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  From Mr Average To Superman Health & Wellbeing
  
  
  From Mr Average … to Superman
  
  
  The journey from an ordinary, everyday life—often referred to colloquially as "Mr Average"—to the pinnacle of physical fitness and
  vitality that can be likened to a superhero is both challenging and profoundly rewarding.
  
  It involves deliberate changes across several dimensions:
  nutrition, exercise, mental resilience, and lifestyle habits.
  Below are key strategies that have proven effective for many who aspire to transform themselves.
  
  
  
  
  
  
  1. Nutrition: Fueling the Body Properly
  
  
  
  
  Balanced Macronutrients
  
  
  A diet rich in lean proteins (chicken, fish, legumes), complex carbohydrates (brown rice, quinoa,
  sweet potatoes), and healthy fats (avocado, nuts, olive oil) supports muscle growth, energy levels, and
  recovery.
  
  
  
  Micronutrient Adequacy
  
  
  Micronutrients such as vitamin D, calcium, iron, and magnesium are essential for bone health, immune function, and overall performance.
  Incorporate colorful vegetables, fortified foods, or supplements
  if needed.
  
  
  
  Hydration
  
  
  Aim to drink at least 2–3 liters of water daily.
  Adequate hydration improves digestion, thermoregulation, and cognitive function.
  
  
  1.3 Training Regimen
  
  
  A balanced training plan should include:
  
  
  
  
  
  Resistance Training (4–5 days/week)
  
  
  Compound lifts (squats, deadlifts, bench press) for overall strength, supplemented with accessory work for muscular
  balance.
  
  
  
  Cardiovascular Conditioning (2–3 days/week)
  
  
  HIIT or moderate-intensity steady-state sessions to support metabolic
  health and improve endurance.
  
  
  
  Flexibility & Mobility Work (Daily)
  
  
  Dynamic stretching pre-workout; static stretching post-workout.
  Include foam rolling and mobility drills for
  the hips, shoulders, and thoracic spine.
  
  
  1.4 Expected Outcomes
  
  
  With consistent application of this program:
  
  
  
  
  
  
  Enhanced Core Stability: A stronger transverse abdominis and multifidus will reduce
  lumbar strain during activities.
  
  
  Improved Postural Alignment: Better control over pelvic tilt and thoracic rotation leads to
  more efficient movement patterns.
  
  
  Reduced Pain & Injury Risk: Balanced musculature mitigates compensatory overload on joint structures.
  
  
  
  
  
  
  
  
  2. Comparative Evaluation of Core‑Stability Regimens
  
  
  Below is a side‑by‑side assessment of four prominent core‑stability protocols:
  
  
  
  
  Protocol Key Emphasis Targeted Muscles Common Exercises Pros Cons
  
  
  Swiss Ball (Dynamic) Stability + proprioception Transversus abdominis, multifidus,
  glutes, hip abductors Ball roll‑outs, ball bridges, ball side planks Engages core
  in a moving environment; improves balance. Requires equipment; risk of falling if not controlled.
  
  
  
  Pilates (Static & dynamic) Core strength + flexibility Transversus abdominis, diaphragm,
  pelvic floor Hundred, roll‑up, single leg stretch Low-impact; focuses on breath
  and alignment. May be too slow for high-intensity training.
  
  
  
  TRX Suspension Trainer (Dynamic) Multi-directional core
  stability Transversus abdominis, glutes, shoulders TRX plank, TRX mountain climbers, TRX pike Bodyweight training; adjustable
  difficulty. Requires a sturdy anchor point; learning curve for form.
  
  
  
  Ab Wheel / Stability Ball (Static) Core endurance Rectus
  abdominus, obliques Ab wheel roll‑out, ball crunch Simple equipment; high reps possible.
  Can be risky if technique off; requires core strength first.
  
  
  
  ---
  
  
  
  
  7. Sample Weekly Core‑Focused Plan
  
  
  
  Day Workout Key Movements Sets × Reps
  
  
  Mon Bodyweight Circuit (no equipment) Plank, Side plank, Mountain climbers,
  V‑ups 3×30‑sec each
  
  
  Tue Gym – Upper‑body + Core Bench press, Pull‑ups,
  Hanging leg raise, Cable woodchop 4×8–12 for lifts; 3×15–20 for core
  
  
  Wed Active Recovery / Mobility Yoga flow, foam rolling, light walk N/A
  
  
  
  Thu Gym – Lower‑body + Core Squats, Romanian deadlift, Swiss ball crunches, Pallof press 4×8–12;
  3×15–20
  
  
  Fri Home Circuit Push‑ups, lunges, bicycle crunches, plank variations 3 rounds
  
  
  Sat Optional HIIT / Sport Activity 30 min of interval training
  or a sport (e.g., soccer) N/A
  
  
  Sun Rest Day Light stretching, relaxation N/A
  
  
  The program balances resistance training with core work and recovery days.
  
  Adjust volume/intensity to fit your schedule.
  
  
  
  
  ---
  
  
  
  
  7. Monitoring Progress
  
  
  
  Parameter How to Measure Frequency
  
  
  Core strength Static hold: Plank duration, Dynamic lift: Deadlift or hip‑bridge with weight Every 4–6 weeks
  
  
  Muscle size (optional) Circumference of abdominal area, calipers for muscle thickness Every 8 weeks
  
  
  Body composition Bioimpedance scale or skinfold calipers Every month
  
  
  Functional performance Time to complete a 20‑m sprint
  + back‑to‑back squats Every 4–6 weeks
  
  
  Use the data to adjust volume, load, and exercise selection. If core strength stagnates,
  increase load or add an extra set; if you’re not seeing hypertrophy,
  ensure progressive overload and adequate
  protein intake (≈1.8 g kg⁻¹ body weight).
  
  
  
  ---
  
  
  
  
  3. Sample 4‑Week Program
  
  
  
  Day Warm‑up & Mobility Strength Phase Conditioning
  
  
  Mon 10 min rowing; dynamic hip circles; 2×15 s banded
  side steps Progressive overload – squat + deadlift (3–4 sets, 5–6 reps) HIIT on treadmill:
  30 sec sprint/90 sec walk ×8
  
  
  Tue 5 min jump rope; scapular push‑ups; band pull‑apart Upper‑body focus – bench + rows (3–4 sets,
  6–7 reps) 15 min steady‑state bike at 70% HRmax
  
  
  Wed Rest or gentle mobility: foam roll quads, hamstrings, thoracic spine Active recovery
  – light yoga flow, deep breathing 30 min brisk walk
  
  
  Thu Dynamic warm‑up: inchworms, lunges, side‑step squats
  Lower‑body focus – squats + deadlifts (3–4 sets, 6–7 reps) 20 min hill repeats on treadmill
  
  
  Fri Warm‑up with jump rope, band pulls, and shoulder circles Full‑body
  conditioning – kettlebell swings, push‑ups, box jumps (2 rounds)
  30 min HIIT circuit
  
  
  Sat Light jog or bike ride to promote active recovery Stretching session + foam rolling for muscle relaxation Rest &
  prepare for next week
  
  
  Sun Optional yoga flow or gentle walk in nature Reflect
  on progress, set goals for the upcoming week
  
  
  Feel free to adjust each day’s activities to match
  your energy levels and schedule. The key is consistency,
  so choose workouts that keep you motivated!
  ????
  
  
  
  ---
  
  
  
  
  2️⃣ Quick Tips: How to Keep a Fitness Routine
  Going
  
  
  
  
  Set SMART Goals
  
  
  Specific → "Run 3 miles in 30 minutes"
  
  Measurable → Track with an app or journal
  
  Achievable → Start at your current level, then progress
  
  Relevant → Align with what you want (health, confidence)
  
  Time‑Bound → Finish by a certain date
  
  
  
  
  
  Schedule It
  
  
  Put workouts in the calendar like meetings.
  
  
  
  
  Prep Ahead
  
  
  Pack gym bag or lay out workout clothes the night before.
  
  
  
  
  Mix It Up
  
  
  Change exercises every 4–6 weeks to avoid plateaus and boredom.
  
  
  
  
  Track Progress
  
  
  Keep a log of weights, reps, heart rate, or how you feel.
  
  
  
  
  
  
  Reward Yourself
  
  
  Celebrate small wins with something enjoyable (favorite snack, movie).
  
  
  
  
  
  
  How I Use My "Assistant" to Stay on Track
  
  
  
  Task Assistant Feature Example
  
  
  Plan a week’s workouts Daily prompts & calendar sync "Create a balanced training plan for Monday–Friday."
  
  
  Track reps/weights Quick log entry "Add 10x12 at bench press"
  
  
  Get motivation Reminders & affirmations "You’ve trained consistently for 3 weeks! Keep going!"
  
  
  Adjust due to schedule changes Re‑scheduling tool "Move Thursday’s session to Friday because of a meeting."
  
  
  ---
  
  
  
  
  Bottom Line
  
  
  Your body is your most valuable asset. Even if you can’t hit the gym as often as you’d like, regular movement—whether
  it’s a brisk walk, a home circuit, or office stretches—is far better than inactivity.
  By treating each day’s activity as a step toward a healthier future, you’ll preserve muscle, boost metabolism, and enjoy
  mental clarity—all of which make your eventual return to
  the gym feel stronger and more rewarding.
  
  
  
  Take the next step today: commit to at least 30 minutes of movement
  this week, track it, and watch how small consistency builds lasting momentum.
  Your future self will thank you.
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  A Double-blind Crossover Trial Of Methandienone Dianabol, CIBA
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  **Title:**
  *Effect of a Novel β‑Blocker on Cardiac Output
  and Systemic Vascular Resistance in Patients with Heart Failure: A Randomized, Double‑Blind Study*
  
  ---
  
  ## 1️⃣ Abstract
  
  A double‑blind, randomized, placebo‑controlled trial was conducted to evaluate the cardiovascular
  effects of **β‑Blocker X**, a newly developed selective β₁‑adrenergic antagonist.
  Eighty adults (age 35–75 yr) with NYHA class II–III heart failure
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  **Primary endpoint:** change in left ventricular end‑diastolic dimension (LVEDD) measured by transthoracic echocardiography at baseline and month 12.
  **Secondary endpoints:** changes in LV mass, LVEF, exercise capacity (6‑minute walk test), BNP levels, and adverse events.
  
  At 12 months, mean LVEDD decreased by 4.8 mm (p  0.001) in the β‑Blocker X group versus a 1.2 mm increase (p = 0.07)in placebo (between‑group difference: −6.0 mm; p  0.001). LV mass decreased by 12 g/m² (p  0.01) and LVEF improved from 45 % to 52  (p  0.005). BNP fell by 35 %, an exercise capacity ncreased by 15 %. Adverse events were mild: dizziness in 8 % of patients, transient hypotension in 5 %. No hospitalizations or deaths occurred during the study.
  
  The trial demonstrates that a structured, supervised aeroic program can reverse structural remodeling, enhance systolic function, and improve functional status in patients with HCM. These findings support incorporating cardiac rehabilitation into routine management for eligible individuals, particularly those with preserved exercise capacity but symptomatic diastolic dysfunction. The authors call for larger multicenter trials to confirm benefits across diverse phenotypes and to assess long‑term safety, including arrhythmic risk.
  
  ---
  
  ### 3. Comparative Table of the Two Articles
  
  | Feature | Article 1 (Meta‑analysis) | Article 2 (RCT) |
  |---------|---------------------------|-----------------|
  | **Design** | Systematic review + meta‑analysis of RCTs | Single‑center, prospective, randomized controlled trial |
  | **Sample Size** | 12 000+ participants across 25 trials | 300 participants total (150 per arm) |
  | **Intervention(s)** | Standardized anti‑arrhythmic drugs (class I/III), compared to placebo or other drugs | Novel anti‑arrhythmic agent versus standard therapy in high‑risk atrial fibrillation |
  | **Primary Outcome** | Composite of all‑cause mortality, arrhythmia recurrence, hospitalisation | 30‑day composite of major adverse cardiac events (death, MI, stroke) |
  | **Secondary Outcomes** | Cardiovascular morbidity, drug toxicity, quality of life | Long‑term rhythm control, biomarker changes, neurocognitive scores |
  | **Statistical Analysis** | Meta‑analysis with random effects; subgroup analysis by age, comorbidities; risk ratios (RR) and 95% CIs | Kaplan‑Meier survival curves; Cox proportional hazards models; intention‑to‑treat principle |
  | **Key Findings** | Statistically significant reduction in mortality with certain antiarrhythmic agents; heterogeneity across studies | No significant difference in primary endpoint, but improved rhythm control and fewer hospitalizations in the treatment arm |
  
  ---
  
  ## 1. Background & Rationale
  
  ### 1.1 Disease Burden
  - **Cardiovascular diseases (CVD)** are the leading cause of mortality worldwide (~17.9 million deaths annually).
  - A significant proportion of CVD deaths are attributable to *arrhythmogenic conditions* such as atrial fibrillation, ventricular tachycardia, and sudden cardiac death.
  
  ### 1.2 Existing Evidence
  - Numerous observational studies suggest that specific antiarrhythmic medications reduce morbidity and mortality.
  - However, the magnitude of benefit remains uncertain due to heterogeneity in study designs, populations, and endpoints.
  
  ---
  
  ## 2. Objectives
  
  ### Primary Objective
  - **To determine whether treatment with antiarrhythmic medication reduces all-cause mortality compared to placebo or standard care in patients with arrhythmogenic cardiac conditions.**
  
  ### Secondary Objectives
  1. Evaluate the impact on *cardiovascular-specific* outcomes (e.g., heart failure hospitalizations, arrhythmia recurrence).
  2. Assess safety and tolerability profiles.
  3. Examine subgroup effects by age, sex, comorbidities, and baseline arrhythmia severity.
  
  ---
  
  ## 3. Study Design
  
  - **Type:** Multicenter, randomized, double-blind, placebo-controlled trial.
  - **Duration:** Minimum follow-up of 2 years (median 24 months).
  - **Population:**
  - Inclusion Criteria:
  - Adults ≥18 years with documented arrhythmia (e.g., atrial fibrillation, ventricular tachycardia) confirmed by ECG/monitoring.
  - Stable baseline medication regimen for at least 4 weeks.
  - Exclusion Criteria:
  - Severe uncontrolled heart failure (NYHA III-IV), active ischemic disease, or other major comorbidities limiting life expectancy **Dr. Maria Santos (Ethics Reviewer):**
  
  > "While I appreciate the rigorous design, there are several ethical concerns. First, the exclusion of participants with comorbidities may limit generalizability and potentially deny individuals who could benefit from the intervention access to participation. Second, the 24-month follow-up requires sustained participant commitment; we must ensure robust retention strategies and consider incentives that do not coerce."
  
  > **Dr. Luis Almeida (Statistical Consultant):**
  > "From a statistical standpoint, stratifying by sex is sound, but we should also account for baseline disease severity as a covariate in the analysis to reduce residual confounding. Additionally, we need to predefine subgroup analyses and adjust for multiple comparisons."
  
  > **Dr. Marta Silva (Ethics Officer):**
  > "The informed consent process must clearly explain potential risks and benefits, especially since this is a randomized trial with no standard-of-care control arm. We should also establish an independent data monitoring committee to oversee safety."
  
  ---
  
  ### 4. "What If" Scenarios
  
  #### Scenario A: Inclusion of Patients Without Diabetes
  - **Impact on Trial Design**: Removing the diabetes requirement would broaden eligibility, potentially
  increasing enrollment speed but also introducing
  heterogeneity in metabolic status.
  - **Statistical Power**: The sample size calculation must account for increased variability; larger
  cohorts may be needed to maintain power.
  - **Clinical Relevance**: Findings could generalize to all patients with
  COVID‑19 and cardiovascular comorbidities, but confounding by baseline glycemic control would
  need careful adjustment.
  
  #### Scenario B: Alternative Primary Endpoint (Composite
  Cardiovascular Outcome)
  - **Impact on Trial Design**: A composite endpoint (e.g., myocardial infarction, stroke, arrhythmia) may capture more
  events but could dilute the specific effect of a drug on inflammation‑related pathways.
  
  - **Statistical Power**: More events increase
  power; however, heterogeneity in event definitions may complicate interpretation.
  - **Clinical Relevance**: Clinicians might value broader cardiovascular
  risk reduction over singular inflammatory markers.
  
  
  
  ---
  
  ## 3. Comparative Evaluation of Drug Classes
  
  | Drug Class | Mechanism (Pharmacodynamics) | Typical Use in COVID‑19 | Evidence Strength |
  |------------|------------------------------|------------------------|-------------------|
  | **Corticosteroids** (e.g., dexamethasone) | Potent anti‑inflammatory via
  glucocorticoid receptor; suppress cytokine production, stabilize membranes.
  | Standard of care for hospitalized patients requiring oxygen or ventilation. |
  High – RCTs and meta‑analyses demonstrate mortality benefit.
  |
  | **IL‑6 Receptor Antagonists** (tocilizumab) | Block IL‑6 signaling via
  anti‑IL‑6R antibodies; reduce downstream JAK/STAT activation. | Used in severe cytokine storm;
  evidence mixed but some benefit in specific subgroups.
  
  | Moderate – Some RCTs show reduced progression to mechanical ventilation. |
  | **JAK Inhibitors** (baricitinib) | Inhibit intracellular JAK1/2, dampening multiple cytokine pathways (IFN‑γ, IL‑6, IL‑12).
  
  | Shown to reduce viral replication and inflammation when combined with antivirals.
  | Moderate – RCTs indicate improved recovery
  times. |
  | **Anti‑IL‑17A** (secukinumab) | Neutralizes IL‑17A; reduces neutrophil recruitment & cytokine amplification. | Preclinical data suggest attenuation of hyperinflammatory lung
  injury. | Low – No clinical trials yet in COVID‑19. |
  
  ---
  
  ## 3. Suggested Clinical Trial Design
  
  | Element | Recommendation |
  |---------|----------------|
  | **Population** | Adults (≥18 y) with confirmed SARS‑CoV‑2 infection, presenting within 7 days of symptom
  onset, requiring supplemental oxygen but not invasive ventilation. Exclude
  those on chronic immunosuppression or active infections.
  |
  | **Intervention** | Secukinumab 300 mg IV once at baseline
  (dose adapted from rheumatology). |
  | **Comparator** | Placebo IV matched in volume and appearance.
  |
  | **Co‑interventions** | All patients receive standard of care: dexamethasone, anticoagulation, remdesivir if indicated.
  
  No other biologics allowed. |
  | **Primary Endpoint** | Time to clinical improvement defined as a ≥2‑point reduction on the WHO Ordinal Scale for
  Clinical Improvement or discharge from hospital within 28 days.
  |
  | **Secondary Endpoints** | 1) All‑cause mortality at day 60; 2) Duration of mechanical ventilation; 3) Incidence
  of serious adverse events (sepsis, secondary infections); 4)
  Viral clearance by RT‑PCR at days 7 and 14; 5) Levels
  of IL‑6, CRP, ferritin over time. |
  | **Sample Size** | Assuming a hazard ratio of 1.4 for improvement, with α=0.05 (two‑sided) and power 80%, approximately
  280 participants (140 per arm) are needed; inflated to 320 to account for dropout and loss to follow‑up.
  |
  | **Statistical Analysis Plan** | Primary analysis:
  Cox proportional hazards model comparing time to improvement between groups, adjusting for baseline covariates (age, sex, comorbidities).
  Secondary endpoints: mixed‑effects models for
  repeated measures of biomarkers; logistic regression for mortality at 28 days;
  Kaplan–Meier survival curves with log‑rank test. Sensitivity
  analyses will include per‑protocol population and multiple imputation for missing data.
  All tests two‑tailed with α=0.05. |
  
  ---
  
  ### 4. Risk Management
  
  | **Potential Risk** | **Mitigation Strategy** |
  |--------------------|-------------------------|
  | Cytokine release from exosomes may worsen inflammation | Use
  low‑dose pilot phase; monitor cytokines (IL‑6, TNF‑α) hourly; predefine stopping rules for cytokine surge.
  |
  | Viral entry via exosomal membrane or receptor interaction | Confirm absence of ACE2 and TMPRSS2 in final
  product; test for spike protein binding to exosomes in vitro.
  |
  | Contamination with host cell proteins/viral particles | Implement stringent purification,
  endotoxin testing, and virus filtration steps; use size‑exclusion chromatography.
  |
  | Immunogenicity from bovine miRNA or surface proteins
  | Use humanized miRNA cocktail; minimize bovine protein content
  through extensive washing. |
  | Scale‑up failure (aggregation, loss of activity) | Pilot scale batches with monitoring of particle size,
  zeta potential, and antiviral potency before large‑scale production.
  |
  
  ---
  
  ## 4. Suggested Experimental Workflow for a New Bovine‑Derived Product
  
  1. **Design & Production**
  - Clone miRNA expression cassette into a suitable bovine cell line (e.g., MDBK).
  
  - Transduce cells with lentivirus containing the construct;
  select stable integrants.
  
  2. **Purification of Exosomes**
  - Collect conditioned media, centrifuge at 300 × g → 2 000 × g
  to remove debris.
  - Filter (0.22 µm).
  - Ultracentrifuge at 100 000 × g for 70 min; wash pellet with PBS; repeat spin.
  
  3. **Characterization**
  - Nanoparticle tracking analysis for size distribution and concentration.
  - Western blot for CD63, TSG101, and the specific miRNA (qPCR).
  
  - Electron microscopy to confirm morphology.
  
  4. **Functional Assay**
  - Treat cultured cells with exosomes; measure uptake of
  labeled miRNA by qRT‑PCR.
  - Assess downstream effect (e.g., target gene repression).
  
  
  5. **Optimization**
  - Test alternative isolation methods (ultrafiltration, size‑exclusion chromatography).
  
  - Compare yield and purity across methods.
  
  ---
  
  ## 4. Summary Checklist
  
  | Step | Action | Key Tips |
  |------|--------|----------|
  |1|Define biological question|Choose exosomes or microvesicles?|
  |2|Select isolation method|Ultracentrifugation (gold standard) vs precipitation, chromatography|
  |3|Collect conditioned media|Minimize cell death; use low‑FBS
  medium|
  |4|Sequential centrifugation|300 ×g → 2000 ×g → 10 000 ×g (optional) →
  100 000 ×g|
  |5|Resuspend pellet in PBS or buffer|Keep volume consistent for quantification|
  |6|Characterize vesicles|Nanoparticle tracking, electron microscopy,
  Western blot for CD9/CD63/CD81|
  |7|Store appropriately|-80 °C; avoid freeze–thaw cycles|
  
  ---
  
  ## 4. Example Protocol (10 mL culture)
  
  | Step | Details |
  |------|---------|
  | **Cell preparation** | Grow 3 × 10⁶ cells
  in 10 mL serum‑free medium for 24 h. |
  | **Centrifugation** | 300 g, 10 min (remove cells).
  
  2000 g, 20 min (removing debris).
  100 000 g, 70 min (ultracentrifuge; 4 °C). |
  | **Resuspension** | Wash pellet with 1× PBS; spin again at
  100 000 g for 30 min.
  Resuspend in 200 µL of sterile PBS or buffer of choice. |
  
  ---
  
  ### Practical Tips
  
  * Use a clean, RNase‑free system if you plan to keep the RNA intact (e.g.,
  for qRT‑PCR).
  * Always pre‑cool tubes and centrifuge at 4 °C;
  higher temperatures can degrade RNA.
  * Store isolated material at –80 °C in small aliquots to avoid repeated freeze–thaw cycles.
  
  
  ---
  
  **Bottom line:** The protocol you described (high‑speed spins, short times) is typical for extracting total
  nucleic acids from fungal mycelium while preserving
  RNA integrity. It is well suited for downstream
  applications such as RT‑qPCR or RNA‑seq of *T.
  
  reesei* cultures.
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  Below is a concise "cook‑book" style protocol that you can follow
  in a clinical setting (or even as part of an academic paper)
  to assess whether a patient’s hair‑loss treatment is working.
  The recipe mixes **visual grading**, **objective measurements** and **biomarker data** so that the result is reproducible, statistically meaningful and easy to report.
  
  
  ---
  
  ## Ingredients
  
  | Category | Specifics | Why it matters |
  |----------|-----------|----------------|
  | **Baseline data** | • Age, sex, ethnicity
  • Family history of alopecia
  • Duration & pattern of hair loss (Hamilton–Norwood / Ludwig scale)
  • Current medications/ supplements | Sets the context;
  helps interpret change relative to expected progression |
  | **Visual grading** | • 3‑point or 5‑point scale for
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  • Standardised photographs (frontal, lateral, vertex)
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  | **Hair density** | • Trichoscopy: follicular unit count per
  cm² (or per 3×3 cm area)
  • Hair shaft diameter distribution | Quantifies the number of active follicles; less subjective
  than visual alone |
  | **Hair strength & breakage** | • Hair tensile strength test
  • Scoring of visible hair breakage or thinning | Assesses functional improvement beyond density
  |
  | **Patient‑reported outcomes (PROs)** | • VAS for satisfaction, quality of life questionnaire | Captures patient perspective,
  often correlating with objective findings |
  
  These parameters can be obtained through a combination of
  non‑invasive imaging and simple mechanical tests that
  are affordable and reproducible.
  
  ---
  
  ## 2. Practical Measurement Protocol
  
  Below is a step‑by‑step protocol designed to run in about **30 minutes**
  per patient. All equipment should be calibrated before use.
  
  
  | Step | Time | Method | Equipment |
  |------|------|--------|-----------|
  | **1. Patient preparation** | 3 min | Explain the procedure; obtain written consent;
  ask about medications and recent hair care habits.
  | – |
  | **2. Scalp area selection** | 2 min | Mark a rectangular area (≈ 4 cm × 6 cm) on the scalp where
  the patient has been experiencing thinning. Use a disposable
  marker that can be washed off. | Disposable marker, ruler |
  | **3. Hair density counting** | 5 min | Using a stereomicroscope
  or dermatoscope, count the number of hairs within the marked area at magnification ×10–×20.
  Record as Hairs per cm². | Dermatoscope or stereomicroscope
  with grid overlay |
  | **4. Follicle viability test (optional)** | 7 min | Apply
  a small drop (~0.1 mL) of 1% hydrogen peroxide onto the
  marked area and observe for bubbling within 30 s. Positive bubbling indicates
  viable follicles; record as %Viable. Alternatively, use trypan blue exclusion or histological staining if resources allow. | Hydrogen peroxide solution, pipette |
  | **5. Data entry** | 3 min | Enter Hairs/cm², %Viable, and any notes into a pre-made
  spreadsheet (Excel or Google Sheets). | Computer/tablet with spreadsheet software |
  
  ---
  
  ### Expected Results
  
  | Parameter | Healthy Adult Human Hair | Aged Human Hair (70–80 yrs) |
  |-----------|--------------------------|-----------------------------|
  | Hairs/cm² | 30–35 (±5) | 25–28 (±4) |
  | % Viable | 85–90 % | 65–75 % |
  
  - **Interpretation**: A noticeable decline in both density and viability is expected with advanced
  age, reflecting reduced hair follicle activity and
  increased cellular senescence.
  
  ---
  
  ### Troubleshooting & Common Issues
  
  1. **Low Yield / Poor Density**
  - *Cause*: Inadequate sampling area or uneven scalp pressure.
  
  - *Fix*: Ensure a 10 cm² area; apply uniform pressure with the brush head.
  
  
  2. **Contamination / Bacterial Growth**
  - *Cause*: Improper sterilization of instruments.
  - *Fix*: Autoclave or use sterile disposable brushes; disinfect all surfaces.
  
  
  3. **Uneven Sample Collection**
  - *Cause*: Non‑uniform brush strokes.
  - *Fix*: Practice consistent, gentle strokes in one direction to avoid bias.
  
  
  
  4. **Inconsistent Brush Strokes**
  - *Cause*: Operator fatigue or lack of training.
  - *Fix*: Allow breaks; train staff on standardized brushing technique.
  
  
  
  ---
  
  ### Summary
  
  By following this detailed protocol—comprising meticulous preparation, precise sample collection with a custom‑made sterile brush, controlled
  environmental conditions, and rigorous post‑collection handling—you will obtain high‑quality hair follicle samples suitable for subsequent analyses (e.g., histology,
  immunohistochemistry, DNA/RNA extraction). Adhering to these best practices ensures reproducibility, minimizes contamination, and preserves the integrity of both tissue
  and genetic material.
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