Discovery of dual-target natural antimalarial agents against DHODH and PMT of Plasmodium falciparum: pharmacophore modelling, molecular docking, quantum mechanics, and molecular dynamics simulations.

Malaria is a lethal disease that claims thousands of lives worldwide annually. The objective of this study was to identify new natural compounds that can target two P. falciparum enzymes; P. falciparum Dihydroorotate dehydrogenase (PfDHODH) and P. falciparum phosphoethanolamine methyltransferase (PfPMT). To accomplish this, e-pharmacophore modelling and molecular docking were employed against PfDHODH. Following this, 1201 natural compounds with docking scores of ≤ -7 kcal/mol were docked into the active site of the second enzyme PMT. The top nine compounds were subjected to further investigation using MM-GBSA free binding energy calculations and ADME analysis. The results revealed favourable free binding energy values better than the references, as well as acceptable pharmacokinetic properties. Compounds ZINC000013377887, ZINC000015113777, and ZINC000085595753 were scrutinized to assess their interaction stability with the PfDHODH enzyme, and chemical stability reactivity using molecular dynamics (MD) simulation and density functional theory (DFT) calculations. These findings indicate that the three natural compounds are potential candidates for dual PfDHODH and PfPMT inhibitors for malaria treatment.

The dichotomy of the systemic and local cytokine responses of mucosal leishmaniasis patients in Sudan: A pilot study.

Sudanese mucosal leishmaniasis (ML) is a rare clinical form of leishmaniasis and characterized by persistent ulcer of the oral and/or the nasal mucous membranes caused by Leishmania donovani. No data is available about the systemic and local immune responses in mucosal leishmaniasis. This study aimed to measure the systemic and the local cytokines responses of Sudanese ML patients compared to cured cutaneous leishmaniasis patients (Leishmanin skin test positive, LST+ve) and unexposed healthy controls (Leishmanin skin test negative, LST-ve). Six parasitological confirmed ML patients, 7 LST+ve, and 6 LST-ve were enrolled. Systemic Th-1 (IFN-γ and TNF-α), Th-2 (IL-10 and IL-13), Treg (TGF-ß1), and inflammatory cytokines IL-6 and IL-8 concentration were measured in the supernatant of whole blood samples following stimulation with live L. donovani promastigotes using ELISA. Local intralesion IL-10, IFN-γ, and IL-13 expression was measured using Real Time PCR. A significant high concentrations of IFN-γ, TNFα, IL-10, TGFß, IL-6, and IL-8 were detected in the supernatant of stimulated whole blood samples of ML patients compared with the LST+ve and LST-ve controls. Using Real Time-PCR and primers for various cytokines, a significant high expression of TH2 cytokines IL-10 and IL-13 mRNA was detected in contrast to a low TH1 cytokine IFN-γ mRNA in the mucosal lesion. There is a clear dichotomy in the cytokine response during Mucosal leishmaniasis. A significantly high TH1, inflammatory and Treg cytokines response is produced systemically, in contrast to a significant high TH2 cytokines response in the mucosal lesion.

The dichotomy of the systemic and local cytokine responses of mucosal leishmaniasis patients in Sudan: A pilot study

@#Sudanese mucosal leishmaniasis (ML) is a rare clinical form of leishmaniasis and characterized by persistent ulcer of the oral and/or the nasal mucous membranes caused by Leishmania donovani. No data is available about the systemic and local immune responses in mucosal leishmaniasis. This study aimed to measure the systemic and the local cytokines responses of Sudanese ML patients compared to cured cutaneous leishmaniasis patients (Leishmanin skin test positive, LST+ve) and unexposed healthy controls (Leishmanin skin test negative, LST-ve). Six parasitological confirmed ML patients, 7 LST+ve, and 6 LST-ve were enrolled. Systemic Th-1 (IFN-γ and TNF-α), Th-2 (IL-10 and IL-13), Treg (TGF-β1), and inflammatory cytokines IL-6 and IL-8 concentration were measured in the supernatant of whole blood samples following stimulation with live L. donovani promastigotes using ELISA. Local intralesion IL-10, IFN-γ, and IL-13 expression was measured using Real Time PCR. A significant high concentrations of IFN-γ, TNFα, IL-10, TGFβ, IL-6, and IL-8 were detected in the supernatant of stimulated whole blood samples of ML patients compared with the LST+ve and LST-ve controls. Using Real Time-PCR and primers for various cytokines, a significant high expression of TH2 cytokines IL-10 and IL-13 mRNA was detected in contrast to a low TH1 cytokine IFN-γ mRNA in the mucosal lesion. There is a clear dichotomy in the cytokine response during Mucosal leishmaniasis. A significantly high TH1, inflammatory and Treg cytokines response is produced systemically, in contrast to a significant high TH2 cytokines response in the mucosal lesion.

Identification and characterization of Leishmania amastigote and axenic form antigens.

This study was aimed to identify and characterize Leishmania amastigote, and axenic form antigens. Two in vitro techniques were used to change leishmania parasite isolates from promastigote form to amastigotes and amastigote like (axenic) forms. The main strategy relied upon in vitro infection of murine macrophages cell line J774 with leishmania promastigote, at 37°C with 5% CO2, while the second technique relied upon the culture of promastigote at 37°C with low pH (5.5), and 5-10% CO2. Proteins were extracted and fractionated utilizing 12% Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS PAGE). Antigens were recognized using both immune dot blot and western blot procedures. PCR was performed for recognition of leishmania parasites in infected J774 macrophages. L. major was quicker in infectivity of macrophages cell line than L. donovani. Shared proteins ranging from 26-116 kDa were identified by SDS PAGE in all stages. Immune Dot-blot method showed positive outcomes, while western blot identified an exceptional antigen band of 16 kDa in amastigote, this unique band could be of value in diagnosis and vaccination of leishmaniasis. PCR results confirmed presence of both isolates demonstrating that coinfection is conceivable, and no indications of hereditary recombination at kinetoplast DNA (kDNA) were identified in macrophages simultaneously infected by L. major and L. donovani.

Identification and characterization of Leishmania amastigote and axenic form antigens

@#This study was aimed to identify and characterize Leishmania amastigote, and axenic form antigens. Two in vitro techniques were used to change leishmania parasite isolates from promastigote form to amastigotes and amastigote like (axenic) forms. The main strategy relied upon in vitro infection of murine macrophages cell line J774 with leishmania promastigote, at 37°C with 5% CO2, while the second technique relied upon the culture of promastigote at 37°C with low pH (5.5), and 5-10% CO2. Proteins were extracted and fractionated utilizing 12% Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS PAGE). Antigens were recognized using both immune dot blot and western blot procedures. PCR was performed for recognition of leishmania parasites in infected J774 macrophages. L. major was quicker in infectivity of macrophages cell line than L. donovani. Shared proteins ranging from 26-116 kDa were identified by SDS PAGE in all stages. Immune Dot-blot method showed positive outcomes, while western blot identified an exceptional antigen band of 16 kDa in amastigote, this unique band could be of value in diagnosis and vaccination of leishmaniasis. PCR results confirmed presence of both isolates demonstrating that coinfection is conceivable, and no indications of hereditary recombination at kinetoplast DNA (kDNA) were identified in macrophages simultaneously infected by L. major and L. donovani.

Continuous intravenous infusion of ketamine for maintenance sedation.

Ketamine HCl is a rapidly acting general anesthetic with sedative and analgesic properties that has been reported to have favorable effects on the cardiovascular and pulmonary systems. The goal of this review is to determine the hemodynamic and pulmonary effects of continuous intravenous (IV) ketamine infusion in mechanically ventilated patients, and to determine whether sufficient evidence exists to support its use as an agent for maintenance anesthesia. PubMed/Medline, EMBASE, and Index Medicus databases as well as relevant bibliographies were searched. Studies were independently evaluated for inclusion and exclusion criteria, as well as study parameters, by two evaluators. Any discrepancy was resolved by a third evaluator. Twenty studies (281 patients) met the inclusion criteria for this review including 11 prospective studies (250 patients). Data suggests that ketamine decreases airway resistance, improves dynamic compliance, and preserves functional residual capacity, minute ventilation and tidal volume, while retaining protective pharyngeal and laryngeal reflexes. In patients with refractory bronchospasm, continuous infusion of intravenous ketamine decreases audible wheeze, bronchodilator requirements, and hypercarbia. It also improves respiratory rate and oxygenation, and does not promote respiratory depression. Additionally, ketamine does not result in significant perturbations in blood pressure, heart rate, or vascular resistance. Ketamine may be a safe and effective tool for maintenance sedation of mechanically ventilated patients, however a large prospective clinical trial is warranted.

Identification of Leishmania donovani as a cause of cutaneous leishmaniasis in Sudan.

Eight patients with cutaneous ulcers were referred to the Institute of Endemic Diseases, Khartoum, Sudan, from June 2000 to March 2002 for the diagnosis of suspected cutaneous leishmaniasis (CL). Diagnosis was confirmed parasitologically by both positive Giemsa-stained smears and successful culture of Leishmania promastigotes in NNN medium. The eight parasite isolates were shown to belong to the Leishmania donovani complex by kDNA PCR. Isoenzyme typing of three isolates revealed that they were identical to the L. donovani MON-82 reference strain, and the gp63 PCR-RFLP profile showed similar patterns to a reference strain of MON-82. CL is endemic in most regions of Sudan and has been reported previously as being caused by L. major MON-74. The results of this study suggest that L. donovani is also a cause of CL in Sudan and that further study of isolates from Sudanese patients with cutaneous ulcers is warranted to ascertain whether L. donovani or L. major is the causative agent.

Lipoblastoma in a four-year-old African child.

A four-year old Sudanese child presented with a growing mass in the medial aspect of the right thigh. The mass appeared during the neonatal period. On clinical examination a diagnosis of lipoblastoma was entertained on the basis of the patient's age and the clinical features of the mass. The tumor was completely excised surgically. The clinical diagnosis of lipoblastoma was confirmed pathologically. Follow-up of the patient for 6 months postoperatively showed no evidence of recurrence.

Uncommon clinical presentations of cutaneous leishmaniasis in Sudan.

Cutaneous leishmaniasis in Sudan is caused by Leishmania major zymodeme LON1. Self-healing usually occurs within 1 year but occasionally its duration is prolonged and treatment is required. The clinical forms are ulcers, nodules and noduloulcerative lesions. Here we describe seven patients with uncommon lesions that were difficult to recognize as Leishmania infections. These included mycetoma-like lesions, lesions that resembled L. tropica infection and others. One HIV/AIDS patient had Kaposi's sarcoma with Leishmania parasites in the Kaposi lesions. Most of these uncommon clinical forms were difficult to treat. The diagnosis depended on a high degree of suspicion and the demonstration of parasites in smears or culture. PCR was used to characterize parasites from the patients described here. Leishmania major was found by kDNA PCR in all patients, except one, who had a leishmanioma due to L. donovani. In three patients, including one with a L. tropica like-lesion, the parasites were confirmed as L. major by gp63 PCR-RFLP.

Is daily mvi required in patients receiving parenteral nutrition?

Between November 1996 and January 1999 there was a nationwide shortage of intravenous multivitamins (MVI) in the US. We investigated whether patients who received parenteral nutrition (PN) with MVI three times per week instead of daily developed clinical manifestations of vitamin deficiency during that period. Five patients were enrolled in the study All received home PN as their sole source of nutrition for at least six months. None of these patients showed clinical manifestations of vitamin deficiency. In addition, compared to daily MVI administration, supplementation with MVI three times per week saved $1,000 per patient per year. This preliminary study raises the question of how much MVI is needed to prevent vitamin deficiency. Using MVI three days a week instead of daily would substantially reduce the cost of treatment and might reduce the risk of infections for these patients.

Recent advances in pulmonary embolism diagnosis and management.

Pulmonary embolism remains a major problem in clinical practice. Pulmonary angiography remains the most accurate diagnostic procedure. Standard therapy includes heparin then warfarin, and now low molecular weight heparin. The role of prophylactic measures cannot be over emphasized.

Isolated hypogonadotropic hypogonadism in a man with a history of head trauma during childhood.

OBJECTIVE: To report a case of isolated hypogonadotropic hypogonadism as a delayed consequence of major head injury. METHODS: We present the relevant history, findings on physical examination, and results of laboratory evaluation, and supportive data from appropriate reports in the literature are reviewed. RESULTS: A 37-year-old man with progressive loss of libido, impotence, and gradual loss of facial and axillary hair had sustained severe trauma to the head (sufficient to cause blindness in one eye) in a fall from a tree at 7 years of age. Physical examination and laboratory investigations substantiated the presence of isolated hypogonadotropic hypogonadism. Other causes of hypogonadotropic hypogonadism, including Kallmann's syndrome, were excluded. CONCLUSION: Severe head injury should be considered in the differential diagnosis of isolated hypogonadotropic hypogonadism. Long-term follow-up and endocrine assessment may be important in patients who have sustained severe head injury.

Calcifying panniculitis with renal failure: a new management approach.

A 60-year-old woman with chronic renal failure, hyperphosphatemia, normocalcemia and hyperparathyroidism presented with skin and subcutaneous tissue necrosis over the upper lateral aspects of both thighs which became progressively worse with subsequent ulceration over the period of 1 week. A biopsy of the affected skin and subcutaneous tissues demonstrated discrete foci of calcification compatible with the diagnosis of calcifying panniculitis. While it was possible to control the patient's hyperphosphatemia using magnesium carbonate, local ulcer care, including daily whirlpool and repeated local surgical excision, failed to stop the progression of the surrounding skin and subcutaneous tissue necrosis with subsequent ulceration. A trial of oral prednisone for a total of 10 days followed by oral cimetidine for 3 months resulted in complete healing at the incision sites without recurrence after 9 months. In this case of calcifying panniculitis the conditions for calciphylaxis were present, and a new management approach was applied.