85 µmol/min/mL).Catalase activity in the control group recorded 0.48 µmol/min/mL c. In com450 activity in the plasma of male rats treated with the tested pesticides. The control treatment recorded 0.136 nmol/min/mL, a value which was significantly higher (p ≤ 0.01) than those recorded for MET (0.038 nmol/min/mL), ABM (0.037 nmol/min/mL), MET+ABM (0.027 nmol/min/mL), and MET+ABM +Zn (0.083 nmol/min/mL). Supplementation of Zn in conjunction with MET or ABM limited the enzyme decline to some extent (p ≤ 0.05).4), were determined in the plasma of male rats treated with MET, ABM, and their mixture (MET+ABM), either with or without zinc administration. Testosterone hormone (T) measured in plasma of control rats was found to be 4.86 ng/mL (p ≤ 0.01) in MET, ABM, MET+ABM, and MET+ABM+Zn treatments, where the estimated values equaled 1.83, 1.99, 1.69, and 2.78 ng/mL, respectively. Co-administration of Zn with MET or ABM limited such decrease (p ≤ 0.05) to a noticeable extent.Two hormones, testosterone (T) and thyroxine (T86 ng/mL b. Its le4) showed a similar pattern to that of testosterone where MET, ABM, MET+ABM, and MET+ABM+Zn treatments recorded a severe decline in the level of T4 compared with control (3.89 µg/dL). Co-administration of Zn with MET or ABM limited such a decline to a noticeable extent , testosterone (T), and thyroxine (T4).4 in MET treatments. The percent of changes in GST and CYP450 in MET and ABM treatments were nearly equal.The percent of changes in some biochemical parameters in male rats treated with MET, ABM, and the mixture (MET+ABM), and the ameliorative effect of zinc supplementation, are presented in The effect of zinc supplementation on minimizing the differences between values of the measured biochemical parameters and their corresponding control values was expressed in terms of the “Amelioration Index” (AI). As shown in 50 for the used commercial products of MET and ABM to male rats differed slightly and revealed that MET was more toxic than ABM. Both compounds were tested at an equixotic dose , however, they induced varying degrees of alterations in the tested biochemical parameters, as presented above.The estimated acute oral LDIn toxicity studies, a variety of specific biochemical parameters are measured to evaluate physiological and metabolic functions which affect target organs and tissue injury . The mosAntioxidant defense plays an important role in the response of organisms to environmental pollutants. Several processes enhance the production of reactive oxygen species (ROS) or deplete the antioxidant defense. Such oxidative stress, if not regulated properly, may lead to damage in DNA, proteins, or lipids. On the other hand, ROS are also beneficial as they play an important role in defense against infectious agents. Hence, a delicate balance between antioxidants and free radical formation is required . Studies2O2). However, endogenous (H2O2) may be reduced to H2O either by catalase or glutathione peroxidase, or it may generate the highly-reactive free hydroxyl radical (OH•) by the Fenton reaction, which is believed to be mainly responsible for oxidative damage [2O2 is catalyzed by SOD, while CAT converts H2O2 to water. Therefore, these enzymes have the capability to alleviate the hazards of ROS [In this respect, it may be convenient to demonstrate the role of the antioxidants of relevance to the present study. Lipid peroxidation (LPO) is a chain reaction between polyunsaturated fatty acids and ROS, yielding lipid peroxides and hydrocarbon polymers. These are extremely toxic to the cells. Peroxidation of polyunsaturated fatty acids and related esters produces malondialdehyde (MDA) as an end product. Therefore, MDA serves as a biomarker of LPO . Catalase damage . Glutathe damage . Glutathe damage . It is we damage . These es of ROS .In agreement with the results of the present investigation, many studies have reported elevation of lipid peroxidation (LPO) and CAT and a decline of SOD, as well as the group of glutathioneenzymes, following exposure to organophosphorus (OP) pesticides, such as chlorpyrifos ,15,26. T450 (CYP450) enzyme system plays an important role in the bio-activation of cholinesterase inhibitors, such as organophosphorus (OP) pesticides, through catalyzing oxidation of one-atom molecular oxygen into a substrate  by an electron transport pathway [450. These results are in agreement with those reported by Yamano and Morita [The cytochrome P pathway . In thisd Morita  and may d Morita . Therefod Morita .Testosterone (T) is the main steroid sex-hormone in male rats. It is secreted by Leydigcells of the testes under the control of complex neuroendocrine interactions . The pre4) might be able to regulate the activities of SOD, CAT, and GPx enzymes in lymphoid organs and skeletal muscles [4) level in the treated rats. These findings are supported by the results of several investigators who addressed the thyroid inhibitory nature of OP insecticides [4 in the insecticide-treated groups of rats may refer to some damage in the thyroid gland due to oxidative stress induction and functional impairment of the pituitary-thyroid axis [Thyroid hormones  around 1. Values of AI exceeding 1.0 may refer to either better improvement or negligible experimental errors. The obtained results are supported by our previously published investigations on different pesticides ,62,63.450, testosterone, and thyroxine). However, alterations induced by the mixture were greater than those recorded for each of the individual insecticides, and ABM seemed to be more effective than MET. The joint action analysis, based on the obtained biochemical data, revealed the dominance of antagonistic action among MET and ABM. Zinc supplementation in conjunction with the tested pesticides achieved considerable ameliorative effect expressed in terms of the amelioration index (AI), which was close to 1.0, indicating a maximum amelioration effect of zinc in favor of the most-assessed parameters. In conclusion, the study has focused on the joint analysis of the tested mixture and the role of zinc to alleviate oxidative stress exerted by low doses of individual pesticides and their combination, providing novel data of special concern to individuals who are occupationally exposed daily to low doses of such toxicants.The results of the present study revealed that MET was acutely more toxic than ABM to male rats. The percent of changes in the tested biochemical parameters indicated how much deviation from normal values occurred due to exposure to the pesticides. Treatments with MET and ABM, either individually or in combination, induced significant elevations in lipid peroxidation (in terms of MDA) and catalase levels, while it declined the levels of the other tested parameters (e.g., SOD, GST, GPx, GR, CYP➢The use of pesticide mixtures of “antagonistic action” should be encouraged to protect public health.➢Most current global environmental agencies derive and regulate their pesticide standard values on individual pesticides. This process must take pesticide mixtures into account when developing toxicological and regulatory standards.➢Development of protective agents against pesticide-induced oxidative stress should be among the toxicological standards sponsored by regulatory agencies.Since this study was carried out on rats, more implementation regarding human health, pesticide management, agriculture, industry, and regulatory agencies may take into consideration the following proposals:Hopefully, the highlighted recommendations could make the present study more significant regarding public health, agriculture practice, pesticide management, and global regulatory jurisdictions.


Pseudomonas aeruginosa. IgG4 elevation may be a marker of chronic infection or inflammatory stimulation. The aim of this study was to explore the prevalence of elevated IgG4 levels in CF and its correlation with the major clinical and microbiological features found in CF patients.Serum immunoglobulin (Ig) G4 elevation has been associated with several pathological conditions other than IgG4-related disease (IgG4-RD). In cystic fibrosis (CF), an elevation of specific IgG4 has been associated with colonization and infection by In a cross-sectional study, we analyzed data from a large cohort of adult CF patients attending the CF center of Lyon University Hospital. An elevated IgG4 level was defined as being above the cut-off value of 135 mg/dL.Staphylococcus aureus colonization and higher IgG, IgG1, IgG2 and IgE levels. No significant differences were observed in terms of pulmonary function, colonization with Pseudomonas aeruginosa, or the annual rate of bronchial exacerbations.One hundred and sixty-five CF patients were analyzed. An IgG4 elevation was detected in 43 patients (26%). Compared with the control group (≤ 135 mg/dL), high IgG4 patients exhibited a greater prevalence of An elevated IgG4 serum level was frequently detected in adult CF patients and did not appear to be associated with poor lung function. We suggest that IgG4 elevation is a marker of the activation of tolerance. Its clinical significance remains to be demonstrated. Cystic fibrosis (CF) is the most common severe autosomal recessive genetic disease in Europe. The disease is caused by a mutation in the gene encoding the cystic fibrosis transmembrane conductance regulator protein, a chloride ion channel whose abnormal function leads to the hyperviscosity of exocrine secretions and obstructive damage to many organs, including the lung, pancreas, hepatobiliary tracts, and vas deferens . ImmunogPseudomonas aeruginosa has remained unclear ), presence of allergic bronchopulmonary aspergillosis (ABPA), number of annual exacerbations, bacteriological chronic colonization (identification of the same microorganism during over a period of 6 months in three samplings), the results from sputum collection, digestive clinical status , CFTR-related diabetes, glycosylated hemoglobin levels (HbA1c), osteoporosis, and characteristics and presence of arthralgias. Allergic bronchopulmonary aspergillosis was defined by elevated serum IgE levels , aspergillus specific IgE (> 500 IU/mL) and/or IgG, and recent abnormalities on chest radiography.Data were prospectively collected from medical records between June 2013 and June 2014 during routine annual clinic visits under stable clinical conditions that included a sputum sample collected for bacterial and fungal examination. The clinical data extracted included the patient’s age, weight, height, body mass index (BMI), CF genotype, pulmonary function (forced expiratory volume in one second [FEV® Siemens). Normal ranges were 682–1266 mg/dL for serum IgG, 80–254 mg/dL for IgA, 71–201 mg/dL for IgM. Subclasses of IgG levels were determined by immunoturbidimetry (SPA Plus® The Binding Site). The normal ranges were 382.4–928.6 mg/dL for IgG1, 241.8–700.3 mg/dL for IgG2, 21.8–176 mg/dL for IgG3, and 3.9–86.4 mg/dL for IgG4. The patients were divided into either the IgG4high group (IgG4 > 135 mg/dL) or the IgG4norm group (IgG4 ≤ 135 mg/dL) according to their IgG4 levels. The patients’ serum IgE levels were determined by a fluorescence enzyme immunoassay (ImmunoCAP 250® ThermoFisher), with a normal range < 150 UI/mL.Immunologic parameters  were recorded. Total serum IgG, IgA and IgM serum levels were measured by immunonephelometric assay  attending the Lyon University adult CF center were included during the one-year period.According to our definition, an IgG4 elevation was detected in 43 (26.1%) of the patients. The mean serum IgG4 level in the IgG4high group was 228 mg/dL ± 68.7 mg/dL. The mean age and BMI were not significantly different between the IgG4high and IgG4norm groups .1 was similar between the two groups (63.2% ± 25.3% in the IgG4high group and 63% ± 23.4% in the IgG4norm group). The only difference observed between the 2 groups was the prevalence of Staphylococcus aureus (Sa), which was higher in the IgG4high patients than in the IgG4norm patients . No statistically significant difference were observed for methicillin resistant Staphylococcus aureus (MRSA), Pseudomonas aeruginosa, other non-fermenters , nontuberculous Mycobacterium, Aspergillus fumigatus colonization, ABPA and the number of annual exacerbations. Despite no difference in terms of Aspergillus fumigatus colonization and ABPA, the mean IgE level was 463 ±116 UI/mL in the IgG4high group versus 189 ±31 UI/mL in the IgG4norm group (p = 0.002). None of the patients in the IgG4high group had chronic respiratory insufficiency. Details are shown in The mean FEVThe prevalence of diabetes, treatment with insulin, and HbA1c values did not differ between the two groups. A majority of the patients had received supplementation for exocrine pancreatic insufficiency since childhood; there was not an observed difference between the two groups. No difference in the occurrence of cirrhosis, requirement of ursodeoxycholic acid treatment, or number of distal intestinal obstruction syndromes was noted. The occurrence of arthralgia did not differ between the 2 groups. The presence of osteoporosis was not correlated to the patients’ IgG4 levels.The elevation of IgG4 was strongly associated with hypergammaglobulinemia p = 0.0001) . The IgG . The IgStaphylococcus aureus colonization and between hypergammaglobulinemia and high IgG1, IgG2, and IgE values.To our knowledge, this study is the first to report the frequency of high IgG4 values in a large cohort of adult CF patients. In our CF cohort, the prevalence of IgG4 values > 135 mg/dL was 26%. This study did not suggest an association between the elevation of serum IgG4 levels and the clinical features evocative of an inflammatory pattern similar to that of IgG4normRD. Our results showed an association between high IgG4 value and 1, whereas hypogammaglobulinemia was associated with stable pulmonary function and a better prognosis. Only one study included adult CF patients [1, the IgG levels may represent a hyperimmune response that is ineffective against infection and possibly destructive of the airway. Overall, an elevated serum IgG level is clearly a marker of an increased inflammatory response to pathogens, and its correlation with lung function in CF patients supports the link between inflammation and deterioration in respiratory function in CF. In our study, we demonstrated that an elevation in serum IgG4 levels was associated with hypergammaglobulinemia, particularly the elevation of IgG1 and IgG2. Nevertheless, we did not find a correlation between a decline in FEV1 and an elevation in the IgG4 levels. Thus, our results suggest a neutral effect of IgG4 as an anti-inflammatory immunoglobulin in patients with hypergammaglobulinemia and without the progression of pulmonary disease. Furthermore, these results suggest that hypergammaglobulinemia may be promoted by a non-specific immune activation, which would explain the high frequency of autoantibodies in adult CF patients reported in our group [Several studies have evaluated the immunologic profile and IgG in pediatric CF patients –15. Thespatients . In thisur group .Staphylococcus aureus colonization. There was no association with MRSA or other types of bacterial colonization  or with the number of exacerbations. Staphylococcus aureus colonization in the adult CF patients was frequent, and as high as 83% in the IgG4high group versus 65% in the IgG4norm group. It is the most common species colonizing the CF respiratory tract. Moreover, chronic Pseudomonas aeruginosa pulmonary infection is associated with a decline in lung function, increased exacerbations and reduced survival in people with CF [Pseudomonas aeruginosa can induce an inflammatory response that leads to tissue breakdown and severe lung damage. Many studies have analyzed the inflammatory biomarkers in the sputum and bloodstream of CF patients to characterize pulmonary inflammation [Pseudomonas aeruginosa colonization, as would have been expected to increase in the group with hypergammaglobulinemia [In our study, we highlighted the association between elevated IgG4 values and  with CF ,19. Pseuammation ,21. In oulinemia ,15,22,23Our study addressed the association between the elevation of IgG4 levels and with IgE levels. Our results are in accordance with the literature and can be explained by a common regulation of IgE and IgG4 that is dependent on Th2 cells . In thisOur study suffered from several limitations. First, this study was conducted over one year and the evolution of the IgG4 levels was not assessed. Second, our hypothesis was not supported by experimental evidence illustrating the relation between IgG4 levels and a putative anti-inflammatory effect in CF. Third, our observations need to be interpreted with caution because of the risk of the inflation of false positive tests related to multiple comparisons.Staphylococcus aureus colonization; hypergammaglobulinemia; and IgG1, IgG2, and IgE concentrations. High IgG4 values were not associated with lung function decline. The elevation of IgG4 levels may be a consequence of chronic B-cell stimulation of gammaglobulin production associated with the activation of tolerance induction. Further studies are needed to understand the clinical significance of high IgG4 values in CF.In conclusion, high IgG4 serum values were detected in 26% of the adult CF patients. We found an association between high IgG4 values and S1 Table(XLSX)Click here for additional data file.


Hwangtoh, a type of red clay broadly deposited around the world, has traditionally been considered an eco-friendly construction material, with bonus advantages of having health and cost benefits. Presently, Hwangtoh is not commonly used as a modern construction material due to properties such as low strength and high rates of shrinkage cracking. Recent studies, however, have shown that Hwangtoh can be used as a mineral admixture to improve the strength of concrete. In addition, polyethylene terephthalate (PET) fibers recycled from PET bottle waste can be used to control shrinkage cracks in Hwangtoh concrete. Therefore, in this study, performance verification is conducted on newly developed Hwangtoh concrete mixed with short recycled PET fibers. The results show that Hwangtoh concrete has compressive strength, elastic modulus, and pH properties that are similar to these features in ordinary cement concrete. The properties of carbonation depth and creep strain of Hwangtoh concrete, however, are larger and smaller, respectively, than in ordinary cement concrete. According to flexural tests, reinforced concrete (RC) specimens cast with Hwangtoh admixtures (with and without PET fibers) possess similar or better capacities than ordinary RC specimens. The addition of PET fibers significantly improves the structural ductility of RC specimens under normal environmental conditions. However, the implementations of the concrete in aggressive environment must be carefully considered, since a previous study result indicates degradation of its durability performance in aggressive environments, such as seawater [In order to reduce carbon dioxide (COseawater . The res In addition, practices related to concrete construction exploit natural resources due to a lack of careful planning, which has triggered an urgent need to find alternative construction materials to mitigate environmental damage [2 emissions and energy consumption in the international community, construction industries all over the world are trying to reduce cement usage, find alternative materials, and use recycled materials for future construction projects [Concrete is the most widely used construction material in the world due to its low cost, high availability, and simple constructability. However, the use of cement is a main contributor to high-energy usage, COl damage . In terml damage ,4,5,6,7.projects ,9,10,11.In order to develop more optimal eco-friendly concrete and reduce cement usage, research on mineral admixtures made from industrial by-products, such as fly ash, blast furnace slag, and silica fume, is being widely conducted ,12,13,14i.e., a type of red clay) is receiving substantial interest as a possible eco-friendly construction material. Accordingly, Hwangtoh is the subject of much industry research [Hwangtoh  of plain Hwangtoh concrete, as well as the structural properties  of RC beams cast with Hwangtoh concrete and recycled PET fibers are examined.Previous research on Hwangtoh has mostly consisted of studies that investigate the basic material and mechanical properties of Hwangtoh, together with mixing proportions and procedures, to evaluate its constructability ,27,28,292) forms stable pozzolan products in Hwangtoh by combining with cement hydrate Ca(OH)2 at room temperature, thus acting as a pozzolan material, such as fly ash or silica fume [Hwangtoh has traditionally been considered an eco-friendly construction material for use in plastering walls and ondol  in Korea. The main advantages of Hwangtoh are its high heat absorption capacity, auto-purification properties, antibiotic properties, and infrared ray emission capability . Howeverica fume . The chePolyethylene terephthalate (PET) is the most widely used plastic material in the world for manufacturing secondary products such as beverage bottles. However, PET bottles are commonly thrown away after a single use, which contributes to significant environmental problems. Therefore, methods to recycle PET bottle waste have been examined in order to improve PET recycling applications. The use of PET plastic fibers recycled from PET bottle waste in construction projects has been actively studied. Short PET fibers can be utilized to effectively reduce shrinkage strain and to enhance the ductility of concrete, especially to mitigate the inherent shrinkage problem of Hwangtoh concrete ,36,41,42Our experiments aimed to reduce the use of ordinary Portland cement (OPC) in concrete mixes as much as possible by replacing OPC with Hwangtoh and blast furnace slag powders. Before the actual casting of the specimens, trial tests were performed to find the optimal replacement ratio of Hwangtoh and blast furnace slag powders. In a previous study on the use of Hwangtoh powder as an admixture in a concrete mix, serious degradations in concrete performance were observed when the Hwangtoh to cement substitution ratio exceeded 20%. Therefore, a substitution ratio exceeding 20% is generally not recommended in concrete mix proportions ,28. SignTrial mix proportion tests were carried out by setting the water-to-cement ratio (W/C) as a constant, while varying the Hwangtoh and slag powder replacement ratios. The results from the trial tests are shown in In this study, replacement ratios of 20% and 30% were used for the Hwangtoh and slag powders, respectively, for a 50% total cement replacement ratio in the concrete mix. In order to improve the high rate of shrinkage in Hwangtoh concrete, a 0.5% volume of short plastic fibers made from recycled PET plastic bottles was also added to the mix. From the trial tests, it was found that the optimal PET fiber volume percentage to control crack formation in the concrete was approximately 0.5%.3. Elastic modulus, tensile strength, and ultimate elongation percentage of the fiber were 1.2 × 104 MPa, 420.7 MPa, and 11.2%, respectively. The final mixture proportions selected for the experiments are shown in The material properties of the Hwangtoh used in this study are shown in A total of 27 specimens were used for compression and elastic modulus tests. Hwangtoh concrete cylinders with dimensions of 100 mm × 200 mm were compression tested at 28 days from initial casting. These cylindrical specimens were tested with a universal testing machine (UTM) with a maximum load capacity of 2000 kN. The compressive strength of each specimen type was determined by averaging three specimen strength values. After casting, the cylindrical specimens were covered with plastic wrap for 24 h, after which the forms were removed. The specimens were then stored in a humidity chamber at a temperature of 20 ± 1 °C and a humidity of 60% ± 5% until testing. Compressive strength and elastic modulus experiments were performed according to Korean Standards 2405 and 2438, respectively ,45. A phIn order to measure the pH of Hwangtoh cement paste, Hwangtoh specimens with W/C of 55% were prepared. For comparative purposes, additional specimens made exclusively from Korean and Chinese Hwangtoh  were prepared. The formwork was removed after 24 hours of casting. In order to prevent evaporation of moisture from the specimens, the specimens were wrapped with plastic wrap before being put into a curing chamber with a temperature of 20 ± 2 °C and a relative humidity of 64% for 200 days. The loss of alkali residuals in the powder (sieve size 300 μm) was minimized by drying the power in a drying chamber for 24 h prior to testing. A volume of 3.0 g of the dried power was subsequently mixed with 40 mL of distilled water. After stirring the wet powder for 10 min, the pH was measured using a pH-measuring device. The pH correction factor was used to set the initial pH values at 4.01, 7.00, and 10.01 prior to measurement ,47.2 conditions of 20 ± 2 °C, 60% ± 5%, and 5% ± 0.2%, respectively. Carbon depth was measured based on the location of phenolphthalein color change at testing times of one, four, eight, 12, and 25 weeks, following the guidelines of KS F 2596 [In order to evaluate the effect of Hwangtoh on the carbonation properties of Hwangtoh concrete, carbonation depths at various curing ages were measured. The accelerated carbonation test was performed following the test guidelines of KS F 2584 . The speS F 2596 .Currently, there are no guidelines available for the creep testing of recycled PET fiber-added Hwangtoh concrete. Accordingly, the concrete creep test guidelines of ASTM C 512 were used for this study . A cylinTo evaluate the drying shrinkage cracking capacity of Hwangtoh concrete, free and restrained drying shrinkage strain experiments were performed according to Korean Standard 2595, which details the drying shrinkage cracking test method for restrained concrete . The speAll specimens were tested for flexural strength 28 days after casting. The specimens were reinforced with three D13 rebars (with a diameter of 13 mm) as tension reinforcement and D10 rebars (with a diameter of 10 mm) with 150 mm spacing as shear reinforcement. The dimensions and details of an RC flexural test specimen are shown in The compressive strength test results for the Hwangtoh concrete are shown in The elastic modulus test results for the various Hwangtoh concrete specimens are shown in The compressive failure modes of the cylindrical specimens are shown in 2) is approximately 12.5 [Measurement data on pH values are shown in ely 12.5 . Thus, t2 gas into the concrete. In this way, the interior microstructure compactness of concrete dictates its carbonation resistance. In addition, interior defects or interfaces decrease the carbonation resistance of concrete. Based on these fundamental mechanics, it is logical that slag and Hwangtoh-added concrete would have less carbonation resistance, because lesser amount of calcium oxide are produced in these specimens than in OPC specimens. Additionally, the addition of PET fibers increases the damage-like interfaces in the engineered specimens, resulting in lower carbonation resistance.Carbonation depth data are shown in −6), slag (780 × 10−6), to Hwangtoh-slag-PET fiber (871 × 10−6) concrete, with Hwangtoh-slag concrete having the lowest creep strain at 120 days. The creep strain test results demonstrate that Hwangtoh is less susceptible to creep strain than slag cement, because the proportion of cement is reduced by the addition of the Hwangtoh admixture in the Hwangtoh-slag concrete mix relative to the slag concrete mix. In addition, the addition of recycled PET fibers increased the interfaces in the concrete, resulting in significant creep strain from the closing of interfaces as the duration of loading increased.The creep strain test results for slag concrete, Hwangtoh concrete, and Hwangtoh-slag-PET fiber concrete measured up to 120 days are shown in The measured free drying shrinkage strain in the test specimens is shown in For the restrained drying shrinkage strain tests, the cracking times from the initial casting for all specimens are listed in The load-deflection test results for different admixture-added RC specimens are shown in The ductility index is used to evaluate the energy absorbing capacity of reinforced concrete members, which, along with the strength, is one of the most important factors in evaluating the safety of a structural member. In this study, the ductility index (Δu/Δy), which is defined as the ratio of deflections at the ultimate load, Δu, to the deflections at the time of tension rebar yielding, Δy, is used to evaluate the safety of a member see . As calci.e., the energy absorbing capacity). These results validate the results of a previous study by Lin et al. [The recycled PET fiber reinforced specimens exhibited better ductility indices than the non-reinforced specimens. As shown in n et al. , whereini.e., KHSP and CHSP) exhibited significantly larger deflections than the deflections of the Hwangtoh concrete specimens without recycled PET fibers . These findings validate the addition of recycled PET fibers in Hwangtoh concrete to improve the crack-controlling capacity of RC members.The failure modes of the RC specimens are shown in (1)From the trial tests, the optimum replacement ratio of Hwangtoh powder to cement is 20%. In addition, without using a chemical water-reducing admixture, desired workability can be achieved through the addition of blast furnace slag powder. By replacing cement with Hwangtoh and blast furnace slag powders in proportions of up to 50%, eco-friendly Hwangtoh concrete with material properties equivalent to the properties of OPC concrete can be developed.(2)According to the results of the tests herein, Hwangtoh concrete exhibits a slightly lower compressive strength and elastic modulus than plain concrete. However, the differences (less than 2%–6%) are minute, thus, validating the possibility of using Hwangtoh concrete as a structural construction material.(3)The free drying shrinkage strain of Hwangtoh concrete is approximately 1.5 to 2.0 times greater than the free drying shrinkage strain of plain concrete. The cracking time of Hwangtoh concrete is approximately five days earlier than the cracking time of plain concrete. However, the cracking time of the recycled PET fiber-reinforced Hwangtoh concrete is similar to the cracking time of plain concrete. Accordingly, these finding suggest that the crack-prone characteristics of Hwangtoh concrete can be mitigated by adding short plastic fibers, such as PP or PET fibers, to the material.(4)Hwangtoh concrete measures slightly lower pH values than the pH values of ordinary cement concrete. The carbonation depth of Hwangtoh concrete is larger than the carbonation depth of ordinary cement concrete due to the decreased production of calcium oxide caused by the replacement of ordinary cement with Hwangtoh. Creep strain is highest in Hwangtoh-PET fiber concrete, followed by slag cement concrete and Hwangtoh concrete. These findings show that there is a tradeoff between drying shrinkage strain resistance and creep strain resistance, which comes with the addition of PET fibers.(5)The flexural failure behavior of Hwangtoh concrete RC beams is very similar to the flexural failure behavior control specimens herein. The fiber-bridging capability of members reinforced with recycled PET fibers in the tensile region improves the flexural capacity of the RC members by delaying ultimate failure.(6)This study’s evaluation of the ductility index reveals almost no difference between Hwangtoh and plain concrete without recycled PET fibers. RC specimens with recycled PET fibers, however, have relative ductility indices that are approximately twice as great as the ductility indices of concrete without recycled PET fibers. Therefore, the ductility of RC members cast with Hwangtoh concrete is significantly improved by the addition of recycled PET fibers.(7)Since all of the performance evaluations are conducted under normal environments in dry condition, additional durability tests of the material in a variety of aggressive environments, such as seawater, must be addressed in future works.This study evaluates the material, durability, and structural capacities of Hwangtoh concrete. Plain and RC specimens cast with Korean and Chinese Hwangtoh admixtures and short recycled PET fibers are considered herein. The results of basic material property tests and RC structural member capacity tests are summarized as follows.


Syst. Appl. 18:251-264, et al. (Abstr. Appl. Anal. 2014:852324, In the present study, we work on the problem of the existence of positive solutions of fractional integral equations by means of measures of noncompactness in association with Darbo’s fixed point theorem. To achieve the goal, we first establish new fixed point theorems using a new contractive condition of the measure of noncompactness in Banach spaces. By doing this we generalize Darbo’s fixed point theorem along with some recent results of (Aghajani  Fractional calculus is the study of integrals and derivatives of an arbitrary order. Fractional calculus seeks to find the integrals and derivatives of a real or even complex order using the Gamma function, Euler’s generalization of the factorials. In modern times differential/integral equations with nonintegral order have drawn the attention of numerous researchers due to their wide applications in several fields of science and engineering. The need for fractional order differential/integral equations stems in part from the fact that many phenomena cannot be modeled by differential/integral equations with integer derivatives. Analytical and numerical techniques have been implemented to study such equations.H defined by the formula I . We areers  given in [Throughout this paper, we assume that given in .E if it satisfies the following conditions: ∘)(1the family ∘)(2∘)(3∘)(4∘)(5∘), represents the kernel of the MNC β and sinceThe subfamily kerβ an MNC and C to be a nonempty, bounded, closed and convex subset of a Banach space E.From now on we denote by Darbo’s fixed point theorem (DFPT) is a very important generalization of Schauder’s fixed point theorem and Banach’s fixed point theorem.Banaś and Goebel LetQbe a self-continuous operator onCand assume ∃ to be a constantsuch thatfor any nonempty subsetThenQhas at least one fixed point inC.Following the trend of Lemma To complete the proof, we need the following notions.For n, then For  We denote by ϒ a pair Let Take Let Δ be a set of functions (i)(ii)F is continuous,(iii)Let We are now in a position to state and prove a new DFPT theorem.LetTbe a self-continuous operator onCsuch thatfor any subsetXofC, whereandis a continuous function. ThenThas at least one fixed point inC.N such that ∘) of Definition C. Besides we know that β. So T. Consequently, Schauder’s fixed point theorem implies this result in C. □We start with Also, by , we haveTaking LetTbe a self-continuous operator onCsuch thatfor any subsetXofC, whereandis a continuous function. ThenThas at least one fixed point inC.Take Take Take Set entclass1pt{minimaTaking LetTbe a self-continuous operator onCsuch thatfor any subsetXofC, whereandis a continuous function, whereis a function such thatforifthenforwithiffor alln, thenThenThas at least one fixed point inC.Take Following Proposition 9 (see ) and TheLetTbe a self-continuous operator onCsuch thatfor any subsetXofC, whereis a continuous function, andare two nondecreasing and continuous functions satisfyingif and only ifThenThas at least one fixed point inC.Take entclass1pt{minimaTake LetTbe a self-continuous operator onCsuch thatfor any subsetXofC, whereandThenThas at least one fixed point inC.Take mentclass2pt{minimHere we derive some new coupled fixed point (CFP) results by means of the MNC.An element The first CFP result is the following.Suppose thatis continuous operator such that, forfor allwhereis a continuous sub-additive function, andare sub-additive functions. ThenGhas at least one CFPĜ is continuous due to the continuity of G. Following [X. Without loss of generality, we assume M is a nonempty subset of ∘) of Definition Ĝ has at least one fixed point in G has a CFP. □Consider the map ollowing , we defiondition  and usinThe second outcome of this section is the following.Suppose thatis a continuous operator such that, forfor allwhereandis a continuous function. ThenGhas at least one CFPĜ is continuous due to the continuity of G. Following [β̂ as a new MNC in the space M. Without loss of generality, we take M, a nonempty subset of ∘) of Definition Ĝ has at least one fixed point in G has a CFP. □Consider the map ollowing , we exprondition  and usinφ.We can derive some new CFP results from Theorems X be a nonempty and bounded subset of X are nondecreasing on I if and only if Let β of the family β is a MNC in the space Further, we define the function shown in  that theI,the function φ is superadditive, i.e., the function H generated by the function x the conditionthe superposition operator the function the function the inequalityWe consider the following assumptions: Under assumptions (equation (has a positive solutionwhich belongs to the spaceequation  has a poT defined on the space g on the set f on the set I, we have T maps the ball i.e., T is continuous on g in T is continuous on T on the subset T transforms the set T is continuous on X be a nonempty subset of X, we obtaing on the set h on the set a on I, we have β, we obtainFor Finally, we present an illustrative example for Theorem Assumption Assumption Assumption Assumption Assumption Assumption equality  has the  Thus, from all the above mentioned observations, it is clear that equation (Consider the integral equation of the formequation . In thisIn this work we studied the problem of the existence of positive solutions of fractional integral equations by means of the MNC in the association with DFPT. For this purpose we first established a new DFPT and its coupled version that generalized some existing results. To support our results, an illustrative example is provided.


New fossils from the latest Pliocene portion of the Tatrot Formation exposed in the Siwalik Hills of northern India represent the first fossil record of a darter (Anhingidae) from India. The darter fossils possibly represent a new species, but the limited information on the fossil record of this group restricts their taxonomic allocation. The Pliocene darter has a deep pit on the distal face of metatarsal trochlea IV not reported in other anhingids, it has an open groove for the m. flexor perforatus et perforans digiti II tendon on the hypotarsus unlike New World anhingid taxa, and these darter specimens are the youngest of the handful of Neogene records of the group from Asia. These fossil specimens begin to fill in a significant geographic and temporal gap in the fossil record of this group that is largely known from other continents and other time periods. The presence of a darter and pelican  in the same fossil-bearing horizon strongly indicates the past presence of a substantial water body  in the interior of northern India in the foothills of the Himalayan Mountains. Overal century ,6,7). Ne century ,4,8.Anhinga melanogaster) occurs in across much of India, excluding the Himalayas and Northwestern-most portions of India, in freshwater and coastal habitats [Anhinga are known from adjacent Pakistan [Recent fieldwork by individuals at Panjab University (India) is adding new taxa and additional specimens to the Indian Neogene record from the Siwalik Hills in northern India  with three subspecies, and a single New World species (A. anhinga) comprising two subspecies  [A. melanogaster rufa. Attempts at taxonomic and phylogenetic resolution based on morphological data [The taxonomy of the extant species level taxa of Anhingidae is divided into two opinions. One scenario groups all of the Old World individuals into a single species  . Howeverralasia) , where tcal data ,15 have cal data  supportscal data  in his rAnhinga pannonica (from eastern Europe) has had material from Africa [There are many known Neogene fossils of darters in the Old World, and they include specimens from the Miocene of Thailand , the Miom Africa ,23, Pakim Africa , and elsm Africa ,27. The m Africa  was idenm Africa , but them Africa ,30).Anhinga pannonica from the late Miocene of Pakistan [Anhinga cf. pannonica from the Miocene of Thailand [Anhinga sp. from the late Miocene of Abu Dhabi [A. pannonica [Anhinga in Asia have not produced additional tarsometatarsi. Furthermore, the Miocene specimens from Kenya, Tunisia, Chad, and Ethiopia do not include a tarsometatarsus [Anhinga hadarensis from the Pliocene of Ethiopia [Despite that seemingly broad assemblage, there appears to be only three or four published records of anhingids from Asia. There are specimens referred to Pakistan , AnhingaThailand , Anhingabu Dhabi , and thebu Dhabi ,29. All annonica  includesannonica , but thetatarsus ,21,23, aEthiopia . AdditioEthiopia  also lacInstitutional/collection abbreviations: KP/KK/BS–Khetpurali/Kewal Krishan/Bahadur Singh, housed in the Department of Anthropology, Panjab University, India; MVZ–Museum of Vertebrate Zoology, University of California, Berkeley, USA; UCMP–University of California Museum of Paleontology, Berkeley, USA; VPL/RP-KPB–Vertebrate Palaeontology Laboratory /Rajeev Patnaik-Khetpurali Bird.Osteological terminology follows Baumel and Witmer  with somAnhingidaeAnhinga anhinga leucogaster MVZ 85509Anhinga melanogaster novaehollandiae MVZ 143017Anhinga melanogaster novaehollandiae (papua race) MVZ 149268PhalacrocoracidaePhalacrocorax auritus auritus MVZ 151575Phalacrocorax capillatus MVZ 124049Phalacrocorax carbo novaehollandiae MVZ 143257Phalacrocorax melanoleucos melanoleucos MVZ 156698Phalacrocorax gaimardi MVZ 157988Phalacrocorax harrisi MVZ 140913Phalacrocorax brasilianus mexicanus MVZ 46167Phalacrocorax pelagicus resplendens MVZ 19089Phalacrocorax penicillatus MVZ 46809Phalacrocorax sulcirostris sulcirostris MVZ 143250Phalacrocorax punctatus punctatus MVZ 164582Phalacrocorax varius hypoleucos MVZ 143267Phalacrocorax urile MVZ 154268SulidaeMorus bassanus UCMP 131057Morus sp. UCMP 123175 and 137455Sula leucogaster MVZ 54774Sula sula MVZ 132906Order PELECANIFORMES Sharpe, 1891 Family ANHINGIDAE Lesson, 1831 ANHINGA Brisson, 1760 [Genus on, 1760 ANHINGA sp.VPL/RP-KPB1 is a distal left tarsometatarsus , KP/KK/BThe specimens were collected from a bone-bearing horizon exposed in the upper part of the Tatrot Formation  in the Siwalik Hills of Northern India that is latest Pliocene in age (~2.6 Ma), and the locality produced a previously published pelican specimen see ; Fig 1)..Fig 1). On the plantar side, the shaft specimen KP/KK/BS/102  exhibitsA. anhinga, shorter than that of A. walterbolesi, and within the size range of A. melanogaster , a proximal plantar edge of the lateral trochlear rim of metatarsal trochlea IV that forms a hook with a concave proximal margin , a very pointed intercotylar eminence (as compared to the blunter shape in extant and extinct anhingids), and a distinct very deep pit on the distal face of metatarsal trochlea IV . The siHarrison ). Howevechlea IV  that is The preserved portions of the tarsometatarsus  derive from a tarsometatarsus that would have been approximately 46 mm long. That length was derived from a measurement of the total length of the three specimens when their overlapping morphologies are placed in line. The description below proceeds from the distal specimen through the midshaft specimen, to the proximal end fragment.Specimen VPL/RP-KPB1 preserves the most distal portion of a left tarsometatarsus and is broken at its proximal end through the metatarsal I facet leaving only its distal tip preserved . The latMetatarsal trochlea II extends distal to trochlea III, and that extends distal to trochlea IV. The lateral edge of the bone is flush with the lateral surface of metatarsal trochlea IV  meaning that metatarsal trochlea IV is in line with the tarsometatarsal shaft . In contrast, metatarsal trochlea II is distinctly projected mediodistally with the medial margin of the bone being concave . Metatarsal trochlea II is asymmetrical with a single large bulbous portion dorsally and a concave plantar face. The medial face of metatarsal trochlea II has a large collateral ligament pit that extends proximally making a notch in the proximal medial edge of the trochlea. In addition, there is a very large plantar medial flange on metatarsal trochlea II that extends far medially with its apex near the proximodistal midpoint of the trochlea, and the apex is slightly worn. The plantar edge of metatarsal trochlea II also is positioned plantar to the other trochleae. The lateral edge of metatarsal trochlea II is concave, and the lateral edge continues proximally (plantar face) as a raised ridge that ends at the same proximodistal level as the apex of the medial plantar flange. The intertrochlear incisions are approximately equal in width, but the lateral incision extends more proximally (and is rather short). Metatarsal trochlea III has a furrow extending from its proximal dorsal end to its proximal plantar end. The area just proximal to the dorsal proximal end of metatarsal trochlea III is slightly concave, and the proximal end of the plantar surface of the trochlea is rounded and symmetrical. The medial and lateral sides of metatarsal trochlea III are nearly flat. Metatarsal trochlea IV is asymmetric with a large plantar flange and a deep pit on the distal face. The lateral surface of metatarsal trochlea IV has a collateral ligament pit in the plantar half of that face , and there is a smaller concave area dorsal to the collateral ligament pit . The proximal edge of the lateral flange of metatarsal trochlea IV is concave with a distinct notch. The pit on the distal face is deepest dorsally and extends plantarly  as a concave area on the plantar face of the trochlea. The dorsal surface of metatarsal trochlea IV is flat to slightly convex. The dorsal part of the medial face of metatarsal trochlea IV is concave with the dorsal medial edge of the trochlear surface overhanging the intertrochlear space . There is a tendinal groove proximal to the dorsal opening of the distal vascular foramen. Lateral to that groove, the dorsal face of the shaft is relatively flat and more plantar in relation to the area medial to the groove that is convex over its surface.Specimen KP/KK/BS/102 preserves most of the mid-shaft of the tarsometatarsus from the metatarsal I facet proximally to the distal end of the hypotarsus, and it is 27.0 mm long . The facThe tarsometatarsus fragment KP/KK/BS/101 is the proximal portion broken near the distal end of the dorsal infracotylar fossa, and the medial hypotarsal crest is mostly missing . The speThe dorsal infracotylar fossa is deep proximally, and the lateral rim of the fossa extends distal to the medial rim. The proximal vascular foramina are near the medial and lateral edges of the dorsal infracotylar fossa and close to the same proximodistal level as the ridges for the attachment of the extensor retinaculum . The pair of ridges for the extensor retinaculum appear to be highly asymmetrical with the lateral one forming the thin proximal medial edge of the dorsal infracotylar fossa , and the medial one seems to be a short more proximally positioned ridge. However, the area distal to the preserved medial attachment of the extensor retinaculum is damaged, and the ridge may have extended more distally. The area between the two retinacular attachments is concave. The medial rim of the extensor retinaculum distal to the broken area near the retinacular attachment is concave medially, and the proximal end of the broad notch for the m. extensor hallucis longus muscle is preserved near its distal end. The medial margin of the bone just proximal to the proximal end of the broad notch for the m. extensor hallucis longus is shallowly concave. There is a distinct lateral tuberosity for the m. tibialis cranialis near the mediolateral midpoint of the extensor fossa distal to the proximal vascular foramina, and a thin distally elongate ridge in the more medial portion of the fossa. The medial tuberosity for the m. tibialis cranialis is as distinct as the lateral one.The medial plantar face of the tarsometatarsus is a shallow  fossa that extends distally slightly less distal as compared to the medial rim of the extensor fossa. The plantar opening of the medial proximal vascular foramen is near the proximodistal midpoint of that fossa. The lateral edge of that fossa is formed by a (damaged) ridge that extends distally from the medial hypotarsal crest. The area distal to the two main preserved tendinal canals (of the hypotarsus) in the central portion of the plantar tarsometatarsus shaft is slightly concave over its surface and raised above the adjacent medial and lateral faces. The lateral face of the plantar shaft is a proximodistally elongate fossa for m. abductor digiti IV (that extends onto the other specimens described above) adjacent to the thin lateral edge of the bone. The lateral proximal vascular foramen opens plantarly near the border between the fossa for the m. abductor digiti IV and the central raised area, and the two proximal foramina are at approximately the same proximodistal level.There is no large plantar ridge  of the hypotarsus preserved . In thoerbolesi ), it appure 1 in ).A. melanogaster and the Indian specimen above), but its intercotylar eminence is rounded, in contrast to the pointed condition in the Siwalik Hills fossil. The Khetpurali proximal tarsometatarsus also has a narrower proximal mediolateral width (11.0 mm) than that of the A. cf. pannonica specimen from Thailand (12.4 mm) and A. grandis (13.8 mm) from the Miocene of the USA [The anhingid tarsometatarsus from the Miocene of Thailand reportedly has the laterally open groove for the m. flexor perforans et perforatus digiti II tendon  in the Indian specimen. Additionally, the impression of the lateral collateral ligament just distal to the lateral cotyle projects more laterally than the state in the Indian specimen  than the more medioplantarly directed process in A. walterbolesi. Anhinga walterbolesi also lacks the notch in the proximal plantar lateral rim of metatarsal trochlea IV , and the enlarged pit on the distal face of metatarsal trochlea IV  further precludes the likelihood of them being conspecific.The Indian specimens differ from the Oligo-Miocene ure 2 in ). In add18.1 mm; ; Table 1ure 2 in ; Fig 4).g flange . There ig flange . While ture 2 in ). The grAnhinga hadarensis [A. hadarensis is 10.0 mm, close in size to that of extant species [A. hadarensis.The tarsometatarsus is not known from Pliocene African darensis , and thu species . With thMacranhinga and Meganhinga have an enclosed tendinal canal for m. flexor perforans et perforatus digiti II set within the medial hypotarsal crest, consistent with the state in A. anhinga  fossil taxa [There are many extinct species of anhingas in South America that range broadly in size with many of the species much larger than the Indian specimen and some species not known from a tarsometatarsus ,30. Thosure 1 in ). That sure 1 in . Therefoure 1 in  and the ambashag . If thatsil taxa .A. melanogaster such as the laterally open tendinal groove for the m. flexor perforans et perforatus digiti II. Metatarsal trochlea II extends distal to trochlea III  in Anhinga anhinga leucogaster (MVZ 85509) and Anhinga melanogaster novaehollandiae  have an at least partially enclosed canal for that tendon, demonstrating variation in that character that is perhaps ontogenetic. The enclosed canal also is present in the phalacorcoracoid outgroup Sulidae  [The Khetpurali specimens differ from the extant material examined as well, while sharing some features with  131057) .Anhinga anhinga leucogaster and A. melanogaster novaehollandiae, similar to, but perhaps not quite as pronounced as in the Khetpurali material. The size of that projection varies among the cormorants examined with it larger in some species  and smaller in others , but that feature does not approach the extremely large state in the Indian fossil or that in extant anhingas . In all cormorants examined , metatarsal trochlea II is more plantar than the state in the fossil and extant anhingids. The plantar proximal end of metatarsal trochlea III is relatively wide and symmetrical in the fossil. It shares that state with Anhinga anhinga leucogaster, and differs from the asymmetrical and narrowed proximal end in P. carbo novaehollandiae, P. capillatus, P. auritus auritus, P. melanoleucus melanoleucus, P. gairmardi, P. brasilianus mexicanus, P. punctatus punctatus, and P. urile. In P. harrisi, the proximal end narrows, but is rounded, the state in P. penicillatus, P. varius hypoleucos and P. sulcirostris sulcirostris is narrow and symmetric, and the state is more similar to the fossil in P. pelagicus resplendens. Anhinga melanogaster varies a bit from a broad, but slightly asymmetric end, to a narrowed, but symmetrical shape. The fossil displays a notch in the proximal plantar base of metatarsal trochlea IV . That notch is absent in A. anhinga leucogaster, A. melanogaster novaehollandiae , and all cormorants examined, but one specimen of A. melanogaster novaehollandiae (MVZ 149268) has a slightly broader notch than the state in the fossil.There is a relatively large plantar medial projection from metatarsal trochlea II in anhingas . The dorA. pannonica and from the extant species. The Indian material cannot be directly compared to A. hadarensis from the Pliocene of Ethiopia [Anhinga (Old World and New World taxa), it is possible that the African (Hadar) and Indian  specimens could be conspecific. Furthermore, the features of the Indian fossil taxon  might have a greater variability within species than currently realized, and that potential variability may mean that these specimens could be conspecific with a known extinct taxon  or represents an extinct lineage within A. melanogaster. It also is possible that these fossils could represent an anagenetic lineage leading to one of the extant species/subspecies, but that cannot be determined until a larger sample of specimens is found. It should be noted that specimens allocated to Anhinga melanogaster have been published from the Miocene, Pliocene, and Pleistocene of Africa suggesting the antiquity of the species lineage [It would appear based on the comparisons available that these new Asian anhingid specimens might represent a new species. The material is younger (latest Pliocene versus Miocene) than the other specimens from Asia. At a minimum, they appear to differ from the material referred to Ethiopia  because  lineage , but no Phalacrocorax carbo, but admitted that he did not do extensive comparisons. In the illustration of the fossil [A. pannonica). This cormorant specimen from an unknown Neogene horizon in the Siwalik Hills sequence needs to be reexamined in order to determine its phylogenetic affinities, though it most likely is distinctly older  than the Kherpurali material (latest Pliocene).The proximal tarsometatarsus identified as a cormorant from the Siwalik Hills  originale fossil , it can e fossil  or visibThe presence of a pelican and multiple anhingid individuals at the same locality/stratigraphic horizon in the Khetpurali section of the Siwalik Hills all point to a strong aquatic influence on the site that is today over 1000 km from the nearest part of the Indian Ocean. The associated fauna of this bone-bearing horizon in the Tatrot Formation includes crabs, crocodiles, and fish, in addition to the avian fossils ,40, and A. anhinga can be excluded as a possibility), although fossils allocated to A. melanogaster are known from older and younger sediments in Africa [A. pannonica), likely deriving from a new taxon or lineage  that may be an extinct part of the A. melanogaster grouping of Old World individuals. Other workers have supported the evolution of the A. melanogaster group by the Miocene in Africa, but early representatives of that group have not yet been reported in Asia (see [Similar to the pelican taxon from the same locality, the anhingid specimens cannot be placed clearly into any extant species (though n Africa . These Ksia (see  for a lisia (see  slightlysia (see . Given t


Many musculoskeletal pain conditions are characterized by hypersensitivity, which is induced by central sensitization (CS). A questionnaire, the Central Sensitization Inventory (CSI), was recently developed to help clinicians identify patients whose presenting symptoms may be related to central sensitivity syndrome (CSS). The aims of the present study were to examine criterion validity and construct validity of the Japanese version of the CSI (CSI-J), and to investigate prevalence rates of CS severity levels in patients with musculoskeletal disorders.Translation of the CSI into Japanese was conducted using a forward-backward method. Two hundred and ninety patients with musculoskeletal pain disorders completed the resultant CSI-J. A subset of the patients (n = 158) completed the CSI-J again one week later. The relationships between CSI and clinical symptoms, EuroQol 5-dimension (EQ-5D) and Brief Pain Inventory (BPI), were examined for criterion validity. EQ-5D assesses Health-related QOL and BPI measures pain intensity and pain interference. The psychometric properties were evaluated with analyses of construct validity, factor structure and internal consistency, and subsequently investigate the prevalence rates of CS severity levels.The CSI-J demonstrated high internal consistency (Cronbach’s α = 0.89) and test-retest reliability was excellent value (ICC = 0.85). The CSI-J was significantly correlated with EQ-5D (r = −0.44), pain intensity (r = 0.42), and pain interference (r = 0.48) . Ten percent of the participants were above the cutoff “40”. The exploratory factor analysis resulted in 5-factor model.This study reported that the CSI-J was a useful and psychometrically sound tool to assess CSS in Japanese patients with musculoskeletal disorders. The finding of the prevalence rates of CS severity levels in patients with musculoskeletal disorders may help clinicians to decide strategy of treatment. Central sensitization (CS) is defined as increased responsiveness of nociceptive neurons in the central nervous system to normal or subthreshold afferent input by the International Association for the Study of Pain ; it is oCentral sensitivity syndromes (CSS) are an overlapping and similar group of syndromes that are bound by the common mechanisms of CS, which lead to hypersensitivity including hyperalgesia and allodynia. CSS conditions are not confined to one specific region of the body. This is the reason why CSS conditions are disorders of pain processing in the central nervous system. CS is likely to play a causative role in CSS, probably with other risk factors . The preDirect measures of CS are often conducted by Quantitative Sensory Testing (QST), which consists of static and dynamic psychophysical tests to quantify somatosensory function in response to the controlled stimuli . AlthougTranslation and validation studies of the CSI have been completed in several different languages, including Dutch , French The Japanese version was linguistically validated through the general cross-cultural adaptation process: forward-translation, back-translation, and cognitive debriefing. First, the Japanese speakers  translated the original CSI items from English to Japanese. Second, the revised Japanese version was back-translated from Japanese to English by a native English speaker. Third, the back-translation was checked and approved by the developer of the original CSI, and a provisional version of the CSI-J was created. Finally, the provisional CSI-J was administered to six native Japanese patients with musculoskeletal disorders, who provided feedback on comprehensibility and completeness of the content and time exposure. This final pre-testing revealed ambiguity about the answer choices. Therefore, we altered the expression of them, and we developed a final version of the CSI-J  39]..39].A total of 290 patients were recruited from an orthopedic clinic, of which those who were aged between 20 and 80 years and suffered from musculoskeletal pain, such as neck, shoulder, low back, hip, knee, or ankle, were included. Exclusion criteria included patients diagnosed with cancer, brain or spinal cord injury, neurological disease, dementia, and poor Japanese language comprehension. Ethical approval was obtained from the Institutional Ethics Committee of Konan Women’s University. Informed consent was obtained from all subjects prior to the study. The study was conducted in accordance with the Declaration of Helsinki.Demographic , CSI-J, and four pain-related outcomes  were assessed in all participants. A test-retest reliability of the CSI-J was determined with a time interval of 1 week. These domains were selected because patients whose presenting symptoms may be related to a CSS  showed significant relationships between CS outcome and QOL, pain intensity, and disability , 40–42.The CSI-J consists of two parts: A and B. Part A is a 25-item self-report questionnaire designed to assess health-related symptoms that are common to CSSs. Each item is rated on a 5-point Likert-type scale , with total scores of 0–100. Part B (which is not scored) is designed to determine whether one or more specific disorders, including seven separate CSSs, have been previously diagnosed .Health-related QOL was measured using EuroQol 5-dimension (EQ-5D) . EQ-5D wPain intensity and pain interference were measured using the Brief Pain Inventory (BPI) ,47. It c2,1 values ≤0.40 were considered to indicate fair reliability, 0.41–0.60 moderate reliability, 0.61–0.80 substantial reliability, and ≥0.81 almost perfect reliability . An analysis of individual item scores revealed that 21 of 25 items showed an ICC >0.60 (range 0.61–0.82). Items 2 (0.48), 3 (0.48), 11 (0.57), and 19 (0.38) showed an ICC <0.60.Of the all participants, 158 patients answered the questionnaire twice. There was an excellent agreement between the test and retest total scores, with an ICCWhile the CSI-J was not related with the duration of pain , it was significantly correlated with EQ-5D (r = −0.44), pain intensity (r = 0.42), and pain interference (r = 0.48)  indicated subclinical, 44 patients (15.17%) indicated mild, and 32 patients (11.00%) indicated moderate or higher severity. No patients had been diagnosed with fibromyalgia and multiple CSs . As showThe EFA produced a five-factor model, of which three factors “Emotional distress”, “Urological and general symptoms”, and “Headache/Jaw symptoms” were similar in comparison to the categorization of items in the original English version . Factor The aim of this study was to validate and reveal the underlying structure of the CSI-J in a sample of Japanese patients with musculoskeletal disorders. Our results showed that the CSI-J had excellent internal consistency and test-retest reliability, as well as significant positive associations with two pain-related scales of BPI. Additionally, there was a significant negative association between the CSI-J and EQ-5D. Factor analysis revealed that the CSI-J had a 5-factor structure, which contrasts with the English , Dutch , SpanishThe internal consistency of the CSI-J was 0.89, below the accepted 0.95 threshold for item redundancy . It was The ICC score was 0.85, indicating that the CSI-J has excellent reliability, corroborating earlier reports on the English (0.82) , Dutch  before knee arthroplasty reported more severe postsurgical pain intensity , and patThere were some limitations to the present study. First, we did not measure QST as a direct measure of CS. Further research is needed to examine the validation of the CSI-J by using QST. Second, we did not investigate the sensitivity of the CSI-J to changes in clinical status; hence, we cannot conclude about causation, predictive validity, or response to intervention. Third, the number of CSS was assessed using self-report questionnaire, which potentially involved response bias. Therefore, no patient reported the comorbidity of fibromyalgia because participants might be biased toward responding “no”. Such issues would seem to be appropriate suggestive steps for research on the CSI-J.In conclusion, this study reported that the CSI-J was a useful and psychometrically sound tool to assess CSS in Japanese patients with musculoskeletal disorders. The finding of the prevalence rates of CS severity levels in patients with musculoskeletal disorders may help clinicians to decide strategy of treatment.S1 Table(DOCX)Click here for additional data file.


IgG4-related hypophysitis is a rare, inflammatory process of the pituitary that mimics more commonly seen pituitary tumors. We report a case of IgG4-related hypophysitis in a 16-year-old female with diabetes insipidus who was found to have IgG4-related hypophysitis based on tissue diagnosis. This entity has not been previously described in a pediatric patient. Recognition of certain inflammatory processes of the pituitary may lead to alternative means of diagnosis and medical management without a biopsy. IgG4-related disease (IgG4-RD) is a rare, newly recognized, multi-organ disease characterized by a tendency to form tumefactive lesions in multiple sites throughout the body. Pituitary involvement, IgG4 hypophysitis, is less commonly reported and, when described, is usually associated with IgG4-RD elsewhere in the body. In this case report, we present a pediatric patient who presented with diabetes insipidus and an isolated suprasellar mass which was subsequently resected revealing a pathologic diagnosis of IgG4 hypophysitis. This is the first report of IgG4 hypophysitis in a pediatric patient. Given that the treatment of IgG4 hypophysitis is drastically different than treatment for other more common pituitary tumors within this population, it is important to consider this entity in the differential in order to prevent unnecessary invasive procedures and subsequent morbidity.A 16-year-old female presented to our endocrinology, otolaryngology, and neurosurgery clinics for an evaluation of a suprasellar lesion. She had presented three months previous to an outside endocrinologist for excessive polyuria and polydipsia. She was found to have a serum osmolality of 292 mOsm/kg and a urine osmolality of 50 mOsm/kg, and was given a diagnosis of diabetes insipidus. During the workup, magnetic resonance imaging revealed a 2 cm peripherally enhancing, partially cystic sellar/suprasellar mass causing compression of the optic chiasm  (Wegener's) were not present. Because of the possibility of mass effect on the optic chiasm and radiologic findings consistent with craniopharyngioma, namely the solid/cystic appearance and the peripheral enhancement of the lesion, an endoscopic transsphenoidal resection of the pituitary was performed. The lesion was found to be encapsulated and consisted of a white-tan diffluent material. The lesion was completely resected from the sella and sent, in aggregate, for pathologic evaluation.Sections of the lesion showed extensive infiltration of mature plasma cells intermixed with lymphocytes Figure . SectionPostoperatively, the patient experienced a small CSF fluid leak which was repaired surgically without issue. She was discharged from the hospital on steroid replacement and DDAVP. Given the diagnosis, the patient was referred to Rheumatology for follow-up. She was found to have no obvious sign or symptoms of systemic involvement of IgG4-RD. An IgG4 level was performed and was normal; however, the patient was receiving chronic steroid replacement therapy at that time. Unfortunately, there have been no follow-up visits at our institution since that time.e.g.Churg-Strauss syndrome, multicentric Castleman’s disease, sarcoidosis, Sjogren’s syndrome) by additional histopathological examination. Granulomatosis with polyangiitis (GPA) (Wegener's) may mimic IgG4-related disease and is an important consideration in the differential diagnosis [IgG4-related disease (IgG4-RD) is a newly recognized, fibroinflammatory disease that has been shown to affect almost every organ of the body, with preference for the pancreas, salivary gland, orbital tissue, lymph node, lung, and kidney. It is characterized by tumorous swelling of affected organs, prominent lymphoplasmacytic infiltrate rich in IgG4-positive plasma cells, and high serum IgG4 concentrations. IgG4-RD can also cause diffuse infiltrative lesions presenting without constitutional symptoms, which usually makes the diagnosis incidental and based upon radiographic findings or examination of pathological specimens. Spontaneous improvement is very rare, and the majority of cases show slow and indolent progression. IgG4-RD affects mostly middle-aged and elderly men, but the male-female ratio is balanced in patients with predominantly head and neck involvement. It is important to differentiate IgG4-RD from malignancies with mass-forming lesions and some immune-mediated diseases  and several more rare forms . Interestingly, while lymphocytic hypophysitis is also pathologically characterized by a lymphoplasmacytic infiltrate, there is a predominance of lymphocytes (B and T cells) rather than plasma cells . AdditioA review of the 18 reported case reports of IgG4-related hypophysitis -24 showsDemographic details of the 18 reported cases demonstrated a mean age of 62.5 years (range 27-77) and a male predominance 13/18 (72%). The majority of the cases have been described in patients of Asian descent; however, this may be reflective of the increased awareness of IgG4-RD within the community. Each of these cases had other organ manifestations either synchronously or metachronously except two cases having hypophysitis as the sole manifestation in a 74-year-old female  and 55-yDiagnosis of IgG4-related hypophysitis can be made a variety of ways. Pituitary biopsy is the most definitive; however, a biopsy can be invasive and unnecessary in certain cases. Diagnosis of IgG4-related hypophysitis can also be suggested based on biopsy-proven IgG4-related disease in other organs when classic imaging findings of hypophysitis are present, including homogeneously enhancing sellar mass/thickened pituitary stalk. A guideline for a histologic diagnosis of IgG4-related disease has been previously described in a consensus statement on the entity and includes a dense lymphoplasmacytic infiltrate, storiform fibrosis, obliterative phlebitis, and an increased number of plasma cells Table .For some organs, obliterative phlebitis and storiform fibrosis are uncommon, and consensus histologic criteria for these features with pituitary involvement have not been described . The numWhile there is no standardized regimen for steroid therapy for this disease, a dose of 0.6 mg/kg/day of prednisolone is most often used. This should be continued for 1-2 months, with a taper of 5 mg per week. If relapse should occur, the physician may consider a maintenance dose for an extended period of time, up to three years, or a combination of immunosuppressants -27.The present case differs from the reported cases of IgG4-related hypophysitis in several important aspects. The first aspect which makes this case interesting is the lack of convincing systemic features. Most patients with this disease have concurrent or previous involvement of other organs. Her lack of systemic involvement made it difficult to diagnose hypophysitis without tissue examination. Imaging studies were also misleading in this patient, with features suggestive of a neoplastic process, particularly solid and cystic components that would be most commonly seen in craniopharyngioma.Another unusual aspect is the age of the patient. There have been no reported cases of IgG4-related hypophysitis in a patient less than 27 years old. While rare, IgG4-RD has been described in the pediatric population, usually within the context of autoimmune pancreatitis. Often these patients have additional systemic manifestations including retroperitoneal fibrosis, sialadenitis, and mediastinal adenopathy . It is lIgG4-related hypophysitis is a rare condition not previously reported in a pediatric age group. The clinical presentation is similar to that for other inflammatory conditions of the pituitary. The decision to biopsy a pituitary mass in the teenage population is often straight forward for cases of a tumor; however, inflammatory processes which create a mass effect can sometimes be treated without a biopsy. This relies on the physician’s ability to recognize the possibility of rare, inflammatory processes such as IgG4 hypophysitis and having a knowledge of alternative ways to confirm the diagnosis. Avoiding a biopsy can decrease morbidities associated with the procedure and may avoid the need for long-term hormone replacement. Because this disease process has not yet been described in the pediatric population, and due to lack of systemic organ involvement or serum IgG4 levels to indicate a possible IgG4-related disease process, a biopsy was necessary to rule out a more common pituitary tumor and to achieve the diagnosis of IgG4-related hypophysitis. Recent literature has suggested that not infrequently there are cases of hypophysitis with IgG4 plasma cells that are not part of the systemic syndrome. Although it appears likely that this patient is not at risk for further involvement of multiple sites, a definitive biopsy has allowed the patient to seek close clinical follow-up by a rheumatologist.