Over the past few decades, there has been an increase in the number of reports about newly recognized (atypical or unusual) manifestations of Graves' disease (GD), that are related to various body systems. One of these manifestations is sometimes the main presenting feature of GD. Some of the atypical manifestations are specifically related to GD, while others are also similarly seen in patients with other forms of hyperthyroidism. Lack of knowledge of the association between these findings and GD may lead to delay in diagnosis, misdiagnosis, or unnecessary investigations. The atypical clinical presentations of GD include anemia, vomiting, jaundice, and right heart failure. There is one type of anemia that is not explained by any of the known etiological factors and responds well to hyperthyroidism treatment. This type of anemia resembles anemia of chronic disease and may be termed GD anemia. Other forms of anemia that are associated with GD include pernicious anemia, iron deficiency anemia of celiac disease, and autoimmune hemolytic anemia. Vomiting has been reported as a presenting feature of Graves' disease. Some cases had the typical findings of hyperthyroidism initially masked, and the vomiting did not improve until hyperthyroidism has been detected and treated. Hyperthyroidism may present with jaundice, and on the other hand, deep jaundice may develop with the onset of overt hyperthyroidism in previously compensated chronic liver disease patients. Pulmonary hypertension is reported to be associated with GD and to respond to its treatment. GD-related pulmonary hypertension may be so severe to produce isolated right-sided heart failure that is occasionally found as the presenting manifestation of GD.

The acute toxicity of four selected heavy metals to juvenile crayfish Orconectes immunis (Hagen) (1-2 g wet body wt. each) at room temperature increased in the following order: cadmium (x3) < copper (x10) < zinc (x2) < lead. The toxicity of these metals to crayfish acclimated at 17, 20, 23/24, and 27 degrees C increased with temperature (by 7-20% between 20 and 24 degrees C and 14-26% between 20 and 27 degrees C) as judged by the lowering of LT(50) (time to kill 50% of test animals at a fixed concentration) values. A 4 degrees C rise in temperature (from 20 to 24 degrees C), which increased the toxicity of copper by about 7%, increased the rate of oxygen consumption by about 34%. Heavy metals inhibited the rate of oxygen consumption at all temperatures. In 20 degrees C-acclimated crayfish, copper caused about 17% inhibition of oxygen consumption compared to about 7-12% by other metals including the most toxic cadmium. A 3-4 degrees C rise in temperature tripled the inhibitory effect of copper (20%), cadmium and zinc (26 and 18%, respectively), but not of lead, on oxygen consumption. A 7 degrees C-rise in temperature (from 20 to 27 degrees C) increased the inhibitory effect of heavy metals, including lead, on oxygen consumption by up to 54% in the case of copper. The data indicate that rising global temperatures (currently 0.60 degrees C) associated with climate change can have the potential to increase the sensitivity of aquatic animals to heavy metals in their environment.

Phyllanthus niruri L. is a widespread tropical plant which is used in Ayurvedic system for liver and kidney ailments. The present study aims at specifying the most active hepatoprotective extract of P. niruri and applying a bio-guided protocol to identify the active compounds responsible for this effect. P. niruri aerial parts were extracted separately with water, 50%, 70% and 80% ethanol. The cytoprotective activity of the extracts was evaluated against CCl 4 -induced hepatotoxicity in clone-9 and Hepg2 cells. Bioassay-guided fractionation of the aqueous extract (AE) was accomplished for the isolation of the active compounds. Antioxidant activity was assessed using DPPH (1, 1-diphenyl-2-picrylhydrazyl) radical scavenging method and ferric reducing antioxidant power (FRAP). The in vivo hepatoprotective activity of AE was evaluated in CCl 4 -induced hepatotoxicity in rats at different doses after determination of its LD 50 . Pretreatment of clone-9 and Hepg2 with different concentrations of AE (1, 0.1, 0.01 mg/ml) had significantly reduced the levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) against CCl 4 injures, and restored the activity of the natural antioxidants; glutathione (GSH) and superoxide dismutase (SOD) towards normalization. Fractionation of AE gave four fractions (I-IV). Fractions I, II, and IV showed a significant in vitro hepatoprotective activity. Purification of I, II and IV yielded seven compounds; corilagin C1, isocorilagin C2, brevifolin C3, quercetin C4, kaempferol rhamnoside C5, gallic acid C6, and brevifolin carboxylic acid C7. Compounds C1, C2, C5, and C7 showed the highest (p< 0.001) hepatoprotective potency, while C3, C4, and C6 exhibited a moderate (p< 0.001) activity. The AE exhibited strong antioxidant DPPH (IC 50 11.6 ± 2 μg/ml) and FRAP (79.352 ± 2.88 mM Ferrous equivalents) activity. In vivo administration of AE in rats (25, 50, 100 and 200 mg/kg) caused normalization of AST, ALT, alkaline phosphatase (ALP), lactate dehydrogenase (LDH), total cholesterol (TC), triglycyrides (TG), total bilirubin (TB), glucose, total proteins (TP), urea and creatinine levels which were elevated by CCl 4 . AE also decreased TNF-α, NF-KB, IL-6, IL-8, IL10 and COX-2 expression, and significantly antagonizes the effect of CCl 4 on the antioxidant enzymes SOD, catalase PLOS ONE | https://doi.org/10.1371/journal.pone.0226185 January 15, 2020 1 / 23 OPEN ACCESS Citation: Ezzat MI, Okba MM, Ahmed SH, El-Banna HA, Prince A, Mohamed SO, et al. (2020) In-depth hepatoprotective mechanistic study of Phyllanthus niruri: In vitro and in vivo studies and its chemical characterization. PLoS ONE 15(1): e0226185.

Earthworms (Lumbricus terrestris) acclimated at 2 degrees C above their habitat temperature (10-12 degrees C) showed about 5% increase in basal rate of oxygen consumption, which increased to about 38% in 14-16 degrees C- and 40% in 16-18 degrees C-, but decreased by 84% in 20-22 degrees C-acclimated worms. Temperature also increased the blood hemoglobin (Hb) concentration, which decreased slightly in 20-22 degrees C-acclimated worms. The worms acclimated at 20-22 degrees C showed their blood to be hypovolemic than that of 10-12 degrees C worms indicating dehydration. Pre-exposure of 10-14 degrees C-acclimated worms to sublethal concentrations of zinc, copper, and lead did not significantly affect the rate of respiration. However, at higher temperatures all these metals inhibited oxygen consumption; zinc, lead, and cadmium by approximately 11% and copper by approximately 18% of that at 14-16 degrees C. At 20-22 degrees C, the respiration was further inhibited, 36% by copper, 18% by cadmium, and approximately 10% by lead and zinc. Copper, lead, and zinc decreased the temperature-enhanced increase in blood Hb concentration at all temperatures. In 20-22 degrees C-acclimated worms heavy metal exposure slightly lowered the oxygen affinity of Hb as well as caused shifts in carbon monoxide difference spectra. The acute toxicity of these metals was not affected by a 2 degrees C rise in acclimation temperature but increased by 17% (lead), 33% (copper), and 5% (zinc) in 14-16 degrees C- and by 40% (lead), 149% (copper), and 132% (zinc) in 20-22 degrees C-acclimated worms. The increase in toxicity of metals caused by high temperatures may be due to limiting the scope of aerobic metabolism (oxygen extraction, transport, and utilization) via quantitative and qualitative effects on Hb. This terrestrial species appears to be tolerant of slight increases in habitat temperature, such as that expected with current global climate change.

Contribution/ OriginalityThis study is emphasizing the adverse effects of the synthetic food colorants on human health as previously reported in the literature.