Validation of QTLs for plant ideotype, earliness and growth habit traits in pigeonpea (Cajanus cajan Millsp.).

Pigeonpea productivity is greatly constrained by poor plant ideotype of existing Indian cultivars. Enhancing pigeonpea yield demands a renewed focus on restructuring the ideal plant type by using more efficient approaches like genomic tools. Therefore, the present study aims to identify and validate a set of QTLs/gene(s) presumably associated with various plant ideotype traits in pigeonpea. A total of 133 pigeonpea germplasms were evaluated along with four checks in the augmented design for various ideotype traits i.e. initiation of flowering (IF), days to 50% flowering (DFF), days to maturity (DM), plant height (PH), primary branches (PB), seeds per pod (SP) and pod length (PL). We observed significant genetic diversity in the germplasm lines for these traits. The genetic control of IF, DFF, DM and PH renders these traits suitable for detection of marker trait associations. By using residual maximum likelihood algorithm, we obtained appropriate variance-covariance structures for modeling heterogeneity, correlation of genetic effects and non-genetic residual effects. The estimates of genetic correlations indicated a strong association among earliness traits. The best linear unbiased prediction values were calculated for individual traits, and association analysis was performed in a panel of 95 diverse genotypes with 19 genic SSRs. Out of five QTL-flanking SSRs used here for validation, only ASSR295 could show significant association with FDR and Bonferroni corrections, and accounted for 15.4% IF, 14.2% DFF and 16.2% DM of phenotypic variance (PV). Remaining SSR markers (ASSR1486, ASSR206 and ASSR408) could not qualify false discovery rate (FDR) and Bonferroni criteria, hence declared as false positives. Additionally, we identified two highly significant SSR markers, ASSR8 and ASSR390 on LG 1 and LG 2, respectively. The SSR marker ASSR8 explained up to 22 and 11% PV for earliness traits and PB respectively, whereas ASSR390 controlled up to 17% PV for earliness traits. The validation and identification of new QTLs in pigeonpea across diverse genetic backgrounds brightens the prospects for marker-assisted selection to improve yield gains in pigeonpea.

Identification of cyclic lipopeptides produced by Bacillus vallismortis R2 and their antifungal activity against Alternaria alternata.

AIM: To identify the different antifungal biomolecules produced by isolate Bacillus vallismortis R2 and analyse their effect on Alternaria alternata, a common pathogen causing black point disease of wheat. METHODS AND RESULTS: The different antifungal molecules produced by isolate R2 were purified by column chromatography. The liquid chromatography-mass spectrometry of purified fractions confirmed the ability of R2 to produce biomolecules putatively similar to three different families of cyclic lipopeptides (CLPs) viz. surfactins, iturins and fengycins. The synergistic interaction among CLPs was evident from significant increase in the antifungal activity of mixture of purified fractions as compared to that of the individual fractions by agar well diffusion assay. The evaluation of antifungal activity of purified fractions by 96-well microtitre plate assay showed that the fengycin-like molecules supported significantly higher antifungal activity against A. alternata than iturin A and no antifungal activity was supported by surfactin. CONCLUSIONS: The isolate R2, producing different CLPs, can be used as an environmental friendly alternative to chemical fungicides. Among the three different CLPs viz. surfactin, iturin A and fengycin produced by R2, the fengycins were the most active lipopeptides. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report on co-production of three different types of CLPs by the cells of B. vallismortis.

Early Postmarket Results with EmboTrap II Stent Retriever for Mechanical Thrombectomy: A Multicenter Experience.

BACKGROUND AND PURPOSE: EmboTrap II is a novel stent retriever with a dual-layer design and distal mesh designed for acute ischemic stroke emergent large-vessel occlusions. We present the first postmarket prospective multicenter experience with the EmboTrap II stent retriever. MATERIALS AND METHODS: A prospective registry of patients treated with EmboTrap II at 7 centers following FDA approval was maintained with baseline patient characteristics, treatment details, and clinical/radiographic follow-up. RESULTS: Seventy patients were treated with EmboTrap II (mean age, 69.9 years; 48.6% women). Intravenous thrombolysis was given in 34.3%, and emergent large-vessel occlusions were located in the ICA (n = 18), M1 (n = 38), M2 or M3 (n = 13), and basilar artery (n = 1). The 5 × 33 mm device was used in 88% of cases. TICI ≥ 2b recanalization was achieved in 95.7% (82.3% in EmboTrap II-only cases), and first-pass efficacy was achieved in 35.7%. The NIHSS score improved from a preoperative average of 16.3 to 12.1 postprocedure and to 10.5 at discharge. An average of 2.5 [SD, 1.8] passes was recorded per treatment, including non-EmboTrap attempts. Definitive treatment was performed with an alternative device (aspiration or stent retriever) in 9 cases (12.9%). Some hemorrhagic conversion was noted in 22.9% of cases, of which 4.3% were symptomatic. There were no device-related complications. CONCLUSIONS: Initial postmarket results with the EmboTrap II stent retriever are favorable and comparable with those of other commercially available stent retrievers. Compared with EmboTrap II, the first-generation EmboTrap may have a higher first-pass efficacy; however, data are limited by retrospective case analysis, incomplete clinical follow-up, and small sample size, necessitating future trials.

Real world use of loteprednol etabonate ophthalmic gel 0.5% in cases representative of comorbid pathologies responding to minimally invasive glaucoma surgery.

PURPOSE: With the increasing use of minimally invasive surgical techniques for intraocular pressure (IOP) lowering in glaucoma patients, there is a need to examine best practices regarding the postoperative management of these patients. Corticosteroids, though effective in controlling postoperative ocular pain and inflammation, present distinct challenges in glaucoma surgery patients, as their use can be associated with IOP elevation. Loteprednol etabonate (LE) is an ocular corticosteroid designed to have an improved safety profile relative to other corticosteroids. METHODS: We report here a representative selection of cases in which patients were successfully treated with LE ophthalmic gel 0.5% (LE gel) following a variety of minimally invasive glaucoma surgery (MIGS) procedures. Cases included patients undergoing various procedures including a Trabectome combined with cataract surgery; micro-stent surgery (iStent) combined with cataract surgery; supraciliary CyPass Micro-Stent placement combined with cataract surgery; Kahook Dual Blade goniotomy; and ab interno canaloplasty using the iTrack catheter. OBSERVATIONS: In all cases, use of LE gel during the postoperative period appeared effective and safe in reducing inflammation and controlling pain. No adverse events or IOP elevations were noted, even in those patients continuing use of LE gel past the postoperative period for longer than six months with documented follow-up. In two cases, patients with elevated IOP using either prednisolone or difluprednate postoperatively were switched to LE gel, with a subsequent reduction in IOP. CONCLUSIONS: This selection of cases involving patients undergoing MIGS suggests that LE gel may be an effective and safe option for treating postoperative inflammation and pain following such procedures with minimal to no effect on IOP or other negative sequalae.

A host defense role for a natural antiviral substance in the nervous system.

The pathogenesis of virus infections of the nervous system (NS) is regulated by host defenses. The defensive role of a major constitutive antiviral substance was studied by determining its distribution in the human nervous system, its concentration and the ability of this viral inhibitor to protect mice against viral infection. The 4000 kDa inhibitor complex in the human nervous system was detected in brain gray and white matter, spinal cord, and sciatic nerve but not in human cerebrospinal fluid. The inhibitor was found in the extracellular medium incubated with minced murine brain. The inhibitory titer ranged from approximately 50 to 200 antiviral units per gram against polio 1, Semliki Forest, Banzi, mengo, Newcastle disease and herpes simplex 1 viruses. The inhibitor is composed of lipid and essential protein and carbohydrate moieties as determined by enzymatic inactivation. Protection of inhibitor-treated mice was demonstrated against both an alphavirus and a picornavirus. Thus a natural defensive role for the broadly antiviral inhibitor is suggested by its constitutively high concentration, wide distribution in nervous system tissues, presence in extracellular fluid and its ability to provide protection in infected mice.

Further characterization of a broad-spectrum antiviral substance in human serum.

A broadly active antiviral glycoprotein (UTI beta) occurs naturally in human sera at an average antiviral titer of 50 U/ml. This inhibitor is active against all virus families tested to date, including representative poxviruses, herpesviruses, enteroviruses, paramyxoviruses, alpha-viruses, flaviviruses, bunyaviruses, and rhabdoviruses. It is a glycoprotein of approximately 60,000 +/- 10,000 Da, which is stable at pH 2 to 10 and at 80 degrees C for up to 10 min. Mild oxidation with NaIO4 and treatment with glycosidases inactivates the material. Proteolytic degradation of the inhibitor molecule releases small active components of < 1000 Da, which retain antiviral activity. This activity of the small components has increased heat stability (120 degrees C for 15 min) and is inactivated by glycosidases. The antiviral activity thus appears to reside mainly in the oligosaccharide moiety of the glycoprotein. The inhibitor does not neutralize virions, but prevents attachment of most viruses to cells. These properties occur also in highly purified preparations. These findings indicate that human serum contains significant concentrations of a broadly active antiviral glycoprotein, which is distinct from interferon and other antiviral substances naturally found in human body fluids and tissues.

Innate gastrointestinal immunity: characterization of broadly active viral inhibitors.

Innate viral inhibitors that are broadly active have been characterized in the serum and the nervous system, but incompletely characterized in the gastrointestinal (GI) tract. GI preparations from porcine gastric mucosa, mouse intestine, and in neuramide (a pharmaceutical product), were examined for broad antiviral activity, molecular size and mechanism of action for comparison with the previously characterized, innate inhibitors in the serum and nervous system. The GI inhibitors were found to be active in high titers against RNA and DNA viruses, resistant to proteolysis, glycolysis, lipid extraction and possessed differing mechanisms of action. The mouse intestinal inhibitor prevented virus attachment to cells, and neuramide acted at an early post-attachment stage of virus multiplication. The porcine mucosal inhibitor acted as late as 6 h after initiation of the multiplication cycle. These broadly active GI inhibitors differed from the previously described serum inhibitor (UTI beta) high density lipoproteins (HDL) and the nervous system (NS) inhibitor by being smaller (600 +/- 400 kDa) and resistant to proteinase K, glycosidases and organic solvents. The mouse intestinal inhibitor acts similarly to UTI beta and NS inhibitor by preventing attachment of virus to the cells. In comparison, the neuramide and the porcine mucosal inhibitor, like HDL, acted after attachment to the target cells. The innate nonspecific, broadly-active virus inhibitors, based on high titers and location, are considered important initial immune defense mechanisms against viral infections and thus potentially useful in medical applications.

Vertebrate brains contain a broadly active antiviral substance.

Brain tissue extracts from vertebrates were examined for non-specific, broad-spectrum virus inhibitors, previously identified and characterized from other body tissues and fluids. An antiviral activity found in human, bovine, ovine, porcine, lapine, murine and piscine brain tissues shares some properties with a contact blocking-virus inhibitor, which was previously found only in cell culture supernatants. The inhibitor was active against (in order of sensitivity to inhibitor) Banzi, Sindbis, Bunyamwera, Newcastle disease, herpes simplex I, Semliki forest, polio I, mengo, vaccinia and vesicular stomatitis viruses. It is approximately 4000 kDa and possesses a complex structure containing protein, carbohydrate and lipid moieties. The inhibitor does not directly neutralize virus or induce an antiviral state in cells, but appears to act early in the replication cycle, most likely by preventing virus attachment to target cells. Its occurrence in concentrations sufficient to reduce virus yield in cell cultures at least 30-fold may indicate a role in limiting viral infections of the central nervous system.

Lipoproteins account for part of the broad non-specific antiviral activity of human serum.

Several antiviral substances have been detected in human serum but few have been shown to possess broad antiviral activity. These broadly active antiviral molecules could be of significance as innate defense mechanisms. We have previously identified and characterized a broadly antiviral glycoprotein, UTI3, which accounts for 50 antiviral units/ml of human and mammalian sera. In addition there are reports of antiviral activity of human serum apolipoprotein A-1 (apo A-1), an important constituent of high density lipoprotein (HDL), against human immunodeficiency virus (HIV) and herpesvirus. Therefore we investigated (1) whether HDL is broadly antiviral, (2) how much of the broad antiviral activity of serum is due to HDL, and (3) the mechanism(s) of HDL's antiviral action. In this paper we report that (1) HDL does have broad antiviral activity, (2) HDL accounts for a modest but significant portion of the antiviral activity of serum, and (3) HDL acts by preventing virus penetration. Overall, HDL may be one of the broadly antiviral defences in the bloodstream.

Broad antiviral activity in tissues of crustaceans.

Innate antiviral substances occur in vertebrates and may function as host defenses. Virus infections are common among invertebrates, but little is known about the ability of invertebrates to control viral infections. Pre-existing antiviral substances may be particularly important, since invertebrates lack the antiviral defense conferred by specific immunity. In our study, we found that tissue extracts of blue crab (Callinectes sapidus), shrimp (Penaeus setiferus), and crayfish (Procambarus clarkii) contained antiviral activities that inhibit a variety of DNA and RNA viruses, i.e. Sindbis virus (SB), vaccinia virus (VAC), vesicular stomatitis virus (VS), mengo virus (MENGO), banzi virus (BANZI) and poliomyelitis (POLIO). The concentration of inhibitory activity was relatively high, ranging from 102 to 216 U/g tissue for Sindbis virus, using the various tissue extracts. The other viruses were somewhat less sensitive to the inhibitor. The main antiviral activity in the inhibitor preparation from blue crab resided in an approximately 440 kDa fraction. It was inactivated significantly by lipid extraction, but not by proteinase K or glycosidases. The antiviral mechanism of the inhibitor from the blue crab was inhibition of virus attachment to eukaryotic cells, as evidenced by inhibitory activity at 4 degrees C. These studies are among the first to show the existence of broadly active antiviral activities in aquatic crustaceans. These antiviral substances may function as innate host defenses in these species that lack specific antibody immunity and, therefore, merit further study.

Treatment of intracranial alphavirus infections in mice by a combination of specific antibodies and an interferon inducer.

Finding an effective treatment for viral infections that cause encephalitis remains an important problem. A model of human alphavirus infections, Semliki Forest virus, causes lethal encephalitis in weanling mice. Mice are viremic within 24 hr of an intraperitoneal challenge with the equivalent of three 75% lethal doses of Semliki Forest virus. Virus reaches the brain by 48 hr, and mortality results in all mice in 5-7 days. Introduction of virus intracranially accelerates the course of the infection. Neither anti-Semliki Forest virus hyperimmune serum nor the potent interferon inducer poly I:CLC given intraperitoneally are protective when used therapeutically after an intracranial virus infection, but a combination of 1,000 U hyperimmune serum and 80 micrograms/mouse of poly I:CLC results in a 50% survival rate. This combination treatment of intracranial Semliki Forest virus infection eliminates detectable viremia and reduces virus load in the brain over the course of the infection. These data show that when combined, specific antibody and an interferon inducer can interact synergistically to protect mice from alphavirus infections of the central nervous system even when given after the virus is replicating in the target organ.

Generalized occurrence of the broadly antiviral substance UTI beta in mammalian sera.

This study compares the characteristics of naturally occurring antiviral activities in nonhuman mammalian sera to UTI beta (University of Texas virus inhibitor beta), an innate, non-specific viral inhibitor found in human serum. The antiviral agent in sera from four different species appears to possess properties similar to those of UTI beta, e.g. molecular weight of 60 +/- 10 kDa, broad spectrum antiviral activity, glycoprotein structure with antiviral oligosaccharide moiety(s) attached to a carrier protein, and inhibition of most viruses by preventing their attachment to target cells. These findings suggest that UTI beta or UTI beta-like virus inhibitors are a normal component of the non-specific immune defenses of mammals.

Late audiovestibular consequences of radical radiotherapy to the parotid.

It has been claimed that the inner ear is generally resistant to the effects of radiotherapy schedules commonly used for treating head and neck cancer. A study was made of 28 patients previously irradiated for parotid pleomorphic adenoma to assess the effect on the audiovestibular apparatus. The dose most commonly used was 50 Gy in 15 daily fractions over 20 days and the median interval between radiotherapy and assessment was 14 years. Of 28 patients 15 had significant hearing deficit on the irradiated side on audiometry and this was predominantly sensorineural in type. In addition seven of these 15 patients had semicircular canal paresis. The reason for this morbidity may be arithmetic (total biological dose; dose per fraction) or geometric (upper extent; depth of target volume). Patients are now routinely warned of the possibility of late audiovestibular effects following radical radiotherapy to the parotid.

Differentiation of avian infectious bronchitis virus isolates by thermal sensitivity.

Six isolates of avian infectious bronchitis virus (IBV) at different passage levels in the chicken embryo were tested for sensitivity at 56 C, with the following results: 1) isolates differed in thermal sensitivity; 2) virus was inactivated in a first-order exponential-kinetics 2-component fashion indicative of a heterogeneous population with a preponderance, 98% or more, of thermal-sensitive (S) virions over thermal-resistant (R) virons; 3) R virions 2-component populations were inactivated; 4) recovered R virions could be maintained in continuous passage in the chicken embryo as a one-component population by the limiting-dilution technique, although there was a progressive parallel increase in thermal sensitivity associated directly with the continuous passage; 5) growth curves of isolated one-component populations of R virions were quantitatively and in time sequence similar to curves of 2-component populations of continuous-passage virus.

Nasopharyngeal Corynebacterium ulcerans: a different diphtheria.

A case of toxigenic Corynebacterium ulcerans infection is presented. The diagnosis was delayed and no anti-toxin administered. A nasopharyngeal biopsy was complicated by severe haemorrhage necessitating a post nasal pack. A brief review of the pathology and treatment of Corynebacterium ulcerans is given.

Experimental pathogenesis of buffalo pox virus in rabbits: clinico-pathological studies.

Buffalo pox virus produced typical pock lesions on the skin of rabbits at the site of primary inoculation following an incubation period of 48-72 hr. Gross lesions in internal organs, characterized by focal or diffuse necrotic areas on lung, liver and spleen were seen from day 5 post-inoculation (p. i.). Isolated lesions of approximately 2 mm diameter appeared in skin, stomach, intestine and uterus from day 7 p. i. Histopathological changes, i. e. intra-alveolar and intra-bronchial haemorrhages were seen in lungs and severe fatty changes were found in the liver. Multinuclear cells were detected in liver during recovery. Virus particles were demonstrated by electron microscopy in skin, lung, liver and spleen lesions.

Tissue changes induced by enterotoxic preparations of Salmonella weltevreden in ligated rabbit gut segments.

Culture filtrates and cell lysates of two strains of S. weltevreden which caused dilation of ligated rabbit gut segments (characteristic associated with the enterotoxic activity) induced mild to severe architectural changes in the test segments of intestine. The dilated segments contained thick, bloody and mucoid exudates. The results suggested that besides invasiveness and enterotoxigenicity, S. weltevreden possibly possessed factor (s) that damaged intestinal tissue and played part in the pathogenesis of Salmonella gastroenteritis.

Falciparum malaria--present day problems. An experience with 425 cases.

Clinical details and present day problems encountered in 425 cases of falciparum malaria (PF) are reported. 10.11% had taken chloroquine prior to reporting to us. Parasitic count done in 23.05% cases lacked correlation with severity of disease. Pattern of fever varied markedly but 5.4% were afebrile throughout and presented only with bodyache and malaise. Apyrexial spell was noted in 5.64%. 28.70% had typical facial looks of anaemia and sallow complexion. Cerebral symptoms were noted in 3.05%. Other symptoms were severe headache 33.4%, pain abdomen 3.29%, gastroenteritis 5.64%, jaundice 2.58% and bronchitis in 7.50%. We encountered subconjunctival haemorrhages with purpura and/or urticaria in four cases, symptoms suggestive of shock lung in 3, pulmonary oedema in 2, severe anaemia (HB less than 4 g%) in seven pregnant ladies, extrapyramidal symptoms in follow up period in 5 and congenital malaria in 2 cases. 83.25% were cured with chloroquine and oxytetracycline. 8.47% (who deteriorated despite the above treatment) were treated with quinine for 6 days. 5.17% (with severe disease) were also given quinine as first line drug. 2.82% (unresponsive to chloroquine and oxytetracycline but with mild disease) were treated with pyrimethamine-sulphamezathine combination for 5 days. One case who did not respond to quinine was treated with quinidine. Recrudescence was seen in 3.67% of patients treated with chloroquine and oxytetracycline. There was no case with renal failure, haemolysis due to G6PD deficiency and black water fever. There was only one death (0.23%) in our series. Self-medication, haphazard therapy and the slogan "Fever may be malaria-take chloroquine" can lead to problems in falciparum malaria.

Cardiovascular system in leprosy.

Involvement of cardiovascular system (CVS) in 50 multibacillary (MB) and 20 paucibacillary (PB) cases of leprosy was evaluated. 20 age and sex matched controls were also studied. In addition to detailed clinical examination and resting electro-cardiogram, Master's two step exercise test (DMT) was also carried out to find out the occult and asymptomatic cardiac involvement. We have not found any significant symptomatic or electrocardiographic evidence of CVS involvement in various groups of leprosy.

Experience with multidrug therapy in paucibacillary leprosy.

Eighty paucibacillary leprosy cases were randomly put on two different multidrug regimens for 6 months followed by dapsone monotherapy. Regimen I was according to WHO (1982) recommendations consisting of Dapsone and six once a month rifampicin. In regimen II in addition to above two constituents, clofazimine was added 100 mg on alternate days. Dapsone thereafter was continued in both the regimens upto one year. The efficacy, acceptability and side effects of multidrug regimens were observed for a period of one year. Histopathological assessment was done on completion of multidrug therapy in all cases. A comparative evaluation of effect of two multidrug regimens in paucibacillary leprosy patients is reported. Addition of clofazimine over WHO (1982) recommended regimen appears to have no added benefit. The duration of WHO (1982) recommended regimens was found to be inadequate in many cases.