Tariq Asim

𝐊𝐚𝐫𝐲𝐨𝐭𝐲𝐩𝐢𝐧𝐠: 𝐀 𝐂𝐨𝐦𝐩𝐫𝐞𝐡𝐞𝐧𝐬𝐢𝐯𝐞 𝐆𝐮𝐢𝐝𝐞

A karyotype is a preparation of the complete set of metaphase chromosomes in the cells of a species or in an individual organism, sorted by length, centromere location and other features and for a test that detects this complement or counts the number of chromosomes. Karyotyping is the process by which a karyotype is prepared from photographs of chromosomes, in order to determine the chromosome complement of an individual, including the number of chromosomes and any abnormalities.

Karyotypes describe the chromosome count of an organism and what these chromosomes look like under a light microscope. Attention is paid to their length, the position of the centromeres, banding pattern, any differences between the s*x chromosomes, and any other physical characteristics. The preparation and study of karyotypes is part of cytogenetics.

The basic number of chromosomes in the somatic cells of an individual or a species is called the somatic number and is designated 2n. In the germ-line (the s*x cells) the chromosome number is n (humans: n = 23). [3]p28 Thus, in humans 2n = 46. So, in normal diploid organisms, autosomal chromosomes are present in two copies.

Types of Karyotyping:-
* Spectral Karyotyping
* Virtual Karyotyping

Application of Karyotyping:-
* Genetic Disorders
* Oncology
* Personalized Medicine, others.

The semen released by a healthy man after in*******se with a woman contains 400 million s***ms.
So, logically, if that amount of s***m found its way into the uterus, 400 million babies would be born!
While these 400 million s***m race like mad towards the mother's uterus, only 300 or 500 survive the race.
And the rest? They die of exhaustion or defeat on the way. It is 300-500 s***matozoa, which manage to reach the o**m. Only one of them, yes only a very strong s***matozoon, enters the o**m and fertilizes it, or reaches the o**m and makes its seat.
Do you know who is the lucky, victorious and strongest s***m?
That lucky s***m is you, me, or all of us.
Have you ever thought about this great war?
When you ran "there were no eyes, no hands, no feet, no head, yet you won!"
When you ran, you didn't have the certificates, you didn't have the brains, but you still won!
When you ran, you were not educated, nobody helped you but you lived.
Your intuition only had the destination in sight when you ran and you ran with one mind, your determination was only that destination and you won in the end.
After that, many children were lost in the mother's womb. But you stayed, you completed your 9 months.
And today...
You panic when something happens, you get frustrated, but why? Why do you think you lost? Why have you lost confidence? Now you have friends, siblings, certificates, everything. There are hands and feet, there is education, there is a great brain to plan, there are people to help, yet you have given up hope.
Why do you break up when something happens?
Why do you say I don't want to live?
Why did you say I lost?
It is possible to highlight thousands of such things, but why are you disappointed?
Why did you get frustrated? You win at the beginning, you win at the end, you win in the middle.

D-zone test performed by using Disc diffusion method. 💛

▪️An agar plate containing an isolate of a staphylococcus or streptococcal bacteria is inoculated, and a 2-μg clindamycin disc(lincosamide) is placed next to Erythromycin(Macrolide). The plate is then left to incubate for the entire night.

▪️The growth of the organism up to the edges of the disc, flattening of the clindamycin zone shows (positive D test) near the erythromycin disc (resistant).

▪️Positive D test indicates the presence of macrolide-inducible resistance to clindamycin produced by an inducible methylase that alters the common ribosomal binding site for macrolides, clindamycin and the group B streptogrammins.

▪️The cross-resistance, called the MLS-B phenotype, results from enzymatic methylation of an adenine residue of the 23S component of the 50S ribosomal subunit that these 3 drug groups bind to. The methylase is encoded by a plasmid-borne gene called ERM gene.

▪️While clindamycin may still be effective in some patients with this phenotype, the working assumption is that the isolate is “presumed” to be Clindamycin resistant based upon detection of inducible clindamycin resistance.

▪️Since clindamycin and streptogramin are among the few drugs of choice in the treatment of methicillin resistant S. aureus (MRSA) infections, knowing the resistance to these antibiotics is crucial.
C🔗

𝐄𝐧𝐯𝐢𝐫𝐨𝐧𝐦𝐞𝐧𝐭𝐚𝐥 𝐌𝐨𝐧𝐢𝐭𝐨𝐫𝐢𝐧𝐠 𝐚𝐧𝐝 𝐁𝐞𝐲𝐨𝐧𝐝: 𝐓𝐡𝐞 𝐕𝐞𝐫𝐬𝐚𝐭𝐢𝐥𝐢𝐭𝐲 𝐨𝐟 𝐃𝐢𝐠𝐢𝐭𝐚𝐥 𝐏𝐂𝐑

Digital PCR (dPCR) is a sophisticated molecular biology technique used to quantify and analyze the amount of a specific DNA or RNA target sequence in a given sample. It is a more precise and sensitive method for measuring nucleic acid concentrations compared to traditional PCR (polymerase chain reaction), particularly when dealing with low abundance or rare targets.

📢 𝐇𝐞𝐫𝐞'𝐬 𝐡𝐨𝐰 𝐝𝐢𝐠𝐢𝐭𝐚𝐥 𝐏𝐂𝐑 𝐰𝐨𝐫𝐤𝐬 𝐚𝐧𝐝 𝐢𝐭𝐬 𝐤𝐞𝐲 𝐟𝐞𝐚𝐭𝐮𝐫𝐞𝐬:

✅ Sample Partitioning: In digital PCR, the sample containing the DNA or RNA to be quantified is divided into thousands or even millions of tiny partitions. Each partition is a micro-scale reaction vessel.

✅ Amplification: The DNA or RNA in each partition undergoes PCR amplification. This means that if the target sequence is present in a partition, it will be duplicated exponentially through repeated cycles of heating and cooling (denaturation, annealing, and extension).

✅ Endpoint Analysis: After the amplification is complete, each partition is analyzed to determine whether it contains the target sequence. This is usually done by detecting a signal, such as fluorescence, that is associated with the presence of the target.

✅ Quantification: The number of partitions with a positive signal is counted. This count is then used to calculate the concentration of the target sequence in the original sample. Because each partition is considered independently, digital PCR provides absolute quantification rather than relative quantification, which is typical in traditional real-time PCR.

📢𝐊𝐞𝐲 𝐅𝐞𝐚𝐭𝐮𝐫𝐞𝐬 𝐚𝐧𝐝 𝐀𝐝𝐯𝐚𝐧𝐭𝐚𝐠𝐞𝐬 𝐨𝐟 𝐃𝐢𝐠𝐢𝐭𝐚𝐥 𝐏𝐂𝐑:

✅ Higher Precision: Digital PCR offers greater precision and accuracy, especially when quantifying low-abundance or rare targets. It is less affected by factors like PCR inhibitors and sample variations.

✅ Absolute Quantification: Unlike real-time PCR, which provides relative quantification, dPCR provides absolute quantification of the target molecule. This is essential in applications like clinical diagnostics and quantifying rare mutations.

✅ Robustness: Digital PCR is robust against the presence of impurities and contaminants in the sample, making it suitable for a wide range of sample types.

📢 𝐀𝐩𝐩𝐥𝐢𝐜𝐚𝐭𝐢𝐨𝐧𝐬 𝐨𝐟 𝐃𝐢𝐠𝐢𝐭𝐚𝐥 𝐏𝐂𝐑:

✅ Cancer Research
✅ Disease Diagnostics
✅ Environmental Monitoring
✅ Food Safety
✅ Genetic Testing

Digital PCR is a powerful tool in molecular biology, particularly when precise quantification of nucleic acids is required, and it has become increasingly important in fields such as clinical diagnostics and genomics research.

𝐓𝐞𝐜𝐡𝐧𝐢𝐪𝐮𝐞𝐬 𝐔𝐬𝐞𝐝 𝐢𝐧 𝐂𝐞𝐥𝐥 𝐀𝐧𝐚𝐥𝐲𝐬𝐢𝐬

𝐀𝐜𝐜𝐞𝐬𝐬 𝐒𝐚𝐦𝐩𝐥𝐞 𝐏𝐃𝐅: https://lnkd.in/dmgjgqhw

𝐌𝐢𝐜𝐫𝐨𝐬𝐜𝐨𝐩𝐲:
𝐋𝐢𝐠𝐡𝐭 𝐌𝐢𝐜𝐫𝐨𝐬𝐜𝐨𝐩𝐲: This includes techniques like bright-field, phase-contrast, & fluorescence microscopy, which allow for the visualization of cell morphology, organelles, & fluorescently labeled molecules within cells.

𝐄𝐥𝐞𝐜𝐭𝐫𝐨𝐧 𝐌𝐢𝐜𝐫𝐨𝐬𝐜𝐨𝐩𝐲: Transmission electron microscopy (TEM) and scanning electron microscopy (SEM) provide high-resolution images of cellular structures & ultrastructure.

𝐅𝐥𝐨𝐰 𝐂𝐲𝐭𝐨𝐦𝐞𝐭𝐫𝐲:
Flow cytometry is used to analyze and sort cells based on various characteristics such as size, shape, & fluorescence properties.

𝐈𝐦𝐦𝐮𝐧𝐨𝐜𝐲𝐭𝐨𝐜𝐡𝐞𝐦𝐢𝐬𝐭𝐫𝐲 𝐚𝐧𝐝 𝐈𝐦𝐦𝐮𝐧𝐨𝐡𝐢𝐬𝐭𝐨𝐜𝐡𝐞𝐦𝐢𝐬𝐭𝐫𝐲:
These techniques use antibodies labeled with fluorophores or enzymes to detect specific proteins or antigens within cells.

𝐂𝐞𝐥𝐥 𝐕𝐢𝐚𝐛𝐢𝐥𝐢𝐭𝐲 𝐚𝐧𝐝 𝐏𝐫𝐨𝐥𝐢𝐟𝐞𝐫𝐚𝐭𝐢𝐨𝐧 𝐀𝐬𝐬𝐚𝐲𝐬:
Techniques like MTT assays, MTS assays, & trypan blue exclusion are used to assess cell viability & proliferation.

𝐂𝐞𝐥𝐥 𝐒𝐨𝐫𝐭𝐢𝐧𝐠:
Flow cytometry and magnetic-activated cell sorting (MACS) are used for isolating specific cell populations based on surface markers or other characteristics.

𝐂𝐞𝐥𝐥 𝐂𝐮𝐥𝐭𝐮𝐫𝐞 𝐚𝐧𝐝 𝐂𝐞𝐥𝐥 𝐋𝐢𝐧𝐞𝐬:
Establishing and maintaining cell cultures is essential for many cell-based assays. This includes primary cell cultures & established cell lines.

𝐃𝐍𝐀 𝐚𝐧𝐝 𝐑𝐍𝐀 𝐀𝐧𝐚𝐥𝐲𝐬𝐢𝐬:
Techniques like PCR, qPCR, RT-PCR, & RNA sequencing are used to study gene expression, mutations, & other genetic aspects of cells.

𝐏𝐫𝐨𝐭𝐞𝐨𝐦𝐢𝐜𝐬:
Mass spectrometry-based techniques can identify and quantify proteins within cells, enabling the study of protein expression, post-translational modifications, & interactions.

𝐌𝐞𝐭𝐚𝐛𝐨𝐥𝐨𝐦𝐢𝐜𝐬:
Metabolomic techniques analyze the small molecules present in cells, providing insights into cellular metabolism & metabolic pathways.

𝐋𝐢𝐯𝐞-𝐂𝐞𝐥𝐥 𝐈𝐦𝐚𝐠𝐢𝐧𝐠:
Real-time monitoring of cellular processes using time-lapse microscopy, which can be coupled with fluorescence markers to track dynamic changes within cells.

𝐄𝐥𝐞𝐜𝐭𝐫𝐨𝐩𝐡𝐲𝐬𝐢𝐨𝐥𝐨𝐠𝐲:
Patch clamping and voltage-clamp techniques are used to study the electrical properties of cells, particularly neurons & muscle cells.

𝐌𝐢𝐜𝐫𝐨𝐚𝐫𝐫𝐚𝐲 𝐀𝐧𝐚𝐥𝐲𝐬𝐢𝐬:
DNA microarrays and gene expression arrays allow for the simultaneous analysis of thousands of genes, helping researchers understand gene regulation and expression profiles in cells.

𝐒𝐢𝐧𝐠𝐥𝐞-𝐂𝐞𝐥𝐥 𝐀𝐧𝐚𝐥𝐲𝐬𝐢𝐬:
This emerging field focuses on studying individual cells within a heterogeneous population, providing insights into cellular heterogeneity and rare cell populations.

⚠️ metabolites and play an important role in .

❇️The hypothesis that the gut microbiota may affect pediatric migraine by directly or indirectly regulating tryptophan metabolism.

❇️Tryptophan is an essential amino acid metabolized in the human body in three main pathways:

1️⃣Microbiota-related indole pathways

2️⃣The serotonin (5-hydroxyptamine,5-HT)

3️⃣Kynurenine (KYN)

❇️Metabolites produced in the serotonin and kynurenine pathways are related to migraine in adults.

❇️Gut microbiota can metabolize tryptophan, through
1️⃣the serotonin pathway
2️⃣the kynurenine pathway, resulting in the production of metabolites such as serotonin, kynurenine, kynurenic acid ( ), and quinolinic acid (QUIN).

❇️QUIN is known as a pro-oxidant inflammatory molecule that activates NMDA receptors and regulates glutamate release or uptake to facilitate migraine progression.

❇️Impaired tryptophan metabolism was found in children with migraine, as we observed a significant decrease in the plasma concentrations of KYNA, while 5- and QUIN showed a significant increase in migraine children compared to healthy children.

❇️A sudden increase in 5-HT release may be a triggering event that culminates in migraine attacks or a response elevation to the migraine attack.

❇️Decreased concentrations of Kynurenic acid (KYNA) in adults with chronic migraine or cluster headaches.

❇️KYNA has been shown to prevent trigeminovascular activation and cortical spreading depression in animal models of migraine.

❇️KYNA also interacts with endogenous N-methyl-D-aspartic acid (NMDA) receptors to inhibit the glutamate pathway, potentially preventing migraine-causing sensitization processes.

❇️Beta diversity differences and variations in twenty-three genera were found between adults with migraine and healthy adults.

❇️The high- diet-induced increase in the abundance of the phylum was attributed to the increased bioavailability of respiratory electron acceptors of the host, such as oxygen and nitrate, which fueled the phylum Proteobacteria proliferation.

❇️The increase in Proteobacteria levels in migraine children might further support the opinion that microbial is closely associated with the pathogenesis of pediatric migraine.

❇️They identified a total of twenty-five bacterial genera that exhibited distinctive abundance patterns between the two groups, suggesting an association between migraine and alterations in GM composition.

📌To sum up, their study suggests that the gut microbiota can regulate tryptophan metabolism, particularly the kynurenine pathway, to be involved in the pathogenesis of migraine. These findings contribute to the understanding of the role of the gut-brain axis in migraine and provide a potential mechanism for further exploration of migraine.

🖇Plz read more on the topic here👉🏻: https://lnkd.in/dDRRkV3F
🔗

🟨🟪 𝘼𝙙𝙫𝙖𝙣𝙘𝙚𝙨 𝙞𝙣 𝙙𝙚𝙣𝙙𝙧𝙞𝙩𝙞𝙘 𝙘𝙚𝙡𝙡 𝙫𝙖𝙘𝙘𝙞𝙣𝙖𝙩𝙞𝙤𝙣 𝙩𝙝𝙚𝙧𝙖𝙥𝙮 𝙤𝙛 𝙘𝙖𝙣𝙘𝙚𝙧

👉

https://lnkd.in/de7HWdJR

🔶️ (DCs) are classified into plasmacytoid DC (pDC) and conventional DC (cDC), with the latter further subdivided into cDC1s and cDC2s.
DCs represent a promising means for cancer vaccines to strengthen the anti-cancer immune response to overcome the immunosuppressive

👉🔹️ DC vaccination acts through enhancement of antitumor immune responses.

🔶️ DCs are the most potent antigen presenting cells that play essential roles in tumor immunotherapies through induction of CD8+ T cell immunity.

🔶️ Accordingly, various strategies have been tested to employ DCs as therapeutic vaccines for exploiting their activity against tumor cells.

🔶️ Application of whole tumor cells or purified/recombinant antigen peptides are the most common approaches for pulsing DCs, which are then injected back into the patients.

🔶️ Although some hopeful results are reported for a number of DC vaccines tested and clinical trials of cancer patients, such approaches are still inefficient and require optimization.

🔶️ Failure of DC vaccination is postulated due to:

👉Immunosuppressive tumor microenvironment

👉Overexpression of checkpoint proteins

👉 Suboptimal avidity of tumor-associated antigen (TAA)-
specific T lymphocytes

👉 and lack of appropriate adjuvants

👉🔹️So researchers have chosen strategies to improve vaccine potential including,
Combination with immune checkpoint inhibitors that increases antitumor efficacy of DC vaccination.

🔶️ is a promising approach in the fight against cancer, recent advances in this field including:

🔹️ Recent studies have explored DC vaccination in the treatment of hematological malignancies, including acute myeloid leukemia and myelodysplastic syndromes, as well as other non-leukemia malignancies.

🔹️ DCs-based vaccines are considered an alternative therapeutic approach to treat esophageal and ovarian cancer.

🔹️The combination of dendritic cell with other treatments, such as
ICIs or chemotherapy, can be effective in treating certain types of solid cancers.

🔹️For example, a combination therapy of pemetrexed with DCs was effective as well as well-tolerated in advanced esophageal cancer patients.

🔹️ Additionally, DCs vaccination therapy has been shown to inhibit further spread of metastatic tumors or tumor recurrence in mice after surgery.

🔶️ Some of the latest clinical trials include:

🔹️ combination therapy for the treatment of , including :




(RCC)

.


🔶️ So dendritic cell-based cancer therapy is an exciting area of research with promising developments and potential for improving cancer treatment.

𝐍𝐞𝐱𝐭 𝐆𝐞𝐧𝐞𝐫𝐚𝐭𝐢𝐨𝐧 𝐃𝐍𝐀 𝐒𝐞𝐪𝐮𝐞𝐧𝐜𝐢𝐧𝐠 (𝐍𝐆𝐒):
𝐀 𝐂𝐨𝐦𝐩𝐫𝐞𝐡𝐞𝐧𝐬𝐢𝐯𝐞 𝐆𝐮𝐢𝐝𝐞

𝐏𝐃𝐅: - https://lnkd.in/df92V472

Next generation sequencing services include whole-genome sequencing, genome assembly and annotation, screening for antimicrobial resistance & virulence factors, secondary metabolite prediction, microbial community analysis (metagenomics), comparative genomics, strain differentiation and strain identification.

The term "next-generation sequencing" refers to a number of different technologies that all have one thing in common, they produce very large volumes of sequence data and have opened up a whole new set of applications in microbiology. Our next generation sequencing platforms can provide data on tens to hundreds of samples in a single run, with millions of individual DNA sequences produced per run. The increase in sequencing throughput with these next-generation sequencing platforms has delivered reduced costs and increased accuracy per base.

The method of determining the sequence of nucleotides in a piece of DNA is known as next-generation sequencing, or high-throughput sequencing. Sequencing by synthesis (SBS), ion semiconductor sequencing, nanopore sequencing, and single-molecule real-time (SMRT) sequencing are some of the techniques used. It is a cost-effective solution that provides precise, high-accuracy, and fast outcomes.

T𝐡𝐞 𝐂𝐨𝐦𝐩𝐥𝐞𝐭𝐞 𝐀𝐧𝐭𝐢𝐛𝐢𝐨𝐭𝐢𝐜 𝐂𝐡𝐚𝐫𝐭

Antibiotic classification refers to the categorization of antibiotics based on their chemical structure, mechanism of action, and spectrum of activity. Antibiotics are drugs that are used to treat bacterial infections by killing or inhibiting the growth of bacteria.

𝐓𝐡𝐞𝐫𝐞 𝐚𝐫𝐞 𝐬𝐞𝐯𝐞𝐫𝐚𝐥 𝐰𝐚𝐲𝐬 𝐭𝐨 𝐜𝐥𝐚𝐬𝐬𝐢𝐟𝐲 𝐚𝐧𝐭𝐢𝐛𝐢𝐨𝐭𝐢𝐜𝐬, 𝐛𝐮𝐭 𝐨𝐧𝐞 𝐜𝐨𝐦𝐦𝐨𝐧 𝐦𝐞𝐭𝐡𝐨𝐝 𝐢𝐬 𝐛𝐚𝐬𝐞𝐝 𝐨𝐧 𝐭𝐡𝐞𝐢𝐫 𝐦𝐞𝐜𝐡𝐚𝐧𝐢𝐬𝐦 𝐨𝐟 𝐚𝐜𝐭𝐢𝐨𝐧. 𝐓𝐡𝐢𝐬 𝐜𝐥𝐚𝐬𝐬𝐢𝐟𝐢𝐜𝐚𝐭𝐢𝐨𝐧 𝐢𝐧𝐜𝐥𝐮𝐝𝐞𝐬:

1. Cell wall synthesis inhibitors: These antibiotics target the bacterial cell wall, which is essential for bacterial growth and survival. Examples of antibiotics in this class include penicillins, cephalosporins, carbapenems, and monobactams.

2. Protein synthesis inhibitors: These antibiotics target the ribosome, which is responsible for synthesizing proteins in bacteria. Examples of antibiotics in this class include macrolides, tetracyclines, aminoglycosides, and chloramphenicol.

3. DNA synthesis inhibitors: These antibiotics target the enzymes responsible for replicating bacterial DNA. Examples of antibiotics in this class include quinolones and metronidazole.

4. RNA synthesis inhibitors: These antibiotics target the enzymes responsible for transcribing bacterial RNA. Rifampin is an example of an antibiotic in this class.

5. Metabolic inhibitors: These antibiotics target the metabolic pathways that bacteria use to synthesize essential molecules. Examples of antibiotics in this class include sulfonamides and trimethoprim.

6. Precision Therapeutics Leading the Way: The landscape of Dystonia drugs is evolving with precision therapies. Imagine treatments tailored to the individual's genetic makeup, offering more targeted and effective relief.