Membrane Protein-Lipid Interactions Probed Using Mass Spectrometry.

Membrane proteins that exist in lipid bilayers are not isolated molecular entities. The lipid molecules that surround them play crucial roles in maintaining their full structural and functional integrity. Research directed at investigating these critical lipid-protein interactions is developing rapidly. Advancements in both instrumentation and software, as well as in key biophysical and biochemical techniques, are accelerating the field. In this review, we provide a brief outline of structural techniques used to probe protein-lipid interactions and focus on the molecular aspects of these interactions obtained from native mass spectrometry (native MS). We highlight examples in which lipids have been shown to modulate membrane protein structure and show how native MS has emerged as a complementary technique to X-ray crystallography and cryo-electron microscopy. We conclude with a short perspective on future developments that aim to better understand protein-lipid interactions in the native environment.

Human Antibodies that Slow Erythrocyte Invasion Potentiate Malaria-Neutralizing Antibodies.

The Plasmodium falciparum reticulocyte-binding protein homolog 5 (PfRH5) is the leading target for next-generation vaccines against the disease-causing blood-stage of malaria. However, little is known about how human antibodies confer functional immunity against this antigen. We isolated a panel of human monoclonal antibodies (mAbs) against PfRH5 from peripheral blood B cells from vaccinees in the first clinical trial of a PfRH5-based vaccine. We identified a subset of mAbs with neutralizing activity that bind to three distinct sites and another subset of mAbs that are non-functional, or even antagonistic to neutralizing antibodies. We also identify the epitope of a novel group of non-neutralizing antibodies that significantly reduce the speed of red blood cell invasion by the merozoite, thereby potentiating the effect of all neutralizing PfRH5 antibodies as well as synergizing with antibodies targeting other malaria invasion proteins. Our results provide a roadmap for structure-guided vaccine development to maximize antibody efficacy against blood-stage malaria.

Structures of DPAGT1 Explain Glycosylation Disease Mechanisms and Advance TB Antibiotic Design.

Protein N-glycosylation is a widespread post-translational modification. The first committed step in this process is catalysed by dolichyl-phosphate N-acetylglucosamine-phosphotransferase DPAGT1 (GPT/E.C. 2.7.8.15). Missense DPAGT1 variants cause congenital myasthenic syndrome and disorders of glycosylation. In addition, naturally-occurring bactericidal nucleoside analogues such as tunicamycin are toxic to eukaryotes due to DPAGT1 inhibition, preventing their clinical use. Our structures of DPAGT1 with the substrate UDP-GlcNAc and tunicamycin reveal substrate binding modes, suggest a mechanism of catalysis, provide an understanding of how mutations modulate activity (thus causing disease) and allow design of non-toxic "lipid-altered" tunicamycins. The structure-tuned activity of these analogues against several bacterial targets allowed the design of potent antibiotics for Mycobacterium tuberculosis, enabling treatment in vitro, in cellulo and in vivo, providing a promising new class of antimicrobial drug.

Structural Basis of Tail-Anchored Membrane Protein Biogenesis by the GET Insertase Complex.

Membrane protein biogenesis faces the challenge of chaperoning hydrophobic transmembrane helices for faithful membrane insertion. The guided entry of tail-anchored proteins (GET) pathway targets and inserts tail-anchored (TA) proteins into the endoplasmic reticulum (ER) membrane with an insertase (yeast Get1/Get2 or mammalian WRB/CAML) that captures the TA from a cytoplasmic chaperone (Get3 or TRC40, respectively). Here, we present cryo-electron microscopy reconstructions, native mass spectrometry, and structure-based mutagenesis of human WRB/CAML/TRC40 and yeast Get1/Get2/Get3 complexes. Get3 binding to the membrane insertase supports heterotetramer formation, and phosphatidylinositol binding at the heterotetramer interface stabilizes the insertase for efficient TA insertion in vivo. We identify a Get2/CAML cytoplasmic helix that forms a "gating" interaction with Get3/TRC40 important for TA insertion. Structural homology with YidC and the ER membrane protein complex (EMC) implicates an evolutionarily conserved insertion mechanism for divergent substrates utilizing a hydrophilic groove. Thus, we provide a detailed structural and mechanistic framework to understand TA membrane insertion.

Effect of salinity stress and surfactant treatment with zinc and boron on morpho-physiological and biochemical indices of fenugreek (Trigonella foenum-graecum).

Micronutrient application has a crucial role in mitigating salinity stress in crop plants. This study was carried out to investigate the effect of zinc (Zn) and boron (B) as foliar applications on fenugreek growth and physiology under salt stress (0 and 120 mM). After 35 days of salt treatments, three levels of zinc (0, 50, and 100 ppm) and two levels of boron (0 and 2 ppm) were applied as a foliar application. Salinity significantly reduced root length (72.7%) and shoot length (33.9%), plant height (36%), leaf area (37%), root fresh weight (48%) and shoot fresh weight (75%), root dry weight (80%) and shoot dry weight (67%), photosynthetic pigments (78%), number of branches (50%), and seeds per pod (56%). Fenugreek's growth and physiology were improved by foliar spray of zinc and boron, which increased the length of the shoot (6%) and root length (2%), fresh root weight (18%), and dry root weight (8%), and chlorophyll a (1%), chlorophyll b (25%), total soluble protein content (3%), shoot calcium (9%) and potassium (5%) contents by significantly decreasing sodium ion (11%) content. Moreover, 100 ppm of Zn and 2 ppm of B enhanced the growth and physiology of fenugreek by reducing the effect of salt stress. Overall, boron and zinc foliar spray is suggested for improvement in fenugreek growth under salinity stress.

Structure of BRCA1-BRCT/Abraxas Complex Reveals Phosphorylation-Dependent BRCT Dimerization at DNA Damage Sites.

BRCA1 accumulation at DNA damage sites is an important step for its function in the DNA damage response and in DNA repair. BRCA1-BRCT domains bind to proteins containing the phosphorylated serine-proline-x-phenylalanine (pSPxF) motif including Abraxas, Bach1/FancJ, and CtIP. In this study, we demonstrate that ionizing radiation (IR)-induces ATM-dependent phosphorylation of serine 404 (S404) next to the pSPxF motif. Crystal structures of BRCT/Abraxas show that phosphorylation of S404 is important for extensive interactions through the N-terminal sequence outside the pSPxF motif and leads to formation of a stable dimer. Mutation of S404 leads to deficiency in BRCA1 accumulation at DNA damage sites and cellular sensitivity to IR. In addition, two germline mutations of BRCA1 are found to disrupt the dimer interface and dimer formation. Thus, we demonstrate a mechanism involving IR-induced phosphorylation and dimerization of the BRCT/Abraxas complex for regulating Abraxas-mediated recruitment of BRCA1 in response to IR.

Pathological Effects of SARS-CoV-2 Associated with Hematological Abnormalities.

The SARS coronavirus 2 (SARS-CoV-2) is the causative agent of the 2019 coronavirus disease (COVID-19) pandemic that has claimed the lives of 6.9 million people and infected over 765 million. It has become a major worldwide health problem and is also known to cause abnormalities in various systems, including the hematologic system. COVID-19 infection primarily affects the lower respiratory tract and can lead to a cascade of events, including a cytokine storm, intravascular thrombosis, and subsequent complications such as arterial and venous thromboses. COVID-19 can cause thrombocytopenia, lymphopenia, and neutrophilia, which are associated with worse outcomes. Prophylactic anticoagulation is essential to prevent complications and death rates associated with the virus's effect on the coagulation system. It is crucial to recognize these complications early and promptly start therapeutic anticoagulation to improve patient outcomes. While rare, COVID-19-induced disseminated intravascular coagulation (DIC) exhibits some similarities to DIC induced by sepsis. Lactate dehydrogenase (LDH), D-dimer, ferritin, and C-reactive protein (CRP) biomarkers often increase in serious COVID-19 cases and poor prognosis. Understanding the pathophysiology of the disease and identifying risk factors for adverse outcomes is critical for effective management of COVID-19.

Dynamics of an LPS translocon induced by substrate and an antimicrobial peptide.

Lipopolysaccharide (LPS) transport to the outer membrane (OM) is a crucial step in the biogenesis of microbial surface defenses. Although many features of the translocation mechanism have been elucidated, molecular details of LPS insertion via the LPS transport (Lpt) OM protein LptDE remain elusive. Here, we integrate native MS with hydrogen-deuterium exchange MS and molecular dynamics simulations to investigate the influence of substrate and peptide binding on the conformational dynamics of LptDE. Our data reveal that LPS induces opening of the LptD ß-taco domain, coupled with conformational changes on ß-strands adjacent to the putative lateral exit gate. Conversely, an antimicrobial peptide, thanatin, stabilizes the ß-taco, thereby preventing LPS transport. Our results illustrate that LPS insertion into the OM relies on concerted opening movements of both the ß-barrel and ß-taco domains of LptD, and suggest a means for developing antimicrobial therapeutics targeting this essential process in Gram-negative ESKAPE pathogens.

The threat of honey bee RNA viruses to yellow-legged hornets: Insights from cross-species transmission events.

Viral diseases are a significant challenge in beekeeping, and recent studies have unveiled a potential link between these diseases and the yellow-legged hornets (Vespa velutina), notorious predators of honey bees. However, it remains unclear whether virus diseases are commonly shared between honey bees and hornets or are merely sporadic cross-species transmission events. To address this knowledge gap, we conducted a study utilizing hornet-keeping practices in Yunnan, Southwest China. Our findings demonstrate that deformed wing virus (DWV-A) and Israeli acute paralysis virus (IAPV) can be transmitted from honey bees to yellow-legged hornets. We detected virus replication in various hornet stages, including pupae with IAPV infections, indicating the similarities between infected hornet and honey bee stages. Furthermore, we observed signs and infection intensities of DWV-A and IAPV comparable to those in honey bees. While different polymorphisms were found in the virus isolates from yellow-legged hornets, the sequences remain similar to honey bee counterparts. While our findings suggest that DWV-A and IAPV behave like common diseases, we observed a natural elimination of the viruses in hornet colonies, with minimal alterations in viral sequences. Consequently, these events appear to be cross-species transmission from honey bees, with yellow-legged hornets acting as potential incidental hosts. Further investigations of virus monitoring in hornets promise valuable insights into the disease ecology of bee-infecting viruses.

Comparative evaluation of different moulting methods on productive performance, egg quality, and antibody response of leghorn hens.

The aim of the study was to compare three moulting procedures and their effect on productive performance, egg quality, and antibody response of leghorn hens. For this, a total of 324 laying hens were distributed into three treatment groups having 12 replicates of 9 birds each according to completely randomized design. Treatments consisted of three moulting procedures based on feed and light restriction. The targeted weight at the end of moulting was 1450-1470 g. As soon as the moulting procedure is complete, the comparative analysis of post-moult productive performance (feed intake, egg production, egg weight, egg mass, feed per dozen eggs, feed per kg egg mass, livability), egg quality characteristics (egg weight, egg length and width, shape index, surface area, volume, albumen height, weight, Haugh unit score, yolk width, height, index, egg shell pore number, shell weight, thickness, breaking strength), and antibody response against Newcastle disease and avian influenza (H-9) were evaluated. Birds subjected to moulting procedure 3 (8 days fasting and gradual decrease in light) showed improvement in productive performance, egg geometry and quality traits, and antibody response against Newcastle disease virus. Birds experienced moulting procedure 1 (11 days fasting) had improved feed intake, egg production, and livability. However, birds moulted with procedure 2 (6 days fasting) revealed intermediate result in all the studied parameters. In conclusion, moulting through feed and light restriction with 8 days fasting and gradual reduction in light has potential to improve performance of leghorn hens.

Effects of prenatal dichromatic light exposure on hatching results and post-hatch performance of Japanese quail.

Present study evaluated the effect of lighted incubation on pre- and post-hatch performance of Japanese quail. In a completely randomized design, 1200 eggs were evenly divided into 4 treatments groups having six replicates (each tray was considered as replicate), 50 eggs each. Different dichromatic lights (Green + Red; GR, Green + Blue; GB, and Blue + Red; BR) of 250 lux were provided during incubation for 12 h daily and effects of these lights very evaluated on hatching results and post-hatch growth. After hatch, 600 quail chicks were divided into 4 treatments, 6 replicates, and 25 birds each. Regarding hatching traits, better hatchability was found in the GR group compared to GB, BR, and dark group; while early embryonic mortality was lower in BR, GB, and dark group than GR; mid embryonic mortality was lower in dark group and late embryonic mortality was noted in the GR group than those of other treatment groups. In addition, moisture loss during incubation was minimum in BR and dark groups; however, chick spread was better in the GR group. In terms of growth performance, weight gain was better in the GR group; feed intake in dark, feed conversion ratio in BR, and livability were better in BR and GR group. In morphometrics, keel and shank length were higher in all the colored groups (GB, BR, and GR) whereas body length, wing spread, shank circumference, drumstick length, and circumference were higher in the GR group. Regarding serum chemistry, glucose, albumin, and globulin levels were higher in the GR group. It was concluded that under the experimental conditions, GR light at the prenatal stage to Japanese quail eggs positively influenced hatching performance and post-hatch growth.

IoMT-Based Osteosarcoma Cancer Detection in Histopathology Images Using Transfer Learning Empowered with Blockchain, Fog Computing, and Edge Computing.

Bone tumors, such as osteosarcomas, can occur anywhere in the bones, though they usually occur in the extremities of long bones near metaphyseal growth plates. Osteosarcoma is a malignant lesion caused by a malignant osteoid growing from primitive mesenchymal cells. In most cases, osteosarcoma develops as a solitary lesion within the most rapidly growing areas of the long bones in children. The distal femur, proximal tibia, and proximal humerus are the most frequently affected bones, but virtually any bone can be affected. Early detection can reduce mortality rates. Osteosarcoma's manual detection requires expertise, and it can be tedious. With the assistance of modern technology, medical images can now be analyzed and classified automatically, which enables faster and more efficient data processing. A deep learning-based automatic detection system based on whole slide images (WSIs) is presented in this paper to detect osteosarcoma automatically. Experiments conducted on a large dataset of WSIs yielded up to 99.3% accuracy. This model ensures the privacy and integrity of patient information with the implementation of blockchain technology. Utilizing edge computing and fog computing technologies, the model reduces the load on centralized servers and improves efficiency.

Network Meddling Detection Using Machine Learning Empowered with Blockchain Technology.

The study presents a framework to analyze and detect meddling in real-time network data and identify numerous meddling patterns that may be harmful to various communication means, academic institutes, and other industries. The major challenge was to develop a non-faulty framework to detect meddling (to overcome the traditional ways). With the development of machine learning technology, detecting and stopping the meddling process in the early stages is much easier. In this study, the proposed framework uses numerous data collection and processing techniques and machine learning techniques to train the meddling data and detect anomalies. The proposed framework uses support vector machine (SVM) and K-nearest neighbor (KNN) machine learning algorithms to detect the meddling in a network entangled with blockchain technology to ensure the privacy and protection of models as well as communication data. SVM achieves the highest training detection accuracy (DA) and misclassification rate (MCR) of 99.59% and 0.41%, respectively, and SVM achieves the highest-testing DA and MCR of 99.05% and 0.95%, respectively. The presented framework portrays the best meddling detection results, which are very helpful for various communication and transaction processes.

Effects of age, body weight, semen collection frequency and holding duration on semen traits of broiler breeder reared under different housing systems.

Semen traits play the vital role in determining the fertility of a broiler breeder flock; however, it can be influenced by several factors. This experiment was carried out to assess some of these factors affecting the semen. A total of 89 male birds and 960 hens of 20-week-old broiler breeder (2215 g ± 7.5%) were divided into two main groups; one was kept in cages (AIC) and another group was kept on deep litter floor (AIF), while both these groups were subjected to AI. The male birds of aforementioned groups (44 males and 480 females) were further divided into 4 sub-groups (11 males and 120 females) to execute different semen collection frequencies i.e., 2, 3, 4, and 5th days' interval. The impact of time duration between semen collection and insemination on sperm kinematics was monitored. The quantitative and qualitative analysis of semen including sperm concentration and sperm kinematics of the collected semen was conducted through a computer-assisted sperm analyzer (CASA) and ONGO machine (working on the CASA principle). Resultantly, the data revealed that the studied parameters of semen were deteriorated with the progression of age of male birds, while the group of males with standard body weight produced the best semen quantitatively and qualitatively followed by overweight particularly during the post peak phase (46-65 = 20 weeks). Although the 3rd day, semen collection frequency was found better for quality, the higher quantity of semen was achieved when males were being collected at the intervals of 4th and 5th day respectively regardless of housing systems. Significant decline in sperm kinematics was recorded with the progression of semen holding duration at the temperature of poultry farm. Furthermore, the highest contamination of E. coli, Salmonella, and Mycoplasma gallisepticum was recorded in the reproductive tract of hens and semen of the AIF group as compared to AIC. Thus, conclusion can be settled that the semen properties are significantly affected by age, body weight, and semen collection intervals in both housing systems, while sperm kinematics is being disrupted with the progression of holding duration. Although housing systems could affect the semen insignificantly, yet lesser contamination was recorded in semen and in the reproductive tract of hens of AIC.

Dosimetric sensitivity of leaf width on volumetric modulated arc therapy plan quality: an objective approach.

Background: Several authors investigated a dosimetric impact of leaf width on radiotherapy plan quality subjectively, and concluded that thinner leaf-width multileaf collimators (MLC) are beneficial because of their better coverage of clinically relevant structures. Study aimed to investigate the dosimetric effect of MLC leaf width on volumetric modulated arc therapy plan quality by objective approach. Materials and methods: Twelve of each prostate and head-and-neck patients were planned for volumetric modulated arc therapy (VMAT) treatments for MLC leaf widths of 4 mm and 10 mm. Three different VMAT schemes single-arc, dual-arc and two combined independent single-arcs were optimized. Dose volume histogram and Isodose distribution were used for quantitative and qualitative comparison of the treatment plan, respectively. Dose-volume-indices of the planning target volume, organs at risk and number of delivered monitor units were compared. The 4 mm leaf width being reference over 10 mm and results were noted as statistically significant if p ≤ 0.05 using student t-test. Results: All VMAT schemes for both tumor sites showed a gain in target coverage, similar organs at risk doses and higher monitor units to be delivered, when changing leaf width from 10 mm to 4 mm. The p-values were significant for majority of head-and-neck dose indices. Conclusion: The thinner-leaf MLCs, owing to their better spatial resolution, result in an overall gain for target coverage, while maintaining permissible doses to the organs at risk.

A small molecule inhibitor of HER3: a proof-of-concept study.

Despite being catalytically defective, pseudokinases are typically essential players of cellular signalling, acting as allosteric regulators of their active counterparts. Deregulation of a growing number of pseudokinases has been linked to human diseases, making pseudokinases therapeutic targets of interest. Pseudokinases can be dynamic, adopting specific conformations critical for their allosteric function. Interfering with their allosteric role, with small molecules that would lock pseudokinases in a conformation preventing their productive partner interactions, is an attractive therapeutic strategy to explore. As a well-known allosteric activator of epidermal growth factor receptor family members, and playing a major part in cancer progression, the pseudokinase HER3 is a relevant context in which to address the potential of pseudokinases as drug targets for the development of allosteric inhibitors. In this proof-of-concept study, we developed a multiplex, medium-throughput thermal shift assay screening strategy to assess over 100 000 compounds and identify selective small molecule inhibitors that would trap HER3 in a conformation which is unfavourable for the formation of an active HER2-HER3 heterodimer. As a proof-of-concept compound, AC3573 bound with some specificity to HER3 and abrogated HER2-HER3 complex formation and downstream signalling in cells. Our study highlights the opportunity to identify new molecular mechanisms of action interfering with the biological function of pseudokinases.

Development of a fluorogenic ADAMTS-7 substrate.

The extracellular protease ADAMTS-7 has been identified as a potential therapeutic target in atherosclerosis and associated diseases such as coronary artery disease (CAD). However, ADAMTS-7 inhibitors have not been reported so far. Screening of inhibitors has been hindered by the lack of a suitable peptide substrate and, consequently, a convenient activity assay. Here we describe the first fluorescence resonance energy transfer (FRET) substrate for ADAMTS-7, ATS7FP7. ATS7FP7 was used to measure inhibition constants for the endogenous ADAMTS-7 inhibitor, TIMP-4, as well as two hydroxamate-based zinc chelating inhibitors. These inhibition constants match well with IC50 values obtained with our SDS-PAGE assay that uses the N-terminal fragment of latent TGF-ß-binding protein 4 (LTBP4S-A) as a substrate. Our novel fluorogenic substrate ATS7FP7 is suitable for high throughput screening of ADAMTS-7 inhibitors, thus accelerating translational studies aiming at inhibition of ADAMTS-7 as a novel treatment for cardiovascular diseases such as atherosclerosis and CAD.

Optimisation of user-selectable volumetric modulated arc therapy (VMAT) planning parameters: VMAT arcs for prostate and head-and-neck cancers.

OBJECTIVE: To evaluate different VMAT planning techniques for a multi-leaf collimator (MLC)providing maximum aperture of 21 × 16 cm². METHODS: A VMAT planning study of nine prostate and nine head-and-neck cancer patients was carried out. The patients were previously treated with Intensity Modulated Radiotherapy (IMRT) technique during 2014-15, at radiation oncology SanBortolo Hospital, Vicenza, Italy. Three VMAT plans for each of prostate and head-and-neck cancer patient were optimized using Pinnacle treatment planning system for 6MV photon beam from ElektaSynergyS®Linac system. Single arc, dual arc and combined two independent-single-arcs were optimized for collimator angle 45°. VMAT treatment planning comparison was made among single-arc dual-arc and combined two independent-single-arcs. The student's t-test (two-sided) with p ≤ 0.05 was applied for significant difference between dose volume indices of plans. RESULTS: All VMAT techniques produced clinically acceptable plans for prostate, while for head-and-neck, DA and ISAs significantly improved target coverage compared to SA. Single arc is sufficient for prostate, while inefficient in case of head-and-neck dose-planning. In spite of having different VMAT optimization approach dual arc and two combined independent-single-arcs provided very similar target coverage, while dual arc improved sparing of organs-at-risk. Similar treatment delivery times were observed for DA and ISAs VMAT techniques. CONCLUSIONS: Single arc is sufficient for prostate, while inefficient in case of head-and-neck dose-planning. Dual arc and two combined independent-single-arcs provided similar PTV coverage, while DA provided better sparing of organs at risk.

Impact of light stimulation during incubation on hatching traits and post-hatch performance of commercial broilers.

Light in terms of photo- and scoto-periods is the key ambient factor affecting the physiology of birds through establishing normal biological clock and circadian rhythms. In natural incubation light significantly influences embryonic development, however, at commercial setups eggs are incubated under a dark environment. Presently not a single commercial poultry hatchery is using light during incubation; hence, comprehensive studies are needed to address the industry for considering light as a potential embryonic growth stimulant. In the present study, white Light-emitting diodes (LEDs; 5000 K) were installed in the incubator and 250 lx light intensity was provided for 0, 12, and 24 h per day during the whole incubation period. A total of 900 broiler hatching eggs (Hubbard classic; from 58 weeks old parents) were randomly allocated to 3 treatment groups, having 5 replicates of 60 eggs each, a tray was considered as replicate during incubation and these eggs were incubated under standard incubation protocols. After hatching, a total of 300 chicks were picked and divided into 3 described treatments (0, 12, and 24 h of photo-stimulation to eggs during incubation) having 5 replicates of 20 birds each. The results indicated that incubation of eggs under 12 and 24 h of lighting significantly improved (P ≤ 0.05) hatch window, hatchability % (0.0002), a hatch of fertile % (0.001), and carcass yield % (0.0454). Embryonic mortality, dead germs, and dead in shell embryos were lower in eggs incubated under 12 h light. Significantly better FCR (0.0006), stress susceptibilities such as H/L ratio (0.0227), and physical asymmetry (0.0065) were observed among the birds incubated under 12 h light (P ≤ 0.05). In conclusion, an appropriate light stimuli (12 h) may help to improve hatching traits and post-hatch performance of commercial broiler.

Production analysis of broiler farming in central Punjab, Pakistan.

This study examined the analysis of personal and production performance of broiler farming in 10 cities of central Punjab (Lahore, Sheikhupura, Kasur, Okara, Sargodha, Sahiwal, Gujranwala, Gujrat, Nankana, and Kamalia) of Pakistan. These districts are densely populated and renowned for broiler production. Primary data were collected through a survey of 100 poultry farmers. Personal and production information were collected from the respondents. Descriptive statistics was employed in the form of averages and percentages. The comparison among different districts was made through one-way ANOVA technique followed by Tukey's HSD test. Parameters were observed significantly in experience of farmer, age, number of sheds, capacity of farm, and flock/annuum during the survey of Gujranwala and Kasur, while the high percentage of graduate farmers was observed in Lahore and Sheikhupura regions. Significant results were observed during the production performance feed efficiency and mortality in Okara and Kasur as well as maximum administration of both live and killed vaccine in Lahore through drinking water in Nankana. In the districts of Sheikhupura and Kamalia, farmers due to lake of education left their dead birds on the surface ground. In conclusion, this study pointed out the exact trend of modern broiler farming and its price fluctuation in Punjab (Pakistan) as well as the reason behind the losses to the farmers.