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ИИ помог обойти защиту резистентных бактерий: открыт новый класс перспективных антибиотиков

По прогнозам к 2050 году смертность от инфекционных и бактериальных заболеваний, не поддающихся лечению в связи с устойчивостью возбудителей к антибактериальным препаратам, составит 10 млн человек в год и выйдет на одно из лидирующих мест наряду с сердечно-сосудистыми и онкологическими заболеваниями. Основным объектом данного исследования как раз является один из таких типов бактерий – метициллинрезистентный золотистый стафилококк (MRSA), устойчивый к стандартным лекарственным препаратам, известным и применяемым на практике антибиотикам.

Инфицирование происходит просто и быстро, более того, такого рода заболевания относятся к внутрибольничным инфекциям или инфекциям, связанным с оказанием медицинской помощи (ИСМП), что также является насущной проблемой как для каждого из нас, так и для государства в целом.

Данной проблемой уже долгие годы занимаются ученые и медицинские организации по всему миру, и, наконец, с мертвой точки позволили сдвинуться силы искусственного интеллекта и глубокого обучения, основанного на применении графовых моделей нейронных сетей, знакомых для каждого из нас.

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Ученым из Массачусетского университета удалось осуществить данное исследование за счет использования глубокого обучения. Помимо того, что новый класс антибиотиков способен уничтожать MRSA бактерии, он также обладает очень низкой токсичностью по отношению к клеткам человека, что является безупречным результатом. Ключевым нововведением данного исследования является то, что ученые также смогли выяснить, какие виды информации использовала модель глубокого обучения для прогнозирования эффективности антибиотиков. Эти знания необходимы для дальнейшей разработки дополнительных препаратов, которые могли бы работать даже лучше, чем те, которые были определены моделью.

В данной статье я подробнее расскажу о методах и ходе данного исследования, от зарождения идеи, до ее реализации и практических результатов.

Приятного прочтения! :)

Путь от идеи до реализации исследования

Основной идеей исследователей являлось изучение химических пространств.

Химическое пространство – пространство потенциальных фармакологически активных молекул; его размер оценивается порядка 1060 молекул. Исследование химического пространства возможно путем создания “in silico” (или, проще говоря, “с помощью компьютерного моделирования (симуляции)”) баз данных виртуальных молекул, которые можно визуализировать, проецируя многомерное пространство свойств молекул в меньших измерениях.

Для исследования химических пространств используются подходы машинного обучения. Базовой моделью такого обучения является модель “черного ящика”, которая не обеспечивает понимания всех происходящих химических процессов в данном случае. Общий принцип заключается в том, что на выходе мы получаем готовый результат или принятое решение, однако используемые внутренние процессы и различные взвешенные факторы остаются неизвестными.

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Однако этого было недостаточно, т.к. понимание выборки конкретных химических соединений играло ключевую роль в прогнозировании дальнейших более эффективных структурных классов антибиотиков.

Исследователи разработали объяснимый, основанный на субструктурах подход для рационального исследования химических пространств с помощью глубокого обучения.

Субструктуры относятся к различным компонентам, модулям или слоям моделей глубокого обучения. Они представляют собой части модели, которые обрабатывают и извлекают информацию из входных данных. Субструктуры могут быть разными в зависимости от типа модели, но обычно они включают слои нейронной сети.

Кроме того, они обеспечивают "запоминание" важных шаблонов или связей в данных, что в конечном итоге способствует лучшим результатам прогнозирования или классификации. Это свойство субструктур и является основным в реализации исследования (для непосредственного понимания химических процессов).

Далее, благодаря использованию ансамблей графовых нейронных сетей, удалось предсказать антибиотическую активность и цитотоксичность для 12,076,365 соединений.

Ансамбль графовых нейронных сетей представляет собой комбинацию нескольких графовых нейронных сетей для решения задач анализа графов. Ансамбль графовых нейронных сетей объединяет прогнозы и предсказательные способности отдельных моделей графовых нейронных сетей для получения более надежных и точных результатов. Каждая модель в ансамбле может быть обучена на разных частях графов или с использованием различных гиперпараметров, что приводит к разнообразию в их предсказаниях.

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Таким образом, используя алгоритмы объясняемых графов, удалось на основании субструктур определить необходимые обоснования для выявленных соединений.

После получения обоснований итоговых результатов, эмпирически были протестированы 283 соединения. Обнаружили, что соединения, проявляющие антибиотическую активность, были обогащены предполагаемыми структурными классами, вытекающими из полученных обоснований.

В итоге, из этих структурных классов соединений один является селективным в отношении MRSA и ванкомицин-резистентных энтерококков, избегает значительной резистентности и снижает титры бактерий в мышиных моделях кожной и системной инфекции бедра, вызванной MRSA.

Теперь более детально рассмотрим особенности исследования!

Модели антибиотической активности

Был проверен исходный набор из 39 312 соединений, содержащий большинство известных антибиотиков, природных продуктов и структурно разнообразных молекул с молекулярными массами от 40 до 4200 Da, на ингибирующую рост активность против чувствительного к метициллину штамма S. aureus RN4220.

На графике отображается молекулярно-массовое распределение отобранных соединений. Частота показана в логарифмической шкале.
На графике отображается молекулярно-массовое распределение отобранных соединений. Частота показана в логарифмической шкале.

Исследование проводилось в течение ночи в богатой питательными веществами среде в конечной концентрации 50 мкМ, и их действие было бинаризировано как активное или неактивное при условии 80 % нормализованного подавления роста бактерии (начиная с 80% – активное), в результате чего было получено 512 активных соединений (1,3 % всех соединений).

Непосредственно данные по ингибированию роста S. aureus RN4220, полученные в результате анализа при конечной концентрации 50 мкМ. Данные получены в двух биологических репликах. Активные соединения – те, для которых средний относительный рост составляет менее 0,2.
Непосредственно данные по ингибированию роста S. aureus RN4220, полученные в результате анализа при конечной концентрации 50 мкМ. Данные получены в двух биологических репликах. Активные соединения – те, для которых средний относительный рост составляет менее 0,2.

Далее исследователи использовали Chemprop.

Chemprop - это метод машинного обучения, специально разработанный для задач химинформатики, в особенности для предсказания биологической активности новых соединений, поиска лекарственных препаратов, ранжирования молекул по их свойствам, генерации новых молекул и других задач, связанных с химической разработкой и оптимизацией.

Исследователи обучили ансамбли графовых нейронных сетей осуществлять бинарные классификационные прогнозы о том, будет ли новое соединение подавлять рост бактерий, на основе его химической структуры. Каждая графовая нейронная сеть работает путем формирования шагов свертки, которые зависят от атомов и связей каждой входной химической структуры, которая рассматривается как математический граф с вершинами (атомы) и ребрами (связи).

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После последовательных шагов свертки, объединяющих информацию от соседних атомов и связей, каждая модель генерирует окончательный балл предсказания между 0 и 1, представляющий собой оценку вероятности того, что молекула активна.

Для получения дополнительных данных, которые могут улучшить работу модели, каждой был предоставлен список молекулярных характеристик, рассчитанных с помощью RDKit для каждого входа.

RDKit – это набор программного обеспечения для химинформатики и машинного обучения, написанного на C++ и Python. RDKit позволяет выполнять различные операции над молекулами, включая генерацию, представление и редактирование молекулярных структур, расчет физико-химических свойств, числовых характеристик, описывающих молекулу в химической системе, визуализацию молекулярных структур, анализ и поиск структурных фрагментов, разработку моделей и тд.

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Для повышения надежности прогноза оценки нескольких моделей в ансамбле усреднялись. Каждая модель обучалась и проверялась, а затем тестировалась на одних и тех же фрагментах обучающего набора данных. Для ансамбля из десяти моделей, примененных к закрытым тестовым данным, площадь под кривой AUPRC составила 0,364, что свидетельствует о хорошей производительности при учете дисбаланса активных соединений в обучающих данных.

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Кривая AUPRC (Area Under the Precision-Recall Curve) - это графическое представление и метрика для оценки качества моделей классификации или ранжирования, особенно в задачах с несбалансированными классами или большим количеством ложноположительных результатов. Кривая AUPRC строится на основе двух основных метрик: precision (точность) и recall (полнота). Precision измеряет долю верно классифицированных положительных примеров относительно всех классифицированных положительных примеров. Recall измеряет долю верно классифицированных положительных примеров относительно всех действительно положительных примеров. Чем ближе AUPRC к значению 1, тем лучше модель способна разделять классы.

При этом, наблюдалось снижение производительности, измеряемой по AUPRC для тестового набора у альтернативных моделей, включая ансамбль из десяти моделей Chemprop без использования RDKit.

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В целом, эти результаты показывают, что модели Chemprop с молекулярными характеристиками, вычисленными с помощью RDKit, дают многообещающие прогнозы активности антибиотиков и могут превзойти более простые и поверхностные модели глубокого обучения.

Модели цитотоксичности человеческих клеток

Чтобы лучше определить соединения, селективные против изучаемых бактерий, были разработаны ортогональные модели, предсказывающие цитотоксичность в клетках человека.

Ортогональные модели являются специальным видом моделей, используемых в математике, статистике и анализе данных. Они характеризуются свойством ортогональности, что означает независимость и неперекрываемость компонентов модели. В контексте многомерного пространства, ортогональные модели связаны с ортогональными базисами, где каждая компонента (измерение) модели независима от остальных, и изменение одной компоненты не влияет на другие. Это обеспечивает более простой анализ данных и интерпретацию результатов.

Был проведен контрскрининг обучающего набора из 39,312 соединений на цитотоксичность в клетках карциномы печени человека (HepG2), клетках скелетных мышц человека (HSkMCs) и клетках фибробластов легких человека (IMR-90).

Клетки HepG2 обычно используются для изучения гепатотоксичности и общей цитотоксичности, в то время как клетки HSkMCs и IMR-90 могут лучше моделировать токсичность ”in vivo” (внутри клетки/живого организма), чем бессмертные клеточные линии (это клетки, которые способны продолжать делиться (продолжать жизненный цикл) бесконечно длительное время, в отличие от обычных клеток, которые имеют ограниченное количество делений и в конечном итоге умирают).

Жизнеспособность клеток измеряли после 2-3 дней обработки каждым соединением в концентрации 10 мкМ, которая соответствует и широко используется в культурах клеток человека. Затем активность соединений бинаризировали, используя строгий пороговый уровень жизнеспособности клеток в 90%, в результате чего в общей сложности 3341 (8,5%), 1490 (3,8%) и 3447 (8,8%) соединений были классифицированы как цитотоксичные для клеток HepG2, HSkMCs и IMR-90, соответственно, а из 512 активных антибактериальных соединений 306 оказались нецитотоксичными для всех трех типов клеток.

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Как и выше, эти данные были использованы для обучения бинарных классификационных моделей, предсказывающих вероятность того, является ли новое соединение цитотоксичным для клеток HepG2, HSkMCs или IMR-90, на основе химической структуры соединения. Для ансамблей из 10 моделей Chemprop, обученных и проверенных, а затем протестированных на тех же данных, значения AUPRC для моделей составили 0,176, 0,168 и 0,335, соответственно.

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Это свидетельствует о положительной, но менее предсказуемой эффективности, чем у моделей для антибиотической активности, что может быть обусловлено более строгими критериями для объявления соединений нецитотоксичными.

Фильтрация и визуализация химического пространства

После удовлетворения эффективностью наших моделей, исследователи переобучили ансамбли из 20 моделей Chemprop на всех обучающих наборах данных, в результате чего были получены четыре ансамбля, предсказывающие активность антибиотиков, цитотоксичность HepG2, цитотоксичность HSkMCs и цитотоксичность IMR-90. Ансамбли были применены для прогнозирования антибиотической активности и цитотоксичности 12 076 365 соединений, включая 11 277 225 соединений из базы данных Mcule purchasable, в которой большинство соединений можно легко приобрести, не прибегая к собственному химическому синтезу, а также 799 140 соединений из базы данных Broad Institute.

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Соединения с высокими (>0,4 для базы данных Mcule и >0,2 для базы данных Broad Institute) баллами прогноза антибиотической активности сохраняются, и аналогичные графовые нейронные сети применяются для прогнозирования цитотоксичности этих соединений для клеток HepG2, HSkMCs и клетки IMR-90. Соединения с низкими (<0,2) баллами прогноза цитотоксичности для всех типов клеток отбираются, затем вычислительным путем проверяются на наличие неблагоприятных химических подструктур (PAINS и Brenk substructures).

PAINS (Pan-Assay Interference Compounds) и Brenk substructures – это наборы подструктур, которые могут вызывать нежелательные эффекты или ложноположительные результаты в биологических экспериментах. PAINS является набором химических структурных фрагментов, которые часто обнаруживаются в молекулах, проявляющих побочную активность. Brenk substructures представляют собой схожий набор подструктур, разработанный для выявления химических групп, которые часто связаны с нежелательными свойствами молекул, такими как токсичность, незаурядность или проблемы с разработкой лекарств.

Наконец, оставшиеся соединения фильтруются на предмет структурной новизны, определяемой коэффициентом сходства Танимото – менее 0,5 по отношению к любому активному соединению в обучающем наборе данных.

Коэффициент Танимото является метрикой, используемой для оценки сходства между двумя множествами или наборами данных (преимущественно для молекул и генов). Коэффициент Танимото вычисляется как отношение числа общих элементов между двумя наборами к общему числу элементов в обоих наборах. Он может принимать значения в диапазоне от 0 до 1, где 0 означает полное отсутствие сходства, а 1 обозначает полное совпадение.

Формула для вычисления коэффициента Танимото:

T(A, B) = (A ∩ B) / (|A| + |B| - A ∩ B)

На основе полученных данных исследователи визуализировали химическое пространство, используя метод t-SNE.

t-SNE (t-distributed stochastic neighbor embedding) - это алгоритм машинного обучения, используемый для визуализации высокоразмерных данных в низкоразмерное пространство. Он позволяет отображать сложные многомерные данные на двух- или трехмерную плоскость, сохраняя при этом важные отношения и структуру в данных. Алгоритм t-SNE основывается на концепции близости объектов: близкие объекты в исходном пространстве имеют более высокие шансы быть близкими и после проекции на низкоразмерное пространство.

Диаграмма с использованием алгоритма t-SNE (t-distributed neighbor embedding) показывает соединения с высокими и низкими оценками прогноза антибиотиков, а также соединения из обучающего набора данных при разных пороговых значениях оценки прогноза. Данная диаграмма позволяет визуализировать химическую схожесть или различие представленных соединений. Активные соединения из обучающего набора данных (красные точки) в основном разделяют соединения с высокими оценками прогнозов (зеленые, черные и фиолетовые точки) от соединений с низкими оценками прогнозов (коричневые точки)
Диаграмма с использованием алгоритма t-SNE (t-distributed neighbor embedding) показывает соединения с высокими и низкими оценками прогноза антибиотиков, а также соединения из обучающего набора данных при разных пороговых значениях оценки прогноза. Данная диаграмма позволяет визуализировать химическую схожесть или различие представленных соединений. Активные соединения из обучающего набора данных (красные точки) в основном разделяют соединения с высокими оценками прогнозов (зеленые, черные и фиолетовые точки) от соединений с низкими оценками прогнозов (коричневые точки)

Особое замечание: как указано в t-SNE, несколько найденных веществ структурно отличаются от активных соединений в обучающем наборе, что указывает на то, что разработанные модели обобщаются на невидимые химические пространства.

Обоснование прогнозов классов антибиотиков

Так как графовые нейронные сети делают прогнозы на основе информации, содержащейся в атомах и связях каждой молекулы, исследователи предположили, что соединения с высокими оценками прогноза антибиотической активности содержат подструктуры («обоснования»), которые в значительной степени определяют их оценки.

Выявление таких обоснований обеспечило бы гарантии объяснимости модели для интересующих нас результатов. Оценка прогноза антибиотической активности для любого найденного вещества могла бы быть прямо отнесена к его обоснованию, так, что обоснование (или же рационал), рассматриваемое самостоятельно как молекулярный вход в Chemprop, сразу бы обладало высокой оценкой прогноза антибиотической активности. Возможность классификации таких обоснований сделала бы прогнозы Chemprop более понятными для людей и позволила бы последующий анализ подструктур с помощью машинного обучения.

Учитывая обученные модели Chemprop, удалось вычислить такие обоснования с помощью алгоритмов поиска на основе графов. Эти алгоритмы позволили определить в контексте одной молекулы наименьшее обоснование с заданным пороговым числом атомов, имеющее положительное предсказывающее значение.

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Исследователи стремились найти обоснования, содержащие не менее восьми атомов и проявляющие высокие предсказательные оценки антибиотической активности, превышающие 0,1, с использованием поисковых деревьев методом Монте-Карло.

Поиск методом Монте-Карло включает выбор начальной подструктуры, итеративное обрезание подструктуры и выбор удалений, приводящих к высоким оценкам предсказания, когда подграфы передаются в качестве входных данных в Chemprop.

Итеративное обрезание подструктуры – это метод оптимизации моделей глубокого обучения путем последовательного удаления нейронов или связей с низкой значимостью. В начале процесса все нейроны или связи обучаются на тренировочных данных. Затем производится оценка значимости каждого нейрона или связи - например, на основе их вклада в ошибку модели или значения градиентов. На основе этих оценок выбираются нейроны/связи с наименьшей значимостью и удаляются. После удаления происходит повторное обучение модели, чтобы компенсировать потерю ее емкости. Этот процесс повторяется, пока не достигнута желаемая эффективность модели.

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Примеры расчетов обоснования с использованием поиска по дереву Монте-Карло (Monte-Carlo Tree Search, MCTS):

a) Иллюстрация прямого прохода MCTS с использованием соединения 1. На рисунке показаны три возможных пути поиска от корня путем удаления периферийных связей или колец (выделены оранжевым). Из-за ограничений места показаны только три шага от корня.

b) Иллюстрация полного поискового пути от корня до листового узла (обоснование). Для предсказания активности каждого листового узла используется Chemprop, и эти предсказания используются для обновления статистики каждого промежуточного узла при обратном проходе.

Этот графический поиск выводит обоснование, объясняющее пороговое значение (не менее 0,1) предсказательной оценки соединения, если оно сходится; в противном случае, если обоснование не найдено, то интересующий нас случай нельзя объяснить таким образом.

Применяя этот анализ обоснований к отфильтрованным результатам полной модели, удалось вычислить обоснования для 380 из 3 646 выбранных соединений (10,4%). Как и ожидалось, многие рационалы совпали с известными фрагментами структурных классов. Однако также обнаружили обоснования, которые не были связаны ни с одним известным классом антибиотиков. Поэтому цель заключалась в лучшей фильтрации структурно новых интересных соединений и исследовании их соответствующих обоснований.

Новые фильтрованные подструктуры

Основываясь на выявлении известных классов антибиотиков в результате проведенных анализов, и способности графовых обоснований (то же, что и рационалы) предсказывать подструктуры, диагностирующие высокие показатели антибиотической активности, исследователи попытались выявить структурно новые классы антибиотиков, предсказанные обученными моделями.

Чтобы рассмотреть химические структуры с благоприятными свойствами для медицинской химии, исследователи удалили все найденные подструктуры, содержащие PAINS и Brenk-предупреждения, которые относятся к подструктурам, которые могут проявлять неспецифичную реакцию, мутагенность или иметь неблагоприятную фармакокинетику. Это уменьшило количество предсказанных вариантов с 3 646 до 2 209.

Далее исследователи сосредоточились на поиске соединений, не похожих на те, что были в обучающем наборе. Вычислили максимальное сходство по Танимото каждого соединения с любым активным соединением из обучающего набора и отобрали только с максимальным сходством ≤0,5 в качестве предварительной оценки.

В результате был получен окончательный набор из 1 261 совпадения, из которых 162 были из базы данных Mcule и 1 099 - из базы данных Broad Institute. При расчете обоснований для этого более узкого набора совпадений выяснилось, что 186 совпадений (14,8 %) имеют необходимые обоснования.

Теперь все исследования будут сосредоточены вокруг этих 186 совпадений.

Для того чтобы использовать эти рационалы для ясного прогнозирования структурных классов, предположили, что изучение химических основ, общих для рассматриваемых обоснований, позволит выделить наиболее значимые прогнозы структурных классов. Это особенно полезно для уменьшения объема выборки, поскольку типичные рационалы содержат большое количество атомов (более 17) и отличаются друг от друга незначительными модификациями. Были вычислены химические основы с хотя бы 12 атомами, которые сохранялись во всех обоснованиях.

С использованием этого подхода обнаружили, что 16 из 186 вариантов с нужными обоснованиями (8,6%) могут быть объединены с использованием 5 различных основ – G1-G5 – каждая группа содержит как минимум 2 варианта с соответствующими обоснованиями. Следует отметить, что три из пяти основ содержат хлор, что свидетельствует о том, что модели рассматривают присутствие атома хлора в этих химических контекстах как важный фактор, влияющий на антибиотическую активность.

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Здесь 16 соединений с необходимыми обоснованиями были связаны с пятью каркасами G1-G5.

Поскольку количество совпадений, оставшихся после фильтрации и анализа, было достаточно большим, исследователи напрямую проверили предсказания модели, получив девять совпадений, связанных с обоснованиями в группах G1-G5. Для сравнения также получили 45 совпадений (из отфильтрованных 1 261) с вычисленными обоснованиями, не связанными с G1-G5, 187 совпадений (из отфильтрованных 1 261) без вычисленных обоснований и 30 структурно несхожих соединений с низкими (менее 0,1) баллами предсказания. В результате такого подхода был получен набор из 283 соединений, которые экспериментально проверили.

Ранжированные оценки антибактериальной активности для 253 соединений с высокими (>0,2) оценками антибиотической активности и 30 соединений с низкими (<0,1) оценками антибиотической активности были запрошены для экспериментального тестирования.
Ранжированные оценки антибактериальной активности для 253 соединений с высокими (>0,2) оценками антибиотической активности и 30 соединений с низкими (<0,1) оценками антибиотической активности были запрошены для экспериментального тестирования.

Структурный класс антибиотиков из рационалов

При тестировании на ингибирование роста исследователи обнаружили, что 4 из 9 закупленных соединений (44%), связанных с группами G1-G5, проявляют активность против S. aureus с минимальными ингибирующими концентрациями (МИК) ≤ 32мкг/мл-1.

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Ни одно из 45 закупленных соединений с рационалами, не связанными с G1-G5, и 17 из 187 закупленных соединений без рационала (9,1%) не проявляли активности. Рабочие верные уровни обнаружения, связанные с всеми испытанными структурно новыми соединениями с рационалами (7,4%) и по всем испытанным структурно новым соединениям (8,7%), были выше доли активных соединений в обучающем наборе (1,3%), что указывает на полезность данного подхода. Эти значения указывают на то, что усилия по испытанию соединений могут быть так же продуктивными, как испытание случайных находок, когда они сосредоточены на структурных классах, предсказанных моделями глубокого обучения. Кроме того, в соответствии с низким уровнем ложного опускания для модели, ни одно из 30 закупленных соединений с низкими оценками предсказания не ингибировало рост S. aureus.

В итоге, из четырех найденных активных соединений, проявляющих активность против S. aureus и связанных с G1–G5, ни одно из них ранее не изучалось по отношению к рассматриваемым патогенам, и вместе они связаны с тремя группами рационалов — G1, G2 и G5. Стоит отметить, что G2 ассоциирована с двумя подтвержденными (активными) находками, что указывает на то, что эта группа рационалов может представлять активный структурный класс.

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*тут важно сконцентрировать внимание на соединениях 1 и 2, связанных с G2.

Дополнительные свойства, включая физико-химические наблюдения О'Ши и Мозера для антибиотиков, могут еще больше сузить химическое пространство и послужить основой для последующей разработки, особенно при рассмотрении кандидатов из больших библиотек. Однако исследователи не фильтровали находки на основе этих или других физико-химических свойств, при этом отмечается, что подтвержденные находки были меньше и менее полярны, чем типичные антибиотики для грамположительных бактерий.

Проведение дополнительных экспериментов по ингибированию роста показало, что соединения 1 и 2, а также почти все другие структурно новые подтвержденные находки также обладали активностью против MRSA USA300, сравнимым с его метициллиночувствительными аналогами. При контрсортировке их на цитотоксичность по отношению к клеткам HepG2, HSkMCs и IMR-90 мы обнаружили, что 20 из 21 структурно новых подтвержденных находок были нецитотоксичны при концентрации 10 мкМ. Соединения 1 и 2 проявляли значение полумаксимальной ингибирующей концентрации (IC50) ≥ 128 мкг/мл-1 для всех типов клеток, что указывает на прочную селективность против S. aureus.

В качестве окончательного эмпирического фильтра исследователи измерили МИК (минимальную ингибирующую концентрацию) S. aureus подтвержденных находок, связанных с G1-G5, в среде с добавлением 10% фетальной телячьей сыворотки в качестве контроля связывания соединений с белками сыворотки. Обнаружили, что МИК соединений 1 и 2 возросли в 4-8 раз до 16 мкг/мл-1, но оставались существенно ниже их значений IC50 для клеток человека. В отличие от этого, МИК других двух соединений увеличилась до ≥64 мкг/мл-1 в сыворотке. Вместе с их выгодными значениями МИК в среде без сыворотки (≥64-кратно меньше их значений IC50 для клеток человека), эти наблюдения указывали на то, что соединения 1 и 2 были наиболее селективными из всех подтвержденных находок и требовали дальнейшего изучения.

Механизм действия и резистентность

Для дальнейшего изучения отобранных соединений ученые дополнительно исследовали механизмы действия этих соединений с помощью эволюции устойчивых мутантов. Последовательно пассировали S. aureus RN4220, обработанный каждым из соединений 1 и 2 в жидкой культуре, и обнаружили, что их МИК оставались практически неизменными через 30 дней. Кроме того, исследователи высевали S. aureus RN4220 с высокой инокуляцией на твердую среду в присутствии сверхмикроскопических уровней соединений 1 и 2 и обнаружили, что через 5 дней колонии росли при 4×, но не 8×, что свидетельствует о низком уровне резистентности (устойчивости; частота резистентности при 4× МИК, ~10-8).

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Для дальнейшего изучения фенотипических эффектов (наблюдаемых свойств или характеристик) соединений 1 и 2 исследователи объединили микроскопические наблюдения с измерениями клеточной физиологии.

Сначала провели визуализацию одноклеточных; здесь исследователи сосредоточились на B. subtilis, чья палочковидная форма демонстрирует более заметные морфологические изменения, чем S. aureus. Одноклеточная визуализация показала, что клетки, обработанные соединением 1 или 2 лизировались (клетки разрушились, их мембраны разорвались или проникли вещества, приводящие к гибели клеток), что согласуется с бактерицидной активностью этих соединений и свидетельствует о механизме действия, направленном на клеточную оболочку.

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Кроме того, ученые обнаружили, что оба соединения были активны против 40 изолятов различных видов бактерий, выделенных Центром по контролю и профилактике заболеваний США (CDC), с различными факторами устойчивости, включая устойчивость к ванкомицину, аминогликозидам/тетрациклинам и оксазолидинону.

Следует отметить, что оба соединения были активны в отношении ванкомицинрезистентных энтерококков (VRE), представляющих серьезную угрозу устойчивости к антимикробным препаратам. Более того, эксперименты по уничтожению бактерий по времени показали, что оба соединения были эффективны против персистирующих (длительно существующих в организме хозяина) B. subtilis, что приводило к уничтожению культуры в лог-фазе после обработки канамицином. Эти результаты свидетельствуют о том, что соединения 1 и 2 способны преодолевать общие детерминанты устойчивости и толерантности к антибиотикам у грамположительных бактерий.

Свойства и эффективность in vivo

Исследователи обнаружили, что соединения 1 и 2 не являются гемолитическими, не хелатируют железо, не генотоксичны, химически стабильны в растворах с различным pH и нетоксичны при местном нанесении (1%) на кожу человека и внутрибрюшинном введении (80 мг кг−1 ) мышам.

Вывод

Основная цель исследования была достигнута – удалось выявить структурный класс антибиотиков, эффективный как при местном, так и при системном лечении MRSA в мышиных моделях инфекции.

Кроме того, эта работа представляет подход глубокого обучения, который систематически основывается на прогнозах отдельных составных соединений и позволяет эффективно исследовать огромные химические пространства на основе подструктур. В дополнение к уменьшению основного химического пространства, полезной особенностью данного подхода является возможность автоматизированного определения структурных мотивов. Эта возможность представляет собой источник химической новизны, который может предложить химические пространства для исследования и продуктивно дополнить существующие процессы открытия препаратов, например, путем генерации интересных химических фрагментов.

Исследователи предполагают, что лучшее понимание предсказаний на основе графовой обоснованности может помочь в открытии и проектировании дополнительных, крайне необходимых классов антибиотиков, например, таких, которые активны против грамотрицательных бактерий, а также классов препаратов, нацеленных на другие биологические процессы и заболевания, включая антивирусные и противораковые лекарства.

Таким образом, идея и необходимость ухода от “черного ящика” стала движущей силой для новых проектов в области биоинформатики.

Для меня исключительной важностью обладает внимание ученых, работающих с ИИ и моделями глубокого обучения, к насущным проблемам медицины и здравоохранения. Во многих вопросах мы оказываемся слишком бессильны, применяя стандартные методы исследования. Такие проекты помогают находить инновационные решения, без которых дальнейшая работа над решением проблем попросту невозможна или займет непростительно много времени. Можно с уверенностью сказать, что искусственный интеллект на страже нового здорового поколения! :)

На этом всё!

Будем ждать Вас в комментариях! Спасибо за прочтение :)

Автор: Черенковская Анастасия

Источник

  • 22.06.26 21:51 kimberlyhebert786

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  • 24.06.26 01:25 Fraddy Pual

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    I never thought it would happen to me—but I lost $256,100 in Bitcoin through a shady investment deal. I was shattered, panicked, and convinced my savings were gone forever. Just when I was about to give up, I stumbled upon reviews for FUNDSRETRIEVER, a cyber recovery team with a solid reputation. I decided to give it a shot, and to my absolute shock, they recovered every single cent in record time. Working with them was a lifesaver. If you've been tricked by fake investment platforms, don't lose hope—FUNDSRETRIEVER can help. Contact them here: Email: [email protected] | WhatsApp: +16035121448 | Telegram: @Fundsretriever

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    I never thought it would happen to me—but I lost $256,100 in Bitcoin through a shady investment deal. I was shattered, panicked, and convinced my savings were gone forever. Just when I was about to give up, I stumbled upon reviews for FUNDSRETRIEVER, a cyber recovery team with a solid reputation. I decided to give it a shot, and to my absolute shock, they recovered every single cent in record time. Working with them was a lifesaver. If you've been tricked by fake investment platforms, don't lose hope—FUNDSRETRIEVER can help. Contact them here: Email: [email protected] | WhatsApp: +16035121448 | Telegram: @Fundsretriever.

  • 24.06.26 01:58 robertalfred175

    CRYPTO SCAM RECOVERY SUCCESSFUL – A TESTIMONIAL OF LOST PASSWORD TO YOUR DIGITAL WALLET BACK. My name is Robert Alfred, Am from Australia. I’m sharing my experience in the hope that it helps others who have been victims of crypto scams. A few months ago, I fell victim to a fraudulent crypto investment scheme linked to a broker company. I had invested heavily during a time when Bitcoin prices were rising, thinking it was a good opportunity. Unfortunately, I was scammed out of $120,000 AUD and the broker denied me access to my digital wallet and assets. It was a devastating experience that caused many sleepless nights. Crypto scams are increasingly common and often involve fake trading platforms, phishing attacks, and misleading investment opportunities. In my desperation, a friend from the crypto community recommended Capital Crypto Recovery Service, known for helping victims recover lost or stolen funds. After doing some research and reading multiple positive reviews, I reached out to Capital Crypto Recovery. I provided all the necessary information—wallet addresses, transaction history, and communication logs. Their expert team responded immediately and began investigating. Using advanced blockchain tracking techniques, they were able to trace the stolen Dogecoin, identify the scammer’s wallet, and coordinate with relevant authorities to freeze the funds before they could be moved. Incredibly, within 24 hours, Capital Crypto Recovery successfully recovered the majority of my stolen crypto assets. I was beyond relieved and truly grateful. Their professionalism, transparency, and constant communication throughout the process gave me hope during a very difficult time. If you’ve been a victim of a crypto scam, I highly recommend them with full confidence contacting: Email: [email protected] Telegram: @Capitalcryptorecover Contact: [email protected] Call/Text: +1 (336) 390-6684 Website: https://recovercapital.wixsite.com/capital-crypto-rec-1

  • 24.06.26 01:58 robertalfred175

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  • 24.06.26 14:16 Universina da Mota

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  • 24.06.26 22:01 robertalfred175

    CRYPTO SCAM RECOVERY SUCCESSFUL – A TESTIMONIAL OF LOST PASSWORD TO YOUR DIGITAL WALLET BACK. My name is Robert Alfred, Am from Australia. I’m sharing my experience in the hope that it helps others who have been victims of crypto scams. A few months ago, I fell victim to a fraudulent crypto investment scheme linked to a broker company. I had invested heavily during a time when Bitcoin prices were rising, thinking it was a good opportunity. Unfortunately, I was scammed out of $120,000 AUD and the broker denied me access to my digital wallet and assets. It was a devastating experience that caused many sleepless nights. Crypto scams are increasingly common and often involve fake trading platforms, phishing attacks, and misleading investment opportunities. In my desperation, a friend from the crypto community recommended Capital Crypto Recovery Service, known for helping victims recover lost or stolen funds. After doing some research and reading multiple positive reviews, I reached out to Capital Crypto Recovery. I provided all the necessary information—wallet addresses, transaction history, and communication logs. Their expert team responded immediately and began investigating. Using advanced blockchain tracking techniques, they were able to trace the stolen Dogecoin, identify the scammer’s wallet, and coordinate with relevant authorities to freeze the funds before they could be moved. Incredibly, within 24 hours, Capital Crypto Recovery successfully recovered the majority of my stolen crypto assets. I was beyond relieved and truly grateful. Their professionalism, transparency, and constant communication throughout the process gave me hope during a very difficult time. If you’ve been a victim of a crypto scam, I highly recommend them with full confidence contacting: Email: [email protected] Telegram: @Capitalcryptorecover Contact: [email protected] Call/Text: +1 (336) 390-6684 Website: https://recovercapital.wixsite.com/capital-crypto-rec-1

  • 24.06.26 22:01 robertalfred175

    CRYPTO SCAM RECOVERY SUCCESSFUL – A TESTIMONIAL OF LOST PASSWORD TO YOUR DIGITAL WALLET BACK. My name is Robert Alfred, Am from Australia. I’m sharing my experience in the hope that it helps others who have been victims of crypto scams. A few months ago, I fell victim to a fraudulent crypto investment scheme linked to a broker company. I had invested heavily during a time when Bitcoin prices were rising, thinking it was a good opportunity. Unfortunately, I was scammed out of $120,000 AUD and the broker denied me access to my digital wallet and assets. It was a devastating experience that caused many sleepless nights. Crypto scams are increasingly common and often involve fake trading platforms, phishing attacks, and misleading investment opportunities. In my desperation, a friend from the crypto community recommended Capital Crypto Recovery Service, known for helping victims recover lost or stolen funds. After doing some research and reading multiple positive reviews, I reached out to Capital Crypto Recovery. I provided all the necessary information—wallet addresses, transaction history, and communication logs. Their expert team responded immediately and began investigating. Using advanced blockchain tracking techniques, they were able to trace the stolen Dogecoin, identify the scammer’s wallet, and coordinate with relevant authorities to freeze the funds before they could be moved. Incredibly, within 24 hours, Capital Crypto Recovery successfully recovered the majority of my stolen crypto assets. I was beyond relieved and truly grateful. Their professionalism, transparency, and constant communication throughout the process gave me hope during a very difficult time. If you’ve been a victim of a crypto scam, I highly recommend them with full confidence contacting: Email: [email protected] Telegram: @Capitalcryptorecover Contact: [email protected] Call/Text: +1 (336) 390-6684 Website: https://recovercapital.wixsite.com/capital-crypto-rec-1

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  • 25.06.26 21:25 Emilie Safi

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  • 26.06.26 01:04 robertalfred175

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  • 26.06.26 01:04 robertalfred175

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  • 06.07.26 16:20 Olga Ognjanović

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  • 06.07.26 16:31 Joseph Weigl

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  • 06.07.26 16:33 Jaran Løvlien

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  • 07.07.26 18:00 robertalfred175

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  • 07.07.26 18:01 robertalfred175

    CRYPTO SCAM RECOVERY SUCCESSFUL – A TESTIMONIAL OF LOST PASSWORD TO YOUR DIGITAL WALLET BACK. My name is Robert Alfred, Am from Australia. I’m sharing my experience in the hope that it helps others who have been victims of crypto scams. A few months ago, I fell victim to a fraudulent crypto investment scheme linked to a broker company. I had invested heavily during a time when Bitcoin prices were rising, thinking it was a good opportunity. Unfortunately, I was scammed out of $120,000 AUD and the broker denied me access to my digital wallet and assets. It was a devastating experience that caused many sleepless nights. Crypto scams are increasingly common and often involve fake trading platforms, phishing attacks, and misleading investment opportunities. In my desperation, a friend from the crypto community recommended Capital Crypto Recovery Service, known for helping victims recover lost or stolen funds. After doing some research and reading multiple positive reviews, I reached out to Capital Crypto Recovery. I provided all the necessary information—wallet addresses, transaction history, and communication logs. Their expert team responded immediately and began investigating. Using advanced blockchain tracking techniques, they were able to trace the stolen Dogecoin, identify the scammer’s wallet, and coordinate with relevant authorities to freeze the funds before they could be moved. Incredibly, within 24 hours, Capital Crypto Recovery successfully recovered the majority of my stolen crypto assets. I was beyond relieved and truly grateful. Their professionalism, transparency, and constant communication throughout the process gave me hope during a very difficult time. If you’ve been a victim of a crypto scam, I highly recommend them with full confidence contacting: Email: [email protected] Telegram: @Capitalcryptorecover Contact: [email protected] Call/Text: +1 (336) 390-6684 Website: https://recovercapital.wixsite.com/capital-crypto-rec-1

  • 09.07.26 19:06 Toivo Walli

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  • 09.07.26 19:10 Misty Alexander

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  • 12.07.26 03:30 Kora Baltacha

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  • 12.07.26 03:33 Pahal Mathew

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  • 13.07.26 23:49 [email protected]

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  • 15.07.26 11:53 Sarah Green

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  • 16.07.26 21:38 patricialovick86

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  • 16.07.26 21:38 patricialovick86

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  • 17.07.26 19:24 laimqq90

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  • 17.07.26 20:12 martinsjude080

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  • 18.07.26 17:12 Malthe Larsen

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  • 18.07.26 17:42 پریا رضایی

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  • 18.07.26 17:46 Adem Akışık

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  • 18.07.26 23:51 bernalzenaida

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  • 19.07.26 04:10 Fraddy Pual

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  • 19.07.26 19:53 kimberlyhebertt6877

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  • 19.07.26 19:53 kimberlyhebertt6877

    I invested in bitcoin trading After losing $78.4 USDT) linked to a romance fraud scam worth of cryptocurrency through an online investment platform and later discovered it was a scam. After extensive research for recovery options, I contacted CAPITAL CRYPTO RECOVER based on positive client reviews and recommendations. Their professional security team guided me through the recovery process using advanced technology, and I was able to recover my lost cryptocurrency successfully. I am truly grateful for their support and assistance during such a difficult experience. I will advise you to contact CAPITAL CRYPTO RECOVER helped me recover my funds. For anyone facing similar issues, Website: https://recovercapital.wixsite.com/capital-crypto-rec-1 Email: [email protected] Telegram: @Capitalcryptorecover Contact: [email protected] Call/Text Number: +1 (336) 390-6684

  • 19.07.26 19:54 kimberlyhebertt6877

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  • 30.07.26 00:04 Ahmed

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  • 30.07.26 17:27 wendytaylor015

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  • 30.07.26 17:27 wendytaylor015

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  • 31.07.26 16:21 rssllhrnsb

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TEXT : (tatianasorina06 at G.Ma IL ..c 0 m ) Before committing money to any investment, take time to verify the legitimacy of the platform, confirm any relevant regulatory authorisations, and avoid investing more than you can comfortably afford to lose.

  • 01.08.26 15:05 keithwilson9899

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  • 01.08.26 15:05 keithwilson9899

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  • 03.08.26 20:05 Philip

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  • 03.08.26 20:06 Philip

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  • 04.08.26 11:05 Kisnoles

    Excellent analysis, thank you. I was particularly struck by how rigidly the skill sets the order: data processing and form come first, with color coming last. This really cures the habit of "first a pretty palette, then we'll figure it out." A palette validator using OKLCH + color blindness + WCAG is something most chart generators lack. And regarding the boundaries of competence, it's very clear. Where there's a self-checking loop (color), you delegate freely. Where there's a heuristic (form, data interpretation), you remain the final filter. This is a universal principle, not just for /dataviz. By the way, when you look at trading dashboards (including those of decent platforms like ExpertOption), it's immediately clear who thought about readability and who just threw in rainbow lines. Tools like this skill could greatly improve the quality of analytics. Thanks again for the detailed analysis – saved.

  • 04.08.26 12:37 kimberlyhebertt6877

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  • 23.08.26 20:02 leslieyee

    Excellent analysis by /dataviz! I was particularly struck by how strictly the order is defined: "data meaning first, color last" and how the validator actually adjusts the palette according to OKLCH, color blindness, and WCAG standards. These rules greatly improve the quality of charts. By the way, a similar approach to clean and legible charts is highly valued in trading—for example, on the ExpertOption platform, the interface and price visualization are designed to ensure information is read instantly and without unnecessary noise.

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  • 11.09.26 03:23 kimberlyhebertt6877

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  • 15.09.26 03:35 elioduncan

    I fell victim to a freelance web development contract scam that ultimately cost me CAD 4,000. It all started when I came across a job posting on Indeed Canada, a popular online job portal. The position seemed perfect: a freelance web developer role for an established company looking for someone to design and build a fully functional e-commerce website. The job promised a high payment of CAD 20,000 upon successful completion, which sounded like a great opportunity for me to gain experience and earn decent pay.The employer, who introduced himself as a project manager from a "well-known" tech company, was very persuasive and professional in our initial communication. He explained that the project involved developing a user-friendly, responsive online store for their client and that they had a strict timeline to meet. He assured me that the payment would be made promptly after completing the tasks. However, before starting the work, he told me that I would need to pay an upfront fee of CAD 4,000 to cover certain software tools and licensing fees required for the project. This was supposedly a part of their company policy for freelance contractors, ensuring access to their premium resources. The idea of working on a professional project, coupled with the promise of a substantial payout, convinced me to pay the upfront fee.As soon as I made the payment, the project manager became increasingly difficult to reach. He initially responded to my emails and provided some vague instructions on what the project would entail, but as time went on, communication slowed to a complete halt. I never received the required tools or any proper project details. My emails went unanswered, and any attempts to contact the company were met with silence. After waiting for weeks, I realized that I had been scammed.Desperate to recover my money, I turned to TechY Force Cyber Retrieval, a service that specializes in helping victims of online scams. They helped me track the payment and took legal steps to pursue the fraudsters. Through their guidance, I was able to recover the full CAD 4,000. TechY Force Cyber Retrieval worked with my bank to reverse the transaction and liaised with the authorities to trace the scammer's details.This was a hard lesson, and I now know to be highly cautious about online job offers that require upfront payments. It is crucial to research companies thoroughly and avoid any job that seems too good to be true, especially when it involves paying money upfront. WhatsApp https://wa.link/2x6ktp Mail. [email protected]

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  • 15.09.26 15:45 lydiassmith567

    HIRE A HACKER YOUR STOLEN CRYPTO RECOVERY / BTC / USDT / ETH WITH THE HELP OF CAPITAL CRYPTO RECOVER. I want to share my experience publicly regarding cryptocurrency wallet recovery, I highly recommend you to contact CAPITAL CRYPTO RECOVER, a professional private investigator in the Bitcoin world. They have a certified expert security team specializing in Bitcoin Recovery Services and have helped many people worldwide recover their lost funds. My wife and I were defrauded by an online manipulator posing as an experienced crypto investment professional. We lost $9.2 Million Stolen BTC in cryptocurrency and were left feeling homeless. After spending hours searching for a reliable crypto recovery service, I discovered CAPITAL CRYPTO RECOVER online. By patiently explaining my situation to their team, I was able to recover all my funds. Remarkably, my money was returned to my wallet in less than 24 hours. I am extremely grateful to CAPITAL CRYPTO RECOVER for their excellent assistance—they truly were a godsend in my difficult situation. If you have fallen victim to a cryptocurrency scam, you can reach CAPITAL CRYPTO RECOVER through the following channels Email: [email protected] OR Call/WhatsApp: +1 (336) 390-6684 Contact: [email protected] Website: https://recovercapital.wixsite.com/capital-crypto-rec-1

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