Comparison of Soft Tissue Chin Thickness in Adult Patients With Various Mandibular Divergence Patterns.

Objective This study aimed to compare the soft tissue chin (STC) thickness at different levels in patients presenting for orthodontic treatment with different vertical facial types. Materials and methods This comparative cross-sectional study was conducted at Sharif Medical & Dental College, Lahore, Pakistan, on 195 subjects. Patients presenting for orthodontic treatment, both genders, aged from 18 to 32 years, and Pakistani nationals were included. Patients with any craniofacial deformity, syndrome, cleft lip and palate, previous orthodontic or orthognathic treatment, and multiple missing teeth and prostheses in edentulous areas were excluded. Vertical facial patterns and STC thickness were recorded from pre-treatment lateral cephalograms. One-way analysis of variance (ANOVA) was applied to compare STC among various vertical facial patterns. Post-hoc analysis was done using the Tukey test. Results There were 126 females (64.62%) and 69 males (35.38%). The mean age was 21.66 ± 3.44 years. All three soft tissue chin thickness distances significantly differed among vertical facial patterns (p<0.001). Multiple comparisons show that the distance between soft and hard tissue pogonion was insignificant between low and normal angle facial heights (p=0.5). Similarly, no significant difference was observed for the distance between soft and hard tissue menton in low and normal angle subjects (p=0.4). The rest of the multiple comparisons were statistically significant (p<0.05). Conclusion The STC thickness is significantly associated with vertical facial divergence. While planning orthognathic surgery or genioplasty of the mandible, due consideration should be given to vertical divergence of the face to avoid unwanted facial changes.

Correction: Abbas et al. Bioactive Compounds, Antioxidant, Anti-Inflammatory, Anti-Cancer, and Toxicity Assessment of Tribulus terrestris-In Vitro and In Vivo Studies. Antioxidants 2022, 11, 1160.

In the original publication [...].

Effect of hesperidin on growth, photosynthesis, antioxidant systems and uptake of cadmium, copper, chromium and zinc by Celosia argentea plants.

Rapid industrialization and extensive agricultural practices are the major causes of soil heavy metal contamination, which needs urgent attention to safeguard the soils from contamination. However, the phytotoxic effects of excessive metals in plants are the primary obstacle to efficient phytoextraction. The present study evaluated the effects of hesperidin (HSP) on metals (Cu, Cd, Cr, Zn) phytoextraction by hyperaccumulator (Celosia argentea L.) plants. For this purpose, HSP, a flavonoid compound with strong antioxidant potential to assist metal phytoextraction was used under metal stress in plants. Celosia argentea plants suffered significant (P ≤ 0.001) oxidative damage due to the colossal accumulation of metals (Cu, Cd, Cr, Zn). However, HSP supplementation notably (P ≤ 0.001) abated ROS generation (O2•‒, •OH, H2O2), lipoxygenase activity, methylglyoxal production, and relative membrane permeability that clearly indicated HSP-mediated decline in oxidative injury in plants. Exogenous HSP improved (P ≤ 0.001) the production of non-protein thiol, phytochelatins, osmolytes, and antioxidant compounds. Further, HSP enhanced (P ≤ 0.001) H2S and NO endogenous production, which might have improved the GSH: GSSG ratio. Consequently, HSP-treated C. argentea plants had higher biomass alongside elevated metal accumulation mirrored as profound modifications in translocation factor (TF), bioaccumulation coefficient (BAC), and bioconcentration factor (BCF). In this context, HSP significantly enhanced TF of Cr (P ≤ 0.001), Cd (P ≤ 0.001), and Zn (P ≤ 0.01), while BAC of Cr (P ≤ 0.001), Cd (P ≤ 0.001), and Zn (P ≤ 0.001). Further, BCF was significant (P ≤ 0.05) only in plants grown under Cr-spiked soil. Overall, HSP has the potential for phytoremediation of metals by C. argentea, which might be a suitable strategy for metal-polluted soils.

On analysis of iron (II) chloride via graph entropy measures and statistical models.

The crystalline material that is greenish-white and dissolves in water is iron chloride. It is utilized in sewage treatment, dyeing, and medicine. Graph entropy plays a significant role in measuring the complexity of atoms, molecules, and structures in nature. It has specific chemical applications in biology, neuroscience, and chemistry. A compound's molecular structure consists of many atoms. Particularly, hydrocarbons are a chemical combination of hydrogen and carbon atoms. In this article, we discuss the entropy of the chemical structure Iron (II) Chloride. Additionally, we discuss the idea of degree-based indices and compute the Shannon entropy(ENT) using these indices. The linear regression(LR) of various indices and entropies for iron chloride, FeCl2, is also discussed. Also, we link the degree-based indices and entropies via line fit.

Anti-microbial host factor HDAC6 is antagonised by the influenza A virus through host caspases and viral PA.

Histone deacetylase 6 (HDAC6), through the repertoire of its substrate proteins, plays a critical role in human physiology, and an aberrant function of HDAC6 contributes to various pathophysiological conditions. HDAC6 is also known to be an anti-microbial host factor and has been implicated in restricting or clearing the infection of various human viral and bacterial pathogens. However, the state and the mechanisms of its antagonism in infected cells are not understood. Here, we demonstrate that influenza A virus (IAV) antagonises HDAC6 by recruiting both viral and host components. We found that HDAC6 mRNA expression, and consequently, the HDAC6 polypeptide expression is downregulated in human lung epithelial cells during early stage of IAV infection but can be rescued by depleting the expression of viral polymerase acidic (PA) protein, a subunit of IAV RNA polymerase. In addition, during later stage of the infection, the HDAC6 polypeptide undergoes caspase-mediated cleavage at two sites, generating two cleaved fragments. Both these fragments disappeared when the expression of caspase 3 was depleted in infected cells, whereas only second fragment disappeared when the expression of caspase 6 was depleted. But both fragments disappeared and the level of full-length HDAC6 polypeptide was rescued when the expression of PA was depleted in infected cells. Collectively, these data indicated that IAV antagonises the HDAC6 by decreasing its expression level in infected cells, both at mRNA and polypeptide level via PA gene, which has been implicated in auxiliary functions like degradation of host mRNA and induction of apoptosis.

Bioactive Compounds, Antioxidant, Anti-Inflammatory, Anti-Cancer, and Toxicity Assessment of Tribulus terrestris-In Vitro and In Vivo Studies.

Tribulus terrestris L. belongs to the family Zygophyllaceae and integral part of various ancient medicinal systems including Chinese, Indian, and European to combat various health ailments. The aim of the present study was to assess the phytochemical constituents, in vitro antioxidant activity using DPPH, FRAP, and H2O2 assays, in vitro anticancer activity using MTT assay, and in vitro and in vivo anti-inflammatory properties of T. terrestris. The acute and sub-acute toxicity of extracts exhibiting most biological potential was examined using murine models. Liquid-liquid partitioning followed by RP-HPLC sub-fraction of crude extract was performed. After that, ESI-MS/MS analysis was done for the timid identification of bioactive metabolites responsible for bioactivities of sub-fractions and HPLC analysis to quantify the compounds using external standards. Among all extracts, T. terrestris methanol extract was noted to hold maximum phenolic (341.3 mg GAE/g) and flavonoid (209 mg QE/g) contents, antioxidant activity in DPPH (IC50 71.4 µg/mL), FRAP (35.3 mmol/g), and H2O2 (65.3% inhibition) assays, anti-inflammatory activities in vitro at 400 µg/mL (heat-induced hemolysis, % inhibition 68.5; egg albumin denaturation, % inhibition 75.6%; serum albumin denaturation, % inhibition 80.2), and in vivo at 200 mg/kg (carrageenan-induced paw edema, % inhibition 69.3%; formaldehyde-induced paw edema, % inhibition 71.3%) and anticancer activity against breast cancer cell (MCF-7) proliferation (IC50 74.1 µg/mL). Acute and sub-acute toxicity studies recorded with no change in body weight, behavior, hematological, serum, and histopathological parameters in treated rats with T. terrestris methanol extracts when compared to control group. Fraction B obtained through liquid-liquid partitioning resulted in more bioactive potential as compared to the parent methanol extract. RP-HPLC analysis of fraction B resulted with four sub-fractions (TBTMF1-TBTMF4), wherein TBTMF3 delineated notable bioactive capabilities as compared to other fractions and parent methanol extract. ESI-MS/MS analysis of TBTMF3 resulted with tentative identification of myricetin, rutin, liquitrigenin, physcion, and protodioscin. It can be stated that T. terrestris is a potential bearing herb and findings of current study further verify the claims made in ancient medicinal systems. However, after investigation of each identified compound, it must be considered for drug discovery.

Dengue virus-free defective interfering particles have potent and broad anti-dengue virus activity.

Dengue virus (DENV) is spread from human to human through the bite of the female Aedes aegypti mosquito and leads to about 100 million clinical infections yearly. Treatment options and vaccine availability for DENV are limited. Defective interfering particles (DIPs) are considered a promising antiviral approach but infectious virus contamination has limited their development. Here, a DENV-derived DIP production cell line was developed that continuously produced DENV-free DIPs. The DIPs contained and could deliver to cells a DENV serotype 2 subgenomic defective-interfering RNA, which was originally discovered in DENV infected patients. The DIPs released into cell culture supernatant were purified and could potently inhibit replication of all DENV serotypes in cells. Antiviral therapeutics are limited for many viral infection. The DIP system described could be re-purposed to make antiviral DIPs for many other RNA viruses such as SARS-CoV-2, yellow fever, West Nile and Zika viruses.

Hepatoprotective Effect Of Prazosin Is Comparable To N-Acetylcysteine In Acetaminophen Induced Hepatotoxicity In Mice.

BACKGROUND: Autonomic nervous system modulates acetaminophen induced hepatotoxicity. The purpose of the study was to determine the hepatoprotective effect of α1 antagonist (prazosin) and ß2 agonist (salbutamol) on acetaminophen induced hepatotoxicity in mice. METHODS: This experimental study was conducted at Post Graduate Medical Institute, Lahore in which 50 adult mice were divided in to five groups. With the exception of normal control, hepatotoxicity was induced in all other study groups by giving single intraperitoneal injection of acetaminophen 300 mg/ kg. First and second groups served as normal and toxic control were given distilled water 6 ml/ kg while third, fourth and fifth experimental groups were given N-acetylcysteine (300 mg/ kg), prazosin (0.18 mg/ kg) and salbutamol (0.35 mg/kg) intraperitoneally at 2,4 and 8 hours after acetaminophen injection. Serum liver enzymes were analysed at 0 and 72 hours while histopathological finding were assessed at the end of study by using SPSS-20. RESULTS: All the groups treated with toxic dose of acetaminophen showed significant increase in serum ALT, i.e., B (Toxic control 3372%), C (NAC treated 282%), D (Prazosin treated 582%), E(Salbutamol treated 3297%) and AST levels, i.e., B (Toxic control 2750% ), C (NAC treated 230% ), D (Prazosin treated 280%), E (Salbutamol treated 828%) with p-value ˂0.001. When this increase was compared between groups, the lowest increase in serum ALT and AST levels was observed in Nacetylcysteine and prazosin group with no significant difference. Similarly, experimental animals receiving prazosin and N-acetylcysteine had the lowest inflammation, degeneration and necrosis scores than the toxic control group in histopathological analysis of the liver with p-value <0.001. CONCLUSIONS: The hepatoprotective effect of prazosin is comparable to N- acetylcysteine against acetaminophen induced hepatotoxicity in mice.

RNA activation in insects: The targeted activation of endogenous and exogenous genes.

RNA activation (RNAa) is a newly emerging area of research in which dsRNA targeting promoter regions can induce the expression of the target gene. Although still in its infancy, it is already having significant impacts in several research areas in particular as cancer therapeutics. So far, the scope of RNAa has been limited to mammals and Caenorhabditis elegans with no indication of its prevalence in insects. In this study, we aimed to demonstrate the presence of RNAa in the insect dengue vector Aedes aegypti. Furthermore, we looked to uncover some details surrounding the involvement of host factors in order to present this as a new technique for insect research. The outcomes of this study provide new opportunities to further research into arthropod-borne diseases and insect biology in the same way as RNA interference.

The Host GTPase Arf1 and Its Effectors AP1 and PICK1 Stimulate Actin Polymerization and Exocytosis To Promote Entry of Listeria monocytogenes.

Listeria monocytogenes is a foodborne bacterium that causes gastroenteritis, meningitis, or abortion. Listeria induces its internalization (entry) into some human cells through interaction of the bacterial surface protein InlB with its host receptor, the Met tyrosine kinase. InlB and Met promote entry through stimulation of localized actin polymerization and exocytosis. How actin cytoskeletal changes and exocytosis are controlled during entry is not well understood. Here, we demonstrate important roles for the host GTPase Arf1 and its effectors AP1 and PICK1 in actin polymerization and exocytosis during InlB-dependent uptake. Depletion of Arf1 by RNA interference (RNAi) or inhibition of Arf1 activity using a dominant-negative allele impaired InlB-dependent internalization, indicating an important role for Arf1 in this process. InlB stimulated an increase in the GTP-bound form of Arf1, demonstrating that this bacterial protein activates Arf1. RNAi and immunolocalization studies indicated that Arf1 controls exocytosis and actin polymerization during entry by recruiting the effectors AP1 and PICK1 to the plasma membrane. In turn, AP1 and PICK1 promoted plasma membrane translocation of both Filamin A (FlnA) and Exo70, two host proteins previously found to mediate exocytosis during InlB-dependent internalization (M. Bhalla, H. Van Ngo, G. C. Gyanwali, and K. Ireton, Infect Immun 87:e00689-18, 2018, https://doi.org/10.1128/IAI.00689-18). PICK1 mediated recruitment of Exo70 but not FlnA. Collectively, these results indicate that Arf1, AP1, and PICK1 stimulate exocytosis by redistributing FlnA and Exo70 to the plasma membrane. We propose that Arf1, AP1, and PICK1 are key coordinators of actin polymerization and exocytosis during infection of host cells by Listeria.

Endoplasmic Reticulum Stress Induces Macrophages to Produce IL-1ß During Mycobacterium bovis Infection via a Positive Feedback Loop Between Mitochondrial Damage and Inflammasome Activation.

Mycobacterium bovis, the causative agent of tuberculosis in cattle and humans, infects host macrophages and induces endoplasmic reticulum stress (ERS), mitochondrial damage, and interleukin (IL)-1ß production. The relationship between these phenotypes is yet to be elucidated. In this study, we investigated the role of ERS in mitochondrial damage and IL-1ß production in macrophages during infection with a virulent M. bovis strain. We found that ERS activates the inflammasome via NOD-like receptor family, pyrin domain-containing 3 (NLRP3)-caspase-8 and that IFN-inducible protein absent in melanoma 2 (AIM2) triggered mitochondrial damage. ERS increased reactive oxygen species (ROS), which promoted translocation of the inflammasome to the mitochondria. NLRP3, but not AIM2, was involved in the ERS-induced cleavage of caspase-8 and Bid, leading to mitochondrial damage, which was required for the production of mature IL-1ß. Our data suggest that ERS induces macrophages to produce mature IL-1ß during infection with virulent M. bovis through a positive feedback loop between mitochondrial damage and inflammasome activation. To the best of our knowledge, this is the first evidence of the involvement of ERS and mitochondrial damage in inflammasome activation during M. bovis infection.

Pure Tethered Cervical Cord and Review of Literature.

Tethering of the spinal cord in the lumbosacral region with myelomeningocele is a well-known phenomenon. Only sporadic cases of tethering along the rest of the neuraxis, including the hindbrain, cervical, and thoracic spinal cord have been documented, always along with some associated congenital malformations (hydrocephalus, Chiari malformation, myelomeningocele, meningocele, hamartomatous stalk, spina bifida occulta, intramedullary lipoma, intradural fibrous adhesions, the fusion of the sixth and seventh cervical vertebrae, split cord malformation, or low-lying cord). In this report, 14-year-old male developed symptoms related to tethering of the cervical spinal cord, but without any associated congenital malformations, that is the pure tethered cervical cord. This causes his moribund status and makes the manuscript unique and contributes to the hitherto literature. The authors discuss the diagnosis, treatment, and postoperative course of this entity. The uniqueness in treatment is that we have operated the case without the help of intraoperative somatosensory evoked potentials and motor evoked potential from posterolateral approach under local anesthesia.

Thirty Years Cancer Incidence Data for Lahore, Pakistan: Trends and Patterns 1984-2014

This research was conducted to generate trends and patterns of most common male and female cancers from 1984-2014 for the city population of Lahore Pakistan. Cancer incidence data gathered for different organs were processed through cleaning, integration, transformation, reduction and mining for ultimate representation. Risk of cancer appeared to be continuously increasing among both males and females. Overall, lymphomas and breast cancer are the most common neoplasm in males and females, respectively, in Lahore with almost the highest rates in the Asian Pacific region. The incidence of head and neck, brain, and lung cancers, as well as leukemia have rapidly increased among males, whereas, ovarian, cervix, head and neck and lymphomas have become more common among females. The present communication should be helpful for adequate strategic planning, identification of risk factors and taking appropriate prevention and control measures at the national level.

Experimental investigations on a diesel engine operated with fuel blends derived from a mixture of Pakistani waste tyre oil and waste soybean oil biodiesel.

The waste tyre and waste cooking oils have a great potential to be used as alternative fuels for diesel engines. The aim of this study was to convert light fractions of pyrolysis oil derived from Pakistani waste vehicle tyres and waste soybean oil methyl esters into valuable fuel and to reduce waste disposal-associated environmental problems. In this study, the waste tyre pyrolysis liquid (light fraction) was collected from commercial tyre pyrolysis plant and biodiesel was prepared from waste soybean oil. The fuel blends (FMWO10, FMWO20, FMWO30, FMWO40 and FMWO50) were prepared from a 30:70 mixture of waste tyre pyrolysis liquid and waste soybean oil methyl esters with different proportions of mineral diesel. The mixture was named as the fuel mixture of waste oils (FMWO). FT-IR analysis of the fuel mixture was carried out using ALPHA FT-IR spectrometer. Experimental investigations on a diesel engine were carried out with various FMWO blends. It was observed that the engine fuel consumption was marginally increased and brake thermal efficiency was marginally decreased with FMWO fuel blends. FMWO10 has shown lowest NOx emissions among all the fuel blends tested. In addition, HC, CO and smoke emissions were noticeably decreased by 3.1-15.6%, 16.5-33.2%, and 1.8-4.5%, respectively, in comparison to diesel fuel, thereby qualifying the blends to be used as alternative fuel for diesel engines.

Intraoperative Magnetic Resonance Imaging for Cranial and Spinal Cases Using Preexisting "C" Shaped Three Side Open 0.2 Tesla Magnetic Resonance Imaging.

BACKGROUND: The existing Intraoperative MRI (IMRI) of developed countries is too costly to be affordable in any developing country and out of the reach of common and poor people of developing country at remote areas. We have used the pre-existing (refurbished) 3 side open "C" shaped 0.2 Tesla MRI for IMRI in a very remote area. MATERIALS AND METHODS: In this technique the 0.2 Tesla MRI and the operating theatre were merged. MRI table was used as an operation table. We have operated 36 cases via IMRI from November 2005 to till date. First case operated was on 13th nov 2005. RESULTS: Low (0.2) Tesla open setup costs very low (around Rs 40 lakhs) so highly affordable to management and thus to patients, used for diagnostic and therapeutic purposes both, the equipments like Nitrous, oxygen and suction is outside the MRI room so no noise inside operative room, positioning the patient didn't take much time due to manual adjustments, no special training to nurses and technicians required because of low (0.2) Tesla power of magnet and same instruments and techniques, sequencing took only 1.31 mints per sequence and re registration is not required since we always note down the two orthogonal axis in x and y axis in preoperative imaging and we were able to operate on posterior fossa tumors as well because of no head fixation except with leucoplast strap. Moreover the images we got intraoperative are highly acceptable. CONCLUSION: Three side open 0.2 Tesla MRI system, if used for intraoperative guidance, is highly affordable and overcomes the limitations of western setup of IMRI. Postoperative MRI images were highly acceptable and also highly affordable too.

Histone Deacetylase 2 Is a Component of Influenza A Virus-Induced Host Antiviral Response.

Host cells produce variety of antiviral factors that create an antiviral state and target various stages of influenza A virus (IAV) life cycle to inhibit infection. However, IAV has evolved various strategies to antagonize those antiviral factors. Recently, we reported that a member of class I host histone deacetylases (HDACs), HDAC1 possesses an anti-IAV function. Herein, we provide evidence that HDAC2, another class I member and closely related to HDAC1 in structure and function, also possesses anti-IAV properties. In turn, IAV, like HDAC1, dysregulates HDAC2, mainly at the polypeptide level through proteasomal degradation to potentially minimize its antiviral effect. We found that IAV downregulated the HDAC2 polypeptide level in A549 cells in an H1N1 strain-independent manner by up to 47%, which was recovered to almost 100% level in the presence of proteasome-inhibitor MG132. A further knockdown in HDAC2 expression by up to 90% via RNA interference augmented the growth kinetics of IAV in A549 cells by more than four-fold after 24 h of infection. Furthermore, the knockdown of HDAC2 expression decreased the IAV-induced phosphorylation of the transcription factor, Signal Transducer and Activator of Transcription I (STAT1) and the expression of interferon-stimulated gene, viperin in infected cells by 41 and 53%, respectively. The role of HDAC2 in viperin expression was analogous to that of HDAC1, but it was not in the phosphorylation of STAT1. This indicated that, like HDAC1, HDAC2 is a component of IAV-induced host innate antiviral response and performs both redundant and non-redundant functions vis-a-vis HDAC1; however, IAV dysregulates them both in a redundant manner.

Development of Global Consensus of Dengue Virus Envelope Glycoprotein for Epitopes Based Vaccine Design.

Dengue virus (DENV) is the member of Flaviviridae and causative agent of Dengue Haemorrhagic Fever and Dengue Shock Syndrome. Every year, around 70% of the world population is at risk, due to epidemic episodes orchestrated by one or more of its serotypes. So, a tetravalent DENV vaccine is needed which may induce the immune response against all four DENV serotypes. In this study, B-cell and T-cell epitopes have been predicted from the DENV envelope glycoprotein (Eg) using a consensus based approach in complement with the physico-chemical property (PCP) conservancy analysis. Through DENV-Eg analysis, a total of 7 PCP conserved, water soluble, in vitro and in vivo stable epitopes were predicted which may induce the B-cell and T-cell mediated anti-viral immune response.

De Novo modeling of Envelope 2 protein of HCV isolated from Pakistani patient and epitopes prediction for vaccine development.

Hepatitis C virus (HCV) is a universal health issue and a significant risk factor leading to hepatocellular carcinoma. HCV has infected approximately 170 million individuals worldwide. It is a member of Flaviviridae with positive sense RNA genome. In the absence of any effective vaccine against HCV, pegylated interferon with ribavirin is the standard of treatment against HCV infection. In this study, sequence and structural analysis of envelope 2 (E2) protein was performed which was isolated from patients of HCV genotype 3a in Pakistan. Then, epitopes were predicted which were specific for both B-cells and T-cells. Later, conservancy of epitopes was checked with the HCV 3a and 1a sequences from different countries. A total of 6 conserved epitopes were found from extra-membranous regions of E2 protein. Presence of conserved epitopes in E2 protein generates the possibility that these epitopes can be used to elicit the immune response against HCV.

Cloning and characterization of a Dim1-like mitosis gene of Spodoptera frugiperda cells (Sf9) induced by Autographa californica multiple nucleopolyhedrovirus.

Dim1 proteins are evolutionarily highly conserved throughout the eukaryotes and are present in numerous species. These proteins are essential for mitosis and pre-mRNA splicing. In this study, the full-length cDNA of Dim1-like gene from Spodoptera frugiperda cells (Sf9) was obtained. S. frugiperda Dim1 (SfDim1) cDNA is comprised of 975 bp including a 429 bp open reading frame (ORF), 225 bp 5' untranslated region (5' UTR), and 321 bp 3' untranslated region (3' UTR) with a poly A tail. The predicted polypeptide encoded by this gene is 142 aa with a molecular weight of 16.76 kDa and a PI of 5.53. Sequence alignment and phylogenetic analysis showed high similarities with Dim1 of other species. The evolutionary conserved site of Dim1 proteins ((35)Asp-Pro-Thr-Cys(38)) is also present in SfDim1. Silencing of SfDim1 gene decreased cell proliferation at 72 h post-treatment in comparison to mock and control transfected cells. Using RT-PCR, we found relatively higher SfDim1 transcript levels following Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) infection compared to mock-infected cells from 4h post-infection (hpi) up until 24 hpi. The expression level diminished dramatically at 36 hpi up to 120 hpi with no expression detected at 144 hpi. Silencing of SfDim1 resulted in lower levels of virus DNA production in comparison to mock-infected cells, which suggested that SfDim1 might benefit the virus and facilitate viral replication. Overall, the results showed that SfDim1 protein is involved in cell proliferation as well as cell-virus interaction.

Selection of target sequences as well as sequence identity determine the outcome of RNAi approach for resistance against cotton leaf curl geminivirus complex.

Cotton leaf curl disease is caused by a geminivirus complex that involves multiple distinct begomoviruses and a disease-specific DNA satellite, cotton leaf curl Multan betasatellite (CLCuMB), which is essential to induce disease symptoms. Here we have investigated the use of RNA interference (RNAi) for obtaining resistance against one of the viruses, Cotton leaf curl Multan virus (CLCuMV), associated with the disease. Three hairpin RNAi constructs were produced containing either complementary-sense genes essential for replication/pathogenicity or non-coding regulatory sequences of CLCuMV. In transient assays all three RNAi constructs significantly reduced the replication of the virus in inoculated tissues. However, only one of the constructs, that targeting the overlapping genes involved in virus replication and pathogenicity (the replication-associated protein (Rep), the transcriptional activator protein and the replication enhancer protein) was able to prevent systemic movement of the virus, although the other constructs significantly reduced the levels of virus in systemic tissues. In the presence of CLCuMB, however, a small number of plants co-inoculated with even the most efficient RNAi construct developed symptoms of virus infection, suggesting that the betasatellite may compromise resistance. Further analyses, using Rep gene sequences of distinct begomoviruses expressed from a PVX vector as the target, are consistent with the idea that the success of the RNAi approach depends on sequence identity to the target virus. The results show that selection of both the target sequence, as well as the levels of identity between the construct and target sequence, determine the outcome of RNAi-based resistance against geminivirus complexes.