Mutacje i plazmidy w bakteriach

Probiotics in human health and disease: from nutribiotics to pharmabiotics

• Probiotics are live microorganisms, which when consumed in adequate amounts, confer a health effect on the host, according to FAO and WHO.
• Probiotics are delivered orally in foods, food products, food supplements, and medicinal supplements.
• Probiotics are categorized into nutribiotics and pharmabiotics.
• Nutribiotics are probiotics with nutritive claims.
• Pharmabiotics are therapeutic and pharmacological probiotics, with health claims.

Introduction

• Probiotics are discussed in terms of their functions.
• The manner in which to obtain probiotics is examined.
• Categorization of probiotics in terms of their properties (nutritive and therapeutic) is presented.

Definition and categorization of probiotics

• Probiotics are live microorganisms that confer health effects when consumed in adequate amounts.
• Delivery of probiotics in foods, food products, food supplements, and medicinal products is detailed.
• The diagram shows the categories of probiotics: nutribiotics and pharmabiotics.

Nutribiotics producing vitamins

• Microorganisms produce vitamins and contribute to the availability of vitamins in the human host. Examples: B-vitamins.
• Using vitamin-producing nutribiotics is proposed as a natural approach with a lower probability of side effects compared to supplementation with chemically synthesized vitamins.
• Vitamin deficiencies can lead to serious health problems.

Nutribiotics improving nutritional health problems

• Probiotics have been used to treat gastrointestinal conditions, including lactose intolerance.
• Nutribiotics producing health beneficial metabolites are described.

Definition of pharmabiotics

• Pharmabiotics are bacterial cells of human origin, or their products, that have a proven pharmacological role in health or disease.
• This term includes microbes that are alive or dead, components of organisms, or metabolites of microbes that are not covered by the classical definition of probiotics, by FAO/WHO.
• Examples of specific strains/bacteria are given: L. plantarum DSM9843, L. plantarum 299V, L. reuteri, L. helveticus R0052, B. longum R0175 (PF) * B. bifidum + Streptococcus thermophilus.
• The strains listed here are meant for specific treatments: IBS, acute diarrhea/gastroenteritis, anxiety, depression

Conclusion

• Nutraceutical properties of probiotics can improve human health.
• Pharmabiotics are helpful for disease prevention and treatment, but further research is needed.
• The categorization of probiotics is necessary to avoid confusion between foods and medicines.

Mikrobiomy ciała ludzkiego

• Each microbiome encompasses all bacteria, archaea, fungi, viruses, and small eukaryotes inhabiting a particular organ or body region.
• The microbiome includes both physiological microflora and pathogens.
• Metagenome refers to the combined genomic information of bacteria, archaea, fungi, viruses, and small eukaryotes.
• Hologenome refers to the combined genomic information of bacteria, archaea, fungi, viruses, small eukaryotes, and the human genome.

Co oferujemy mikroorganizmom?

• Various conditions in the human body, including temperature, pH, and water content influence the microbiome.
• Different regions of the body have different conditions.
• Exposure to ultraviolet light (UV) also affects the microbiome.

Funkcje niezbędne w utrzymaniu zdrowia człowieka

• Microorganisms play a crucial role in the digestion of foods we cannot digest.
• Breast milk contains oligosaccharides, which support the development of the immune system in babies.
• Biofilms on various surfaces protect tissues.
• The role in the production of essential nutrients, such as ARA and DHA, is detailed.

Rola mikroorganizmów w chorobach cywilizacyjnych

• The role of microbes in the etiology of various diseases is explored (e.g., diarrhea resulting from antibiotic treatment, asthma, autism, tumors, tooth decay).

Metabolizm bakteryjny

• Bacteria require various substances for growth, such as sources of carbon, nitrogen, energy, and water.
• Essential elements for protein synthesis include carbon, oxygen, hydrogen, nitrogen, sulfur, and phosphorus.
• Ions like potassium, sodium, magnesium, calcium, chloride, and various trace elements are important for enzymatic processes.

Żelazo

• Bacteria releases siderophores in order to obtain iron (Fe), which is essential for cell function.
• In the human body, iron is mostly bound, and thus unavailable to bacteria.

Tlen (O2 gazowy)

• Some bacteria cannot grow in the presence of oxygen (obligate anaerobes), while others can grow both in oxygen-free and oxygen-rich conditions.

Autotrofy=litotrofy

• These bacteria can use inorganic compounds as a source of energy and carbon.

Heterotrofy=organotrofy

• These bacteria need organic compounds to survive.

Energia i metabolizm, katabolizm i anabolizm

• Cells need constant energy delivery.
• Energy is stored in the form of ATP (adenosine triphosphate) and GTP (guanosine triphosphate).
• Energy is obtained through the breakdown of organic compounds (carbohydrates, fats, proteins).
• The process involves catabolism, which breaks down molecules, and anabolism, which builds molecules.

Katabolizm białek, polisacharydów i lipidów

The breakdown of proteins, carbohydrates, and fats leads to produce glucose and pyruvate, which then enter the TCA cycle. Energy is produced in the form of ATP and NADH.

Glikoliza i fermentacja

• The Embden-Meyerhof-Parnasa pathway occurs under both aerobic and anaerobic conditions.
• In the breakdown of one glucose molecule, two ATP molecules, two NADH molecules and two molecules of pyruvate.
• Fermentation occurs without oxygen, and the end products depend on the type of organism.
• Lactic acid fermentation involves conversion of pyruvate to lactic acid, a process commonly used in food production, such as making yogurt and sauerkraut.

Cykl kwasów trójkarboksylowych (TCA)

• Pyruvate, created through glycolysis, goes through aerobic metabolism in the TCA cycle to produce a higher amount of energy in the form of CO2, NADH, FADH2, and GTP.

Fosforylacja substratowa

• GTP is produced through substrate level phosphorylation, and ATP is produced through the electron transport chain.
• In the electron transport chain, electrons are transported, along with protons (H+), creating an electrochemical gradient that powers the production of ATP.

Wynik energetyczny

• Organisms without oxygen are less efficient at producing energy compared to oxygen-dependent organisms.

Podsumowanie funkcji cyklu TCA

• The TCA cycle plays a major role in multiple catabolic and anabolic pathways simultaneously.
• The TCA cycle provides intermediates needed for protein, fat, nucleic acid synthesis.

Oddychane beztlenowe, c.d.

• Various substances can serve as final electron acceptors in the absence of oxygen (e.g., nitrates, sulfates, CO2, or iron).
• Anaerobic respiration occurs in environments with low or absent oxygen levels.
• Final electron acceptors in anaerobic respiration are generally inorganic compounds.

Inżynieria genetyczna - technologia rekombinowanego DNA

• The use of tools for purification, replication, and modification of genes, in order to create products of gene expression.
• The main tools in genetic engineering include vectors (for cloning and expression of DNA sequences), amplification & expression systems, restriction enzymes (cutting DNA at specific sites). ligases (joining DNA fragments).
• The plasmids, pBR322, pGEM and other vectors are used for cloning for fragment insertion of up to 20 kb.
• The use of plasmid vectors and phage DNA for cloning larger fragments up to 45kb.

Klonowanie DNA insertowego do wektorów

• DNA is isolated from cells or viruses.
• DNA fragments are amplified by PCR.
• DNA is cut by restriction enzymes and joined by DNA ligase with vectors to create recombinant DNA.
• Recombinant DNA is placed into bacterial cells.
• Successful incorporation is determined via selection based on markers (antibiotic resistance and other phenotypic markers).
• The resulting clones are screened or examined to determine the presence of the DNA fragment.

Biblioteka genomowa i cDNA

• A set of recombinant vectors generated by cloning fragments of the total DNA of an organism.
• It is used to provide a complete genomic representation of that organism
• cDNA is complementary DNA, derived from mRNA, and used to represent the genetic information involved in transcription.

Działania w ramach inżynierii genetycznej

• Isolation of segments of genetic material from cells.
• Modification of genetic information.
• Transfer (cloning) of DNA fragments into cells of another organism.
• Replication of genes and/or entire organisms.

Zastosowanie inżynierii genetycznej

• Production of large quantities of pure immunogens that may be used as vaccine
• Production of proteins such as insulin, interferons, and growth factors.

Grzybiczy element mikrobiomu - Mykobiom

• Fungi components of the human microbiome (Mycobiome).
• Abundance of fungi compared to bacteria.
• Fungi belonging to Ascomycota phylum.
• Examples: Candida, Saccharomyces, Aspergillus and Malassezia.

Królestwo Grzybów (Fungi)

• Fungi differ from other eukaryotic organisms by the presence of chitin in their cell walls, and the absence of cholesterol in their membranes.
• Classification is primarily based on morphology and spore production, with further phylogeny-based updates.
• The simplest morphological classification separates fungi into yeasts and molds.

Drożdże i pleśnie

• Yeasts that reproduce asexually or by budding form round, sticky, or slimy colonies on agar.
• Molds are multicellular organisms and form filaments (hyphae) that extend through or over the medium. Hyphae are often described as filamentous, hairy, or fluffy.

Zarodniki

• Spores are reproductive structures of fungi.
• Two main types of spores in fungi: sporangiospores and conidia.
• Sporangiospores are enclosed in sporangia, whereas conidia are not.
• Conidia are formed in various structures on the fungus.

Interakcje mikrobiota

• Interaction between Pseudomonas aeruginosa and Aspergillus fumigatus in lung infections.

Wszędobylskość grzybów

• Fungi are adaptable to various environmental conditions, including different temperatures, humidity, water content, and pressure.
• Fungi tolerate a wide range of pH values.

Inne cechy grzybów

• Most fungi use aerobic respiration, but some can also use anaerobic respiration.
• Fungi growth cycle is longer than that of bacteria.
• Some fungi produce secondary metabolites including acids, antibiotics, and toxins.

Stadium saprofityczne i pasożytnicze endemicznych grzybów dimorficznych

• Dimorphic fungi exhibit different morphological forms in various environmental conditions.

Identyfikacja grzybów i charakterystyka mykobiomów

• Advanced sequencing techniques can be utilized for microbiome analyses, obviating the limitations of previous methods (18S, 28S, and ITS).
• Shotgun sequencing is detailed as a method for obtaining more complete information compared to amplicon-based sequencing.

Wirusy i wirom jelitowy w IBD

• Viruses in the gut microbiome are classified into eukaryotic viruses and bacteriophages.
• Bacteriophages are more abundant than eukaryotic viruses in the human gut.
• Differences between IBD and healthy people in terms of virus composition are highlighted in the text.

Obecne ograniczenia i przyszłe kierunki badań na wiromami

• Viral identification and classification remain challenging due to the diverse genetic make-up and high mutation rate of viruses.
• Viral abundance in microbiome samples is low.
• Existing tools for culturing viruses are limited, making cultivation difficult.

Cechy, które charakteryzują i definują wirusy

• Viruses are not living organisms in the classic sense.
• Viruses require a host cell for replication and reproduction.
• Viruses have an external protein coat, called a capsid.
• Viruses carry genetic material within the capsid (either DNA or RNA).

Kapsyd i osłonka wirusa

• The capsid and envelope are protective structures surrounding the viral genome.
• The capsid is composed of protein subunits. An envelope surrounding the capsid in some viruses is derived from the host cell membrane.
• Envelopes provide viruses with additional protective features in diverse environments (such as those in the digestive system).

Genom wirusa

• The viral genome may consist of DNA or RNA, which can be either single or double-stranded, linear or circular.
• Positive or negative sense (polarity) is mentioned for RNA viruses.

Wirusy bezosłonkowe (posiadające tylko kapsyd)

• These viruses lack an external envelope.
• Examples of viruses include: Rabies, Tobacco Mosaic, and Bacteriophages.

Replikacja wirusów

• To replicate, viruses rely on the host cell’s machinery;
• The replication process is controlled by the virus's genomes;
• The process of viral reproduction is diverse.

Droga krwi i układu limfatycznego

• Viruses may spread through the bloodstream and lymphatic vessels.

Patogeneza chorób wirusowych

• Mechanisms of viral infection are presented in terms of productive infection, abortive infection, latent infection, and persistent infection.
• The disease process can manifest differently in different individuals.

Rozwój choroby wirusowej

• Viral entry, primary site infection, activation of innate immunity, latent period of viral replication spreading to other sites, host immune response (adaptive immunity), production of virions, release of the virus, transition to latency or resolution.

Prognozowane przyszłe kierunki badań

• Improvement and advancements in techniques for isolating and characterizing viral elements in microbiomes.
• Development and use of computing tools, to aid in the analysis of viral sequences.
• Expansion of databases that contain sequences from different virus strains/species will aid in identification.
• Development of culture methods for viruses, which would allow for better and more accurate study of viral growth patterns.

Quiz ten bada czynniki wpływające na częstość mutacji w komórkach bakterii oraz efekty korzystnych mutacji. Zajmujemy się również rolą plazmidów, ich zastosowaniem oraz wpływem mikroorganizmów na zdrowie ludzkie. Sprawdź swoją wiedzę na temat genetyki bakterii!