Evaluation of CT Scan Diagnostic Value in the Novel Coronavirus Disease and Presenting a Corona CT Severity Index.

BACKGROUND: Several diagnostic methods were proposed and evaluated for the COVID-19 disease. However, published studies have reported different diagnostic values for these methods. AIMS: The present study aimed to evaluate the diagnostic performance and accuracy of CT in the novel coronavirus disease (COVID-19) and developed and presented a Corona CT severity index. METHODS: In this retrospective study, CT diagnostic performance was measured based on sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and diagnostic accuracy, and RT-PCR was regarded as a standard. Patients' CT reports were evaluated by a radiologist, and scoring and calculating of the CT severity index were performed. RESULTS: Of 208 patients, 82 showed positive, and 126 showed negative RT-PCR results with a positive frequency of 39.4% (95% CI, 32.7-46.4). The chest CT scan related to 136 patients indicated COVID- 19, whereas the initial RT-PCR assays of 56 patients were negative. Considering RT-PCR results as the reference standard, the sensitivity, specificity, and accuracy of chest CT to indicate COVID-19 infection were 100%, 55.6% (95% CI 46.4-64.4%), and 72.8% (95% CI 66.2-78.8%), respectively. The severity of pulmonary involvement was assigned with different grades. For 60.7% of patients with severity grades of 2 to 6, who showed the involvement of at least one lung lobe in CT, PCR retrieved negative results. CONCLUSION: CT scan shows an acceptable sensitivity as well as a consistently better specificity to identify COVID-19 pneumonia than PCR. It may be considered a major method to identify COVID-19 in epidemic locations.

The High Potential of Ozone Gas to Inactivate Echinococcus granulosus Protoscoleces During Hydatid Cyst Surgery.

BACKGROUND: In medicine, ozone therapy is effectively used in a broad spectrum of diseases. Reviews have shown that ozone gas demonstrates potent antimicrobial effects against a wide range of pathogenic microorganisms, such as oral bacteria, fungi, viruses, and parasite even in resistant strains. The present investigation was designed to assess the protoscolicidal effects of ozone gas on hydatid cysts protoscoleces in vitro and in vivo. METHODS: Hydatid cyst protoscoleces were acquired from sheep livers that were slaughtered at Kerman slaughterhouse, Iran. The viability of protoscoleces was assessed by the eosin exclusion examination after exposure with ozone gas for 1 to 14 min in vitro and ex vivo. RESULTS: In this study, in vitro assay showed that ozone gas at the concentration of 20 mg/L killed 85 and 100% of hydatid cyst protoscoleces after 4 and 6 min of treatment, respectively. However, in the ex vivo analysis, a longer time was needed to confirm a potent protoscolicidal activity such that ozone gas after an exposure time of 12 min, 100% of the protoscoleces were killed within the hydatid cyst. CONCLUSION: In conclusion, the findings of the present study showed that ozone gas at low concentrations (20 mg/L) and short times (4-6 min) might be used as a novel protoscolicidal drug for use in hydatid cyst surgery. However, more clinical surveys are required to discover the precise biological activity of ozone gas in animal and human subjects.

Transient knockdown of Nucleoside transporter 4 gene expression as a therapeutic target in Leishmania major by antisense RNA: In vitro and in vivo studies.

BACKGROUND & OBJECTIVES: Leishmania parasites cause various clinical symptoms in humans such as cutaneous ulcers and fatal visceral diseases. These parasites cannot synthesize purine rings de novo and must uptake purines from their hosts via salvage. Salvage is regulated by permeases in the cell membrane. There are hundreds of membrane transporter proteins to receive nutrients in Leishmania. Nucleoside transporter 4 (NT4) is one of the purine transporters that is involved in enhancing the uptake of adenine in Leishmania major. They are important new drug targets for the treatment of leishmaniasis because they can be used to transport toxic purine analogs to kill parasitic cells, thus preventing the progression of the infection. The present study was conducted to silence the NT4 nucleobase involved in the salvage pathway to interrupt purine nucleotide membrane transport in the cells of L. major. METHODS: In this study, a 502 bp segment of NT4 gene sequence was selected and designed as antisense transcripts after insertion in the parasite. The NT4 construct was transfected into L. major promastigotes for in vitro study of gene expression. Then, BALB/c mice infected with transgenic Leishmania and wild-type strain along with the number and size of lesions were studied in vivo. RESULTS: The study showed that relative expression of NT4 gene in mutant Leishmania was lower than in the control on Day 3 to 20. The percentages and the number of amastigotes in infected macrophages with wild-type strain L. major were more than infected macrophages with mutant parasites. Infected BALB/c mice with transgenic Leish- mania showed a lower number and size of lesions than the BALB/c mice infected with wild-type strain. INTERPRETATION & CONCLUSION: The results of the study indicated that the use of antisense RNA reduced NT4 gene expression in L. major. Further, studies are needed to ascertain that the use of antisense can be considered as a new treatment for leishmaniasis.

Differential Expression of Hox and Notch Genes in Larval and Adult Stages of Echinococcus granulosus.

This investigation aimed to evaluate the differential expression of HoxB7 and notch genes in different developmental stages of Echinococcus granulosus sensu stricto. The expression of HoxB7 gene was observed at all developmental stages. Nevertheless, significant fold differences in the expression level was documented in the juvenile worm with 3 or more proglottids, the germinal layer from infected sheep, and the adult worm from an experimentally infected dog. The notch gene was expressed at all developmental stages of E. granulosus; however, the fold difference was significantly increased at the microcysts in monophasic culture medium and the germinal layer of infected sheep in comparison with other stages. The findings demonstrated that the 2 aforementioned genes evaluated in the present study were differentially expressed at different developmental stages of the parasite and may contribute to some important biological processes of E. granulosus.

Comparison of ex vivo harvested and in vitro cultured materials from Echinococcus granulosus by measuring expression levels of five genes putatively involved in the development and maturation of adult worms.

Parts of the natural life cycle of Echinococcus granulosus can be retraced in vitro such as the development of protoscoleces into semiadult worms with three or more proglottids, or the redifferentiation of in vitro cultured protoscoleces into metacestode-like cystic structures. Most in vitro generated samples share-at the microscopical level-high similarities with those naturally grown, but developmental differences have also been documented, such as missing egg production in in vitro grown adults or unusual bladder/vesicle formation in protoscoleces cultured into the metacestode direction. The aim of the present study was to explore how far different in vitro generated stage-specific materials/structures match the natural situation on the transcriptome level, based on testing five exemplarily chosen different genes: the frizzled receptor eg-fz4 (posterior marker), the FGF receptor-like factor eg-fgfrl (anterior association), the cell differentiation protein eg-rcd1 (part of the CCR4-NOT complex, a key regulator of eukaryotic gene expression), the rapidly accelerated fibrosarcoma serin/threonin kinase eg-braf (part of the MAPK pathway involved, e.g., in EGF signaling) and the co-smad eg-smadD (downstream factor of TGFß/BMP2/activin signaling). These genes-tested via qPCR-were selected such as to allow a discussion on their potential role in the development of E. granulosus into the adult stage. Thus, testing took place with three ex vivo isolated samples, namely (i) egg-containing adult worms, (ii) invaginated protoscoleces, and (iii) protoscolex-free germinal layer tissue. Respective data were compared (a) with in vitro generated metacestode-like microcysts developed from protoscolices, and (b) different development stages of protoscoleces in vitro cultured toward adult maturation. As a finding, only eg-smadD and partially eg-fz4 showed high expression similarities between ex vivo harvested and in vitro cultured E. granulosus, thus suggesting a putative role in adult maturation. Conclusively, the fact of using "only" five genes did not allow answering the question if ex vivo and in vitro materials are similar on the transcriptome level. Another experimental restriction arises from different growth conditions of the in vitro cultured materials, and comparing these to the ex vivo harvested ones. Future experiments may solve the problems by using fully standardized E. granulosus sample collection and fully standardized culture conditions.

Chemical composition, efficacy and safety of Pistacia vera (var. Fandoghi) to inactivate protoscoleces during hydatid cyst surgery.

At present, various scolicidal agents have been used for inactivation of protoscoleces during hydatid cyst surgery, however, they are associated with serious adverse side effects including sclerosing colangititis (biliary tract fibrosis), liver necrosis and methaemoglobinaemia. This investigation was designed to evaluate the chemical composition and in vitro scolicidal effects of Pistacia vera (var. Fandoghi) essential oil against protoscoleces of hydatid cysts and also its toxicity in mice model. The components of the P. vera essential oil were identified by gas chromatography/mass spectroscopy (GC/MS) analysis. Protoscoleces were aseptically aspirated from sheep livers having hydatid cysts. Various concentrations of the essential oil (25-200µl/mL) were used for 5-30min. Viability of protoscoleces was confirmed using eosin exclusion test (0.1% eosin staining). In addition, forty male NIH mice were used to determine the acute and sub-acute toxicity of P. vera essential oil for 2 and 14 days, respectively. The main components of P. vera essential oil were limonene (26.21%), α-pinene (18.07%), α-thujene (9.31%) and α-terpinolene (9.28%). Findings of the present study demonstrated that the P. vera essential oil at the concentrations of 100 and 200µl/mL killed 100% protoscoleces after 10 and 5min of exposure, respectively. The LD50 values of intraperitoneal injection of the P. vera essential oil was 2.69ml/kg body weight, and the maximum nonfatal doses were 1.94ml/kg body weight. No significant difference (P>0.05) was observed in the clinical chemistry and hematological parameters following oral administrations of P. vera essential oil at the doses 0.1, 0.2, and 0.4ml/kg for 14 days. The obtained findings demonstrated new chemical composition and promising scolicidal activity of the P. vera with no significant toxicity which might be used as a natural scolicidal agent in hydatid cyst surgery.

The role of GlcNAc-PI-de-N-acetylase gene by gene knockout through homologous recombination and its consequences on survival, growth and infectivity of Leishmania major in in vitro and in vivo conditions.

At present, there are no efficacious vaccines or effective drugs against leishmaniasis; therefore new and innovative control methods are urgently required. One way to achieve this important goal is through using reverse genetic engineering to evaluate important enzymes, proteins and macromolecules. One of the most important enzymes for Glycosylphosphatidylinositol (GPI) biosynthetic pathways is GlcNAc-PI-de-N-acetylase (GPI12). The molecular constructs were cloned in Escherichia coli strain Top 10 and confirmed by molecular methods and were transfected by electroporation into Leishmania major. We demonstrated that two alleles of the GPI12 gene in L. major were successfully removed and enabling the generation of a null mutant, which supports the idea that GPI12 is not an essential gene for the growth and survival of Leishmania and the homozygous knockouts of Leishmania are able to survive. We were able to produce a mutant parasite that caused no damaged to the host. Further investigations are essential to check the safety profile in laboratory animals.

Efficacy of Myrtus communis L. to Inactivate the Hydatid Cyst Protoscoleces.

PURPOSE: The present study aims to investigate the scolicidal effects of Myrtus communis L. essential oil against protoscoleces of hydatid cysts and also its toxicity in mice model. MATERIALS AND METHODS: Protoscoleces were aseptically aspirated from sheep livers having hydatid cysts. Various concentrations of the essential oil (12.5-100 µl/ml) were used for 5-30 min. Viability of protoscoleces was confirmed using eosin exclusion test (0.1% eosin staining). Moreover, 48 male NMRI mice were used to determine the acute and sub-acute toxicity of M. communis essential oil. One-way ANOVA with Tukey's post-hoc test was used to assess differences between experimental groups. RESULTS: Findings of the present study demonstrated that the M. communis essential oil at the concentration of 100 µl/ml after 5 min of exposure killed 100% protoscoleces. Similarly, the mean mortality rate of protoscoleces after 10 min of exposure to concentration of 50 µl/ml was 100%. However, lower concentrations (12.5 and 25 µl/ml) of M. communis essential oil provoked a delayed protoscolicidal effects. The LD50 values of intraperitoneal injection of the M. communis essential oil was 2.23 mL/kg body wt. No significant difference (p > .05) was observed in the clinical chemistry and hematological parameters following oral administrations of M. communis essential oil at the doses 0.05, 0.1, 0.2, and 0.4 mL/kg for 14 days. CONCLUSION: The results showed potent scolicidal activity of M. communis with no significant toxicity, which might be used as a natural scolicidal agent in hydatid cyst surgery.

Antileishmanial and cytotoxic effects of essential oil and methanolic extract of Myrtus communis L.

Plants used for traditional medicine contain a wide range of substances that can be used to treat various diseases such as infectious diseases. The present study was designed to evaluate the antileishmanial effects of the essential oil and methanolic extract of Myrtus communis against Leishmania tropica on an in vitro model. Antileishmanial effects of essential oil and methanolic extract of M. communis on promastigote forms and their cytotoxic activities against J774 cells were evaluated using MTT assay for 72 hr. In addition, their leishmanicidal activity against amastigote forms was determined in a macrophage model, for 72 hr. Findings showed that the main components of essential oil were α-pinene (24.7%), 1,8-cineole (19.6%), and linalool (12.6%). Findings demonstrated that M. communis, particularly its essential oil, significantly (P<0.05) inhibited the growth rate of promastigote and amastigote forms of L. tropica based on a dose-dependent response. The IC50 values for essential oil and methanolic extract was 8.4 and 28.9 µg/ml against promastigotes, respectively. These values were 11.6 and 40.8 µg/ml against amastigote forms, respectively. Glucantime as control drug also revealed IC50 values of 88.3 and 44.6 µg/ml for promastigotes and amastigotes of L. tropica, respectively. The in vitro assay demonstrated no significant cytotoxicity in J774 cells. However, essential oil indicated a more cytotoxic effect as compared with the methanolic extract of M. communis. The findings of the present study demonstrated that M. communis might be a natural source for production of a new leishmanicidal agent.

Evaluation of antileishmanial activity and cytotoxicity of the extracts of Berberis vulgaris and Nigella sativa against Leishmania tropica.

BACKGROUND & OBJECTIVES: Leishmaniasis is a major public health problem, and the alarming spread of parasite resistance underlines the importance of discovering new therapeutic products. The present study aims to investigate the in vitro antileishmanial activity and cytotoxicity of the ethanolic extract of Berberis vulgaris fruits and chloroform extract of Nigella sativa seeds against Leishmania tropica. METHODS: In this study, antileishmanial activity of B. vulgaris and N. sativa extracts on promastigote and amastigote stages of L. tropica in comparison to meglumine antimoniate (MA) was evaluated, using MTT assay and macrophage model, respectively. MTT test was also used to assess the cytotoxicity of extracts on murine macrophages. The significance of differences was determined by analysis of variances (ANOVA) and student's t-test using SPSS software. RESULTS: The results showed that ethanolic extract of B. vulgaris (IC50 4.83 µg/ml) and chloroform extract of N. sativa (IC50 7.83 µg/ml) significantly reduced the viability of promastigotes of L. tropica in comparison to MA (IC50 11.26 µg/ml). Furthermore, extracts of B. vulgaris (IC50 24.03 µg/ml) and N. sativa (IC50 30.21 µg/ml) significantly decreased the growth rate of amastigotes in each macrophage as compared with positive control (p <0.05). Our findings also revealed that extracts of B. vulgaris and N. sativa had no significant cytotoxicity against murine macrophages. CONCLUSION: The B. vulgaris and N. sativa extracts exhibited an effective leishmanicidal activity against L. tropica on in vitro model. Further, works are required to evaluate the exact effect of these extracts on Leishmania species using a clinical setting.

Scolicidal effects of black cumin seed (Nigella sativa) essential oil on hydatid cysts.

Surgery remains the preferred treatment for hydatid cyst (cystic echinococcosis, CE). Various scolicidal agents have been used for inactivation of protoscolices during surgery, but most of them are associated with adverse side effects. The present study aimed to evaluate the in vitro scolicidal effect of Nigella sativa (Ranunculaceae) essential oil and also its active principle, thymoquinone, against protoscolices of hydatid cysts. Protoscolices were aseptically aspirated from sheep livers having hydatid cysts. Various concentrations of the essential oil (0.01-10 mg/ml) and thymoquinone (0.125-1.0 mg/ml) were used for 5 to 60 min. Viability of protoscolices was confirmed by 0.1% eosin staining. Furthermore, the components of the N. sativa essential oil were identified by gas chromatography/mass spectroscopy (GC/MS). Our study revealed that the essential oil of N. sativa at the concentration of 10 mg/ml and its main component, thymoquinone, at the concentration of 1 mg/ml had potent scolicidal activities against protoscolices of Echinococcus granulosus after 10 min exposure. Moreover, thymoquinone (42.4%), p-cymene (14.1%), carvacrol (10.3%), and longifolene (6.1%) were found to be the major components of N. sativa essential oil by GC/MS analysis. The results of this study indicated the potential of N. sativa as a natural source for production of a new scolicidal agent for use in hydatid cyst surgery. However, further studies will be needed to confirm these results by checking the essential oil and its active component in in vivo models.