Prolonged Survival of the Patient with Head and Neck Cancer with Squamous Cell Carcinoma treated with Cetuximab: A Case Report

Introduction: Cetuximab, an epidermal growth factor (EGFR) inhibitor has radio sensitizing activity for locoregionally, advanced head and neck cancer (LAHNC). The drug is either used in combination with radiotherapy or in combination with other chemotherapeutic drugs.Case Presentation: A 35-year-old male patient diagnosed with cancer of tongue atloco regional stage who had previously undergone hemiglossectomy was treated with TPF (docetaxel, cisplatin and 5-fluorouracil) as induction therapy(ICT)followed by concurrent local radiation therapy (RT) and cetuximab for 6weeks.The first recurrence appeared 4 years after the surgery, which was treated with RT along with standard treatment with cetuximab for 6weeks.Second recurrence was treated with cetuximab in combination with cisplatin and docetaxel. The patient survived for 8 years since he began his treatment.Conclusion: Cetuximab in combination with RT and other chemotherapeutic drugs can offer a longer survival in patients with both recurrent, locally advanced and metastatic squamous cell carcinoma of head and neck cancer

Profile of Indian adults consulting nurses/midwives for healthcare

The article aims to explore and characterize Indian adults who seek nursing consultations. Nursing professionals constitute two-fifth of the Indian healthcare workforce, but their consultation with patients is limited. Hence investigating the profile of Indian adults who consult nurses is necessary. The study used Secondary data of 72,250 nationally representative adults from the first wave of the Longitudinal Ageing Study of India, which was analyzed for nursing consultation sought by the participants during the 12 months preceding the survey using appropriate statistical methods and weights. Out of 257.1 million adult’s ≥45 years old and their spouses, 69.3% had consulted one or more healthcare workers during the last 12 months preceding the survey, and only 1.30% of adults had consulted nurses/midwives. Those, who consulted nurses/midwives, 16.7%, 26.2%, and 13.3% had consulted at community health facilities such as sub-centers, primary-health-centers, and community-health-centres respectively. Rural residency, female gender, adults with non-chronic diseases, caste, religion, education, and wealth status were found to be significant influencers of nursing consultation. The study concluded that nursing consultation is under-utilized, but serves the need of vulnerable population groups

Interaction between Nbp35 and Cfd1 proteins of cytosolic Fe-S cluster assembly reveals a stable complex formation in Entamoeba histolytica.

Iron-Sulfur (Fe-S) proteins are involved in many biological functions such as electron transport, photosynthesis, regulation of gene expression and enzymatic activities. Biosynthesis and transfer of Fe-S clusters depend on Fe-S clusters assembly processes such as ISC, SUF, NIF, and CIA systems. Unlike other eukaryotes which possess ISC and CIA systems, amitochondriate Entamoeba histolytica has retained NIF & CIA systems for Fe-S cluster assembly in the cytosol. In the present study, we have elucidated interaction between two proteins of E. histolytica CIA system, Cytosolic Fe-S cluster deficient 1 (Cfd1) protein and Nucleotide binding protein 35 (Nbp35). In-silico analysis showed that structural regions ranging from amino acid residues (P33-K35, G131-V135 and I147-E151) of Nbp35 and (G5-V6, M34-D39 and G46-A52) of Cfd1 are involved in the formation of protein-protein complex. Furthermore, Molecular dynamic (MD) simulations study suggested that hydrophobic forces surpass over hydrophilic forces between Nbp35 and Cfd1 and Van-der-Waal interaction plays crucial role in the formation of stable complex. Both proteins were separately cloned, expressed as recombinant fusion proteins in E. coli and purified to homogeneity by affinity column chromatography. Physical interaction between Nbp35 and Cfd1 proteins was confirmed in vitro by co-purification of recombinant Nbp35 with thrombin digested Cfd1 and in vivo by pull down assay and immunoprecipitation. The insilico, in vitro as well as in vivo results prove a stable interaction between these two proteins, supporting the possibility of its involvement in Fe-S cluster transfer to target apo-proteins through CIA machinery in E. histolytica. Our study indicates that initial synthesis of a Fe-S precursor in mitochondria is not necessary for the formation of Cfd1-Nbp35 complex. Thus, Cfd1 and Nbp35 with the help of cytosolic NifS and NifU proteins can participate in the maturation of non-mitosomal Fe-S proteins without any apparent assistance of mitosomes

Purification of recombinant Cfd1 and Nbp35 protein.

<p>The recombinant Cfd1 protein was purified through Ni⁺²-NTA column. A) Lane M-marker, lane 1-total lysate, lane 2- supernatant, lane 3-flow through, lane 4-,5-, 6-, & 7- wash (10, 20, 35, & 50 mM imidazole), lane 8- eluate (100 mM imidazole), lane 9- eluate (200 mm imidazole), lane10- eluate (300 mM imidazole). B) Immunoblot was probed with anti-Cfd1 antibody. Lane M- represents molecular weight proteins marker, lane 11- purified rCfd1 protein was probed with anti-His monoclonal antibodies, lane 12- total <i>E. histolytica</i> cell lysate, and lane-13- supernatant of <i>E. histolytica</i> lysate C) Similarly, the recombinant Nbp35 protein was purified through Ni⁺²-NTA column Lane M-marker, lane 1- total lysate, lane 2- supernatant, lane 3- flow through, lane 4-,5-, 6-, & 7- wash (10, 20, 35, & 50 mM imidazole), lane 8- eluate (100 mM imidazole) lane 9- (200 mM imidazole), lane10- eluate (300 mM imidazole). D) Immunoblot was probed with anti-Nbp35 antibody. Lane M represents –molecular weight proteins marker, lane 11- purified rNbp35 protein was probed with anti-His monoclonal antibodies, lane 12- total <i>E. histolytica</i> cell lysate, and lane-13- supernatant of <i>E. histolytica</i> lysate.</p

Nbp35-Cfd1 complex structure showing the regions involved in interaction.

<p>(A) Complex structure of Nbp35-Cfd1 where the molecular contacts are highlighted (blue–Nbp35) and (green-Cfd1) with mentioned residues involved in interaction. (B) Porcupine plots for Nbp35-Cfd1 complex showing backbone fluctuation from simulation time course. The regions involved in contact are highlighted with dotted region.</p

Interaction study between Cfd1 and Nbp35 proteins.

<p>(A) Co-purification using recombinant Nbp35 and thrombin digested Cfd1; Immunoblots of fractions obtained from co-purification experiment as described in material and methods probed with different antibodies (i) Anti-Nbp35 (ii) Anti-Cfd1 (iii) Anti-His antibodies for interaction between recombinant Nbp35 and thrombin digested Cfd1 proteins. Lane M- represents molecular weight proteins marker, lane 1- <i>E. coli</i> lysate overexpressing rNbp35, lane 2- purified rNbp35, lane 3- undigested rCfd1, lane 4- digested rCfd1, lane 5- wash (60 mM imidazole), lane 6 final wash did not show any proteins in all blots, lane 7- eluate (200 mM imidazole), lane 8- eluate (500 mM imidazole). (B) Pull down assay using rNbp35 and <i>E. histolytica</i> lysate: Ni⁺²-NTA bound rNbp35 was incubated with <i>E. histolytica</i> lysate as described in material and methods for pull down assay and eluate (500 mM imidazole) fraction was subjected to immunoblot analysis proved different antibodies (upper panel) Anti-His (middle panel) Anti-Nbp35 (lower panel) Anti-Cfd1 antibodies for rNbp35 and endogenous amoebic Cfd1 interaction. Lane M- represents molecular weight proteins marker, lane 1- total <i>E. coli</i> lysate expressed rNbp35, lane 2- supernatant rNbp35, lane 3- total amoebic lysate, lane 4- amoebic lysate flow through, lane 5 final wash did not show any proteins in all blots, lane 6- eluate (500 mM imidazole).</p

RMSD and RMSF profile of Nbp35-Cfd1 complex.

<p>(A) RMSD profile of Nbp35-Cfd1 complex for all-atoms (red), backbone atoms (blue) and side chain (green) plotted as a function of time (B) Root mean square fluctuation for Nbp35 and Cfd1 plotted against time from MD simulation.</p