COVID-19: Clinical laboratory diagnosis and monitoring of novel coronavirus infected patients using molecular, serological and biochemical markers: A review.

COVID-19, a novel coronavirus disease, has provoked a variety of health and safety concerns, and socioeconomic challenges around the globe. The laboratory diagnosis of SARS-CoV-2 was quickly established utilizing nucleic acid amplification techniques (NAAT) after the disease causing virus has been identified, and its genetic sequence has been determined. In addition to NAAT, serological tests based on antibodies testing against SARS-CoV-2 were introduced for diagnostic and epidemiologic studies. Other biochemical investigations include monitoring of peripheral blood cells count, platelets/lymphocyte ratio, coagulation profile, cardiac, and inflammatory markers such as cytokines storm are also crucial in combating COVID-19 pandemic. Further, accurate and reliable laboratory results for SARS-CoV-2 play very important role in the initiation of early treatment and timely management of COVID-19 patients, provide support in clinical decision-making process to control infection, and detection of asymptomatic cases. The Task Force on Coronavirus-19 constituted by International Federation of Clinical Chemistry and Laboratory Medicine (IFCC) has recognized informational framework for epidemiology, pathogenesis, and recommended the PCR-based analysis, serological and biochemical assays for analysis, monitoring, and management of disease. This literature review provides an overview of the currently used diagnostic techniques in clinical laboratories for the diagnosis, treatment monitoring, and management of COVID-19 patients. We concluded that each assays differ in their performance characteristics and the utilization of multiple techniques is necessary for the accurate diagnosis and management of SARS-CoV-2 infection.

Bromocriptine therapy: Review of mechanism of action, safety and tolerability.

Bromocriptine is a sympatholytic dopamine D2 receptor agonist with remarkable bioactivities. It has been used clinically as a prescription drug for more than 30 years to treat hyperprolactinemia associated conditions, Parkinson's disease, acromegaly, prolactinomas and other pituitary hormone dependent adenomas and recently, diabetes mellitus as well as various other disorders. Long-term treatment with bromocriptine has minimal or no harmful effects on renal, hepatic, cardiac or hematologic functions. This review article was planned to study the hypothetical and proposed mechanism of action as well as provide a brief discussion about its safety issues and tolerability. Bromocriptine represents an attractive option with high efficacy and safety profile for hyperprolactinemia-associated conditions, acromegaly, parkinsonism, type 2 diabetes mellitus and various other diseases in a variety of dosage forms for best possible beneficial effects. It appeared to be an effective and safe addition to the pharmacopoeia of drugs for the treatment of a vast variety of diseases as monotherapy or in combination with other drugs.

Prevalence of transfusion-transmitted infections in multiple blood transfusion-dependent thalassemic patients in Asia: A systemic review.

BACKGROUND: Thalassemia is a hereditary hemolytic anemia marked by a defect in synthesizing one or more globin chains in hemoglobin. In Pakistan, approximately 10,000 patients with thalassemia are primarily dependent on blood transfusions. The ß-thalassemia patients require blood transfusions and iron chelation therapy. Patients who need blood transfusions are at an increased risk of contracting transfusion-transmitted infections (TTIs) such as hepatitis B and C viruses (HBV and HCV, respectively), as well as the human immunodeficiency virus (HIV). OBJECTIVE: This systemic review aims to assess the prevalence of TTIs in transfusion-dependent ß-thalassemia patients in Asia. METHODS: The data for the systematic review were gathered from PubMed, Google Scholar, the Directory of Open Access Journals (DOAJ), and ScienceDirect using the following keywords: "prevalence, HBV, HCV, HIV, thalassemia, and transfusion-transmitted infections (TTIs)," and so on. This review includes the research articles that address the prevalence of viral infections in thalassemic patients following blood transfusion. RESULTS: A preliminary search of various databases identified 231 potential studies. 157 duplicate studies were eliminated, and the eligibility of 59 full-length articles was determined. Only 43 studies met the inclusion criteria. Among the 43 studies analyzed, 11 reported a high prevalence of HCV alone in thalassemic patients, while 21 reported a high prevalence of HCV and HBV infection in thalassemic patients. Eight studies reported the prevalence of all three TTIs examined, namely, HCV, HBV, and HIV, in patients with transfusion-dependent thalassemia. CONCLUSION: Preventable transfusion-transmitted infections occur frequently, and robust national policies and hemovigilance are required to detect and mitigate the infection risk.

Withania somnifera Chemical Constituents' In Vitro Antioxidant Potential and Their Response on Spermatozoa Parameters.

Background: Phytomedicine is becoming more acceptable as an alternative medicinal approach in the modern era. Objectives: The current study examined the antioxidant capacity and in vitro response of phytochemical constituents of Withania somnifera (Ashwagandha) on standard parameters of healthy volunteer semen. Methods: The phytochemicals and their pharmacological response in a hydroethanolic (30:70 v/v) extract of W. somnifera roots were determined using standard protocols. Results: The constituents included flavonoids, phenolic acids, alkaloids, and terpenoids were reported. High-performance liquid chromatography and Fourier-transform infrared spectroscopy determined a diverse array of biologically active chemical constituents in the extract. The extract of W. somnifera exhibits substantial antioxidant properties, including total antioxidant capacity, 2,2-diphenyl-1-picrylhydrazyl inhibition, H2O2 scavenging, and Fe3+ reducing potential (P < .05). The analysis of essential natural minerals explored adequate levels determined using atomic absorption spectrophotometer. Cytotoxic studies revealed significant thrombolytic, RBC membrane stabilization, and DNA damage protection activity (P < .05) while remaining non-mutagenic against Salmonella typhi TA98 and TA100. The best protective response of W. somnifera extract on human semen parameters (n = 30), such as total motility, progressive motility, and viability, demonstrated a significant (P < .05) improvement, particularly at the dose of 25 µg/mL and 50 µg/mL. Conclusion: The study concludes that W. somnifera possesses favorable in vitro characteristics that could aid in the preservation of sperm during intrauterine insemination and in vitro fertilization.

Molecular targets for the management of cancer using Curcuma longa Linn. phytoconstituents: A Review.

Medicinal plants are being used for therapeutic purposes since the dawn of human civilization. The therapeutic efficacy of medicinal plants is due to the presence of wide range phytochemical constituents or secondary metabolites. The medicinal plants are traditionally used for several types of ailments. Even in those pathological conditions where other methods of treatment fail to work. Curcuma longa Linn is very common ingredient used as spice in foods as preservative and coloring material in different part of the world. It has been used as a home remedy for a variety of diseases. Curcuma longa and its isolated constituent curcumin are widely evaluated for anticancer activity. Curcumin possesses broad remedial potential due to its multi-targeting effect against many different carcinoma including leukemia, genitourinary cancers, gastrointestinal cancers and breast cancer etc. Hence, Curcumin has potential for the development of new medicine for the treatment of several diseases.

Hantavirus diseases pathophysiology, their diagnostic strategies and therapeutic approaches: A review.

Hantaviruses are enveloped negative (-) single-stranded RNA viruses belongs to Hantaviridae family, hosted by small rodents and entering into the human body through inhalation, causing haemorrhagic fever with renal syndrome (HFRS) and hantavirus pulmonary syndrome (HPS) also known as hantavirus cardiopulmonary syndrome (HCPS). Hantaviruses infect approximately more than 200 000 people annually all around the world and its mortality rate is about 35%-40%. Hantaviruses play significant role in affecting the target cells as these inhibit the apoptotic factor in these cells. These viruses impair the integrity of endothelial barrier due to an excessive innate immune response that is proposed to be central in the pathogenesis and is a hallmark of hantavirus disease. A wide range of different diagnostic tools including polymerase chain reaction (PCR), focus reduction neutralization test (FRNT), enzyme-linked immunosorbent assay (ELISA), immunoblot assay (IBA), immunofluorescence assay (IFA), and other molecular techniques are used as detection tools for hantavirus in the human body. Now the availability of therapeutic modalities is the major challenge to control this deadly virus because still no FDA approved drug or vaccine is available. Antiviral agents, DNA-based vaccines, polyclonal and monoclonal antibodies neutralized the viruses so these techniques are considered as the hope for the treatment of hantavirus disease. This review has been compiled to provide a comprehensive overview of hantaviruses disease, its pathophysiology, diagnostic tools and the treatment approaches to control the hantavirus infection.

Therapeutic Response of Epimedium gandiflorum's Different Doses to Restore the Antioxidant Potential and Reproductive Hormones in Male Albino Rats.

Current study was planned to explore the therapeutic response of different doses of hydroethanolic extract of Epimedium grandiflorum leaves in male albino rats. Phytochemical analysis, HPLC and FTIR spectroscopy results revealed the presence of wide range of phenolic compounds and functional groups, respectively. Further, extract not induced significant hemolysis (7.56 ± 1.297%) against PBS (3.65 ± 0.35%) as negative control; while have significant clot lysis (44 ± 5.2%) potential, exhibited DPPH (78.87 ± 5.427%) scavenging, H2O2 (31.82 ± 3.491%) scavenging, antioxidant and reducing power activities. In vivo experimentation in albino male rats' revealed that administration of different doses (50, 100, 200 mg/Kg b.w.) of extract orally for 42 days after CCl4 intoxication significantly (P < 0.05) restore the selected parameters including liver enzymes, renal profiles, and stress markers and significantly (P < 0.05) increased reproductive hormones like testosterone, luteinizing hormone, follicle stimulating hormone and prolactin while significantly (P < 0.05) decreased progesterone and estradiol toward normal in dose dependent manner. Significant (P < 0.05) improvement in the structural architecture of testicular tissue particularly in high dose group (200 mg/Kg b.w.) was also observed. Results revealed E. grandiflorum has significant therapeutic response to address the healthcare problems particularly of impotency.

REVIEW - Advancements in combination therapy of Hepatitis C virus.

Hepatitis C virus (HCV) is a primary cause of chronic liver disease along with various complications like liver cirrhosis and malignancy which leads to death. It has infected more than 185 million people worldwide. There is no congruence established for the treatment of various genotypes of HCV infection owing to diversity in prevalence globally. Assessment of affected individuals with HCV by polymerase chain reaction (PCR), viral load of HCV and liver enzyme levels (i.e., ALT and AST) are the foundation to evaluate the safety and efficacy of HCV therapies. The antiviral efficacy has been greatly improved and sustained viral response (SVR) rates increased from 6% with interferon monotherapy to 50-80% with PEG-interferon/ribavirin combination therapy to >95% after the approval of all interferon free oral direct acting antiviral agents. The main objective of this review article is to compile data from reference sources regarding the old and current therapeutic strategies used to manage HCV infection. It is accepted that chronic HCV infection increases patient's thrombocytopenia and neutropenia risk and complications increased in co-morbid disorders like in tuberculosis, HIV, diabetes etc. In past treatment associated side effects were the major consequences and many patients have to stop the treatment. But after the approval of direct acting antiviral drugs create a revolution in the treatment of HCV infection. So, it could be concluded that current combination therapies are a promising hope to eradicate and to control HCV but some safety concerns required more considerations Therefore, this review focus on the available latest combination therapies and their effectiveness to eradicate HCV infection.

Naegleria fowleri: Sources of infection, pathophysiology, diagnosis, and management; a review.

Naegleria fowleri, a thermophilic flagellate amoeba known as a "brain-eating" amoeba, is the aetiological agent of a perilous and devastating waterborne disease known as primary amoebic meningoencephalitis (PAM), both in humans as well as in animals. PAM is a rare but fatal disease affecting young adults all around the world, particularly in the developed world but recently reported from developing countries, with 95%-99% mortality rate. Swimmers and divers are at high risk of PAM as the warm water is the most propitious environment adapted by N. fowleri to cause this infection. Infective amoeba in the trophozoite phase enter the victim's body through the nose, crossing the cribriform plate to reach the human brain and cause severe destruction of the central nervous system (CNS). The brain damage leads to brain haemorrhage and death occurs within 3-7 days in undiagnosed cases and maltreated cases. Though the exact pathogenesis of N. fowleri is still not known, it has exhibited two primary mechanisms, contact-independent (brain damage through different proteins) and contact-dependent (brain damage through surface structures food cups), that predominantly contribute to the pathogen invading the host CNS. For the management of this life-threatening infection different treatment regimens have been applied but still the survival rate is only 5% which is ascribed to its misdiagnosis, as the PAM symptoms closely resembled bacterial meningitis. The main objectives of this review article are to compile data to explore the sources and routes of N. fowleri infection, its association in causing PAM along with its pathophysiology; latest techniques used for accurate diagnosis, management options along with challenges for Pakistan to control this drastic disorder.

Hexose monophosphate shunt, the role of its metabolites and associated disorders: A review.

The hexose monophosphate (HMP) shunt acts as an essential component of cellular metabolism in maintaining carbon homeostasis. The HMP shunt comprises two phases viz. oxidative and nonoxidative, which provide different intermediates for the synthesis of biomolecules like nucleotides, DNA, RNA, amino acids, and so forth; reducing molecules for anabolism and detoxifying the reactive oxygen species during oxidative stress. The HMP shunt is significantly important in the liver, adipose tissue, erythrocytes, adrenal glands, lactating mammary glands and testes. We have researched the articles related to the HMP pathway, its metabolites and disorders related to its metabolic abnormalities. The literature for this paper was taken typically from a personal database, the Cochrane database of systemic reviews, PubMed publications, biochemistry textbooks, and electronic journals uptil date on the hexose monophosphate shunt. The HMP shunt is a tightly controlled metabolic pathway, which is also interconnected with other metabolic pathways in the body like glycolysis, gluconeogenesis, and glucuronic acid depending upon the metabolic needs of the body and depending upon the biochemical demand. The HMP shunt plays a significant role in NADPH2 formation and in pentose sugars that are biosynthetic precursors of nucleic acids and amino acids. Cells can be protected from highly reactive oxygen species by NADPH 2 . Deficiency in the hexose monophosphate pathway is linked to numerous disorders. Furthermore, it was also reported that this metabolic pathway could act as a therapeutic target to treat different types of cancers, so treatments at the molecular level could be planned by limiting the synthesis of biomolecules required for proliferating cells provided by the HMP shunt, hence, more experiments still could be carried out to find additional discoveries.

Antiviral potential of medicinal plants against HIV, HSV, influenza, hepatitis, and coxsackievirus: A systematic review.

Viral infections are being managed therapeutically through available antiviral regimens with unsatisfactory clinical outcomes. The refractory viral infections resistant to available antiviral drugs are alarming threats and a serious health concern. For viral hepatitis, the interferon and vaccine therapies solely are not ultimate solutions due to recurrence of hepatitis C virus. Owing to the growing incidences of viral infections and especially of resistant viral strains, the available therapeutic modalities need to be improved, complemented with the discovery of novel antiviral agents to combat refractory viral infections. It is widely accepted that medicinal plant heritage is nature gifted, precious, and fueled with the valuable resources for treatment of metabolic and infectious disorders. The aims of this review are to assemble the facts and to conclude the therapeutic potential of medicinal plants in the eradication and management of various viral diseases such as influenza, human immunodeficiency virus (HIV), herpes simplex virus (HSV), hepatitis, and coxsackievirus infections, which have been proven in diverse clinical studies. The articles, published in the English language since 1982 to 2017, were included from Web of Science, Cochrane Library, AMED, CISCOM, EMBASE, MEDLINE, Scopus, and PubMed by using relevant keywords including plants possessing antiviral activity, the antiviral effects of plants, and plants used in viral disorders. The scientific literature mainly focusing on plant extracts and herbal products with therapeutic efficacies against experimental models of influenza, HIV, HSV, hepatitis, and coxsackievirus were included in the study. Pure compounds possessing antiviral activity were excluded, and plants possessing activity against viruses other than viruses in inclusion criteria were excluded. Hundreds of plant extracts with antiviral effect were recognized. However, the data from only 36 families investigated through in vitro and in vivo studies met the inclusion criteria of this review. The inferences from scientific literature review, focusing on potential therapeutic consequences of medicinal plants on experimental models of HIV, HSV, influenza, hepatitis, and coxsackievirus have ascertained the curative antiviral potential of plants. Fifty-four medicinal plants belonging to 36 different families having antiviral potential were documented. Out of 54 plants, 27 individually belong to particular plant families. On the basis of the work of several independent research groups, the therapeutic potential of medicinal plants against listed common viral diseases in the region has been proclaimed. In this context, the herbal formulations as alternative medicine may contribute to the eradication of complicated viral infection significantly. The current review consolidates the data of the various medicinal plants, those are Sambucus nigra, Caesalpinia pulcherrima, and Hypericum connatum, holding promising specific antiviral activities scientifically proven through studies on experimental animal models. Consequently, the original research addressing the development of novel nutraceuticals based on listed medicinal plants is highly recommended for the management of viral disorders.

Acidic and enzymatic saccharification of waste agricultural biomass for biotechnological production of xylitol.

BACKGROUND: The plant biomass and agro-industrial wastes show great potential for their use as attractive low cost substrates in biotechnological processes. Wheat straw and corn cob as hemicellulosic substrates were acid hydrolyzed and enzymatically saccharified for high xylose production. The hydrolysate was concentrated and fermented by using Saccharomyces cerevisiae and Kluyveromyces for production of xylitol. RESULTS: Acid hydrolysis of wheat straw and corn cob in combination with enzymatic hydrolysis showed great potential for production of free sugars from these substrates. Kluyveromyces produced maximum xylitol from acid treated wheat straw residues with enzymatic saccharification. The percentage xylitol yield was 89.807 g/L and volumetric productivity of 0.019 g/L/h. Kluyveromyces also produced maximum xylitol from corn cob acid hydrolyzed liquor with xylitol yield 87.716 g/L and volumetric productivity 0.018 g/L/h. CONCLUSION: Plant and agro-industrial biomass can be used as a carbohydrate source for the production of xylitol and ethanol after microbial fermentation. This study revealed that wheat straw acid and enzyme hydrolyzed residue proved to be best raw material for production of xylitol with S. cerevisiae. The xylitol produced can be utilized in pharmaceuticals after purification on industrial scale as pharmaceutical purposes.

Drug resistant strains of Mycobacterium tuberculosis identified through PCR-RFLP from patients of Central Punjab, Pakistan.

The study was carried out to determine, by PCR-RFLP, the magnitude of drug resistance in Mycobacterium tuberculosis The study was carried out on 221 random sputum samples collected from patients and 120 suspected cases of drug resistance. Genetic variation in drug-resistant strains was evaluated through PCR-RFLP for isoniazid, ethambutol, streptomycin, and ofloxacin. Out of 341 patients, 91.5% were confirmed as M. tuberculosis complex infected on the basis of PCR. The random samples revealed resistance in 8.2% cases, while 73.3% of those with suspected drug resistance were found resistant. Among drug-resistant isolates, 56.1% were resistant to a single drug, 33.3% to two drugs, and 10.6% to more than two drugs. Ofloxacin resistance was observed along with isoniazid, ethambutol, and streptomycin in 6.5% cases. Resistance to isoniazid was observed in 61% cases, to ethambutol in 50.4%, and to streptomycin in 43.1% cases. It was concluded that PCR-RFLP is a useful molecular technique for the rapid detection of mutations in drug-resistant tuberculosis patients and may be used to diagnose drug resistance at the earliest.

Identification of ofloxacin-resistant Mycobacterium tuberculosis by PCR-RFLP and Sequencing.

This study was planned to verify the resistance frequency of Ofloxacin (OFX) against Mycobacterium tuberculosis by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) technique and sequencing. Total 366 clinical samples of suspected TB patients were collected from various localities of central Punjab. All of them were found positive by ZN (Zeihl-Nelsen) staining method. Among them, 108 (29.5%) were found negative and 258 (70.5%) positive on PCR based study. The cases not responding to ATT were further characterized by proportion method and by PCR-RFLP to establish the drug resistance. Selected drug resistant case were further sequenced to confirm the results of amplified RFLP. The results showed that out of 118 drug resistant cases, 06 (5.08%), 03 (2.54%) were found resistant to OFX by drug susceptibility testing and PCR-RFLP respectively. The two strains were selected for sequencing procedure. The strain-79 showed point mutation at four points, at codon 70, 71, 76 and 78. The sequence of strain- 81 showed mutation at codon 95.PCR-RFLP is a useful molecular technique for the rapid detection of mutations and may be used to diagnose drug resistance but it should be confirmed by sequencing before starting 2(nd) and 3(rd) generation treatment because the restriction site is the cornerstone of PCR-RFLP and mutation may be occurring elsewhere.

Impact of reactive oxygen species on antioxidant capacity of male reproductive system.

The present research work was aimed to study the mutual interaction of reactive oxygen species (ROS) and basal cells antioxidant capacity in the male reproductive system and to further establish the association between selected heavy metals and stress markers. Total oxidant status (TOS) and total antioxidant status (TAS) of serum and seminal plasma were determined by automated photometric methods. The concentrations of Selenium (Se), Lead (Pb), and Cadmium (Cd) were determined by using atomic absorption spectrophotometer. The TOS was increased significantly (P <0.05) in seminal plasma as well as in the serum of the infertile group when compared with the fertile group. On the other hand, the TAS of the infertile group was found to be noticeably decreased (P <0.05) when compared with the TAS of the fertile group. Among the heavy metals, a noticeably lower concentration of Se was detected in the infertile group whereas markedly elevated levels of Cd and Pb were observed in the infertile group compared with the fertile group. Among the infertile group a significant inverse correlation (r = -0.521, P <0.05) was observed between Se and TOS and between Cd and Pb (r = -0.407, P <0.05). Contrarily among the infertile group a considerable positive relationship was established between Se and TAS (r = 0.507, P <0.05). It was concluded that the oxidant stress reduces the antioxidant activity in infertile men by elevating the production of ROS. A lower concentration of Se and elevated levels of Pb and Cd explain the individual's exposure to these heavy metals. The study also revealed that the heavy metal toxicity contributes significantly to male infertility.

Variation in minerals, phenolics and antioxidant activity of peel and pulp of different varieties of peach (Prunus persica L.) fruit from Pakistan.

Peach (Prunus persica L.), being a potential source of bioactive compounds, has been demonstrated to have medicinal benefits. In this study variation of minerals and antioxidant characteristics (total phenolic contents, total flavonoid contents, reducing power, inhibition of peroxidation using linoleic acid system and DPPH free radical scavenging activity) between peel and pulp parts of different peach varieties, namely Golden, Shireen, and Shahpasand were investigated. The peel and pulp extracts, derived from the varieties analyzed, exhibited an appreciable amount of total phenolics (TP) and total flavonoids (TF), ranging from 1,209.3-1,354.5, 711.7-881.3 mg GAE/100 g and 599.7-785.5, 301.3-499.7 mg CE/100 g on a dry weight basis, respectively. Reducing power of peel and pulp extracts (12.5 mg/mL concentration) ranged from 2.57-2.77 and 1.54-1.99.The inhibition of linoleic acid peroxidation and DPPH scavenging activity of the extracts varied from 70.8-80.9% and 66.8-76.5% in peels, and 51.9-60.1% and 43.4-49.1% in pulps. The mineral analysis revealed that the content of K was highest in both parts of the peach fruit followed by Mg, Ca, Fe, Mn and Zn. The results of our present study indicate that peach peel had significantly higher levels of minerals, antioxidant capacity and phenolics than those of the pulp, suggesting the intake of unpeeled peach as a potential source of high-value components. The peach peel can be a useful as a viable source of natural antioxidants for functional foods and nutraceutical applications.