Status of mannose-binding lectin (MBL) and complement system in COVID-19 patients and therapeutic applications of antiviral plant MBLs.

Coronavirus disease 2019 (COVID-19) is an infectious disease caused by a virus called "Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2)." In the majority of patients, infection with COVID-19 may be asymptomatic or may cause only mild symptoms. However, in some patients, there can also be immunological problems, such as macrophage activation syndrome (CSS) that results in cytokine storm syndrome (CSS) and acute respiratory distress syndrome (ARDS). Comprehension of host-microbe communications is the critical aspect in the advancement of new therapeutics against infectious illnesses. Endogenous animal lectins, a class of proteins, may perceive non-self glycans found on microorganisms. Serum mannose-binding lectin (sMBL), as a part of the innate immune framework, recognizes a wide range of microbial microorganisms and activates complement cascade via an antibody-independent pathway. Although the molecular basis for the intensity of SARS-CoV-2 infection is not generally understood, scientific literature indicates that COVID-19 is correlated with unregulated activation of the complement in terms of disease severity. Disseminated intravascular coagulation (DIC), inflammation, and immune paralysis contribute to unregulated complement activation. Pre-existing genetic defects in MBL and their association with complement play a major role in immune response dysregulation caused by SARS-CoV-2. In order to generate anti-complement-based therapies in Covid-19, an understanding of sMBL in immune response to SARS-CoV-2 and complement is therefore essential. This review highlights the role of endogenous sMBL and complement activation during SARS-CoV-2 infection and their therapeutic management by various agents, mainly plant lectins, since antiviral mannose-binding plant lectins (pMBLs) offer potential applications in the prevention and control of viral infections.

LDH-C4: a target with therapeutic potential for cancer and contraception.

The lactate dehydrogenase (LDH) has been studied widely because it exists in various isozymic forms. The association of A and B subunits of LDH can generate five tetrameric isozymes, but the finding of the sixth isozyme in mature human testis and sperm indicated the presence of an additional subunit of LDH, designated as LDH-X (also termed LDH-C4 due to tetrameric nature of C-subunit). LDH-C4 isozyme is an iso-, allo-, and auto-antigen present in mammalian sperm cells. The synthesis of LDH-C4 in the testis takes place during sexual maturation, and it is the predominant fraction in mature spermatozoa. Though, originally considered to be testis specific, LDH-C or Ldh3 in mice was later detected in the murine oocyte and early embryo. Ldh3 in mouse supports its role in energy production in spermatids that favor lactate as substrate and in spermatozoa with a characteristic aerobic glycolytic path to yield ATP. During last two decades, cancer/testis-associated genes (CTAs) which are expressed only in the germinal epithelium of the testis are also expressed in some cancer cells, but not in non-cancerous somatic tissues. The CTAs are considered promising candidates for diagnosis and immunotherapy of cancer. The sperm-specific Ldh-c gene has been shown to express in a broad spectrum of human tumors, with high frequency in lung cancer, melanoma, and breast cancer; the protein being expressed virtually in all tumor types tested. Accordingly, LDH-C4 is the unique target for contraception in both males and females and offers potential future for immunotherapy of different types of cancers. As LDH-C has a preference for lactate as a substrate, LDH-C activation in cancer may depend on lactate for ATP production. The major aim of this article is to review the salient features of LDH-C subunit and the immune responses of LDH-C4 in homologous and heterologous species in relation to its role in acceptance or rejection of the allograft and its application in contraception and immunotherapy of cancer, directly or indirectly through the regulation of its substrate, the lactate.

The Lactate and the Lactate Dehydrogenase in Inflammatory Diseases and Major Risk Factors in COVID-19 Patients.

Lactate dehydrogenase (LDH) is a terminating enzyme in the metabolic pathway of anaerobic glycolysis with end product of lactate from glucose. The lactate formation is crucial in the metabolism of glucose when oxygen is in inadequate supply. Lactate can also be formed and utilised by different cell types under fully aerobic conditions. Blood LDH is the marker enzyme, which predicts mortality in many conditions such as ARDS, serious COVID-19 and cancer patients. Lactate plays a critical role in normal physiology of humans including an energy source, a signaling molecule and a pH regulator. Depending on the pH, lactate exists as the protonated acidic form (lactic acid) at low pH or as sodium salt (sodium lactate) at basic pH. Lactate can affect the immune system and act as a signaling molecule, which can provide a "danger" signal for life. Several reports provide evidence that the serum lactate represents a chemical marker of severity of disease similar to LDH under inflammatory conditions. Since the mortality rate is much higher among COVID-19 patients, associated with high serum LDH, this article is aimed to review the LDH as a therapeutic target and lactate as potential marker for monitoring treatment response of inflammatory diseases. Finally, the review summarises various LDH inhibitors, which offer potential applications as therapeutic agents for inflammatory diseases, associated with high blood LDH. Both blood LDH and blood lactate are suggested as risk factors for the mortality of patients in serious inflammatory diseases.

Applications of mannose-binding lectins and mannan glycoconjugates in nanomedicine.

Glycosylated nanoparticles (NPs) have drawn a lot of attention in the biomedical field over the past few decades, particularly in applications like targeted drug delivery. Mannosylated NPs and mannan-binding lectins/proteins (MBL/MBP) are emerging as promising tools for delivery of drugs, medicines, and enzymes to targeted tissues and cells as nanocarriers, enhancing their therapeutic benefits while avoiding the adverse effects of the drug. The occurrence of plenty of lectin receptors and their mannan ligands on cell surfaces makes them multifaceted carriers appropriate for specific delivery of bioactive drug materials to their targeted sites. Thus, the present review describes the tethering of mannose (Man) to several nanostructures, like micelles, liposomes, and other NPs, applicable for drug delivery systems. Bioadhesion through MBL-like receptors on cells has involvements applicable to additional arenas of science, for example gene delivery, tissue engineering, biomaterials, and nanotechnology. This review also focuses on the role of various aspects of drug/antigen delivery using (i) mannosylated NPs, (ii) mannosylated lectins, (iii) amphiphilic glycopolymer NPs, and (iv) natural mannan-containing polysaccharides, with most significant applications of MBL-based NPs as multivalent scaffolds, using different strategies. Graphical abstract: Mannosylated NPs and/or MBL/MBP are coming up as viable and versatile tools as nanocarriers to deliver drugs and enzymes precisely to their target tissues or cells. The presence of abundant number of lectin receptors and their mannan ligands on cell surfaces makes them versatile carriers suitable for the targeted delivery of bioactive drugs.

Isolation, properties, immunological specificity and localization of mouse testicular hyaluronidase.

Hyaluronidase (hyaluronate 4-glycanohydrolase, EC 3.2.1.35) was purified from mouse testes by ion-exchange chromatography, Sephadex G-200 filtration and Con A-agarose affinity chromatography. The final preparation had 94-fold purity and 12.2 units spec. act. of the enzyme (unit of specific activity = mumol N-acetylglucosamine released/h per mg protein at 37 degrees C and pH 4.5). Hyaluronidase is relatively heat stable and loses 10-20% of its activity at 50-55 degrees C for 10 min. Ea for eat denaturation of enzyme is 42-45 kcal between 45 an 63 degrees C. The Michaelis constant of mouse testicular hyaluronidase is 1.1 mg/ml hyaluronic acid. Antibodies to the purified enzyme were produced in rabbits and showed a single precipitin line by Ouchterlony gel diffusion. Antiserum to hyaluronidase inhibited enzyme activity by 25%. Immunologically, mouse testicular hyaluronidase is species specific. Tissue extracts of mouse vital organs, except testes and epididymis did not react with the antisera, though nonspecific precipitation occurred between intestinal extracts and anti-hyaluronidase serum. Hyaluronidase was localized in testis sections by indirect immunofluorescence. A specific dark green fluorescence was localized on cell boundaries extending from spermatogonia to spermatids and appeared on the sperm acrosome. Cytoplasm of spermatogonia and spermatocytes showed light green fluorescence, whereas interstitial tissue was devoid of fluorescence.

Isolation and characterization of the major form of beta-glucuronidase from human seminal plasma.

Human seminal plasma contain two forms of beta-glucuronidase (beta-D-glucuronidase glucuronosohydrolase, EC 3.2.1.31) which are present in the ratio of 4:1. The major form of beta-glucuronidase with a slow moving band in electrophoresis was purified to homogeneity as revealed by polyacrylamide gel electrophoresis, double immunodiffusion and immunoelectrophoresis. The major form of beta-glucuronidase shows dual optimum pH at 4.3 and 4.7 with a dip in the activity at pH 4.5. The Km of this form of beta-glucuronidase is dependent on pH and was found to be 0.95, 3.08 and 0.67 mM at pH 4.4, 4.5 and 4.7, respectively. The major form of beta-glucuronidase from seminal plasma is stable at 55 degrees C for 30 min but it denatures at 65 degrees C. Heat denaturation is faster at acidic pH (4.7) than at alkaline pH (7.8). However, the activity of enzyme increased linearly with increase in temperature up to 70 degrees C during incubation with substrate. Cu, Ag, Hg and Ni salts inhibited enzyme activity significantly at 0.1 and 1.0 mM concentration, but the inhibition of HgCl2 was protected by cysteine. 1,4-D-Saccharic acid lactone and ascorbic acid inhibited seminal beta-glucuronidase competitively, yielding Ki values of 1.7 . 10(-3) mM and 10.3 mM, respectively. Though fructose and mannose also showed significant inhibition of beta-glucuronidase at 10-100 mM, glucose did not show any effect. The molecular weight of the major form of beta-glucuronidase was found to be 279 000, and it appears to be composed of four subunits each having a molecular weight of 74 000.

The EEG response to diffuse and patterned photic stimulation during acute untreated alcohol withdrawal.

We studied the responses to diffuse and patterned stroboscopic light stimulation prospectively in 49 individuals during acute alcohol withdrawal prior to pharmacologic treatment. Photomyogenic responses (PMR) occurred in only two (4%) of those tested, and photoparoxysmal responses (PPR) never occurred. These findings suggest that PMR occur far less often during alcohol withdrawal than previously thought and that PPR may not be a direct manifestation of alcohol withdrawal.

Immunological specificity of hexosaminidases from human seminal plasma.

Two isozymes of hexosaminidases (Hex) were purified from human seminal plasma and found to be homogeneous as revealed by immunoelectrophoresis and immunodiffusion experiments. Anti-Hex A antibodies were precipitated with isozyme B and vice versa. Though the precipitin arcs were equally stained for protein in these two antigen and antibody systems, enzyme activity was weakly stained in the reverse systems (ie, Anti-Hex A with Hex B and Anti-Hex B with Hex A). Thus, some common sequential antigenic determinants were indicated in two isozymes of Hex. On the other hand, loss of enzyme activity in the precipitate of reverse antigen-antibody complexes as compared to direct system (Anti-Hex A with Hex A and Anti-Hex B with Hex B) revealed that the two isozymes are present in different sequential and conformational states near the active sites. Moreover, antibodies of both forms of Hex cross-reacted with human sperm and reproductive and other vital organs such as kidney, liver, lung, pancreas, and spleen. Though monkey tissues also cross-reacted with human seminal plasma Hex-antisera, mouse, rat, and goat testis and epididymis extracts failed to show any cross-reaction. It is concluded that human Hex(s) are tissue nonspecific but appear to be specific to the human and the subhuman primates.

Ranking toxicity of industrial dusts by bronchoalveolar lavage fluid analysis.

Female wistar rats were inoculated intratracheally with 10 mg/ml suspensions of various dusts, viz: quartz, fly ash, mica and corundum in physiological saline. Biochemical markers of bronchoalveolar lavage fluid (BALF) were analysed 8 days after the instillation of the dusts. Elevated levels of proteins, sialic acid and phospholipid contents and the activity of lactate dehydrogenase correlated well with the degree of the known fibrogenic potential of different dusts in the lungs in the following order, quartz greater than fly ash greater than mica greater than corundum. beta-glucuronidase activity, was however, only elevated in the quartz inoculated group of rats. It is suggested that biochemical constituents of BALF analysed shortly after the exposure to different dusts can be useful to mirror alterations in the tissue response to mineral dusts.

Cellular and biochemical indices of bronchoalveolar lavage for detection of lung injury following insult by airborne toxicants.

Cellular and biochemical profiles of bronchoalveolar lavage (BAL) material after inhalation or intratracheal exposure to various airborne toxicants clearly reflect that BAL has the potential of being a useful tool for the rapid screening of lung injury. The cellular and biochemical responses not only predict inflammation, extent of tissue damage and toxic nature of the substances, but could also help in understanding the molecular mechanisms of pathogenicity. Depending upon the changes of BAL in animals acutely exposed to a pulmonary toxicant, future in-depth studies along with complete histopathological evaluations could be made. Also, the assessment of macromolecules of pharmacological importance in the lavage, especially the secretory products of alveolar macrophages and other lung cell types, could be very useful in predicting the toxic potential of various airborne substances and could also serve as important indicators of developing chronic lung diseases and, therefore, necessitate further studies.