Table 1. Trial of Isoflavones and Amino Acids for Hot Flashes and Co-occurring Symptoms: Study Population, Design, Interventions, Outcome Measures and Results.

Author, Year, Study Location	Study Population, Sample Size (screened, enrolled, completed, followed)	Study Design	Main Intervention & Comparison/Control	Outcome Measure: Hot Flashes	Outcome Measure: Other symptoms	Results: Hot Flashes	Results: Co-occurring symptoms
Soy Isoflavones
Basaria¹⁵, 2009, Maryland, USA	PM healthy women 46-76y, Tertiary care center, Caucasions 80% Black 10% Other 10% 93 randomized, 7 withdrew, 84 completed	Double blind RCT, 2 groups 12 weeks	Intervention: 20g soy proteinpowder in beverages x1qD 160mg total isoflavones: Genistein-64mg Diadzein-63mg Glycitein-34mg Control: Placebo powder in beverages x1qD (20g whole milk protein)	MENQOL Chi2 and ANOVA 2 sample ttest Wilcoxon sum rank test Linear Regression	MENQOL Chi2 and ANOVA 2 sample ttest Wilcoxon sum rank test Linear Regression Cognition tests: NART Rotating cubes, Identical pictures, Verbal fluency, Trail making A, Trail making B, GPTcDH, GPT c NDH, Grooved Pegboard=	B-group: Vasomotor (no subscale breakdown): Baseline= NS 6 wks= NS 12 wks= NS	B-group: Psychosocial (NA): Baseline= NS 6 wks= NS 12 wks= NS Physical (NA): Baseline=* 6wks=NS 12nwks=NS COGNITION All NS Soy=*** COGNITION All NS (NA)
					GPT Dominant Hand= DH
Hanachi¹⁶, 2008, Iran	PM women 51.3y (4.6) BMI 26.6 (3.9) Ethnicity NR Screened=NR Enrolled=37 Completed=NR Lost to f/u=NR	RCT, 12 weeks	Intervention: 12.5g soy protein in soy milk per day Genistein-13mg Diadzein-4.13mg Comparison: Soymilk + Exercise (1 hour of walking/D) Control specifics =NR	KI: Percent decrease in symptoms	KI: Percent decrease in symptoms: Depression, Insomnia, Forgetful, Muscle/Joint pains	B-group or W-group?: Soymilk=72% ↓* Soymilk + Exercise=83% ↓* Control group=NR % change “after soymilk”	B-group: MOOD (Depression) Soymilk only 58% ↓ NS Soymilk+Exercise= 18% ↓ NS SLEEP (Insomnia) Soymilk only 35% ↓ NS Soymilk+Exercise, 28% ↓ NS COGNITION (Forgetful) Soymilk only 55% ↓ NS Soymilk+Exercise, 20% ↓ NS PAIN (Muscle/Joint) Soymilk only, 33% ↓ NS Soymilk+Exercise, 30% ↓ NS
							Control groups: NR
Jou¹⁷, 2008, Taiwan	Healthy, menopausal women, University Hospital Clinics 49.8- 57.6y BMI= 19.8- 26 kg/m2 120 screened 96 randomized 7 lost to follow-up 89 completed	Double blind RCT 6 months	Intervention: 3g soy germ extract powder 2x qD= 6g total soy protein, or 135mg Isoflavones qD Genistein-17.1mg Diadzein-65.4mg Gycitein-52.5mg Control: Roasted wheat powder	KI, Linear Mixed Model Approach	KI subscores	B-group: 3 months EP * Non-EP NS Control βo NS 6 months EP* Non-EP NS Control βo NS EP= Equol Producing Non-EP= Non Equol Producing	B-group: 3 & 6 months: Sleep (Insomnia) =NS Mood (Melancholia)= NS Pain (Headaches) =NS
Welty¹⁸, 2007, Boston, MA USA	PM + MT women <4.5 HF/d 51.9y (5.5) BMI=27.9 (5.5) >4.5HF/d 54.6y (5.4) BMI=23.9 (4.0)	RCT crossover design 4 wk TLC diet, then 8 wks, then crossover 8wks = 16 weeks total	Intervention: 25g soy protein in soy nuts (aka roasted soy beans, unsalted, ½ cup qD) Genistein-61mg Diadzein-30mg Glycitein-10mg (=101mg isoflavones) Comparison: TLC diet aka	Outcome measure HFs: Daily diary MENQOL Urine Isoflavone HPLC	Outcome measure (Other): MENQOL	Results HFs: B-group: MENQOL scores Soy nut grp: Vms (no breakdown-NA): 19% ↓ *** C: no change in scores W + B group (HFs): Daily diary: C: 22.3(23.5) to 23.3(23.5) HFs/wk [No decrease]	Results (Other): B-group: MENQOL scores Soy nut grp: Psychsocial (NA): 12.9% ↓ ** Physical (NA): 9.7% ↓*
	197 screened 82 enrolled 22 dropped out 60 completed 39/60 had HFs		Therapeutic Lifefstyle Change diet, recommended by Nat'l Cholesterol Ed. Program 30% energy from total fat, 15% protein, 55% carbs, 1200mg Calcium, 2 fatty fish meals q week.			>4.5HF grp: 45% ↓ 7.5(3.6) to 4.1(2.6) * <=4.5HF grp: 41% ↓ 2.2(1.2) to 1.3(1.1) * HF reduction in soy grps noticeable at 2 weeks (53% ↓ in >4.5HF grp and 54% ↓ in <=4.5HF grp)
Lewis¹⁹, 2006, Canada	PM women 45-60y Age: S 53.3(3.1) C 52.9(3.6) F 53.2(2.9) 792 women screened 99 randomized 87 completed	RCT Double blind Intention to tx 3 arm parallel 16 wks	Soy flour muffins (25g soy) Isoflavones 42mg total: Genistein 25.7mg Diadzein 15.5mg Glycitin 0.7mg Comparison: Ground flaxseed (25mg) muffins(F):	MENQOL Hot flash diary	MENQOL Only Vsm + Psych/Social breakdown Vsm= vasomotor	B-group: MENQOL: Vsm (NA) (S, C, F) NS HF diary: Severity (S, C, F) NS Frequency (S, C, F) NS Vsm (NA) (S, C, F) NS	B-group: MENQOL Psych/Social (NA): NS
	S=Soy C=Control F=Flaxseed Some women were taking Black Cohosh during the study.		50mg/d of secoisolariciresinol diglycoside. Wheat flour (C) muffins (all isocaloric)
Other Isoflavones
Evans²⁰, 2011, Canada	PM women (3 were “peri”-menopausal women Genistein: 53.39y (5.05) BMI 25.5 (3.8) C: 53.50y (4.44) BMI 26.4 (3.8) Ethnicity NR 163 screened 84 enrolled 82 completed 3 lost to f/u	RCT Double blind 12 weeks	Intervention: Synthetic Genistein 30mg capsule 1qD Control: Placebo capsule Microcrystalline Cellulose 1 qD	GCS Daily diary	GCS	(Intention to tx data) B-group: HF/d: G: 9.1 to 4.7 51.2%↓ C: 10 to 7.2 Duration (min) of HF: G: 23.63(17.70) to 11.86(11.82) vs. C: 37.95(48.42) to 22.65(22.47) Avg severity: NS Scale 0-3 G: 1.86(0.43) to 1.70(0.58) vs C: 2.06(0.39) to 1.96(0.57) (Per protocol analysis) HF/d: G: 9.8 to 4.8* 51.2% ↓vs. C: 9.9 to 7.1 29.8% ↓ GCS Vsm (NA-no breakdown) NS	GCS: B-group: MOOD: Depression NS
Carmignani²¹, 2010, Brazil	Healthy, symptomatic c PM women 52.4y(3.9) BMI: HT 25.9 Soy 26.4 C 26.6 HT-65% white, 35% non white Soy-40% white! 60% non white C: 70% white, 30% non white 1520 screened, 60 enrolled, 60 completed, none lost to f/u	Double blind, RCT 3 groups 16 weeks	Intervention: 90 mg total isoflavone powder mixed c water: diadzein 16 mg genistein 30mg glycitein 7 mg 1 placebo tab qD+ soy powder 2x qD (45mg isoflavones each dose) HT: 1mg estradiol 0.5mg norethisterone Placebo powder: 40 g maltodextrin 1 tab per day+ placebo powder 2x qD Placebo: 1 placebo tab qD+ placebo powder 2xqD	MRS with some subscale breakdown	MRS with some subscale breakdown	B-group: Somatic subscale: (HFs) HT-Soy=NS HT-C=** Soy-C=*	B-group: Somatic subscale: PAIN (Joint/musc): HT-Soy=NS HT-C=** Soy-C=* SLEEP (Sleep problems-NA)=NS Psych subscale: MOOD (Depression)=NS (Irritability)=NS (Anxiety)=NS
Lee²², 2010, Korea	45-60y PM >6 mos amenorrhea BMI>30 kg/m2	Double blind RCT 2 groups 12 wks Universit	Intervention; Rexflavone: Sophorae fructus extract 20% isoflavones Capsule=175mg	KI	KI	B-groups: Rex-C= NS (0.0542) KI >20: * (12 weeks) KI 8 weeks *	B-groups: SLEEP Insomnia=NS MOOD Melancholia=NS PAIN
	117 recruited, 87 assigned. Rexflavone 43-(4 dropouts) = 39 -4 no complete=3 5. Placebo 44-(3 dropouts)= 41-10 no complete=3 1. 66 completed the study.	y hospital/clinic	Rexflavone Dose=1cap 2xD Control: Placebo capsule=400mg vehicles such as altodextrin, magnesium stearate and microcrystalline cellulose				Joint/pain=NS HA=NS Total KI score=*
Albertazzi²³, 2005, Italy & UK	PM women 44-65y (x=54y) G 53y(3) C 54y(3) BMI G 26(3) C 28(8) All Caucasian 178 screened 121 enrolled 100	RCT Crossove r study Blinded (researchers) 6wks + 6 wks 12 wks total	Intervention: Genistein 90mg 1 capsule qD Placebo: 1 capsule qD	GCS Hot Flush diary Wilcoxon signed rank test Random coeficient's regression model	GCS	B-group: Hot flush score Subgroup flush score >=9 G: * C: NS GCS (Vsm-> NA)	B-group: From GCS: MOOD Anxiety NS Depression NS C: NR (“Compared to placebo”)
	randomized 99 completed
Hildago²⁴, 2005, Ecuador	PM women 51.3y (3.5) BMI 26.6 (3.9) Screened NR 60 enrolled 53 completed	RCT Double blind Crossove r c 7 day washout 12 wks + 12 wks = 24 wks total	Intervention: Commercially available red clover (RC) isoflavone supplement 80mg/d total 2 capsules qD (40mg each Control: Placebo capsule 2 qD	KI % ↓ in sxs	KI % ↓ in sxs	B-group: HFs * Night sweats * C: NS	B-group: SLEEP (NA) Sleeping disorders* MOOD (Dep)* COGNITION (lack of concentration)* PAIN (Arthralgia)* (Headache)*
Jou²⁵, 2005 Taiwan/UK	Women undergoing MT Age NR BMI NR Ethnicity NR 48 recruited Screened NR Randomized NR Completed NR	Randomized trial Single center Prospective	Intervention: Isolflavone 70mg 4 Capsules 325mg containing 17.5mg isoflavones per capsule Comparison: Isoflavone 35mg 2 Capsules 325mg containing 17.5 mg of isoflavones per capsule.	GCS scores Wilcoxon signed rank test	GCS scores Wilcoxon signed rank test	B-group: Vsm (NA) 70mg group* (Compared to 35mg group=C)	B-group: Compare to 35mg group = Control MOOD 35mg (12 weeks): Psychological: Anxiety* Depression* 70mg(12 weeks): Pscyhological: Anxiety* Depression NS (Not used d/t control & tx both significant)+
Uesugi²⁶, 2004 Japan	Climacteric Japanese women Age 58y(7) 44-74y BMI 23.0(3.7) 58 volunteer participants Screened NR	RCT Double blind Crossove r study 2 4-wk periods 8 weeks total	Intervention: Isoflavone (IF) 40mg tablet Daidzein 3.41mg Genistein 0.91mg Glycitein 2.70 mg Control: Placebo tablet (Contents not described)	Questionnaire Interview SMI (Simplified Menopausal Index)	Questionnaire Interview SMI (Simplified Menopausal Index)	B-group: For SMI>26 IF vs. placebo * EQUOL: no HF data	B-group: All other sxs NS Compared to placebo SLEEP Insomnia NS MOOD Depression NS PAIN Joint pain NS EQUOL: no sx data
Combinatio ns
Ishiwata²⁷, 2009, Japan	Symptomatic Japanese women 39 Pre-25 Peri-70 PM 40-59y C: 50.6y (4.9) BMI 22.3(3.0) EQ-1: 50.5y (4.7) BMI	RCT Double blind 12 wks	Interventions: EQ-1: 10mg equol per day EQ—3: 30mg equol per day 1 supplement packet contained: S-equol 10mg Daidzein 0.8mg Genistein 2 mg Glycitein 4.5mg Placebo= C	Menopausal sx Scale for Japanese women questionnaire Daily diaries GCS	Profile of Mood States (POMS) questionnaire 24h urine samples Menopause sx Scale for Japanese women ∼GCS	B-group: EP Menopause Scale: Vasomotor (NA) Non-EP Vasomotor (NA) EQ-3 NS C * EQ-1 NS EQ-3 *	B-group: EP Menopause Scale: MOOD (Depression) NS POMS (NA) Non-EP Menopause Scale: MOOD (Depression) NS POMS: Fatigue-Inertia *anxiety EQ-3
	22.3(3.0) EQ-3: 50.5Y (4.7) BMI 22.0 (3.0) 141 screened 134 randomized 127 completed		Supplement packet containing lactose				Non-EP Meno scale: MOOD (Depression) NS POMS: MOOD Anger-Hostility NS Fatigue-Inertia NS
Mucci²⁸, 2006, Italy	Symptomatic menopausal women 53.8y ES: 53.3y(5.6) Ca+D: 54.4y(6.1) BMI,ES: 25.2(3.5) BMI, Ca+D: 23.2(3.3) Ethnicity NR 89 women 44 ES, 45 Ca+D Screened NR Enrolled NR	RCT Multi-center Longitudi nal, parallel groups 24 wks	Interventions: Magnolia Bark Extract (ES) 60mg and Magnesium 50 mg Isoflavones 60mg: (Genistin 30mg Diadzin 30mg) Lactobacillus 500 million spores Calcium 141 mg Vitamin D3 5μg 1 tab qD of ES Comparison: Calcium 141mg Vit D3 5μg 1tab qD of Ca+D	Chi square Fisher's exact test ANOVA Change vs Baseline % Absence of Sxs (Disappearan ce rate)	Chi square Fisher's exact test ANOVA Change vs Baseline % Absence of Sxs (Disappearance rate)	B-group: Wk 2 Wk 12 *	B-group: SLEEP (Insomnia): Wk 2 Wk 12* MOOD (Irritability, anxiety, depression) Wk 2 Wk 12*
Secreto²⁹, 2004, Italy	PM women Iso+Meloto nin: 53y( 50-57) BMI 23.8 (21.5-26.8) Iso alone: 52y (49-54) BMI 25.0 (22.8-26.0) Melatonin alone: 52y (50-57) BMI 23.6 (21.8-26.8) Control: 52y (50-55) BMI 24.0 (21.9-26.8) 388 screened 262 randomized 232 analyzed 9 lost to f/u	RCT Double blind Multicen ter 2×2 factorial 12 weeks	Interventions: Isoflavones: Diadzein 40mg Glycitein 28mg Genistein 12mg Grp1: Soy isoflavones (80mg)+melatonin (3mg) Grp2: Soy Isoflavones(80mg) + placebo melatonin Grp3: Placebo soy Isoflavones+ melatonin (3mg) Grp4: Placebo soy+ placebo melatonin Placebo=excipient only, sorbitol.	GCS (Only 2 items for HFs questioned by the authors)	GCS	B-group: Vasomotor (NA): Isoflavones alone larger % dec in sxs: Iso 74.6% ↓ NS C 69% ↓ Mel 61.1% ↓ NS Iso+Mel 62.3%↓ No statistical data due to high placebo score. Avg GCS vasomotor baseline=4 Some members (22) had a “zero” score for vasomotor sxs and were not equally distributed among the groups No subscale breakdown	B-group: MOOD (Psych-NA): Iso+Mel: ↑ improvement than others NS No improvement: Iso+mel 18% Iso 29% Mel 30% C 29% No statistical data due to high placebo score. No subscale breakdown
Amino Acids
Guttoso³⁰, 2009, New York	PM women Mean age 54.5/54.1;	Double blind; 2 grps;	Intervention: methionine 2 gm/day (Grp A)	Daily HF diary; 1° outcome	Questionnaire at baseline to wk 12 &	B-group: HF composite & HF Baseline Score = NS	B-group: SLEEP (quality): (PSQI-global) NS
	White (96%/81%) Volunteers TV & newspaper ads; used FSH to dx menopause; all hf; no BSO; hyst (16%/15%); no HRT; 378 screened; 51 randomized into 2 grps; 47 completed	randomized (25/26); concealed; baseline to 12 wks (Phase 1)	Control: placebo (Grp B) Placebo= Cornstarch + microcellulose	measure % chg in HF composite score (severity + freq); 2° outcome measure included % chg HF freq; baseline to 12 wks & baseline to 20 wks Also HFBS (1-10); high=extreme bother	baseline to wk 20 PSQI global score 0-21; high=worse	Grp B 33.4% ↓ Baseline thru 12 wks	(NA)
Guttoso³¹, 2008, New York	PM Mean age 56.1/55.7; White (92%/96%) Volunteers TV & newspaper ads; used FSH to dx menopause; all hf; hyst	2grps (50/50); randomized; concealed; baseline to 12 wks (Phase 1)	Intervention: l-isoleucine 5 mg/day (Grp A) Control: placebo (Grp B) placebo contents not described	Daily HF diary: daily for 1 wk baseline, 1 wk during wks 1-4, 1 wk during wks 8-12, 1 wk during wks 16-17 & 1 wk during wks 21-22; hf sev	Ques at baseline, wk 12 & wk 22 PSQI global score 0-21; high=worse	B-group: Hf freq & hf composite & PPHFS -all NS	B-group: SLEEP (quality) (PSQI-global) NS (NA)
	(30%/36%); no HRT; Phase 1: 100 randomized into 2 grps; 86 completed			recorded when occurred; Also PPHFS

Key: B-group = Between groups

C = control

EP = Equol producing

GCS = Greene Climacteric Scale

HF = Hot flash

HT = Hormone therapy

KI = Kupperman Index

MENQOL = Menopause Quality of Life Scale

MRS = Menopause Rating Scale

MT = Menopausal transition

Non-EP = Non equol producing

NA = Not applicable

NR = Not reported

NS = Not significant

POMS = Profile of Mood States

PPHFS = Patient Perceived Hot Flash Score

PM = Post menopausal

PSQI = Pittsburgh Sleep Quality Index

VMS = Vasomotor symptoms