A comparative study of the epiligament of the medial collateral and anterior cruciate ligaments in the human knee: Immunohistochemical analysis of CD 34, α-smooth muscle actin and vascular endothelial growth factor in relation to epiligament theory.

BACKGROUND: This study evaluated and compared the expression of VEGF, CD34, and α-SMA in the anterior cruciate ligaments and medial collateral ligaments in healthy human knees in order to enrich the epiligament theory regarding ligament healing after injury. METHODS: Samples from the mid-substance of the anterior cruciate ligament and the medial collateral ligament of 12 fresh knee joints were used. Monoclonal antibodies against CD34, α-SMA, and VEGF were used for immunohistochemical analysis. Photomicrographs were analyzed using the ImageJ software. RESULTS: The epiligament of the anterior cruciate ligament showed slightly higher expression of CD34, α-SMA, and VEGF than the epiligament of the medial collateral ligament. Overall, among the tested markers, α-SMA expression was most pronounced in anterior cruciate ligament epiligament images and CD34 dominated in medial collateral ligament epiligament images. The intensity of DAB staining for CD34, α-SMA, and VEGF was higher in vascular areas of the epiligament than in epiligament connective tissue. CONCLUSIONS: The results illustrate that CD34, α-SMA, and VEGF are expressed in the human epiligament. The differences between the epiligament of the investigated ligaments and the fact that CD34, α-SMA, and VEGF, which are known to have a definite role in ligament healing, are predominantly expressed in the main vascular part of the ligament-epiligament complex enlarge the existing epiligament theory. Future investigations regarding better ligament healing should not overlook the epiligament tissue.

The novel epiligament theory: differences in healing failure between the medial collateral and anterior cruciate ligaments.

According to current literature, 90% of knee ligament injuries involve the medial collateral ligament or the anterior cruciate ligament. In contrast to the medial collateral ligament, which regenerates relatively well, the anterior cruciate ligament demonstrates compromised healing. In the past, there were numerous studies in animal models that examined the healing process of these ligaments, and different explanations were established. Although the healing of these ligaments has been largely investigated and different theories exist, unanswered questions persist.Therefore, the aim of this article is 1) to review the different historical aspects of healing of the medial collateral ligament and present the theories for healing failure of the anterior cruciate ligament; 2) to examine the novel epiligament theory explaining the medial collateral ligament healing process and failure of anterior cruciate ligament healing; and 3) to discuss why the enveloping tissue microstructure of the aforementioned ligaments needs to be examined in future studies.We believe that knowledge of the novel epiligament theory will lead to a better understanding of the normal healing process for implementing optimal treatments, as well as a more holistic explanation for anterior cruciate ligament healing failure.

Bacteriophage as an antibacterial agent: a patent perspective.

The present review encompasses a patent landscape on bacteriophage as an antimicrobial agent and one of the alternatives to combat antibiotic resistance in bacteria. This study gives a perspective on use of bacteriophages in various industries such as healthcare, food safety and animal and plant protection. Patenting activity was noted for all the antibiotic-resistant bacterial pathogens listed in the 'critical' category by the WHO. Broadly, claims of the analyzed patents were directed toward bacteriophage, composition/formulation containing phage, phage proteins and various methods of using or producing phage. The challenges to approval of phage therapy in clinical use may be overcome with the help of focused research and modification of the regulatory guidelines for phage therapy.

A patent review on strategies for biological control of mosquito vector.

Mosquitoes are the vectors responsible for transmitting serious and life-threatening diseases such as malaria, dengue, yellow fever, chikungunya and lymphatic filariasis. Very few effective vaccines or drugs have been developed so far to prevent or treat these diseases, highlighting a need for vector control. This paper presents a comprehensive technology overview of patent documents disclosing biological agents for mosquito control. The patent analysis revealed that comparable number of patent documents were filed in two technology categories: non-recombinant agents and genetically modified (GM) agents. In the category of non-recombinant agents, toxic peptides from microbes and biological consortia seemed to be the earliest technology noted right from the year 1965 whereas the patent filings for suppression of mosquito population using genetic modification techniques have emerged from the year 2000 onwards. The United States of America is the leading patent filing jurisdiction followed by China and the Great Britain. Academic institutes have filed higher number of patent applications as compared to private companies. University of Florida was found to be the leading patent filing entity and its patents were focused on suppression of vector population using techniques such as release of insects with dominant lethal (RIDL) and RNA interference (RNAi).

A patent perspective on chikungunya.

Chikungunya is a viral disease caused by chikungunya virus (CHIKV) belonging to the Alphavirus genus and transmitted to humans by mosquitoes of Aedes spp. Nearly 40 countries in Asia, Africa, Europe, and the Americas have documented chikungunya cases. Most recent severe outbreaks have occurred in Indian Ocean islands of Réunion and Mauritius and India. There is no specific drug treatment for the disease, neither is there a standardized vaccine available for prevention of the disease. The present review gives a global perspective on patents filed pertaining to chikungunya. The United States has been the top patent filing jurisdiction followed by China, Europe, and India. The patents have been classified into categories of therapeutics, diagnostics, and vaccines. Maximum patent documents fall under the therapeutics category, in which patent applications are predominantly related to chemically derived drugs. They include nucleic acid analogues, various other host and virus enzyme/protein inhibitors. Patents on biological or plant derived drugs are being filed relatively recently. In the category of diagnostics, immunoassay based tests seemed to be of choice until the year 2005, whereas, patent filings for molecular diagnostics have now surpassed those with immunoassay techniques. In the vaccines category, vaccines based on viral vectors appear to be emerging as the preferred vaccine platform with the majority of patents filed in the years 2014-2017. Corporate sector has the most patent filings to its credit, followed closely by academic institutions. Pasteur Institut along with Chinese Agency for Science, Technology and Research is the top patent filing entity in chikungunya related technology space. Presently, nine vaccine products, three antiviral drugs and one mRNA based gene therapy are under development. Three vaccine products have been given fast track designation by US Food and Drug Administration (FDA) to expedite review and facilitate the development of a vaccine to prevent a serious or life-threatening condition and fill an unmet medical need. Similarly, one vaccine has been given PRIME (Priority Medicines) status by the European Medicines Agency (EMA).

Therapeutic and cosmetic applications of mangiferin: an updated patent review (patents published after 2013).

INTRODUCTION: Mangiferin and its derivatives possess diverse biological activities and desirable drug like properties. In continuation with the authors' previous publication, the present study reviews recently published patents in correlation with efforts of drug development using mangiferin. AREAS COVERED: Patents published after the year 2013 were analyzed for therapeutic and cosmetic applications of mangiferin, its salts, derivatives, and preparations. Patent information pertaining to improving solubility and bioavailability of mangiferin and related bioactives has also been presented. Methods of extraction of mangiferin and poly herbal preparations have not been included. Biological testing data has been put forth to understand the clinical trial status of products comprising mangiferin and/or its derivatives. EXPERT OPINION: Mangiferin has been highlighted as a multitarget bioactive with therapeutic potential. Research efforts in the recent years have contributed to development of new mangiferin derivatives as well as mangiferin formulations with enhanced solubility and bioavailability. Improvements in drug delivery systems for mangiferin might contribute to designing clinical trials to validate its therapeutic potential in humans. Majority of the recent patents have been filed by Chinese inventors claiming second medical use of mangiferin or its related compounds. However, a number of these patents remain in the prosecution stage.

A patent landscape on application of microorganisms in construction industry.

Construction biotechnology includes research and development of construction materials and processes that make use of various microbes. The present technology landscape gives a perspective on how microbes have been used in construction industry as cement and concrete additives by analyzing patents filed in this technology arena. All patents related to the technology of interest published globally to date have been reviewed. The earliest patent filing in this technology domain was recorded in the year 1958 and the patenting activity reached its peak around mid to late 1990s. The early technology was mainly focused on microbial polysaccharides and other metabolic products as additives. Year 2002 onwards, biomineralization has taken precedence over the other technologies with consistent patent filings indicating a shift in innovation focus. Japan has been the global leader with highest number of patents filed on application of microbes in construction industry. Southeast University, China has topped the patent assignee list with maximum number of filings followed by Kajima Corp. and Shin-Etsu Chemical Co., Ltd. Most patent applications have claimed microbe based bio-products. Construction-related microbial technologies are mainly based on activity of different microorganisms such as urease-producing, acidogenic, halophilic, alkaliphilic, nitrate and iron-reducing bacteria. Sporosarcina pasteurii has been the most widely used microbe for biomineralization.

Patents on Therapeutic and Cosmetic Applications of Bioactives of Crocus Sativus L. and their Production through Synthetic Biology Methods: A Review.

BACKGROUND: Saffron (Crocus sativus L.) has a long history of use as a food additive and a traditional medicine for treating a number of disorders. Prominent bioactives of saffron are crocin, crocetin and safranal. OBJECTIVE: The aim of this study was to carry out an extensive patent search to collect information on saffron bioactives and their derivatives as therapeutic and cosmeceutical agents. All patents related to the area of interest published globally till date have been reviewed. Moreover, a recent synthetic biology approach to cost effective and consistent production of saffron bioactives has been highlighted. METHODS: A patent search strategy was designed based on keywords and concepts related to Crocus sativus L. and its bioactives- safranal, crocin and crocetin in combination with different patent classification codes relevant to the technology areas. This search strategy was employed to retrieve patents from various patent databases. The patents which focused on therapeutic or cosmetic applications and claimed compositions comprising crocin, crocetin or safranal as the main active component were selected and analysed. RESULTS: Maximum patenting activity was noticed towards the use of these bioactives in the treatment of neurological disorders followed by multiple uses of the same compound, use in treatment of metabolic disorders and use as cosmeceuticals. Interestingly, there were no patent records related to use of these bioactives in treating infectious disorders. CONCLUSION: Our patent analysis points out the populous and less explored uses of saffron bioactives and areas where there is further scope for research and growth. Recently developed synthetic biology approach is contributory in improving availability, consistency and cost effectiveness of saffron bioactives.

Therapeutic and cosmetic applications of mangiferin: a patent review.

INTRODUCTION: Mangiferin, a natural C-glucoside xanthone [2-C-ß-D-glucopyranosyl-1, 3, 6, 7-tetrahydroxyxanthone], is abundantly present in young leaves and stem bark of the mango tree. The xanthonoid structure of mangiferin with C-glycosyl linkage and polyhydroxy components contributes to its free radical-scavenging ability, leading to a potent antioxidant effect as well as multiple biological activities. AREAS COVERED: An extensive search was carried out to collect patent information on mangiferin and its derivatives using various patent databases spanning all priority years to date. The patents claiming therapeutic and cosmetic applications of mangiferin and its derivatives were analyzed in detail. The technology areas covered in this article include metabolic disorders, cosmeceuticals, multiple uses of the same compound, miscellaneous uses, infectious diseases, inflammation, cancer and autoimmune disorders, and neurological disorders. EXPERT OPINION: Mangiferin has the potential to modulate multiple molecular targets including nuclear factor-kappa B (NF-κB) signaling and cyclooxygenase-2 (COX-2) protein expression. Mangiferin exhibits antioxidant, antidiabetic, antihyperuricemic, antiviral, anticancer and antiinflammatory activities. The molecular structure of mangiferin fulfils the four Lipinski's requisites reported to favor high bioavailability by oral administration. There is no evidence of adverse side effects of mangiferin so far. Mangiferin could thus be a promising candidate for development of a multipotent drug.

Winged bean chymotrypsin inhibitors retard growth of Helicoverpa armigera.

Two putative Kunitz-type chymotrypsin inhibitor genes (WCI2 and WCI5) were isolated from winged bean (Psophocarpus tetragonolobus (L.) DC). While WCI2 has previously been characterized, WCI5 represents a new member of the WCI family. WCI5 was exclusively expressed in winged bean seeds. Theoretical translation of both the genes resulted into polypeptides of 207 amino acids with 86% sequence similarity. The polypeptide sequences contain four half-cysteine residues and a well-conserved Leu(65)-Ser(66) reactive site, typical for chymotrypsin inhibitors. WCI5 and WCI2 were expressed in Pichia pastoris and the recombinant proteins were assayed against various proteinases. Both the inhibitors strongly inhibited commercially available bovine chymotrypsin. More importantly, gut proteinases of Helicoverpa armigera larvae that damage many important crop plants, were inhibited by WCI2 and WCI5. In addition, both proteinase inhibitors demonstrated significant reduction of growth of H. armigera larvae after feeding on inhibitor incorporated artificial diets. The inhibitory effects of WCI2 and WCI5 on activity of proteinases and larval growth make these proteins and their genes promising candidates for enhancing plant defense against H. armigera using transgenic plants.

Characterization of two midgut proteinases of Helicoverpa armigera and their interaction with proteinase inhibitors.

Two serine proteinases from the midgut of Helicoverpa armigera have been partially purified and characterized. One proteinase, HGP-1, was capable of hydrolyzing a synthetic substrate of elastase and was inhibited by elastatinal. The second proteinase, HGP-2, was inhibited by a trypsin inhibitor. Molecular weights of HGP-1 and HGP-2 were approximately 26.0 and 29.0kDa, respectively. Both the proteinases exhibited alkaline pH optima in the range of 10-11. Furthermore, interaction of HGP-1 and HGP-2 with proteinase inhibitors (PIs) from host and non-host plants was studied. HGP-1 was not only insensitive to a PI from chickpea (host) but was also able to degrade it. The same PI from chickpea was able to inhibit over 50% activity of HGP-2. On the contrary, PIs from potato (non-host) showed strong inhibition of both, HGP-1 and HGP-2 and also demonstrated protection of chickpea seed proteins from digestion by both the HGPs. These results could provide important clues in designing strategies for sustainable use of plant PIs in developing insect-tolerant transgenic plants.

Bitter gourd proteinase inhibitors: potential growth inhibitors of Helicoverpa armigera and Spodoptera litura.

Proteinase inhibitors (PIs) from the seeds of bitter gourd (Momordica charantia L.) were identified as strong inhibitors of Helicoverpa armigera gut proteinases (HGP). Biochemical investigations showed that bitter gourd PIs (BGPIs) inhibited more than 80% HGP activity. Electrophoretic analysis revealed the presence of two major proteins (BGPI-1 and-2) and two minor proteins (BGPI-3 and-4) having inhibitory activity against both trypsin and HGP. The major isoforms BGPI-1 and BGPI-2 have molecular mass of 3.5 and 3.0 kDa, respectively. BGPIs inhibited HGP activity of larvae fed on different host plants, on artificial diet with or without added PIs and proteinases excreted in fecal matter. Degradation of BGPI-1 by HGP showed direct correlation with accumulation of BGPI-2-like peptide, which remained stable and active against high concentrations of HGP up to 3 h. Chemical inhibitors of serine proteinases offered partial protection to BGPI-1 from degradation by HGP, suggesting that trypsin and chymotrypsin like proteinases are involved in degradation of BGPI-1. In larval feeding studies, BGPIs were found to retard growth and development of two lepidopteran pests namely Helicoverpa armigera and Spodoptera litura. This is the first report showing that BGPIs mediated inhibition of insect gut proteinases directly affects fertility and fecundity of both H. armigera and S. litura. The results advocate use of BGPIs to introduce insect resistance in otherwise susceptible plants.