Phytochemical screening, cytotoxicity assessment and evaluation of in vitro antiplasmodial and in vivo antimalarial activities of Mentha spicata L. methanolic leaf extract.

ETHNOPHARMACOLOGICAL RELEVANCE: Malaria causes extensive morbidity and mortality, and the decreasing efficacy of artemisinin and its partner drugs has posed a serious concern. Therefore, it is important to identify new antimalarials, and the natural compounds from plants provide a promising platform. Mentha spicata L. representing the Lamiaceae family has been used in traditional medicine for various diseases including malaria. AIM OF THE STUDY: This study was aimed at evaluating the antiplasmodial activity of M. spicata methanolic leaf extract using Plasmodium falciparum (Pf) cultures (Pf3D7 and artemisinin (ART)-resistant PfCam3.IR539T strains) and antimalarial activity using Plasmodium berghei (Pb)-infected mice. Dry leaf powder and methanolic leaf extract were examined for in vivo antimalarial activity and the efficacy of oral versus parenteral administration was compared. MATERIALS AND METHODS: Leaves of M. spicata were collected and extracted using 70% methanol in water (v/v). [3H]-hypoxanthine incorporation assays and Giemsa-stained smears were used to assess the in vitro antiplasmodial activity of M. spicata methanolic extract against Pf3D7 and ART-resistant PfCam3.IR539T strains. Cytotoxicity was evaluated in HeLa and HEK-293T cell lines using MTT assays. Hemolysis assays were performed using red blood cells (RBCs). In vivo antimalarial activities of M. spicata dry leaf powder and methanolic leaf extract were examined in P. berghei-infected mice by Rane's curative test and Peters' 4-day suppressive test. RESULTS: Phytochemical screening of M. spicata methanolic leaf extract indicated the presence of reducing sugars, phenolic compounds, flavonoids, glycosides, sterols, saponins, alkaloids, coumarins, tannins, carbohydrates, and proteins. In vitro studies carried out using Pf cultures showed that M. spicata methanolic leaf extract had significant antiplasmodial activity against Pf3D7 cultures with a 50% inhibitory concentration (IC50) of 57.99 ± 2.82 µg/ml. The extract was also effective against ART-resistant PfCam3.IR539T strain with an IC50 of 71.23 ± 3.85 µg/ml. The extract did not show significant in vitro cytotoxicity, hemolysis, and in vivo toxicity. In vivo studies performed using Pb-infected mice treated with M. spicata dry leaf powder and methanolic leaf extract showed ∼50% inhibition in parasite growth at 1500 mg/kg and 1000 mg/kg doses, respectively. There was also a significant delay in the mortality of treated mice. Parenteral administration was found to be appropriate for the in vivo treatment. CONCLUSIONS: Our in vitro and in vivo findings from Pf and Pb parasites suggested the therapeutic potential of M. spicata leaf extract as an antimalarial. M. spicata leaf extract could also inhibit the growth of ART-resistant Pf strain. Further studies on fractionation and active component analysis of M. spicata leaf extract would be required to identify the bioactive phytochemicals having pharmaceutical and therapeutic values. Such efforts would help us in developing new antimalarials to combat malaria.

[Severe lead poisoning caused by ayurvedic medicine]. / Schwere Bleivergiftung durch ayurvedisches Heilmittel.

HISTORY: We report the case of a young patient who presented to our emergency department with reduced general condition, anemia, and crampy abdominal pain. A previous inpatient workup including abdominal imaging and bone marrow aspiration had not yielded a diagnosis. On inquiry, the patient reported oral ingestion of an Ayurvedic remedy over the course of one month. FINDINGS: 24-year-old circulatory stable patient in reduced general condition with gray skin coloration and a dark gingival margin. Laboratory testing revealed an increase in transaminases and normocytic anemia. A peripheral blood smear showed basophilic stippling of the erythrocytes. Significantly elevated lead levels were detected in the patient's blood and hair. Toxic lead levels were detected in the ingested preparation. DIAGNOSIS: Severe lead poisoning caused by self-medication with an Ayurvedic remedy. Analysis revealed a daily oral lead load of 136 times the maximum permissible dose. THERAPY AND COURSE: By means of chelation therapy, the blood lead levels were significantly reduced, and there was a complete regression of the complaints as well as a normalization of the laboratory findings. CONCLUSION: Lead has toxic effects on all organ systems of the body and is stored in the bone for decades. Symptoms of poisoning are nonspecific; a thorough history and generous indication for measuring lead levels are helpful for the diagnosis.

Abdominal pain caused by lead toxicity due to over the counter herbal medicines: A case series.

In the last three decades, the use of herbal medications has been increasing for the treatment of various chronic disorders. Studies in the past have shown that many of these medicines could contain high levels of heavy metals, including lead. Therefore, we planned this study to evaluate the possibility of lead toxicity as the underlying cause in patients consuming these unnamed herbal medicines among patients presenting with significant abdominal pain. (Unexplained abdominal pain means pain in abdomen in which no etiology could be ascertained after all possible routine and specialized investigations including computerized axial tomography [CT] of the abdomen and upper gastrointestinal [UGI] endoscopy/colonoscopy). This is an observational case series of prospectively maintained data of all patients having unexplained abdominal pain and found to have an elevated blood lead level from 2011 to 2019. Lead toxicity was diagnosed when its blood lead level was >25 µg/dL. Total sixty-six patients with unexplained abdominal pain from 2011 to 2019 were recruited. Out of the sixty-six patients, seventeen had elevated blood lead levels. All seventeen patients had a history of ingestion of herbal medicines for more than 6 months. Among the seventeen patients, eight were taking it for infertility and sexual dysfunction, six for diabetes, two for arthritis and one for hypertension. Basophilic stippling was seen in one patient. Fourteen patients had low hemoglobin with a median value of 9.7 g/dL. Mean serum blood lead level was 87.1 µg/dL. None of them required anti-chelating agent. Lead toxicity owing to herbal medicine is not uncommon cause of unexplained abdominal pain. Most of these patients do not require a chelating agent for treatment. There is a need to bring these herbal medicines under strict regulations for displaying its constituents and their concentrations.

Changes in inflammatory cytokines, antioxidants and liver stiffness after chelation therapy in individuals with chronic lead poisoning.

BACKGROUND: Chronic exposure to lead causes lead to accumulate mainly in the liver. In vivo studies have shown that lead toxicity is related to alterations in the inflammatory response. We aimed to evaluate the association between lead poisoning and liver fibrosis as well as the change in the degree of liver fibrosis, levels of inflammatory mediators and glutathione (GSH) after chelation therapy. METHODS: Workers from a battery factory who were exposed to lead for > 12 months and had a blood lead level (BLL) > 70 µg/dL were enrolled (n = 86) in the study. Participants underwent chelation therapy with intravenous CaNa2EDTA for 2 days followed by treatment with oral D-penicillamine for 90 days. The primary outcome was the change in the degree of liver fibrosis, which was presented as liver stiffness (LS) measured by FibroScan®. Secondary outcomes were the changes in the levels of serum GSH and inflammatory mediators such as tumor necrosis factor-alpha (TNF-α), interleukin-1ß (IL-1ß), and interleukin-6 (IL-6) after chelation therapy. RESULTS: Among the 86 participants, there was a positive correlation between the duration of lead exposure and LS (r = 0.249, p = 0.021). To avoid the confounding effect of obesity-related steatosis, only 70 individuals who had controlled attenuation parameters < 296 dB/m, BMI < 25 kg/m2 and normal waist circumference were included in the interventional analysis. After chelation, the mean LS significantly decreased from 5.4 ± 0.9 to 4.8 ± 1.4 kPa (p = 0.001). Similarly, all of the inflammatory cytokines studied significantly decreased after chelation (p < 0.001); TNF-α decreased from 371.6 ± 211.3 to 215.8 ± 142.7; the levels of IL-1ß decreased from 29.8 ± 1.7 to 25.9 ± 4.3; and the levels of IL-6 decreased from 46.8 ± 10.2 to 35.0 ± 11.9. On the other hand, the mean GSH level increased significantly from 3.3 ± 3.3 to 13.1 ± 3.7 (p < 0.001) after chelation therapy. CONCLUSION: The duration of lead exposure was significantly correlated with the degree of liver fibrosis. Chelation treatment was associated with increased levels of GSH and decreased levels of proinflammatory cytokines and could potentially reduce the degree of LS. TRIAL REGISTRATION: This study was retrospectively registered and approved by the Thai Clinical Trial Registry (TCTR) on 2019-11-07. The TCTR identification number is TCTR20191108001 .

Organ-on-chip models: Implications in drug discovery and clinical applications.

Before a lead compound goes through a clinical trial, preclinical studies utilize two-dimensional (2D) in vitro models and animal models to study the pharmacodynamics and pharmacokinetics of that lead compound. However, these current preclinical studies may not accurately represent the efficacy and safety of a lead compound in humans, as there has been a high failure rate of drugs that enter clinical trials. All of these failures and the associated costs demonstrate a need for more representative models of human organ systems for screening in the preclinical phase of drug development. In this study, we review the recent advances in in vitro modeling including three-dimensional (3D) organoids, 3D microfabrication, and 3D bioprinting for various organs including the heart, kidney, lung, gastrointestinal tract (intestine-gut-stomach), liver, placenta, adipose, retina, bone, and brain as well as multiorgan models. The availability of organ-on-chip models provides a wealth of opportunities to understand the pathogenesis of human diseases and provide a potentially better model to screen a drug, as these models utilize a dynamic 3D environment similar to the human body. Although there are limitations of organ-on-chip models, the emergence of new technologies have refined their capability for translational research as well as precision medicine.

Lead powder use for skin care and elevated blood lead level among children in a Chinese rural area.

To investigate the association between lead powder use, as folk skin care, and blood lead level (BLL) in children, we studied 222 children up to 14-years old living in a Chinese rural area and administered a face to face interview with their parents to collect information on lead powder use and other potential exposure. We measured children's BLL at baseline and 2 years later after an intervention. The children were divided into three categories according to their use of lead powder: regular use, irregular use and never use. We applied multivariate linear regression to determine the association between lead powder use and elevated BLL. The average BLL of all children was 18 µg/dl; 56% of them had BLL of 10 µg/dl or higher. Lead powder use was significantly associated with elevated BLL. After adjusting for potential confounders the BLL of regular and irregular users was higher than non-users by 3.11 µg/dl and 1.47 µg/dl, respectively. Duration of lead powder use was positively associated with BLL, but the time since last use was inversely associated. A significant BLL reduction was observed 2 years later, and the greatest reduction (21 µg/dl) was seen in the youngest group of regular users. This study showed that traditional use of lead powder for a skin care purpose was a major contributor to elevated BLL in these children.

Towards translation of 212Pb as a clinical therapeutic; getting the lead in!

Targeted α-particle therapy offers the potential for more specific tumor cell killing with less damage to surrounding normal tissue than ß-emitters because of the combination of short path length (50-80 µm) with the high linear energy transfer (100 keV µm(-1)) of this emission. These physical properties offer the real possibility of targeted (pre-targeted) α-therapy suitable for the elimination of minimal residual or micrometastatic disease. Targeted and pre-targeted radioimmunotherapy (RIT) using α-emitters such as (212)Bi (T(1/2) = 1.01 h) and (212)Pb (T(1/2) = 10.6 h) has demonstrated significant utility in both in vitro and in vivo model systems. (212)Pb, a promising α-particle emitting source, is the longer-lived parent nuclide of (212)Bi, and serves as an in vivo generator of (212)Bi. The radionuclide has been successfully used in RIT and pre-targeted RIT and demonstrated an enhanced therapeutic efficacy in combination with chemotherapeutics, such as gemcitabine and paclitaxel. The following perspective addresses the modes of radionuclide production, radiolabelling and chelation chemistry, as well as the application of (212)Pb to targeted and pre-targeted radiation therapy.

Internal barium shielding to minimize fetal irradiation in spiral chest CT: a phantom simulation experiment.

PURPOSE: To use a phantom to prospectively examine the attenuating effect of barium sulfate as an internal shield to protect the fetus. MATERIALS AND METHODS: In an adult-size phantom, 1- and 2-cm-thick acrylic slabs containing 315 or 630 mL of water, 2% or 40% barium sulfate suspension, and a 1-mm lead sheet were placed under the diaphragm. In 17 experiments, fetal dose was measured by using thermoluminescent dosimeters that were placed immediately under (near field) and 10 cm below (far field) the water slab (eight experiments), barium sulfate slab (eight experiments), and lead sheet (one experiment). In a pulmonary embolism protocol, the phantom was scanned with single-detector spiral computed tomography (CT) at 130 kVp and 230 mAs. RESULTS: The control radiation dose was 3.60 mSv+/-0.54 (standard deviation) with the water slab at near field, where the uterus dome is at near term, and 0.507 mSv+/-0.07 with the water slab at far field, the uterus position during early gestation. Scattered radiation was attenuated 13% and 21% with 2% barium sulfate and 87% and 96% with 40% barium sulfate, as calculated in the near and far fields, respectively, and 99% with the 1-mm lead sheet. The extrapolated attenuations for 5%-40% barium sulfate suspensions indicated that beyond a 30% suspension, attenuation increased further only slightly. CONCLUSION: Study results in the phantom experiment suggest that fetal irradiation during maternal chest CT can be reduced substantially with barium shielding.

[Basis for the use of a new galena-based substance from plants for prevention and treatment of cholelithic disease]. / Obosnovanie primeneniia novoi galenovoi fitokompozitsii dlia profilaktiki i lecheniia zhelchnokamennoi bolezni.

With the purpose of prevention and treatment of cholelithic disease, a new galena-based phytocomposition having the choleritic, cholekinetic, spasmolytic and hypocholesterolemic effects at the same time was administered to patients along with lithotripsy. Clinical manifestations of cholecystitis disappeared after a daily intake of the phytocomposition during 30 days in 96 patients out of 418 people having the diagnosis of cholelithic disease and high initial concentrations of?-lipoproteins and cholesterol in the blood serum. The level of lipids dropped reliably, and it became much easier to use the lithotripsy method. Due to the decrease of bile viscosity, it was possible to note easier elimination of fragments from the gallbladder. During the monotherapy with the galena-based phytocomposition, full litholisis of single (up to 30 mm) or multiple (up to 50% of the gallbladder volume) calculi was determined in seven patients out of 47 (17%) during the period from two to six months.

Mexican use of lead in the treatment of empacho: community, clinic, and longitudinal patterns.

This paper discusses research designed to investigate community, clinic, and longitudinal patterns in use of lead as a treatment for empacho, a folk illness manifest by gastrointestinal symptoms. The same questionnaire used in a clinic-based study seven years previously in Guadalajara, Mexico, was used to interview a randomly selected community sample: in addition, the study was repeated at the same clinic sites that had been studied previously. The goals were to investigate: (1) What are community wide prevalences of empacho and use of lead based remedies? (2) To what extent are current patterns of use of lead for treatment of empacho in clinic-based samples similar to those seven years ago. The attributable risk to the population as a whole from use of lead based remedies was found to be 11% of the households of Guadalajara. Essentially this same estimate was seen for the 1987 and 1994 clinic populations. Interestingly, while percentages of lead users have declined since 1987, twice as great a percentage of informants reported treating empacho. Other patterns originally identified in 1987 persisted in 1994; lead use continues to be associated with lower levels of parental education and income.

Effect of homeopathic drugs plumbum and opium on experimentally induced lead toxicity in rats.

Homeopathic drugs plumbum 1M and Opium 30 were partially effective in the recovery of delta ALAD activity of the lead (150 mg% lead acetate) intoxicated rats. Plumbum 1M did not exhibit protective effect when dietary lead at high concentrations (> 25 mg% lead acetate) were given concurrently as assessed by blood delta ALAD activity and hemoglobin concentration. However it was partially effective in the recovery of delta ALAD activity and relieving anemia caused by chronic exposure of low doses of lead (below 15 mg% lead acetate).

Pharmacomodulations on new organometallic complexes of Ir, Pt, Rh, Pd, Os: in vitro and in vivo trypanocidal study against Trypanosoma brucei brucei.

New organometallic complexes have been synthesized by association of an active organic molecule with a metallic element such as Pt, Rh, Ir, Pd, Os. Their trypanocidal activity was studied in vitro and in vivo against T. b. brucei. The more active compounds were pentamidine derivatives. The Ir- COD-pentamidine complex, and Iridium (I) cationic and organometallic complex showed and in vitro activity at 60 micrograms/l. Moreover, all infected mice were cured by this compound subcutaneously administered in a single dose at 0.5 mg/kg (0.317 mumol/kg). In the same conditions, pentamidine cured all the mice at 5 mumol/kg. Ir-COD-pentamidine (or P1995) was 16 fold more efficient than pentamidine. Since the chemotherapeutic index of this molecule was 7.5 fold higher than those of pentamidine, P1995 can be considered as a potential trypanocidal drug of the future.

Lead based remedies for empacho: patterns and consequences.

This paper discusses research designed to investigate the patterns and consequences of the use of lead based folk remedies in the treatment of some gastrointestinal problems in Mexico. Use of lead is seen in 35% of the population sampled in Guadalajara who treated the folk illness empacho, and 5% of a similar population in Oaxaca. Lead use seems to be associated with mestizo ethnic background, and with lower levels of parental education and income.

Toxic Mexican folk remedies for the treatment of empacho: the case of azarcon, greta, and albayalde.

This article discusses the availability of three lead-based salts which are being used as folk remedies in Mexico. Distribution systems and geographic availability were determined, and purchased samples were found to be very high in lead content. The findings suggest the need for further research to understand the reasons why such remedies are chosen, and the development of an educational program to discourage their use.